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	<title>bioRxiv Channel: Aligning Science Across Parkinson's (ASAP)</title>
	<link>https://biorxiv.org</link>
	<description>
	This feed contains articles for bioRxiv Channel "Aligning Science Across Parkinson's (ASAP)"
	</description>

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	<prism:publicationName>bioRxiv</prism:publicationName>
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	<title>bioRxiv</title>
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	<link>https://biorxiv.org</link>
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	<item rdf:about="https://biorxiv.org/cgi/content/short/2021.01.08.425958v1?rss=1">
<title>
<![CDATA[
Reconstitution of cargo-induced LC3 lipidation in mammalian selective autophagy 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.01.08.425958v1?rss=1"
</link>
<description><![CDATA[
Selective autophagy of damaged mitochondria, intracellular pathogens, protein aggregates, endoplasmic reticulum, and other large cargoes is essential for health. The presence of cargo initiates phagophore biogenesis, which entails the conjugation of ATG8/LC3 family proteins to membrane phosphatidylethanolamine. Current models suggest that the presence of clustered ubiquitin chains on a cargo triggers a cascade of interactions from autophagic cargo receptors through the autophagy core complexes ULK1 and class III PI 3-kinase complex I (PI3KC3-C1), WIPI2, and the ATG7, ATG3, and ATG12-ATG5-ATG16L1 machinery of LC3 lipidation. This model was tested using giant unilamellar vesicles (GUVs), GST-Ub4 as a model cargo, the cargo receptors NDP52, TAX1BP1, and OPTN, and the autophagy core complexes. All three cargo receptors potently stimulated LC3 lipidation on GUVs. NDP52- and TAX1BP1-induced LC3 lipidation required the ULK1 complex together with all other components, however, ULK1 kinase activity was dispensable. In contrast, OPTN bypassed the ULK1 requirement completely. These data show that the cargo-dependent stimulation of LC3 lipidation is a common property of multiple autophagic cargo receptors, yet the details of core complex engagement vary considerably and unexpectedly between the different receptors.
]]></description>
<dc:creator>Chang, C.</dc:creator>
<dc:creator>Shi, X.</dc:creator>
<dc:creator>Jensen, L.</dc:creator>
<dc:creator>Yokom, A. L.</dc:creator>
<dc:creator>Fracchiolla, D.</dc:creator>
<dc:creator>Martens, S.</dc:creator>
<dc:creator>Hurley, J. H.</dc:creator>
<dc:date>2021-01-09</dc:date>
<dc:identifier>doi:10.1101/2021.01.08.425958</dc:identifier>
<dc:title><![CDATA[Reconstitution of cargo-induced LC3 lipidation in mammalian selective autophagy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-01-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.08.07.234138v1?rss=1">
<title>
<![CDATA[
In situ architecture of neuronal alpha-Synuclein inclusions 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.08.07.234138v1?rss=1"
</link>
<description><![CDATA[
-Synuclein (-Syn) aggregation is a hallmark of devastating neurodegenerative disorders including Parkinsons disease (PD) and multiple systems atrophy (MSA)1,2. -Syn aggregates spread throughout the brain during disease progression2, suggesting mechanisms of intercellular seeding. Formation of -Syn amyloid fibrils is observed in vitro3,4 and fibrillar -Syn has been purified from patient brains5,6, but recent reports questioned whether disease-relevant -Syn aggregates are fibrillar in structure7-9. Here we use cryo-electron tomography (cryo-ET) to image neuronal Lewy body-like -Syn inclusions in situ at molecular resolution. We show that the inclusions consist of -Syn fibrils crisscrossing a variety of cellular organelles such as the endoplasmic reticulum (ER), mitochondria and autophagic structures, without interacting with membranes directly. Neuronal inclusions seeded by recombinant or MSA patient-derived -Syn aggregates have overall similar architecture, although MSA-seeded fibrils show higher structural flexibility. Using gold-labeled seeds we find that aggregate nucleation is predominantly mediated by -Syn oligomers, with fibrils growing unidirectionally from the seed. Our results conclusively demonstrate that neuronal -Syn inclusions contain -Syn fibrils intermixed with cellular membranes, and illuminate the mechanism of aggregate nucleation.
]]></description>
<dc:creator>Trinkaus, V. A.</dc:creator>
<dc:creator>Riera-Tur, I.</dc:creator>
<dc:creator>Martinez-Sanchez, A.</dc:creator>
<dc:creator>Baeuerlein, F. J. B.</dc:creator>
<dc:creator>Guo, Q.</dc:creator>
<dc:creator>Arzberger, T.</dc:creator>
<dc:creator>Baumeister, W.</dc:creator>
<dc:creator>Dudanova, I.</dc:creator>
<dc:creator>Hipp, M. S.</dc:creator>
<dc:creator>Hartl, F. U.</dc:creator>
<dc:creator>Fernandez-Busnadiego, R.</dc:creator>
<dc:date>2020-08-07</dc:date>
<dc:identifier>doi:10.1101/2020.08.07.234138</dc:identifier>
<dc:title><![CDATA[In situ architecture of neuronal alpha-Synuclein inclusions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-08-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.02.17.431620v1?rss=1">
<title>
<![CDATA[
Structural basis for the specificity of PPM1H phosphatase for Rab GTPases 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.02.17.431620v1?rss=1"
</link>
<description><![CDATA[
LRRK2 serine/threonine kinase is associated with inherited Parkinsons disease. LRRK2 phosphorylates a subset of Rab GTPases within their switch 2 motif to control their interactions with effectors. Recent work has shown that the metal-dependent protein phosphatase PPM1H counteracts LRRK2 by dephosphorylating Rabs. PPM1H is highly selective for LRRK2 phosphorylated Rabs, and closely related PPM1J exhibits no activity toward substrates such as Rab8a phosphorylated at Thr72 (pThr72). Here we have identified the structural determinant of PPM1H specificity for Rabs. The crystal structure of PPM1H reveals a structurally conserved phosphatase fold that strikingly has evolved a 110-residue flap domain adjacent to the active site. The flap domain distantly resembles tudor domains that interact with histones in the context of epigenetics. Cellular assays, crosslinking and 3-D modelling suggest that the flap domain encodes the docking motif for phosphorylated Rabs. Consistent with this hypothesis, a PPM1J chimera with the PPM1H flap domain dephosphorylates pThr72 of Rab8a both in vitro and in cellular assays. Therefore, PPM1H has acquired a Rab-specific interaction domain within a conserved phosphatase fold.
]]></description>
<dc:creator>Waschbüsch, D.</dc:creator>
<dc:creator>Berndsen, K.</dc:creator>
<dc:creator>Knebel, A.</dc:creator>
<dc:creator>Alessi, D. R.</dc:creator>
<dc:creator>Khan, A. R.</dc:creator>
<dc:date>2021-02-17</dc:date>
<dc:identifier>doi:10.1101/2021.02.17.431620</dc:identifier>
<dc:title><![CDATA[Structural basis for the specificity of PPM1H phosphatase for Rab GTPases]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-02-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.03.02.433576v1?rss=1">
<title>
<![CDATA[
Pathogenic LRRK2 control of primary cilia and Hedgehog signaling in neurons and astrocytes of mouse brain 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.03.02.433576v1?rss=1"
</link>
<description><![CDATA[
Previously, we showed that cholinergic interneurons of the dorsal striatum lose cilia in mice harboring the Parkinsons disease associated, kinase activating, R1441C LRRK2 mutation (Dhekne et al., 2018). Here we show that this phenotype is also seen in two mouse strains carrying the most common human G2019S LRRK2 mutation. Heterozygous loss of the PPM1H phosphatase that is specific for LRRK2-phosphorylated Rab GTPases (Berndsen et al., 2019) yields the same cilia loss phenotype, strongly supporting a connection between Rab GTPase phosphorylation and cilia loss. In addition, astrocytes throughout the striatum show a ciliation defect in LRRK2 and PPM1H-/+ mutant models. Hedgehog signaling requires cilia, and loss of cilia correlates here with a loss in induction of Hedgehog signaling as monitored by in situ hybridization of Gli1 transcripts. These data support a model in which LRRK2 and PPM1H mutant mice struggle to receive and respond to critical Hedgehog signals in the nigral-striatal pathway.
]]></description>
<dc:creator>Khan, S. S.</dc:creator>
<dc:creator>Sobu, Y.</dc:creator>
<dc:creator>Dhekne, H. S.</dc:creator>
<dc:creator>Tonelli, F.</dc:creator>
<dc:creator>Berndsen, K.</dc:creator>
<dc:creator>Alessi, D. R.</dc:creator>
<dc:creator>Pfeffer, S. R.</dc:creator>
<dc:date>2021-03-02</dc:date>
<dc:identifier>doi:10.1101/2021.03.02.433576</dc:identifier>
<dc:title><![CDATA[Pathogenic LRRK2 control of primary cilia and Hedgehog signaling in neurons and astrocytes of mouse brain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.03.05.434104v1?rss=1">
<title>
<![CDATA[
Multi-Modality Machine Learning Predicting Parkinsons Disease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.03.05.434104v1?rss=1"
</link>
<description><![CDATA[
BackgroundPersonalized medicine promises individualized disease prediction and treatment. The convergence of machine learning (ML) and available multi-modal data is key moving forward. We build upon previous work to deliver multi-modal predictions of Parkinsons Disease (PD).

MethodsWe performed automated ML on multi-modal data from the Parkinsons Progression Marker Initiative (PPMI). After selecting the best performing algorithm, all PPMI data was used to tune the selected model. The model was validated in the Parkinsons Disease Biomarker Program (PDBP) dataset. Finally, networks were built to identify gene communities specific to PD.

FindingsOur initial model showed an area under the curve (AUC) of 89.72% for the diagnosis of PD. The tuned model was then tested for validation on external data (PDBP, AUC 85.03%). Optimizing thresholds for classification, increased the diagnosis prediction accuracy (balanced accuracy) and other metrics. Combining data modalities outperforms the single biomarker paradigm. UPSIT was the largest contributing predictor for the classification of PD. The transcriptomic data was used to construct a network of disease-relevant transcripts.

InterpretationWe have built a model using an automated ML pipeline to make improved multi-omic predictions of PD. The model developed improves disease risk prediction, a critical step for better assessment of PD risk. We constructed gene expression networks for the next generation of genomics-derived interventions. Our automated ML approach allows complex predictive models to be reproducible and accessible to the community.

FundingNational Institute on Aging, National Institute of Neurological Disorders and Stroke, the Michael J. Fox Foundation, and the Global Parkinsons Genetics Program.

RESEARCH IN CONTEXTO_ST_ABSEvidence before this studyC_ST_ABSPrior research into predictors of Parkinsons disease (PD) has either used basic statistical methods to make predictions across data modalities, or they have focused on a single data type or biomarker model. We have done this using an open-source automated machine learning (ML) framework on extensive multi-modal data, which we believe yields robust and reproducible results. We consider this the first true multi-modality ML study of PD risk classification.

Added value of this studyWe used a variety of linear, non-linear, kernel, neural networks, and ensemble ML algorithms to generate an accurate classification of both cases and controls in independent datasets using data that is not involved in PD diagnosis itself at study recruitment. The model built in this paper significantly improves upon our previous models that used the entire training dataset in previous work1. Building on this earlier work, we showed that the PD diagnosis can be refined using improved algorithmic classification tools that may yield potential biological insights. We have taken careful consideration to develop and validate this model using public controlled-access datasets and an open-source ML framework to allow for reproducible and transparent results.

Implications of all available evidenceTraining, validating, and tuning a diagnostic algorithm for PD will allow us to augment clinical diagnoses or risk assessments with less need for complex and expensive exams. Going forward, these models can be built on remote or asynchronously collected data which may be important in a growing telemedicine paradigm. More refined diagnostics will also increase clinical trial efficiency by potentially refining phenotyping and predicting onset, allowing providers to identify potential cases earlier. Early detection could lead to improved treatment response and higher efficacy. Finally, as part of our workflow, we built new networks representing communities of genes correlated in PD cases in a hypothesis-free manner, showing how new and existing genes may be connected and highlighting therapeutic opportunities.
]]></description>
<dc:creator>Makarious, M. B.</dc:creator>
<dc:creator>Leonard, H. L.</dc:creator>
<dc:creator>Vitale, D.</dc:creator>
<dc:creator>Iwaki, H.</dc:creator>
<dc:creator>Sargent, L.</dc:creator>
<dc:creator>Dadu, A.</dc:creator>
<dc:creator>Violich, I.</dc:creator>
<dc:creator>Hutchins, E.</dc:creator>
<dc:creator>Saffo, D.</dc:creator>
<dc:creator>Bandres-Ciga, S.</dc:creator>
<dc:creator>Kim, J. J.</dc:creator>
<dc:creator>Song, Y.</dc:creator>
<dc:creator>Bookman, M.</dc:creator>
<dc:creator>Nojopranoto, W.</dc:creator>
<dc:creator>Campbell, R. H.</dc:creator>
<dc:creator>Hashemi, S. H.</dc:creator>
<dc:creator>Botia, J. A.</dc:creator>
<dc:creator>Carter, J. F.</dc:creator>
<dc:creator>Maleknia, M.</dc:creator>
<dc:creator>Craig, D. W.</dc:creator>
<dc:creator>Keuren-Jensen, K. V.</dc:creator>
<dc:creator>Morris, H. R.</dc:creator>
<dc:creator>Hardy, J. A.</dc:creator>
<dc:creator>Blauwendraat, C.</dc:creator>
<dc:creator>Singleton, A. B.</dc:creator>
<dc:creator>Faghri, F.</dc:creator>
<dc:creator>Nalls, M. A.</dc:creator>
<dc:creator>Accelerating Medicines Program - Parkinsons Disease (AMP PD),</dc:creator>
<dc:creator>Global Parkinsons Genetics Program (GP2),</dc:creator>
<dc:date>2021-03-07</dc:date>
<dc:identifier>doi:10.1101/2021.03.05.434104</dc:identifier>
<dc:title><![CDATA[Multi-Modality Machine Learning Predicting Parkinsons Disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-03-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.12.28.424508v1?rss=1">
<title>
<![CDATA[
Presynaptic autophagy is coupled to the synaptic vesicle cycle via ATG-9 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.12.28.424508v1?rss=1"
</link>
<description><![CDATA[
Autophagy is a cellular degradation pathway essential for neuronal health and function. Autophagosome biogenesis occurs at synapses, is locally regulated and increases in response to neuronal activity. The mechanisms that couple autophagosome biogenesis to synaptic activity remain unknown. In this study we determine that trafficking of ATG-9, the only transmembrane protein in the core autophagy pathway, links the synaptic vesicle cycle with autophagy. ATG-9 positive vesicles in C. elegans are generated from the trans-Golgi network via AP3-dependent budding, and delivered to presynaptic sites. At presynaptic sites, ATG-9 undergoes exo-endocytosis in an activity-dependent manner. Mutations that disrupt endocytosis, including one associated with Parkinsons disease, result in abnormal ATG-9 accumulation at clathrin-rich synaptic foci and defects in activity-dependent presynaptic autophagy. Our findings uncover regulated key steps of ATG-9 trafficking at presynaptic sites, and provide evidence that ATG-9 exo-endocytosis couples autophagosome biogenesis at presynaptic sites with the activity-dependent synaptic vesicle cycle.

HighlightsO_LIIn C. elegans, ATG-9 is delivered to presynaptic sites in vesicles generated from the trans-Golgi network via AP-3-dependent budding
C_LIO_LIATG-9 vesicles undergo activity-dependent exo-endocytosis at presynaptic sites
C_LIO_LIMutations in endocytic proteins, including a mutation associated with Parkinsons disease, result in abnormal ATG-9 accumulation at clathrin-rich foci
C_LIO_LIAbnormal accumulation of ATG-9 at clathrin-rich foci is associated with defects in activity-dependent presynaptic autophagy
C_LI
]]></description>
<dc:creator>Yang, S.</dc:creator>
<dc:creator>Park, D.</dc:creator>
<dc:creator>Manning, L.</dc:creator>
<dc:creator>Hill, S. E.</dc:creator>
<dc:creator>Cao, M.</dc:creator>
<dc:creator>Xuan, Z.</dc:creator>
<dc:creator>Gonzalez, I.</dc:creator>
<dc:creator>Shao, L.</dc:creator>
<dc:creator>Okeke, I.</dc:creator>
<dc:creator>De Camilli, P.</dc:creator>
<dc:creator>Colon-Ramos, D.</dc:creator>
<dc:date>2020-12-28</dc:date>
<dc:identifier>doi:10.1101/2020.12.28.424508</dc:identifier>
<dc:title><![CDATA[Presynaptic autophagy is coupled to the synaptic vesicle cycle via ATG-9]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.10.07.328906v1?rss=1">
<title>
<![CDATA[
Miro recruits VPS13D to mitochondria 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.10.07.328906v1?rss=1"
</link>
<description><![CDATA[
Mitochondria, which are excluded from the secretory pathway, depend on lipid transport proteins for their lipid supply from the ER, where most lipids are synthesized. In yeast, the outer mitochondrial membrane GTPase Gem1 is an accessory factor of ERMES, an ER-mitochondria tethering complex that contains lipid transport domains and that functions, partially redundantly with Vps13, in lipid transfer between the two organelles. In metazoa, where VPS13, but not ERMES, is present, the Gem1 orthologue Miro was linked to mitochondria dynamics but not to lipid transport. Here we show that Miro, including its peroxisome-enriched splice variant, recruits the lipid transport protein VPS13D, which in turn binds the ER in a VAP-dependent way and thus could provide a lipid conduit between the ER and mitochondria. These findings reveal a so far missing link between function(s) of Gem1/Miro in yeast and higher eukaryotes, where Miro is a Parkin substrate, with potential implications for Parkinsons disease pathogenesis.

SummaryVPS13D mutations result in severe mitochondrial defects. Guillen-Samander et al, show that VPS13D binds VAP in the ER, and interacts with Miro on mitochondria and peroxisomes, so that it can provide a bridge for lipid transport between these organelles.
]]></description>
<dc:creator>Guillen-Samander, A.</dc:creator>
<dc:creator>Leonzino, M.</dc:creator>
<dc:creator>De Camilli, P.</dc:creator>
<dc:date>2020-10-07</dc:date>
<dc:identifier>doi:10.1101/2020.10.07.328906</dc:identifier>
<dc:title><![CDATA[Miro recruits VPS13D to mitochondria]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-10-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.12.15.422954v1?rss=1">
<title>
<![CDATA[
powerEQTL: An R package and shiny application for sample size and power calculation of bulk tissue and single-cell eQTL analysis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.12.15.422954v1?rss=1"
</link>
<description><![CDATA[
SummaryGenome-wide association studies (GWAS) have revealed thousands of genetic loci for common diseases. One of the main challenges in the post-GWAS era is to understand the causality of the genetic variants. Expression quantitative trait locus (eQTL) analysis has been proven to be an effective way to address this question by examining the relationship between gene expression and genetic variation in a sufficiently powered cohort. However, it is often tricky to determine the sample size at which a variant with a specific allele frequency will be detected to associate with gene expression with sufficient power. This is particularly demanding with single-cell RNAseq studies. Therefore, a user-friendly tool to perform power analysis for eQTL at both bulk tissue and single-cell level will be critical. Here, we presented an R package called powerEQTL with flexible functions to calculate power, minimal sample size, or detectable minor allele frequency in both bulk tissue and single-cell eQTL analysis. A user-friendly, program-free web application is also provided, allowing customers to calculate and visualize the parameters interactively.

Availability and implementationThe powerEQTL R package source code and online tutorial are freely available at CRAN: https://cran.r-project.org/web/packages/powerEQTL/. The R shiny application is publicly hosted at https://bwhbioinfo.shinyapps.io/powerEQTL/.

ContactXianjun Dong (xdong@rics.bwh.harvard.edu), Weiliang Qiu (weiliang.qiu@sanofi.com)

Supplementary informationSupplementary data are available at Bioinformatics online.
]]></description>
<dc:creator>Dong, X.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Chang, T.-W.</dc:creator>
<dc:creator>Weiss, S. T.</dc:creator>
<dc:creator>Qiu, W.</dc:creator>
<dc:date>2020-12-16</dc:date>
<dc:identifier>doi:10.1101/2020.12.15.422954</dc:identifier>
<dc:title><![CDATA[powerEQTL: An R package and shiny application for sample size and power calculation of bulk tissue and single-cell eQTL analysis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-12-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.04.12.439050v1?rss=1">
<title>
<![CDATA[
ALS and FTD-associated missense mutations in TBK1 differentially disrupt mitophagy 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.04.12.439050v1?rss=1"
</link>
<description><![CDATA[
TANK-binding kinase 1 (TBK1) is a multi-functional kinase with an essential role in mitophagy, the selective clearance of damaged mitochondria. More than 90 distinct mutations in TBK1 are linked to amyotrophic lateral sclerosis (ALS) and fronto-temporal dementia (FTD), including missense mutations that disrupt the ability of TBK1 to dimerize, associate with the mitophagy receptor optineurin (OPTN), auto-activate, or catalyze phosphorylation. We investigated how ALS-associated mutations in TBK1 affect Parkin-dependent mitophagy using imaging to dissect the molecular mechanisms involved in clearing damaged mitochondria. Some mutations cause severe dysregulation of the pathway, while others induce limited disruption. Mutations that abolish either TBK1 dimerization or kinase activity were insufficient to fully inhibit mitophagy, while mutations that reduced both dimerization and kinase activity were more disruptive. Ultimately, both TBK1 recruitment and OPTN phosphorylation at S177 are necessary for engulfment of damaged mitochondra by autophagosomal membranes. Surprisingly, we find that ULK1 activity contributes to the phosphorylation of OPTN in the presense of either WT- or kinase inactive TBK1. In primary neurons, TBK1 mutants induce mitochondrial stress under basal conditions; network stress is exacerbated with further mitochondrial insult. Our study further refines the model for TBK1 function in mitophagy, demonstrating that some ALS-linked mutations likely contribute to disease pathogenesis by inducing mitochondrial stress or inhibiting mitophagic flux. Other TBK1 mutations exhibited much less impact on mitophagy in our assays, suggesting that cell-type specific effects, cumulative damage, or alternative TBK1-dependent pathways such as innate immunity and inflammation also factor into the development of ALS in affected individuals.

SIGNIFICANCE STATEMENTMissense mutations in TANK-binding kinase 1 (TBK1) have various biophysical and biochemical effects on the molecule, and are associated with the neurodegenerative diseases amyotrophic lateral sclerosis (ALS) and fronto-temporal dementia (FTD). TBK1 plays an essential role in clearing damaged mitochondria. Here, we investigate the impact of 10 ALS-linked TBK1 mutations on the critical early stage of mitophagy. We find that both TBK1 recruitment and kinase activity contribute to the clearance of the damaged mitochondria. Furthermore, in neurons, expression of TBK1 mutants alone affects mitochondrial network health. Our investigation utilizes disease-linked mutations to further refine the current model of mitophagy, identifying crosstalk between the regulatory kinases TBK1 and ULK1, and providing new insights into the roles of TBK1 in neurodegenerative pathogenesis.
]]></description>
<dc:creator>Harding, O.</dc:creator>
<dc:creator>Evans, C. S.</dc:creator>
<dc:creator>Ye, J.</dc:creator>
<dc:creator>Cheung, J.</dc:creator>
<dc:creator>Maniatis, T.</dc:creator>
<dc:creator>Holzbaur, E. L. F.</dc:creator>
<dc:date>2021-04-12</dc:date>
<dc:identifier>doi:10.1101/2021.04.12.439050</dc:identifier>
<dc:title><![CDATA[ALS and FTD-associated missense mutations in TBK1 differentially disrupt mitophagy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.05.14.444175v1?rss=1">
<title>
<![CDATA[
Structural Basis for Membrane Recruitment of ATG16L1 by WIPI2 in Autophagy 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.05.14.444175v1?rss=1"
</link>
<description><![CDATA[
Autophagy is a cellular process that degrades cytoplasmic cargo by engulfing it in a double membrane vesicle, known as the autophagosome, and delivering it to the lysosome. The ATG12-5-16L1 complex is responsible for conjugating members of the ubiquitin-like ATG8 protein family to phosphatidylethanolamine in the growing autophagosomal membrane, known as the phagophore. ATG12-5-16L1 is recruited to the phagophore by a subset of the phosphatidylinositol 3-phosphate-binding seven bladed {beta}-propeller WIPI proteins. We determined the crystal structure of WIPI2d in complex with the WIPI2 interacting region (W2IR) of ATG16L1 comprising residues 207-230 at 1.85 [A] resolution. The structure shows that the ATG16L1 W2IR adopts an alpha helical conformation and binds in an electropositive and hydrophobic groove between WIPI2 {beta}-propeller blades 2 and 3. Mutation of residues at the interface reduces or blocks the recruitment of ATG12-5-16L1 and the conjugation of the ATG8 protein LC3B to synthetic membranes. Interface mutants show a decrease in starvation-induced autophagy. Comparisons across the four human WIPIs suggest that WIPI1 and 2 belong to a W2IR-binding subclass responsible for localizing ATG12-5-16L1 and driving ATG8 lipidation, whilst WIPI3 and 4 belong to a second W34IR-binding subclass responsible for localizing ATG2, and so directing lipid supply to the nascent phagophore. The structure provides a framework for understanding the regulatory node connecting two central events in autophagy initiation, the action of the autophagic PI 3-kinase complex on the one hand, and ATG8 lipidation on the other.
]]></description>
<dc:creator>Strong, L. M.</dc:creator>
<dc:creator>Chang, C.</dc:creator>
<dc:creator>Boecker, A.</dc:creator>
<dc:creator>Flower, T. G.</dc:creator>
<dc:creator>Buffalo, C. Z.</dc:creator>
<dc:creator>Ren, X.</dc:creator>
<dc:creator>Stavoe, A. K.</dc:creator>
<dc:creator>Holzbaur, E. L. F.</dc:creator>
<dc:creator>Hurley, J. H.</dc:creator>
<dc:date>2021-05-14</dc:date>
<dc:identifier>doi:10.1101/2021.05.14.444175</dc:identifier>
<dc:title><![CDATA[Structural Basis for Membrane Recruitment of ATG16L1 by WIPI2 in Autophagy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.06.01.446584v1?rss=1">
<title>
<![CDATA[
Subcellular proteomics of dopamine neurons in the mouse brain reveals axonal enrichment of proteins encoded by Parkinson's disease-linked genes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.06.01.446584v1?rss=1"
</link>
<description><![CDATA[
Dopaminergic neurons modulate neural circuits and behaviors via dopamine release from expansive, long range axonal projections. The elaborate cytoarchitecture of these neurons is embedded within complex brain tissue, making it difficult to access the neuronal proteome using conventional methods. Here, we demonstrate APEX2 proximity labeling within genetically targeted neurons in the mouse brain, enabling subcellular proteomics with cell type-specificity. By combining APEX2 biotinylation with mass spectrometry, we mapped the somatodendritic and axonal proteomes of midbrain dopaminergic neurons. Our dataset reveals the proteomic architecture underlying proteostasis, axonal metabolism, and neurotransmission in these neurons. We find a significant enrichment of proteins encoded by Parkinsons disease-linked genes in striatal dopaminergic axons, including proteins with previously undescribed axonal localization. These proteomic datasets provide a resource for neuronal cell biology, and this approach can be readily adapted for study of other neural cell types.
]]></description>
<dc:creator>Hobson, B. D.</dc:creator>
<dc:creator>Choi, S. J.</dc:creator>
<dc:creator>Soni, R. K.</dc:creator>
<dc:creator>Sulzer, D.</dc:creator>
<dc:creator>Sims, P. A.</dc:creator>
<dc:date>2021-06-01</dc:date>
<dc:identifier>doi:10.1101/2021.06.01.446584</dc:identifier>
<dc:title><![CDATA[Subcellular proteomics of dopamine neurons in the mouse brain reveals axonal enrichment of proteins encoded by Parkinson's disease-linked genes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.01.14.425874v1?rss=1">
<title>
<![CDATA[
Integrating protein networks and machine learning for disease stratification in the Hereditary Spastic Paraplegias 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.01.14.425874v1?rss=1"
</link>
<description><![CDATA[
The Hereditary Spastic Paraplegias are a group of neurodegenerative diseases characterized by spasticity and weakness in the lower body. Despite the identification of causative mutations in over 70 genes, the molecular aetiology remains unclear. Due to the combination of genetic diversity and variable clinical presentation, the Hereditary Spastic Paraplegias are a strong candidate for protein-protein interaction network analysis as a tool to understand disease mechanism(s) and to aid functional stratification of phenotypes. In this study, experimentally validated human protein-protein interactions were used to create a protein-protein interaction network based on the causative Hereditary Spastic Paraplegia genes. Network evaluation as a combination of both topological analysis and functional annotation led to the identification of core proteins in putative shared biological processes such as intracellular transport and vesicle trafficking. The application of machine learning techniques suggested a functional dichotomy linked with distinct sets of clinical presentations, suggesting there is scope to further classify conditions currently described under the same umbrella term of Hereditary Spastic Paraplegias based on specific molecular mechanisms of disease.
]]></description>
<dc:creator>Vavouraki, N.</dc:creator>
<dc:creator>Tomkins, J. E.</dc:creator>
<dc:creator>Kara, E.</dc:creator>
<dc:creator>Houlden, H.</dc:creator>
<dc:creator>Hardy, J.</dc:creator>
<dc:creator>Tindall, M. J.</dc:creator>
<dc:creator>Lewis, P. A.</dc:creator>
<dc:creator>Manzoni, C.</dc:creator>
<dc:date>2021-01-16</dc:date>
<dc:identifier>doi:10.1101/2021.01.14.425874</dc:identifier>
<dc:title><![CDATA[Integrating protein networks and machine learning for disease stratification in the Hereditary Spastic Paraplegias]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-01-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.05.28.446215v1?rss=1">
<title>
<![CDATA[
Transcriptional analysis of peripheral memory T cells reveals Parkinson's disease-specific gene signatures 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.05.28.446215v1?rss=1"
</link>
<description><![CDATA[
Parkinsons disease (PD) is a multi-stage neurodegenerative disorder with largely unknown etiology. Recent findings have identified PD-associated autoimmune features including roles for T cells. To further characterize the role of T cells in PD, we performed RNA sequencing on PBMC and peripheral CD4 and CD8 memory T cell subsets derived from PD patients and age-matched healthy controls. When the groups were stratified by their T cell responsiveness to alpha-synuclein (-syn) as a proxy for ongoing inflammatory autoimmune response, the study revealed a broad differential gene expression profile in memory T cell subsets and a specific PD associated gene signature. We identified a significant enrichment of transcriptomic signatures previously associated with PD, including for oxidative stress, phosphorylation, autophagy of mitochondria, cholesterol metabolism and inflammation, and the chemokine signaling proteins CX3CR1, CCR5 and CCR1. In addition, we identified genes in these peripheral cells that have previously been shown to be involved in PD pathogenesis and expressed in neurons, such as LRRK2, LAMP3, and aquaporin. Together, these findings suggest that features of circulating T cells with -syn-specific responses in PD patients provide insights into the interactive processes that occur during PD pathogenesis and suggest potential intervention targets.
]]></description>
<dc:creator>Dhanwani, R.</dc:creator>
<dc:creator>Rodrigues Lima-Junior, J.</dc:creator>
<dc:creator>Sethi, A.</dc:creator>
<dc:creator>Pham, J.</dc:creator>
<dc:creator>Williams, G.</dc:creator>
<dc:creator>Frazier, A.</dc:creator>
<dc:creator>Xu, Y.</dc:creator>
<dc:creator>Amara, A. W.</dc:creator>
<dc:creator>Standaert, D. G.</dc:creator>
<dc:creator>Goldman, J. G.</dc:creator>
<dc:creator>Litvan, I.</dc:creator>
<dc:creator>Alcalay, R. N.</dc:creator>
<dc:creator>Peters, B.</dc:creator>
<dc:creator>Sulzer, D.</dc:creator>
<dc:creator>Lindestam Arlehamn, C. S.</dc:creator>
<dc:creator>Sette, A.</dc:creator>
<dc:date>2021-05-29</dc:date>
<dc:identifier>doi:10.1101/2021.05.28.446215</dc:identifier>
<dc:title><![CDATA[Transcriptional analysis of peripheral memory T cells reveals Parkinson's disease-specific gene signatures]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-05-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.06.08.447593v1?rss=1">
<title>
<![CDATA[
ER-lysosome lipid transfer protein VPS13C/PARK23 prevents aberrant mtDNA-dependent STING signaling 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.06.08.447593v1?rss=1"
</link>
<description><![CDATA[
Mutations in VPS13C cause early onset, autosomal recessive Parkinsons Disease (PD). We have established that VPS13C encodes a lipid transfer protein localized to contact sites between the endoplasmic reticulum (ER) and late endosomes/lysosomes. In the current study, we demonstrate that depleting VPS13C in HeLa cells causes an accumulation of lysosomes with an altered lipid profile, including an accumulation of di-22:6-BMP, a biomarker of the PD-associated leucine-rich repeat kinase 2 (LRRK2) G2019S mutation. In addition, the DNA-sensing cGAS/STING pathway, which was recently implicated in PD pathogenesis, is activated in these cells. This activation results from a combination of elevated mitochondrial DNA in the cytosol and a defect in the degradation of activated STING, a lysosome-dependent process. These results suggest a link between ER-lysosome lipid transfer and innate immune activation and place VPS13C in pathways relevant to PD pathogenesis.
]]></description>
<dc:creator>Hancock-Cerutti, W. F.</dc:creator>
<dc:creator>Wu, Z.</dc:creator>
<dc:creator>Tharkeshwar, A. K.</dc:creator>
<dc:creator>Ferguson, S. M.</dc:creator>
<dc:creator>Shadel, G. S.</dc:creator>
<dc:creator>De Camilli, P.</dc:creator>
<dc:date>2021-06-08</dc:date>
<dc:identifier>doi:10.1101/2021.06.08.447593</dc:identifier>
<dc:title><![CDATA[ER-lysosome lipid transfer protein VPS13C/PARK23 prevents aberrant mtDNA-dependent STING signaling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.04.12.439539v1?rss=1">
<title>
<![CDATA[
Targeted Neuronal Activation of the Gastrointestinal Tract Shapes the Environment of the Gut in Mice 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.04.12.439539v1?rss=1"
</link>
<description><![CDATA[
The gastrointestinal (GI) tract is extensively innervated by intrinsic neurons of the enteric nervous system (ENS) and extrinsic neurons of the central nervous system and peripheral ganglia, which together regulate gut physiology. The GI tract also harbors a diverse microbiome, but interactions between the ENS and the microbiome remain poorly understood. Herein, we activate choline acetyltransferase (ChAT)-expressing or tyrosine hydroxylase (TH)-expressing gut-associated neurons in mice to determine effects on intestinal microbial communities and their metabolites, as well as on host physiology. The resulting multi-omics datasets support broad roles for discrete peripheral neuronal subtypes in shaping microbiome structure, including modulating bile acid profiles and fungal colonization. Physiologically, activation of either ChAT+ or TH+ neurons increases fecal output, while only ChAT+ activation results in increased colonic migrating motor complexes and diarrhea-like fluid secretion. These findings suggest that specific subsets of peripherally-activated ENS neurons differentially regulate the gut microbiome and GI physiology in mice, without involvement of signals from the brain.
]]></description>
<dc:creator>Yoo, B. B.</dc:creator>
<dc:creator>Griffiths, J. A.</dc:creator>
<dc:creator>Thuy-Boun, P.</dc:creator>
<dc:creator>Cantu, V.</dc:creator>
<dc:creator>Weldon, K.</dc:creator>
<dc:creator>Challis, C.</dc:creator>
<dc:creator>Sweredoski, M. J.</dc:creator>
<dc:creator>Chan, K. Y.</dc:creator>
<dc:creator>Thron, T. M.</dc:creator>
<dc:creator>Sharon, G.</dc:creator>
<dc:creator>Moradian, A.</dc:creator>
<dc:creator>Humphrey, G.</dc:creator>
<dc:creator>Zhu, Q.</dc:creator>
<dc:creator>Shaffer, J.</dc:creator>
<dc:creator>Wolan, D. W.</dc:creator>
<dc:creator>Dorrestein, P.</dc:creator>
<dc:creator>Knight, R.</dc:creator>
<dc:creator>Gradinaru, V.</dc:creator>
<dc:creator>Mazmanian, S.</dc:creator>
<dc:date>2021-04-13</dc:date>
<dc:identifier>doi:10.1101/2021.04.12.439539</dc:identifier>
<dc:title><![CDATA[Targeted Neuronal Activation of the Gastrointestinal Tract Shapes the Environment of the Gut in Mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.06.04.447104v1?rss=1">
<title>
<![CDATA[
HyDrop: droplet-based scATAC-seq and scRNA-seq using dissolvable hydrogel beads 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.06.04.447104v1?rss=1"
</link>
<description><![CDATA[
Single-cell RNA-seq and single-cell ATAC-seq technologies are being used extensively to create cell type atlases for a wide range of organisms, tissues, and disease processes. To increase the scale of these atlases, lower the cost, and allow for more specialized multi-ome assays, custom droplet microfluidics may provide complementary solutions to commercial setups. We developed HyDrop, a flexible and generic droplet microfluidic platform encompassing three protocols. The first protocol involves creating dissolvable hydrogel beads with custom oligos that can be released in the droplets. In the second protocol, we demonstrate the use of these beads for HyDrop-ATAC, a low-cost non-commercial scATAC-seq protocol in droplets. After validating HyDrop-ATAC, we applied it to flash-frozen mouse cortex and generated 8,502 high-quality single-cell chromatin accessibility profiles in a single run. In the third protocol, we adapt both the reaction chemistry and the capture sequence of the barcoded hydrogel bead to capture mRNA, and demonstrate a significant improvement in throughput and sensitivity compared to previous open-source droplet-based scRNA-seq assays (Drop-seq and inDrop). Similarly, we applied HyDrop-RNA to flash-frozen mouse cortex and generated 9,508 single-cell transcriptomes closely matching reference single-cell gene expression data. Finally, we leveraged HyDrop-RNAs high capture rate to analyse a small population of FAC-sorted neurons from the Drosophila brain, confirming the protocols applicability to low-input samples and small cells. HyDrop is currently capable of generating single-cell data in high throughput and at a reduced cost compared to commercial methods, and we envision that HyDrop can be further developed to be compatible with novel (multi-) omics protocols.
]]></description>
<dc:creator>De Rop, F. V.</dc:creator>
<dc:creator>Ismail, J. N.</dc:creator>
<dc:creator>Bravo Gonzalez-Blas, C.</dc:creator>
<dc:creator>Hulselmans, G. J.</dc:creator>
<dc:creator>Flerin, C. C.</dc:creator>
<dc:creator>Janssens, J.</dc:creator>
<dc:creator>Theunis, K.</dc:creator>
<dc:creator>Christiaens, V. M.</dc:creator>
<dc:creator>Wouters, J.</dc:creator>
<dc:creator>Marcassa, G.</dc:creator>
<dc:creator>de Wit, J.</dc:creator>
<dc:creator>Poovathingal, S.</dc:creator>
<dc:creator>Aerts, S.</dc:creator>
<dc:date>2021-06-06</dc:date>
<dc:identifier>doi:10.1101/2021.06.04.447104</dc:identifier>
<dc:title><![CDATA[HyDrop: droplet-based scATAC-seq and scRNA-seq using dissolvable hydrogel beads]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-06-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.07.16.452659v1?rss=1">
<title>
<![CDATA[
The extracellular chaperone Clusterin enhances Tau aggregate seeding in a cellular model 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.07.16.452659v1?rss=1"
</link>
<description><![CDATA[
Spreading of aggregate pathology across brain regions acts as a driver of disease progression in Tau-related neurodegeneration, including Alzheimers disease (AD) and frontotemporal dementia. Aggregate seeds released from affected cells are internalized by naive cells and induce the prion-like templating of soluble Tau into neurotoxic aggregates. Here we show in a cellular model system and in neurons that Clusterin, an abundant extracellular chaperone, strongly enhances Tau aggregate seeding. Upon interaction with Tau aggregates, Clusterin stabilizes highly potent, soluble seed species. Tau/Clusterin complexes enter recipient cells via endocytosis and compromise the endolysosomal compartment, allowing transfer to the cytosol where they propagate aggregation of endogenous Tau. Thus, upregulation of Clusterin, as observed in AD patients, may enhance Tau seeding and possibly accelerate the spreading of Tau pathology.
]]></description>
<dc:creator>Yuste-Checa, P.</dc:creator>
<dc:creator>Trinkaus, V. A.</dc:creator>
<dc:creator>Riera-Tur, I.</dc:creator>
<dc:creator>Imamoglu, R.</dc:creator>
<dc:creator>Schaller, T.</dc:creator>
<dc:creator>Wang, H.</dc:creator>
<dc:creator>Dudanova, I.</dc:creator>
<dc:creator>Hipp, M. S.</dc:creator>
<dc:creator>Bracher, A.</dc:creator>
<dc:creator>Hartl, F. U.</dc:creator>
<dc:date>2021-07-16</dc:date>
<dc:identifier>doi:10.1101/2021.07.16.452659</dc:identifier>
<dc:title><![CDATA[The extracellular chaperone Clusterin enhances Tau aggregate seeding in a cellular model]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.07.19.452535v1?rss=1">
<title>
<![CDATA[
Quantitative proteomics reveals the selectivity of ubiquitin-binding autophagy receptors in the turnover of damaged lysosomes by lysophagy 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.07.19.452535v1?rss=1"
</link>
<description><![CDATA[
Removal of damaged organelles via the process of selective autophagy constitutes a major form of cellular quality control. Damaged organelles are recognized by a dedicated surveillance machinery, leading to the assembly of an autophagosome around the damaged organelle, prior to fusion with the degradative lysosomal compartment. Lysosomes themselves are also prone to damage and are degraded through the process of lysophagy. While early steps involve recognition of ruptured lysosomal membranes by glycan-binding Galectins and ubiquitylation of transmembrane lysosomal proteins, many steps in the process, and their inter-relationships, remain poorly understood, including the role and identity of cargo receptors required for completion of lysophagy. Here, we employ quantitative organelle capture and proximity biotinylation proteomics of autophagy adaptors, cargo receptors, and Galectins in response to acute lysosomal damage, thereby revealing the landscape of lysosomal proteome remodeling during lysophagy. Among proteins dynamically recruited to damaged lysosomes were ubiquitin-binding autophagic cargo receptors. Using newly developed lysophagic flux reporters including Lyso-Keima, we demonstrate that TAX1BP1, together with its associated kinase TBK1, are both necessary and sufficient to promote lysophagic flux in both Hela cells and induced neurons (iNeurons). While the related receptor OPTN can drive damage-dependent lysophagy when overexpressed, cells lacking either OPTN or CALCOCO2 still maintain significant lysophagic flux in HeLa cells. Mechanistically, TAX1BP1-driven lysophagy requires its N-terminal SKICH domain, which binds both TBK1 and the autophagy regulatory factor RB1CC1, and requires upstream ubiquitylation events for efficient recruitment and lysophagic flux. These results identify TAX1BP1 as a central component in the lysophagy pathway and provide a proteomic resource for future studies of the lysophagy process.
]]></description>
<dc:creator>Eapen, V.</dc:creator>
<dc:creator>Swarup, S.</dc:creator>
<dc:creator>Hoyer, M. J.</dc:creator>
<dc:creator>Paulo, J. A.</dc:creator>
<dc:creator>Harper, J. W.</dc:creator>
<dc:date>2021-07-19</dc:date>
<dc:identifier>doi:10.1101/2021.07.19.452535</dc:identifier>
<dc:title><![CDATA[Quantitative proteomics reveals the selectivity of ubiquitin-binding autophagy receptors in the turnover of damaged lysosomes by lysophagy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.04.01.438131v1?rss=1">
<title>
<![CDATA[
Global ubiquitylation analysis of mitochondria in primary neurons identifies physiological Parkin targets following activation of PINK1 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.04.01.438131v1?rss=1"
</link>
<description><![CDATA[
Autosomal recessive mutations in PINK1 and Parkin cause Parkinsons disease. How activation of PINK1 and Parkin leads to elimination of damaged mitochondria by mitophagy is largely based on cell culture studies with few molecular studies in neurons. Herein we have undertaken a global proteomic-analysis of mitochondria from mouse neurons to identify ubiquitylated substrates of endogenous Parkin activation. Comparative analysis with human iNeuron datasets revealed a subset of 49 PINK1-dependent diGLY sites upregulated upon mitochondrial depolarisation in 22 proteins conserved across mouse and human systems. These proteins were exclusively localised at the mitochondrial outer membrane (MOM) including, CISD1, CPT1, ACSL1, and FAM213A. We demonstrate that these proteins can be directly ubiquitylated by Parkin in vitro. We also provide evidence for a subset of cytoplasmic proteins recruited to mitochondria that undergo PINK1 and Parkin independent ubiquitylation including SNX3, CAMK2 and CAMK2{beta} indicating the presence of alternate ubiquitin E3 ligase pathways that are activated by mitochondrial depolarisation in neurons. Finally we have developed an online resource to visualise mitochondrial ubiquitin sites in neurons and search for ubiquitin components recruited to mitochondria upon mitochondrial depolarisation, MitoNUb. This analysis will aid in future studies to understand Parkin activation in neuronal subtypes. Our findings also suggest that monitoring ubiquitylation status of the 22 identified MOM proteins may represent robust biomarkers for PINK1 and Parkin activity in vivo.
]]></description>
<dc:creator>Antico, O.</dc:creator>
<dc:creator>Ordureau, A.</dc:creator>
<dc:creator>Stevens, M.</dc:creator>
<dc:creator>Singh, F.</dc:creator>
<dc:creator>Gierlinski, M.</dc:creator>
<dc:creator>Barini, E.</dc:creator>
<dc:creator>Rickwood, M.</dc:creator>
<dc:creator>Prescott, A. R.</dc:creator>
<dc:creator>Toth, R.</dc:creator>
<dc:creator>Ganley, I.</dc:creator>
<dc:creator>Harper, W.</dc:creator>
<dc:creator>Muqit, M.</dc:creator>
<dc:date>2021-04-01</dc:date>
<dc:identifier>doi:10.1101/2021.04.01.438131</dc:identifier>
<dc:title><![CDATA[Global ubiquitylation analysis of mitochondria in primary neurons identifies physiological Parkin targets following activation of PINK1]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.07.30.454065v1?rss=1">
<title>
<![CDATA[
Subcellular and regional localization of mRNA translation in midbrain dopamine neurons 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.07.30.454065v1?rss=1"
</link>
<description><![CDATA[
Local translation within excitatory and inhibitory neurons is involved in neuronal development and synaptic plasticity. Despite the extensive dendritic and axonal arborizations of central monoaminergic neurons, the subcellular localization of protein synthesis is not well-characterized in these populations. Here, we investigated mRNA localization and translation in midbrain dopaminergic (mDA) neurons, cells with enormous axonal and dendritic projections, both of which exhibit stimulation-evoked dopamine (DA) release. Using highly-sensitive ribosome-bound RNA-sequencing and imaging approaches in mDA axons, we found no evidence for axonal mRNA localization or translation. In contrast, mDA neuronal dendritic projections into the substantia nigra reticulata (SNr) contain ribosomes and mRNAs encoding the major components of DA synthesis, release, and reuptake machinery. Surprisingly, we also observed dendritic localization of mRNAs encoding synaptic vesicle-related proteins, including those involved in vesicular exocytic fusion. Our results are consistent with a role for local translation in the regulation of DA release from dendrites, but not from axons. Our translatome data further defined a molecular signature of the sparse mDA neurons resident in the SNr, including enrichment of Atp2a3/SERCA3, an ER calcium pump previously undescribed in mDA neurons.
]]></description>
<dc:creator>Hobson, B. D.</dc:creator>
<dc:creator>Kong, L.</dc:creator>
<dc:creator>Angelo, M. F.</dc:creator>
<dc:creator>Lieberman, O. J.</dc:creator>
<dc:creator>Mosharov, E. V.</dc:creator>
<dc:creator>Herzog, E.</dc:creator>
<dc:creator>Sulzer, D.</dc:creator>
<dc:creator>Sims, P. A.</dc:creator>
<dc:date>2021-08-01</dc:date>
<dc:identifier>doi:10.1101/2021.07.30.454065</dc:identifier>
<dc:title><![CDATA[Subcellular and regional localization of mRNA translation in midbrain dopamine neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-08-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.11.04.467353v1?rss=1">
<title>
<![CDATA[
SHIP164 is a Chorein Motif Containing Lipid Transport Protein that Controls Membrane Dynamics and Traffic at the Endosome-Golgi Interface. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.11.04.467353v1?rss=1"
</link>
<description><![CDATA[
Cellular membranes differ in protein and lipid composition as well as in the protein-lipid ratio. Thus, progression of membranous organelles along traffic routes requires mechanisms to control bilayer lipid chemistry and their abundance relative to proteins. The recent structural and functional characterization of VPS13-family proteins has suggested a mechanism through which lipids can be transferred in bulk from one membrane to another at membrane contact sites, and thus independently of vesicular traffic. Here we show that SHIP164 (UHRF1BP1L) shares structural and lipid transfer properties with these proteins and is localized on a subpopulation of vesicle clusters in the early endocytic pathway whose membrane cargo includes the cation-independent mannose-6-phosphate receptor (MPR) and ATG9. Loss of SHIP164 disrupts retrograde traffic of these organelles to the Golgi complex. Our findings raise the possibility that bulk transfer of lipids to endocytic membranes may play a role in their traffic.
]]></description>
<dc:creator>Hanna, M. G.</dc:creator>
<dc:creator>Suen, P.</dc:creator>
<dc:creator>Wu, Y.</dc:creator>
<dc:creator>Reinisch, K. M.</dc:creator>
<dc:creator>De Camilli, P.</dc:creator>
<dc:date>2021-11-04</dc:date>
<dc:identifier>doi:10.1101/2021.11.04.467353</dc:identifier>
<dc:title><![CDATA[SHIP164 is a Chorein Motif Containing Lipid Transport Protein that Controls Membrane Dynamics and Traffic at the Endosome-Golgi Interface.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-11-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.09.27.461935v1?rss=1">
<title>
<![CDATA[
The activities of LRRK2 and GCase are positively correlated in clinical biospecimens and experimental models of Parkinson's disease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.09.27.461935v1?rss=1"
</link>
<description><![CDATA[
LRRK2 is a kinase involved in different cellular functions, including autophagy, endolysosomal pathways and vesicle trafficking. Mutations in LRRK2 cause autosomal dominant forms of Parkinsons disease (PD). Heterozygous mutations in GBA1, the gene encoding the lysosomal enzyme glucocerebrosidase (GCase), are the most common genetic risk factors for PD. Moreover, GCase function is altered in idiopathic PD and in other genetic forms of the disease. Recent work suggests that LRRK2 kinase activity can regulate GCase function. However, both a positive and a negative correlation have been described. To gain insights into the impact of LRRK2 on GCase, we investigated GCase levels and activity in LRRK2 G2019S knockin mice, in clinical biospecimens from PD patients carrying this mutation and in patient-derived cellular models. In these models we found a positive correlation between the activities of LRRK2 and GCase, which was further confirmed in cell lines with genetic and pharmacological manipulation of LRRK2 kinase activity. Overall, our study indicates that LRRK2 kinase activity affects both the levels and the catalytic activity of GCase.
]]></description>
<dc:creator>Kedariti, M.</dc:creator>
<dc:creator>Frattini, E.</dc:creator>
<dc:creator>Baden, P.</dc:creator>
<dc:creator>Cogo, S.</dc:creator>
<dc:creator>Civiero, L.</dc:creator>
<dc:creator>Ziviani, E.</dc:creator>
<dc:creator>Aureli, M.</dc:creator>
<dc:creator>Kaganovich, A.</dc:creator>
<dc:creator>Cookson, M. R.</dc:creator>
<dc:creator>Stefanis, L. R.</dc:creator>
<dc:creator>Surface, M.</dc:creator>
<dc:creator>Deleidi, M.</dc:creator>
<dc:creator>Di Fonzo, A.</dc:creator>
<dc:creator>Alcalay, R. N.</dc:creator>
<dc:creator>Rideout, H.</dc:creator>
<dc:creator>Greggio, E.</dc:creator>
<dc:creator>Plotegher, N.</dc:creator>
<dc:date>2021-09-28</dc:date>
<dc:identifier>doi:10.1101/2021.09.27.461935</dc:identifier>
<dc:title><![CDATA[The activities of LRRK2 and GCase are positively correlated in clinical biospecimens and experimental models of Parkinson's disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.01.08.475342v1?rss=1">
<title>
<![CDATA[
Intravenous gene transfer throughout the brain of infant Old World primates using AAV 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.01.08.475342v1?rss=1"
</link>
<description><![CDATA[
Adeno-associated viruses (AAVs) can enable robust and safe gene delivery to the mammalian central nervous system (CNS). While the scientific community has developed numerous neurotropic AAV variants for systemic gene-transfer to the rodent brain, there are few AAVs that efficiently access the CNS of higher order primates. We describe here AAV.CAP-Mac, an engineered AAV variant that enables systemic, brain-wide gene delivery in infants of two Old World primate species--the rhesus macaque (Macaca mulatta) and the green monkey (Chlorocebus sabaeus). We identified CAP-Mac using a multi-species selection strategy, initially screening our library in the adult common marmoset (Callithrix jacchus) and narrowing our pool of test-variants for another round of selection in infant macaques. In individual characterization, CAP-Mac robustly transduces human neurons in vitro and Old World primate neurons in vivo, where it targets all lobes of cortex, the cerebellum, and multiple subcortical regions of disease relevance. We use CAP-Mac for Brainbow-like multicolor labeling of macaque neurons throughout the brain, enabling morphological reconstruction of both medium spiny neurons and cortical pyramidal cells. Because of its broad distribution throughout the brain and high neuronal efficiency in infant Old World primates compared to AAV9, CAP-Mac shows promise for researchers and clinicians alike to unlock novel, noninvasive access to the brain for efficient gene transfer.
]]></description>
<dc:creator>Chuapoco, M. R.</dc:creator>
<dc:creator>Flytzanis, N.</dc:creator>
<dc:creator>Goeden, N.</dc:creator>
<dc:creator>Octeau, J. C.</dc:creator>
<dc:creator>Roxas, K. M.</dc:creator>
<dc:creator>Chan, K. Y.</dc:creator>
<dc:creator>Scherrer, J.</dc:creator>
<dc:creator>Winchester, J.</dc:creator>
<dc:creator>Blackburn, R. J.</dc:creator>
<dc:creator>Campos, L. J.</dc:creator>
<dc:creator>Arokiaraj, C. M.</dc:creator>
<dc:creator>Miles, T. F.</dc:creator>
<dc:creator>Jang, M. J.</dc:creator>
<dc:creator>Vendemiatti, J.</dc:creator>
<dc:creator>Deverman, B. E.</dc:creator>
<dc:creator>Pickel, J.</dc:creator>
<dc:creator>Fox, A. S.</dc:creator>
<dc:creator>Gradinaru, V.</dc:creator>
<dc:date>2022-01-09</dc:date>
<dc:identifier>doi:10.1101/2022.01.08.475342</dc:identifier>
<dc:title><![CDATA[Intravenous gene transfer throughout the brain of infant Old World primates using AAV]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/830802v1?rss=1">
<title>
<![CDATA[
Mutations in Auxilin cause parkinsonism via impaired clathrin-mediated trafficking at the Golgi apparatus and synapse 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/830802v1?rss=1"
</link>
<description><![CDATA[
Parkinsons disease (PD) is a common neurodegenerative motor disorder characterized in part by neuropathological lesions in the nigrostriatal pathway. Loss of function mutations in Auxilin, the major neuronal clathrin uncoating protein, cause an aggressive form of juvenile onset PD. How mutations in Auxilin cause PD, is currently not understood. Here, we generated a novel mouse model carrying an endogenous pathogenic Auxilin mutation that phenocopies neurological features observed in patients, including motor impairments and seizures. Unbiased mapping of the Auxilin interactome identified synaptic and Golgi-resident clathrin adaptor proteins as novel interactors. Impaired clathrin-mediated trafficking in mutant Auxilin mice, both at the Golgi and the synapse, results in neuropathological lesions in the nigrostriatal pathway. Collectively, these results provide molecular mechanisms of PD pathogenesis in Auxilin mutation carriers, reinforcing a key role for clathrin-mediated trafficking in PD, and expand our understanding of the cellular function of Auxilin.
]]></description>
<dc:creator>Roosen, D. A.</dc:creator>
<dc:creator>Landeck, N.</dc:creator>
<dc:creator>Conti, M.</dc:creator>
<dc:creator>Smith, N.</dc:creator>
<dc:creator>Saez-Atienzar, S.</dc:creator>
<dc:creator>Ding, J.</dc:creator>
<dc:creator>Beilina, A.</dc:creator>
<dc:creator>Kumaran, R.</dc:creator>
<dc:creator>Kaganovich, A.</dc:creator>
<dc:creator>du Hoffmann, J.</dc:creator>
<dc:creator>Williamson, C.</dc:creator>
<dc:creator>Gershlick, D. C.</dc:creator>
<dc:creator>Bonet-Ponce, L.</dc:creator>
<dc:creator>Sampieri, L.</dc:creator>
<dc:creator>Bleck, C. K. E.</dc:creator>
<dc:creator>Liu, C.</dc:creator>
<dc:creator>Bonifacino, J. S.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Lewis, P. A.</dc:creator>
<dc:creator>Cookson, M. R.</dc:creator>
<dc:date>2019-11-05</dc:date>
<dc:identifier>doi:10.1101/830802</dc:identifier>
<dc:title><![CDATA[Mutations in Auxilin cause parkinsonism via impaired clathrin-mediated trafficking at the Golgi apparatus and synapse]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-11-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.02.14.480397v1?rss=1">
<title>
<![CDATA[
Single-cell transcriptomic and proteomic analysis of Parkinson's disease Brains 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.02.14.480397v1?rss=1"
</link>
<description><![CDATA[
Parkinsons disease (PD) is a prevalent neurodegenerative disorder where recent evidence suggests pathogenesis may be mediated by inflammatory processes. The molecular architecture of the disease remains to be fully elucidated. We performed single-nucleus transcriptomics and unbiased proteomics using postmortem tissue obtained from the prefrontal cortex of 12 individuals with late-stage PD and age-matched controls. We analyzed [~]80,000 nuclei and identified eight major cell types, including brain-resident T cells, each with distinct transcriptional changes in line with the known genetics of PD. By analyzing Lewy body pathology in the same postmortem tissue, we found that -synuclein pathology is inversely correlated with chaperone expression in excitatory neurons. Examining cell-cell interactions, we found a selective abatement of neuron-astrocyte interactions and enhanced neuroinflammation. Proteomic analyses of the same brains identified synaptic proteins in prefrontal cortex that were preferentially downregulated in PD. Strikingly, comparing this dataset to a regionally similar published analysis for Alzheimers disease (AD), we found no common differentially expressed genes in neurons, but identified many shared differentially expressed genes in glial cells, suggesting that disease etiology in PD and AD are likely distinct. These data are presented as a resource for interrogating the molecular and cellular basis of PD and other neurodegenerative diseases.

One Sentence SummaryWe provide an extensive single cell analysis profiling nearly 80,000 brain nuclei from prefrontal cortex of late-stage Parkinsons disease brains, demonstrate that -synuclein pathology is inversely correlated with chaperone expression in excitatory neurons, found a selective abatement of neuron-astrocyte interactions with enhanced neuroinflammation, and augmented the study with proteomic analysis and cross-comparisons with Alzheimers disease datasets, providing valuable insights into the pathways of neurodegeneration and a deep definition of the underlying molecular pathology for Parkinsons disease.
]]></description>
<dc:creator>Zhu, B.</dc:creator>
<dc:creator>Park, J.-M.</dc:creator>
<dc:creator>Coffey, S.</dc:creator>
<dc:creator>Hsu, I.-U.</dc:creator>
<dc:creator>Lam, T. T.</dc:creator>
<dc:creator>Gopal, P. P.</dc:creator>
<dc:creator>Ginsberg, S. D.</dc:creator>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Su, C.</dc:creator>
<dc:creator>Zhao, H.</dc:creator>
<dc:creator>Hafler, D. A.</dc:creator>
<dc:creator>Chandra, S.</dc:creator>
<dc:creator>Zhang, L.</dc:creator>
<dc:date>2022-02-14</dc:date>
<dc:identifier>doi:10.1101/2022.02.14.480397</dc:identifier>
<dc:title><![CDATA[Single-cell transcriptomic and proteomic analysis of Parkinson's disease Brains]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.02.28.482419v1?rss=1">
<title>
<![CDATA[
Synergistic effect of mutations in the Parkinson's disease-linked proteins synaptojanin 1 and auxilin 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.02.28.482419v1?rss=1"
</link>
<description><![CDATA[
Parkinsons disease (PD) is a neurodegenerative disorder characterized by defective dopaminergic (DAergic) input to the striatum. Mutations in two genes encoding synaptically enriched clathrin-uncoating factors, synaptojanin 1 (SJ1) and auxilin, have been implicated in atypical Parkinsonism. SJ1 knock-in (SJ1-KIRQ) mice carrying a disease-linked mutation display neurological manifestations reminiscent of Parkinsonism. Here we report that auxilin knockout (Aux-KO) mice display dystrophic changes of a subset of nigrostriatal DAergic terminals similar to those of SJ1-KIRQ mice. Furthermore, Aux-KO/SJ1-KIRQ double mutant mice have shorter lifespan and more severe synaptic defects than single mutant mice. These include increase in dystrophic striatal nerve terminals positive for DAergic markers and for the PD risk protein SV2C, as well as adaptive changes in striatal interneurons. The synergistic effect of the two mutations demonstrates a special lability of DAergic neurons to defects in clathrin uncoating, with implications for PD pathogenesis in at least some forms of this condition.
]]></description>
<dc:creator>Ng, X. Y.</dc:creator>
<dc:creator>Wu, Y.</dc:creator>
<dc:creator>Lin, Y.</dc:creator>
<dc:creator>Yaqoob, S. M.</dc:creator>
<dc:creator>Greene, L. E.</dc:creator>
<dc:creator>De Camilli, P.</dc:creator>
<dc:creator>Cao, M.</dc:creator>
<dc:date>2022-03-02</dc:date>
<dc:identifier>doi:10.1101/2022.02.28.482419</dc:identifier>
<dc:title><![CDATA[Synergistic effect of mutations in the Parkinson's disease-linked proteins synaptojanin 1 and auxilin]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.01.21.477284v1?rss=1">
<title>
<![CDATA[
Structural basis for Parkinson's Disease-linked LRRK2's binding to microtubules 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.01.21.477284v1?rss=1"
</link>
<description><![CDATA[
Leucine Rich Repeat Kinase 2 (LRRK2) is one of the most commonly mutated genes in familial Parkinsons Disease (PD). Under some circumstances, LRRK2 co-localizes with microtubules in cells, an association enhanced by PD mutations. We report a cryo-electron microscopy structure of the catalytic half of LRRK2, containing its kinase, which is in a closed conformation, and GTPase domains, bound to microtubules. We also report a structure of the catalytic half of LRRK1, which is closely related to LRRK2, but is not linked to PD. LRRK1s structure is similar to LRRK2, but LRRK1 does not interact with microtubules. Guided by these structures, we identify amino acids in LRRK2s GTPase domain that mediate microtubule binding; mutating them disrupts microtubule binding in vitro and in cells, without affecting LRRK2s kinase activity. Our results have implications for the design of therapeutic LRRK2 kinase inhibitors.
]]></description>
<dc:creator>Snead, D. M.</dc:creator>
<dc:creator>Matyszewski, M.</dc:creator>
<dc:creator>Dickey, A. M.</dc:creator>
<dc:creator>Lin, Y. X.</dc:creator>
<dc:creator>Leschziner, A. E.</dc:creator>
<dc:creator>Reck-Peterson, S. L.</dc:creator>
<dc:date>2022-01-22</dc:date>
<dc:identifier>doi:10.1101/2022.01.21.477284</dc:identifier>
<dc:title><![CDATA[Structural basis for Parkinson's Disease-linked LRRK2's binding to microtubules]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.04.01.486724v1?rss=1">
<title>
<![CDATA[
Impact of 98 LRRK2 variants linked to Parkinson's Disease on kinase activity and microtubule binding 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.04.01.486724v1?rss=1"
</link>
<description><![CDATA[
Mutations enhancing the kinase activity of LRRK2 cause Parkinsons disease (PD) and therapies that reduce LRRK2 kinase activity are being tested in clinical trials. Numerous rare variants of unknown clinical significance have been reported, but how the vast majority impact on LRRK2 function is unknown. Here, we investigate 100 LRRK2 variants linked to PD, including previously described pathogenic mutations. We identify 23 LRRK2 variants that robustly stimulate kinase activity, including variants within the N-terminal non-catalytic regions [ARM (E334K, A419V), ANK(R767H), LRR (R1067Q, R1325Q)], as well as variants predicted to destabilise the ROC:CORB interface [ROC (A1442P, V1447M), CORA (R1628P) CORB (S1761R, L1795F)] and COR:COR dimer interface [CORB (R1728H/L)]. Most activating variants decrease LRRK2 biomarker site phosphorylation (pSer935/pSer955/pSer973), consistent with the notion that the active kinase conformation blocks their phosphorylation. We conclude that the impact of variants on kinase activity is best evaluated by deploying a cellular assay of LRRK2-dependent Rab10 substrate phosphorylation, compared to a biochemical kinase assay, as only a minority of activating variants [CORB (Y1699C, R1728H/L, S1761R) and kinase (G2019S, I2020T, T2031S)], enhance in vitro kinase activity of immunoprecipitated LRRK2. Twelve variants including several that activate LRRK2 and have been linked to PD, suppressed microtubule association in the presence of a Type I kinase inhibitor [ARM(M712V), LRR(R1320S), ROC (A1442P, K1468E, S1508R), CORA(A1589S), CORB (Y1699C, R1728H/L) and WD40(R2143M, S2350I, G2385R)]. Our findings will stimulate work to better understand the mechanisms by which variants impact biology and provide rationale for variant carrier inclusion or exclusion in ongoing and future LRRK2 inhibitor clinical trials.
]]></description>
<dc:creator>Kalogeropulou, A. F.</dc:creator>
<dc:creator>Purlyte, E.</dc:creator>
<dc:creator>Tonelli, F.</dc:creator>
<dc:creator>Lange, S. M.</dc:creator>
<dc:creator>Wightman, M.</dc:creator>
<dc:creator>Prescott, A. R.</dc:creator>
<dc:creator>Padmanabhan, S.</dc:creator>
<dc:creator>Sammler, E.</dc:creator>
<dc:creator>Alessi, D. R.</dc:creator>
<dc:date>2022-04-01</dc:date>
<dc:identifier>doi:10.1101/2022.04.01.486724</dc:identifier>
<dc:title><![CDATA[Impact of 98 LRRK2 variants linked to Parkinson's Disease on kinase activity and microtubule binding]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.03.11.484024v1?rss=1">
<title>
<![CDATA[
Structural and biochemical insights into lipid transport by VPS13 proteins. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.03.11.484024v1?rss=1"
</link>
<description><![CDATA[
VPS13 proteins are proposed to function at contact sites between organelles as bridges for lipids to move directionally and in bulk between organellar membranes. VPS13s are anchored between membranes via interactions with receptors, including both peripheral or integral membrane proteins. Here we present the crystal structure of VPS13s adaptor binding domain (VAB) complexed with a Pro-X-Pro peptide recognition motif present in one such receptor, the integral membrane protein Mcp1p, and show biochemically that other Pro-X-Pro motifs bind the VAB in the same site. We further demonstrate that Mcp1p and another integral membrane protein that interacts directly with human VPS13A, XK, are scramblases. This finding supports an emerging paradigm of a partnership between bulk lipid transport proteins and scramblases. Scramblases can re-equilibrate lipids between membrane leaflets as lipids are removed from or inserted into, respectively, the cytosolic leaflet of donor and acceptor organelles in the course of protein-mediated transport.
]]></description>
<dc:creator>Adlakha, J.</dc:creator>
<dc:creator>Hong, Z.</dc:creator>
<dc:creator>Li, P.</dc:creator>
<dc:creator>Reinisch, k. M.</dc:creator>
<dc:date>2022-03-12</dc:date>
<dc:identifier>doi:10.1101/2022.03.11.484024</dc:identifier>
<dc:title><![CDATA[Structural and biochemical insights into lipid transport by VPS13 proteins.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.03.28.486050v1?rss=1">
<title>
<![CDATA[
ggtranscript: an R package for the visualization and interpretation of transcript isoforms using ggplot2 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.03.28.486050v1?rss=1"
</link>
<description><![CDATA[
MotivationThe advent of long-read sequencing technologies has increased demand for the visualisation and interpretation of transcripts. However, tools that perform such visualizations remain inflexible and lack the ability to easily identify differences between transcript structures. Here, we introduce ggtranscript, an R package that provides a fast and flexible method to visualize and compare transcripts. As a ggplot2 extension, ggtranscript inherits the functionality and familiarity of ggplot2 making it easy to use.

Availability and implementationggtranscript is available at https://github.com/dzhang32/ggtranscript, DOI: https://doi.org/10.5281/zenodo.6374061
]]></description>
<dc:creator>Gustavsson, E. K.</dc:creator>
<dc:creator>Zhang, D.</dc:creator>
<dc:creator>Reynolds, R. H.</dc:creator>
<dc:creator>Garcia-Ruiz, S.</dc:creator>
<dc:creator>Ryten, M.</dc:creator>
<dc:date>2022-03-29</dc:date>
<dc:identifier>doi:10.1101/2022.03.28.486050</dc:identifier>
<dc:title><![CDATA[ggtranscript: an R package for the visualization and interpretation of transcript isoforms using ggplot2]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.02.15.480601v1?rss=1">
<title>
<![CDATA[
Highly efficient generation of isogenic pluripotent stem cell models using prime editing 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.02.15.480601v1?rss=1"
</link>
<description><![CDATA[
The recent development of prime editing (PE) genome engineering technologies has the potential to significantly simplify the generation of human pluripotent stem cell (hPSC)-based disease models. PE is a multi-component editing system that uses a Cas9-nickase fused to a reverse transcriptase (nCas9-RT) and an extended PE guide RNA (pegRNA). Once reverse transcribed, the pegRNA extension functions as a repair template to introduce precise designer mutations at the target site. Here, we systematically compared the editing efficiencies of PE to conventional gene editing methods in hPSCs. This analysis revealed that PE is overall more efficient and precise than homology-directed repair (HDR) of site-specific nuclease-induced double-strand breaks (DSBs). Specifically, PE is more effective in generating heterozygous editing events to create autosomal dominant disease-associated mutations. By stably integrating the nCas9-RT into hPSCs we achieved editing efficiencies equal to those reported for cancer cells, suggesting that the expression of the PE components, rather than cell-intrinsic features, limit PE in hPSCs. To improve the efficiency of PE in hPSCs, we optimized the delivery modalities for the PE components. Delivery of the nCas9-RT as mRNA combined with synthetically generated chemically-modified pegRNAs and nicking guide RNAs (ngRNAs) improved editing efficiencies up to 13-fold compared to transfecting the prime editing components as plasmids or ribonucleoprotein particles (RNPs). Finally, we demonstrated that this mRNA-based delivery approach can be used repeatedly to yield editing efficiencies exceeding 60% and to correct or introduce familial mutations causing Parkinsons disease in hPSCs.
]]></description>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Busquets, O.</dc:creator>
<dc:creator>Verma, Y.</dc:creator>
<dc:creator>Syed, K. M.</dc:creator>
<dc:creator>Kutnowski, N.</dc:creator>
<dc:creator>Pangilinan, G. R.</dc:creator>
<dc:creator>Gilbert, L.</dc:creator>
<dc:creator>Bateup, H.</dc:creator>
<dc:creator>Rio, D. C.</dc:creator>
<dc:creator>Hockemeyer, D.</dc:creator>
<dc:creator>Soldner, F.</dc:creator>
<dc:date>2022-02-15</dc:date>
<dc:identifier>doi:10.1101/2022.02.15.480601</dc:identifier>
<dc:title><![CDATA[Highly efficient generation of isogenic pluripotent stem cell models using prime editing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.02.03.479040v1?rss=1">
<title>
<![CDATA[
Multiscale model of primary motor cortex circuits reproduces in vivo cell type-specific dynamics associated with behavior 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.02.03.479040v1?rss=1"
</link>
<description><![CDATA[
Understanding cortical function requires studying multiple scales: molecular, cellular, circuit and behavior. We developed a biophysically detailed multiscale model of mouse primary motor cortex (M1) with over 10,000 neurons and 30 million synapses. Neuron types, densities, spatial distributions, morphologies, biophysics, connectivity and dendritic synapse locations were tightly constrained by experimental data. The model includes long-range inputs from 7 thalamic and cortical regions, as well as noradrenergic inputs from locus coeruleus. Connectivity depended on cell class and cortical depth at sublaminar resolution. The model accurately predicted in vivo layer- and cell type-specific responses (firing rates and LFP) associated with behavioral states (quiet wakefulness and movement) and experimental manipulations (noradrenaline receptor blocking and thalamus inactivation). It also enabled evaluation of multiple mechanistic hypotheses underlying the observed activity. This quantitative theoretical framework can be used to integrate and interpret M1 experimental data and sheds light on the cell type-specific multiscale dynamics associated with a range of experimental conditions and behaviors.
]]></description>
<dc:creator>Dura-Bernal, S.</dc:creator>
<dc:creator>Neymotin, S. A.</dc:creator>
<dc:creator>Suter, B. A.</dc:creator>
<dc:creator>Dacre, J.</dc:creator>
<dc:creator>Schiemann, J.</dc:creator>
<dc:creator>Duguid, I.</dc:creator>
<dc:creator>Shepherd, G. M. G.</dc:creator>
<dc:creator>Lytton, W. W.</dc:creator>
<dc:date>2022-02-06</dc:date>
<dc:identifier>doi:10.1101/2022.02.03.479040</dc:identifier>
<dc:title><![CDATA[Multiscale model of primary motor cortex circuits reproduces in vivo cell type-specific dynamics associated with behavior]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.02.03.479029v1?rss=1">
<title>
<![CDATA[
Large-scale biophysically detailed model of somatosensory thalamocortical circuits in NetPyNE 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.02.03.479029v1?rss=1"
</link>
<description><![CDATA[
The primary somatosensory cortex (S1) of mammals is critically important in the perception of touch and related sensorimotor behaviors. In 2015, the Blue Brain Project developed a groundbreaking rat S1 microcircuit simulation with over 31,000 neurons with 207 morpho-electrical neuron types, and 37 million synapses, incorporating anatomical and physiological information from a wide range of experimental studies. We have implemented this highly-detailed and complex S1 model in NetPyNE, using the data available in the Neocortical Microcircuit Collaboration Portal. NetPyNE provides a Python high-level interface to NEURON and allows defining complicated multiscale models using an intuitive declarative standardized language. It also facilitates running parallel simulations, automates the optimization and exploration of parameters using supercomputers, and provides a wide range of built-in analysis functions. This will make the S1 model more accessible and simpler to scale, modify and extend in order to explore research questions or interconnect to other existing models. Despite some implementation differences, the NetPyNE model preserved the original cell morphologies, electrophysiological responses and spatial distribution for all 207 cell types; and the connectivity properties of all 1941 pathways, including synaptic dynamics and short-term plasticity (STP). The NetPyNE S1 simulations produced reasonable physiological firing rates and activity patterns across all populations. The network generated a 1 Hz oscillation comparable to the original model in vitro-like state. By then reducing the extracellular calcium concentration, the model reproduced the original S1 in vivo-like states with asynchronous activity. These results validate the original study using a new modeling tool. Simulated local field potentials (LFPs) exhibited realistic oscillatory patterns and features, including distance- and frequency-dependent attenuation. The model was extended by adding thalamic circuits, including 6 distinct thalamic populations with intrathalamic, thalamocortical and corticothalamic connectivity derived from experimental data. The thalamic model reproduced single known cell and circuit-level dynamics, including burst and tonic firing modes and oscillatory patterns, providing a more realistic input to cortex and enabling study of thalamocortical interactions. Overall, our work provides a widely accessible, data-driven and biophysically-detailed model of the somatosensory thalamocortical circuits that can be employed as a community tool for researchers to study neural dynamics, function and disease.
]]></description>
<dc:creator>Borges, F. d. S.</dc:creator>
<dc:creator>Moreira, J. V. S.</dc:creator>
<dc:creator>Takarabe, L. M.</dc:creator>
<dc:creator>Lytton, W. W.</dc:creator>
<dc:creator>Dura-Bernal, S.</dc:creator>
<dc:date>2022-02-06</dc:date>
<dc:identifier>doi:10.1101/2022.02.03.479029</dc:identifier>
<dc:title><![CDATA[Large-scale biophysically detailed model of somatosensory thalamocortical circuits in NetPyNE]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.04.05.487195v1?rss=1">
<title>
<![CDATA[
Unaltered T cell responses to common antigens in individuals with Parkinson's disease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.04.05.487195v1?rss=1"
</link>
<description><![CDATA[
Background and ObjectivesParkinsons disease (PD) is associated with a heightened inflammatory state, including activated T cells. However, it is unclear whether these PD T cell responses are antigen specific or more indicative of generalized hyperresponsiveness. Our objective was to measure and compare antigen-specific T cell responses directed towards antigens derived from commonly encountered human pathogens/vaccines in patients with PD and age-matched healthy controls (HC).

MethodsPeripheral blood mononuclear cells (PBMCs) from 20 PD patients and 19 age-matched HCs were screened. Antigen specific T cell responses were measured by flow cytometry using a combination of the activation induced marker (AIM) assay and intracellular cytokine staining.

ResultsHere we show that both PD patients and HCs show similar T cell activation levels to several antigens derived from commonly encountered human pathogens/vaccines in the general population. Similarly, we also observed no difference between HC and PD in the levels of CD4 and CD8 T cell derived cytokines produced in response to any of the common antigens tested. These antigens encompassed both viral (coronavirus, rhinovirus, respiratory syncytial virus, influenza, cytomegalovirus) and bacterial (pertussis, tetanus) targets.

ConclusionsThese results suggest the T cell dysfunction observed in PD may not extend itself to abnormal responses to commonly encountered or vaccine-target antigens. Our study supports the notion that the targets of inflammatory T cell responses in PD may be more directed towards autoantigens like -synuclein (-syn) rather than common foreign antigens.
]]></description>
<dc:creator>Williams, G. P.</dc:creator>
<dc:creator>Muskat, K.</dc:creator>
<dc:creator>Frazier, A.</dc:creator>
<dc:creator>Xu, Y.</dc:creator>
<dc:creator>Mateus, J.</dc:creator>
<dc:creator>Grifoni, A.</dc:creator>
<dc:creator>Antunes, R. d. S.</dc:creator>
<dc:creator>Weiskopf, D.</dc:creator>
<dc:creator>Amara, A. W.</dc:creator>
<dc:creator>Standaert, D. G.</dc:creator>
<dc:creator>Goldman, J. G.</dc:creator>
<dc:creator>Litvan, I.</dc:creator>
<dc:creator>Alcalay, R. N.</dc:creator>
<dc:creator>Sulzer, D.</dc:creator>
<dc:creator>Arlehamn, C. S. L.</dc:creator>
<dc:creator>Sette, A.</dc:creator>
<dc:date>2022-04-08</dc:date>
<dc:identifier>doi:10.1101/2022.04.05.487195</dc:identifier>
<dc:title><![CDATA[Unaltered T cell responses to common antigens in individuals with Parkinson's disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.04.25.489459v1?rss=1">
<title>
<![CDATA[
A Feed-forward Pathway Drives LRRK2 kinaseMembrane Recruitment and Apparent Activation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.04.25.489459v1?rss=1"
</link>
<description><![CDATA[
Activating mutations in the Leucine Rich Repeat Kinase 2 (LRRK2) cause Parkinsons disease and previously we showed that activated LRRK2 phosphorylates a subset of Rab GTPases (Steger et al., 2017). Moreover, Golgi-associated Rab29 can recruit LRRK2 to the surface of the Golgi and activate it there for both auto- and Rab substrate phosphorylation. Here we define the precise Rab29 binding region of the LRRK2 Armadillo domain between residues 360-450 and show that this domain, termed "Site #1", can also bind additional LRRK2 substrates, Rab8A and Rab10. Moreover, we identify a distinct, N-terminal, higher affinity interaction interface between LRRK2 phosphorylated Rab8 and Rab10 termed "Site #2", that can retain LRRK2 on membranes in cells to catalyze multiple, subsequent phosphorylation events. Kinase inhibitor washout experiments demonstrate that rapid recovery of kinase activity in cells depends on the ability of LRRK2 to associate with phosphorylated Rab proteins, and phosphorylated Rab8A stimulates LRRK2 phosphorylation of Rab10 in vitro. Reconstitution of purified LRRK2 recruitment onto planar lipid bilayers decorated with Rab10 protein demonstrates cooperative association of only active LRRK2 with phospho-Rab10-containing membrane surfaces. These experiments reveal a feed-forward pathway that provides spatial control and membrane activation of LRRK2 kinase activity.
]]></description>
<dc:creator>Vides, E. G.</dc:creator>
<dc:creator>Adhikari, A.</dc:creator>
<dc:creator>Lis, P.</dc:creator>
<dc:creator>Purlyte, E.</dc:creator>
<dc:creator>Shumate, J. L.</dc:creator>
<dc:creator>Lasso, E. S.</dc:creator>
<dc:creator>Dhekne, H. S.</dc:creator>
<dc:creator>Alessi, D. R.</dc:creator>
<dc:creator>Pfeffer, S. R.</dc:creator>
<dc:date>2022-04-25</dc:date>
<dc:identifier>doi:10.1101/2022.04.25.489459</dc:identifier>
<dc:title><![CDATA[A Feed-forward Pathway Drives LRRK2 kinaseMembrane Recruitment and Apparent Activation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.04.26.489605v1?rss=1">
<title>
<![CDATA[
Structural basis of human LRRK2 membrane recruitment and activation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.04.26.489605v1?rss=1"
</link>
<description><![CDATA[
Mutations in LRRK2 are the most common genetic cause of late-onset Parkinsons disease (PD). LRRK2 encodes the leucine-rich repeat kinase 2 (LRRK2), whose kinase activity is regulated by Rab29, a membrane-anchored GTPase. However, molecular mechanisms underlying Rab29-dependent recruitment and activation of LRRK2 remain unclear. Here we report cryo-EM structures of LRRK2-Rab29 complexes in three oligomeric states, illustrating snapshots of key steps during LRRK2 membrane recruitment and activation. Rab29 binds to the ARM domain of LRRK2, and disruption at the interface abrogates LRRK2 kinase activity. Activation of LRRK2 is underpinned by the formation of an unexpected Rab29-induced super-assembly containing two central kinase-active and two peripheral kinase-inactive LRRK2 protomers. Central protomers undergo pronounced oligomerization-associated rearrangements and adopt an active conformation. Our work reveals the structural mechanism for LRRK2s spatial regulation controlled by Rab GTPases, provides mechanistic insights into pathogenic mutations and identifies new opportunities to design LRRK2 inhibitors for PD treatment.
]]></description>
<dc:creator>Zhu, H.</dc:creator>
<dc:creator>Tonelli, F.</dc:creator>
<dc:creator>Alessi, D. R.</dc:creator>
<dc:creator>Sun, J.</dc:creator>
<dc:date>2022-04-26</dc:date>
<dc:identifier>doi:10.1101/2022.04.26.489605</dc:identifier>
<dc:title><![CDATA[Structural basis of human LRRK2 membrane recruitment and activation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.11.12.468404v1?rss=1">
<title>
<![CDATA[
Therapeutic Deep Brain Stimulation Disrupts Subthalamic Nucleus Activity Dynamics in Parkinsonian Mice 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.11.12.468404v1?rss=1"
</link>
<description><![CDATA[
Subthalamic nucleus deep brain stimulation (STN DBS) relieves many motor symptoms of Parkinsons Disease (PD), but its underlying therapeutic mechanisms remain unclear. Since its advent, three major theories have been proposed: (1) DBS inhibits the STN and basal ganglia output; (2) DBS antidromically activates motor cortex; and (3) DBS disrupts firing dynamics within the STN. Previously, stimulation-related electrical artifacts limited mechanistic investigations using electrophysiology. We used electrical artifact-free calcium imaging to investigate activity in basal ganglia nuclei during STN DBS in parkinsonian mice. To test whether the observed changes in activity were sufficient to relieve motor symptoms, we then combined electrophysiological recording with targeted optical DBS protocols. Our findings suggest that STN DBS exerts its therapeutic effect through the disruption of STN dynamics, rather than inhibition or antidromic activation. These results provide insight into optimizing PD treatments and establish an approach for investigating DBS in other neuropsychiatric conditions.
]]></description>
<dc:creator>Schor, J. S.</dc:creator>
<dc:creator>Gonzalez Montalvo, I.</dc:creator>
<dc:creator>Spratt, P. W. E.</dc:creator>
<dc:creator>Brakaj, R. J.</dc:creator>
<dc:creator>Stansil, J. A.</dc:creator>
<dc:creator>Bender, K. J.</dc:creator>
<dc:creator>Nelson, A. B.</dc:creator>
<dc:date>2021-11-13</dc:date>
<dc:identifier>doi:10.1101/2021.11.12.468404</dc:identifier>
<dc:title><![CDATA[Therapeutic Deep Brain Stimulation Disrupts Subthalamic Nucleus Activity Dynamics in Parkinsonian Mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-11-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.05.02.490291v1?rss=1">
<title>
<![CDATA[
In situ structural analysis reveals membrane shape transitions during autophagosome formation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.05.02.490291v1?rss=1"
</link>
<description><![CDATA[
Autophagosomes are unique organelles which form de novo as double-membrane vesicles engulfing cytosolic material for destruction. Their biogenesis involves a series of membrane transformations with distinctly shaped intermediates whose ultrastructure is poorly understood. Here, we combine cell biology, correlative cryo-electron tomography (ET) and novel data analysis to reveal the step-by-step structural progression of autophagosome biogenesis at high resolution directly within yeast cells. By mapping individual structures onto a timeline based on geometric features, we uncover dynamic changes in membrane shape and curvature. Moreover, we reveal the organelle interactome of growing autophagosomes, highlighting a polar organization of contact sites between the phagophore and organelles such as the vacuole and the ER. Collectively, these findings have important implications for the contribution of different membrane sources during autophagy and for the forces shaping and driving phagophores towards closure without a templating cargo.
]]></description>
<dc:creator>Bieber, A.</dc:creator>
<dc:creator>Capitanio, C.</dc:creator>
<dc:creator>Erdmann, P. S.</dc:creator>
<dc:creator>Fiedler, F.</dc:creator>
<dc:creator>Beck, F.</dc:creator>
<dc:creator>Lee, C.-W.</dc:creator>
<dc:creator>Li, D.</dc:creator>
<dc:creator>Hummer, G.</dc:creator>
<dc:creator>Schulman, B.</dc:creator>
<dc:creator>Baumeister, W.</dc:creator>
<dc:creator>Wilfling, F.</dc:creator>
<dc:date>2022-05-02</dc:date>
<dc:identifier>doi:10.1101/2022.05.02.490291</dc:identifier>
<dc:title><![CDATA[In situ structural analysis reveals membrane shape transitions during autophagosome formation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.03.31.486371v1?rss=1">
<title>
<![CDATA[
Early Endosome Capture Proteomics and its Application to Amyloid Precursor Protein Intramembrane Processing by β and γ-Secretases 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.03.31.486371v1?rss=1"
</link>
<description><![CDATA[
Degradation and recycling of plasma membrane-localized proteins occurs via the endolysosomal system, wherein endosomes bud into the cytosol from the plasma membrane and subsequently mature into degradative lysosomal compartments. The endolysosomal system also participates in intracellular membrane protein trafficking, signaling, and protein degradation via autophagy. Our understanding of the endolysosomal system has been significantly advanced through the development of methods for selective capture of lysosomes (termed Lyso-IP), but methods for analogous isolation of early intermediates in the endolysosomal system are lacking. Here, we develop an approach for rapid isolation of early/sorting endosomes through affinity capture of the early endosome-associated protein EEA1 (Endo-IP) and provide proteomic and lipidomic snapshots of early endosomes in action. In addition to resident endosomal components, we identify numerous recycling, regulatory and membrane fusion complexes as well as candidate cargo, providing a proteomic landscape of early/sorting endosomes that we distinguished from lysosomes via comparative proteomics. The approach allows capture of internalized transferrin within minutes of its addition to cells, indicating the utility of the approach for dynamic analysis of early/sorting endosomes. Among the endocytic cargo identified by Endo-IP was the amyloid precursor protein (APP) genetically linked with Alzheimers disease. Processing of APP to amyloidogenic A{beta} peptides by {beta}- and {gamma}-Secretases can occur within the endolysosomal system among other compartments, but methods for spatial quantification of A{beta} products in individual organelles are lacking. We combined Endo- and Lyso-IP with targeted proteomics to provide a spatial digital snapshot of A{beta} products. This analysis revealed that products of A{beta} processing by {beta}- and {gamma}-Secretases, and alterations in the specificity of cleavage by small molecule {gamma}-Secretase modulators, can be quantified in both early/sorting endosomes and lysosomes. We anticipate that the Endo-IP approach will facilitate systematic interrogation of the many processes that are coordinated on early endosomes.
]]></description>
<dc:creator>Park, H.</dc:creator>
<dc:creator>Yu, Q.</dc:creator>
<dc:creator>Hundley, F. V.</dc:creator>
<dc:creator>Overmyer, K.</dc:creator>
<dc:creator>Serrano, L.</dc:creator>
<dc:creator>Paulo, J. A.</dc:creator>
<dc:creator>Paoli, J. C.</dc:creator>
<dc:creator>Swarup, S.</dc:creator>
<dc:creator>Coon, J.</dc:creator>
<dc:creator>Gygi, S.</dc:creator>
<dc:creator>Harper, W.</dc:creator>
<dc:date>2022-03-31</dc:date>
<dc:identifier>doi:10.1101/2022.03.31.486371</dc:identifier>
<dc:title><![CDATA[Early Endosome Capture Proteomics and its Application to Amyloid Precursor Protein Intramembrane Processing by β and γ-Secretases]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.05.02.490309v1?rss=1">
<title>
<![CDATA[
Genetic and pharmacological reduction of CDK14 mitigates synucleinopathy 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.05.02.490309v1?rss=1"
</link>
<description><![CDATA[
Parkinsonas disease (PD) is a debilitating neurodegenerative disease characterized by the loss of midbrain dopaminergic neurons (DaNs) and the abnormal accumulation of -Synuclein (-Syn) protein. Currently, no treatment can slow nor halt the progression of PD. Multiplications and mutations of the -Syn gene (SNCA) cause PD-associated syndromes and animal models that overexpress -Syn replicate several features of PD. Decreasing total -Syn levels, therefore, is an attractive approach to slow down neurodegeneration in patients with synucleinopathy. We previously performed a genetic screen for modifiers of -Syn levels and identified CDK14, a kinase of largely unknown function as a regulator of -Syn. To test the potential therapeutic effects of CDK14 reduction in PD, we ablated Cdk14 in the -Syn preformed fibrils (PFF)-induced PD mouse model. We found that loss of Cdk14 mitigates the grip strength deficit of PFF-treated mice and ameliorates PFF-induced cortical -Syn pathology, indicated by reduced numbers of pS129 -Syn-containing cells. In primary neurons, we found that Cdk14 depletion protects against the propagation of toxic -Syn species. We further validated these findings on pS129 -Syn levels in PD patient neurons. Finally, we leveraged the recent discovery of a covalent inhibitor of CDK14 to determine whether this target is pharmacologically tractable in vitro and in vivo. We found that CDK14 inhibition decreases total and pathologically aggregated -Syn in human neurons, in PFF- challenged rat neurons and in the brains of -Syn-humanized mice. In summary, we suggest that CDK14 represents a novel therapeutic target for PD-associated synucleinopathy.
]]></description>
<dc:creator>Parmasad, J.-L. A.</dc:creator>
<dc:creator>Ricke, K. M.</dc:creator>
<dc:creator>Stykel, M. G.</dc:creator>
<dc:creator>Buchner-Duby, B.</dc:creator>
<dc:creator>Lian, E.</dc:creator>
<dc:creator>Nguyen, B.</dc:creator>
<dc:creator>Lengacher, N. A.</dc:creator>
<dc:creator>Geertsma, H. M.</dc:creator>
<dc:creator>Bruce, A.</dc:creator>
<dc:creator>Callaghan, S. M.</dc:creator>
<dc:creator>Joselin, A.</dc:creator>
<dc:creator>Tomlinson, J. J.</dc:creator>
<dc:creator>Schlossmacher, M. G.</dc:creator>
<dc:creator>Stanford, W. L.</dc:creator>
<dc:creator>Brundin, P.</dc:creator>
<dc:creator>Ryan, S. D.</dc:creator>
<dc:creator>Rousseaux, M. W. C.</dc:creator>
<dc:date>2022-05-02</dc:date>
<dc:identifier>doi:10.1101/2022.05.02.490309</dc:identifier>
<dc:title><![CDATA[Genetic and pharmacological reduction of CDK14 mitigates synucleinopathy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.05.03.490522v1?rss=1">
<title>
<![CDATA[
Membrane curvature sensing and stabilization by the autophagic LC3 lipidation machinery 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.05.03.490522v1?rss=1"
</link>
<description><![CDATA[
How the highly curved phagophore membrane is stabilized during autophagy initiation is a major open question in autophagosome biogenesis. Here, we use in vitro reconstitution on membrane nanotubes and molecular dynamics simulations to investigate how core autophagy proteins in the LC3 lipidation cascade interact with curved membranes, providing insight into possible roles in regulating membrane shape during autophagosome biogenesis. ATG12-5-16L1 was up to 100-fold enriched on highly curved nanotubes relative to flat membranes. At high surface density, ATG12-5-16L1 binding increased the curvature of the nanotubes. While WIPI2 binding directs membrane recruitment, the amphipathic helix 2 of ATG16L1 is responsible for curvature sensitivity. Molecular dynamics simulations revealed that helix 2 of ATG16L1 inserts shallowly into the membrane, explaining its curvature-sensitive binding to the membrane. These observations show how the binding of the ATG12-5-16L1 complex to the early phagophore rim could stabilize membrane curvature and facilitate autophagosome growth.
]]></description>
<dc:creator>Jensen, L. E.</dc:creator>
<dc:creator>Rao, S.</dc:creator>
<dc:creator>Schuschnig, M.</dc:creator>
<dc:creator>Cada, A. K.</dc:creator>
<dc:creator>Martens, S.</dc:creator>
<dc:creator>Hummer, G.</dc:creator>
<dc:creator>Hurley, J. H.</dc:creator>
<dc:date>2022-05-04</dc:date>
<dc:identifier>doi:10.1101/2022.05.03.490522</dc:identifier>
<dc:title><![CDATA[Membrane curvature sensing and stabilization by the autophagic LC3 lipidation machinery]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.02.04.479203v1?rss=1">
<title>
<![CDATA[
Dopamine transporter and synaptic vesicle sorting defects initiate auxilin-linked Parkinson's disease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.02.04.479203v1?rss=1"
</link>
<description><![CDATA[
Auxilin participates in the uncoating of clathrin-coated vesicles (CCVs), thereby facilitating synaptic vesicle (SV) regeneration at presynaptic sites. Auxilin (DNAJC6/PARK19) loss-of- function mutations cause early-onset Parkinsons disease (PD). Here, we utilized auxilin-knockout (KO) mice to elucidate the mechanisms through which auxilin deficiency and clathrin-uncoating deficits lead to PD. We demonstrate that auxilin KO mice display the cardinal features of PD, including progressive motor deficits, -synuclein pathology, nigral dopaminergic loss, and neuroinflammation. Through unbiased proteomic and neurochemical analyses, we demonstrate that dopamine homeostasis is disrupted in auxilin KO brains, including via slower dopamine reuptake kinetics in vivo, an effect associated with dopamine transporter misrouting into axonal membrane deformities in the dorsal striatum. We also show that elevated macroautophagy and defective SV protein sorting contribute to ineffective dopamine sequestration and homeostasis, ultimately leading to neurodegeneration. This study advances our knowledge of how presynaptic endocytosis deficits lead to dopaminergic vulnerability and pathogenesis of PD.
]]></description>
<dc:creator>Vidyadhara, D. J.</dc:creator>
<dc:creator>Somayaji, M.</dc:creator>
<dc:creator>Wade, N.</dc:creator>
<dc:creator>Yücel, B.</dc:creator>
<dc:creator>Zhao, H.</dc:creator>
<dc:creator>Shashaank, N.</dc:creator>
<dc:creator>Ribaudo, J.</dc:creator>
<dc:creator>Gupta, J.</dc:creator>
<dc:creator>Lam, T. T.</dc:creator>
<dc:creator>Sames, D.</dc:creator>
<dc:creator>Greene, L. E.</dc:creator>
<dc:creator>Sulzer, D. L.</dc:creator>
<dc:creator>Chandra, S. S.</dc:creator>
<dc:date>2022-02-06</dc:date>
<dc:identifier>doi:10.1101/2022.02.04.479203</dc:identifier>
<dc:title><![CDATA[Dopamine transporter and synaptic vesicle sorting defects initiate auxilin-linked Parkinson's disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.02.18.481043v1?rss=1">
<title>
<![CDATA[
The AAA+ chaperone VCP disaggregates Tau fibrils and generates aggregate seeds 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.02.18.481043v1?rss=1"
</link>
<description><![CDATA[
Amyloid-like aggregates of the microtubule-associated protein Tau are associated with several neurodegenerative disorders including Alzheimers disease. The existence of cellular machinery for the removal of such aggregates has remained unclear, as specialized disaggregase chaperones are thought to be absent in mammalian cells. Here we show in cell culture and in neurons that the AAA+ chaperone VCP is recruited to ubiquitylated Tau fibrils, resulting in their efficient disaggregation. Aggregate clearance depends on the functional cooperation of VCP with Hsp70 and the ubiquitin-proteasome machinery. Inhibition of VCP activity stabilizes large Tau aggregates, and is accompanied by a reduction in the amount of Tau species competent of prion- like aggregate seeding in recipient cells. Thus, disaggregation by VCP generates seeding-active Tau as byproduct. These findings identify VCP as a core component of the machinery for the removal of neurodegenerative disease aggregates and suggest that its activity can be associated with enhanced aggregate spreading in tauopathies.
]]></description>
<dc:creator>Saha, I.</dc:creator>
<dc:creator>Yuste-Checa, P.</dc:creator>
<dc:creator>da Silva Padilha, M.</dc:creator>
<dc:creator>Guo, Q.</dc:creator>
<dc:creator>Körner, R.</dc:creator>
<dc:creator>Holthusen, H.</dc:creator>
<dc:creator>Trinkaus, V. A.</dc:creator>
<dc:creator>Dudanova, I.</dc:creator>
<dc:creator>Fernandez-Busnadiego, R.</dc:creator>
<dc:creator>Baumeister, W.</dc:creator>
<dc:creator>Sanders, D. W.</dc:creator>
<dc:creator>Gautam, S.</dc:creator>
<dc:creator>Diamond, M. I.</dc:creator>
<dc:creator>Hartl, F.-U.</dc:creator>
<dc:creator>Hipp, M. S.</dc:creator>
<dc:date>2022-02-19</dc:date>
<dc:identifier>doi:10.1101/2022.02.18.481043</dc:identifier>
<dc:title><![CDATA[The AAA+ chaperone VCP disaggregates Tau fibrils and generates aggregate seeds]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.05.04.490670v1?rss=1">
<title>
<![CDATA[
The IPDGC/GP2 Hackathon - an open science event for training in data science, genomics, and collaboration using Parkinson's disease data 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.05.04.490670v1?rss=1"
</link>
<description><![CDATA[
BackgroundOpen science and collaboration are necessary to facilitate the advancement of Parkinsons disease (PD) research. Hackathons are collaborative events that bring together people with different skill sets and backgrounds to generate resources and creative solutions to problems. These events can be used as training and networking opportunities.

ObjectiveTo coordinate a virtual hackathon to develop novel PD research tools.

Methods49 early career scientists from 12 countries collaborated in a virtual 3-day hackathon event in May 2021, during which they built tools and pipelines with a focus on PD. Resources were created with the goal of helping scientists accelerate their own research by having access to the necessary code and tools.

ResultsEach team was allocated one of nine different projects, each with a different goal. These included developing post-genome-wide association studies (GWAS) analysis pipelines, downstream analysis of genetic variation pipelines, and various visualization tools.

ConclusionHackathons are a valuable approach to inspire creative thinking, supplement training in data science, and foster collaborative scientific relationships, which are foundational practices for early career researchers. The resources generated can be used to accelerate research on the genetics of PD.
]]></description>
<dc:creator>Leonard, H. L.</dc:creator>
<dc:creator>Murtadha, R.</dc:creator>
<dc:creator>Martinez-Carrasco, A.</dc:creator>
<dc:creator>Muller-Nedebock, A.</dc:creator>
<dc:creator>Gil-Martinez, A.-L.</dc:creator>
<dc:creator>Illarionova, A.</dc:creator>
<dc:creator>Moore, A.</dc:creator>
<dc:creator>Bustos, B. I.</dc:creator>
<dc:creator>Jadhav, B.</dc:creator>
<dc:creator>Huxford, B.</dc:creator>
<dc:creator>Storm, C.</dc:creator>
<dc:creator>Towns, C.</dc:creator>
<dc:creator>Vitale, D.</dc:creator>
<dc:creator>Chetty, D.</dc:creator>
<dc:creator>Yu, E.</dc:creator>
<dc:creator>Jama, F.</dc:creator>
<dc:creator>Grenn, F. P.</dc:creator>
<dc:creator>Salazar, G.</dc:creator>
<dc:creator>Rateau, G.</dc:creator>
<dc:creator>Iwaki, H.</dc:creator>
<dc:creator>Elsayed, I.</dc:creator>
<dc:creator>Foote, I.</dc:creator>
<dc:creator>van Rensburg, Z. J.</dc:creator>
<dc:creator>Kim, J. J.</dc:creator>
<dc:creator>Yuan, J.</dc:creator>
<dc:creator>Lake, J.</dc:creator>
<dc:creator>Brolin, K.</dc:creator>
<dc:creator>Senkevich, K.</dc:creator>
<dc:creator>Wu, L.</dc:creator>
<dc:creator>Tan, M. M. X.</dc:creator>
<dc:creator>Perinan, M. T.</dc:creator>
<dc:creator>Makarious, M. B.</dc:creator>
<dc:creator>Ta, M.</dc:creator>
<dc:creator>Pillay, N. S.</dc:creator>
<dc:creator>Betancor, O. L.</dc:creator>
<dc:creator>Reyes-Perez, P. R.</dc:creator>
<dc:creator>Jerez, P. A.</dc:creator>
<dc:creator>Saini, P.</dc:creator>
<dc:creator>al-Ouran, R.</dc:creator>
<dc:creator>Sivakumar, R.</dc:creator>
<dc:creator>Real, R.</dc:creator>
<dc:creator>Reynolds, R. H.</dc:creator>
<dc:creator>Hu, R.</dc:creator>
<dc:creator>Abrahams, S.</dc:creator>
<dc:creator></dc:creator>
<dc:date>2022-05-10</dc:date>
<dc:identifier>doi:10.1101/2022.05.04.490670</dc:identifier>
<dc:title><![CDATA[The IPDGC/GP2 Hackathon - an open science event for training in data science, genomics, and collaboration using Parkinson's disease data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.01.06.896241v1?rss=1">
<title>
<![CDATA[
Regulation of mitophagy by the NSL complex underlies genetic risk for Parkinson's disease at Chr16q11.2 and on the MAPT H1 allele. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.01.06.896241v1?rss=1"
</link>
<description><![CDATA[
Parkinsons disease (PD) is a common incurable neurodegenerative disease. The identification of genetic variants via genome-wide association studies (GWAS) has considerably advanced our understanding of the PD genetic risk. Understanding the functional significance of the risk loci is now a critical step towards translating these genetic advances into an enhanced biological understanding of the disease. Impaired mitophagy is a key causative pathway in familial PD, but its relevance to idiopathic PD is unclear. We used a mitophagy screening assay to evaluate the functional significance of risk genes identified through GWAS. We identified two new regulators of PINK1-mitophagy, KAT8 and KANSL1, previously shown to modulate lysine acetylation. We show that KAT8 and KANSL1 modulate PINK1 gene expression and subsequent PINK1-mitophagy. These findings suggest PINK1-mitophagy is a contributing factor to idiopathic PD. KANSL1 is located on chromosome 17q21 where the risk associated gene has long been considered to be MAPT. While our data does not exclude a possible association between the MAPT gene and PD, it provides strong evidence that KANSL1 plays a crucial role in the disease. Finally, these results enrich our understanding of physiological events regulating mitophagy and establish a novel pathway for drug targeting in neurodegeneration.
]]></description>
<dc:creator>Soutar, M.</dc:creator>
<dc:creator>Melandri, D.</dc:creator>
<dc:creator>Annuario, E.</dc:creator>
<dc:creator>Monaghan, A.</dc:creator>
<dc:creator>Welsh, N.</dc:creator>
<dc:creator>D'Sa, K.</dc:creator>
<dc:creator>Guelfi, S.</dc:creator>
<dc:creator>Zhang, D.</dc:creator>
<dc:creator>Pittman, A.</dc:creator>
<dc:creator>Trabzuni, D.</dc:creator>
<dc:creator>Pan, K.</dc:creator>
<dc:creator>Kia, D. A.</dc:creator>
<dc:creator>Bictash, M.</dc:creator>
<dc:creator>Gandhi, S.</dc:creator>
<dc:creator>Houlden, H.</dc:creator>
<dc:creator>Cookson, M. R.</dc:creator>
<dc:creator>Wood, N.</dc:creator>
<dc:creator>Singleton, A.</dc:creator>
<dc:creator>Hardy, J.</dc:creator>
<dc:creator>Whiting, P.</dc:creator>
<dc:creator>Blauwendraat, C.</dc:creator>
<dc:creator>Whitworth, A.</dc:creator>
<dc:creator>Manzoni, C.</dc:creator>
<dc:creator>Ryten, M.</dc:creator>
<dc:creator>Lewis, P.</dc:creator>
<dc:creator>Plun-Favreau, H.</dc:creator>
<dc:date>2020-01-07</dc:date>
<dc:identifier>doi:10.1101/2020.01.06.896241</dc:identifier>
<dc:title><![CDATA[Regulation of mitophagy by the NSL complex underlies genetic risk for Parkinson's disease at Chr16q11.2 and on the MAPT H1 allele.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-01-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.06.09.495448v1?rss=1">
<title>
<![CDATA[
PKC isoforms activate LRRK1 kinase by phosphorylating conserved residues (Ser1064, Ser1074 and Thr1075) within the CORB GTPase domain 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.06.09.495448v1?rss=1"
</link>
<description><![CDATA[
Leucine-rich-repeat-kinase 1 (LRRK1) and its homologue LRRK2 are multidomain kinases possessing a ROC-CORA-CORB containing GTPase domain and phosphorylate distinct Rab proteins. LRRK1 loss of function mutations cause the bone disorder osteosclerotic metaphyseal dysplasia, whereas LRRK2 missense mutations that enhance kinase activity cause Parkinsons disease. Previous work suggested that LRRK1 but not LRRK2, is activated via a Protein Kinase C (PKC)-dependent mechanism. Here we demonstrate that phosphorylation and activation of LRRK1 in HEK293 cells is blocked by PKC inhibitors including LXS-196 (Darovasertib), a compound that has entered clinical trials. We show multiple PKC isoforms phosphorylate and activate recombinant LRRK1 in a manner reversed by phosphatase treatment. PKC unexpectedly does not activate LRRK1 by phosphorylating the kinase domain, but instead phosphorylates a cluster of conserved residues (Ser1064, Ser1074 and Thr1075) located within a region of the CORB domain of the GTPase domain. These residues are positioned at the equivalent region of the LRRK2 DK helix reported to stabilize the kinase domain C-helix in the active conformation. Thr1075 represents an optimal PKC site phosphorylation motif and its mutation to Ala, blocked PKC-mediated activation of LRRK1. A triple Glu mutation of Ser1064/Ser1074/Thr1075 to mimic phosphorylation, enhanced LRRK1 kinase activity ~3-fold. From analysis of available structures, we postulate that phosphorylation of Ser1064, Ser1074 and Thr1075 activates LRRK1 by promoting interaction and stabilization of the aC-helix on the kinase domain. This study provides new fundamental insights into the mechanism controlling LRRK1 activity and reveals a novel unexpected activation mechanism.
]]></description>
<dc:creator>Malik, A. U.</dc:creator>
<dc:creator>Karapetsas, A.</dc:creator>
<dc:creator>Nirujogi, R. S.</dc:creator>
<dc:creator>Chatterjee, D.</dc:creator>
<dc:creator>Phung, T. K.</dc:creator>
<dc:creator>Wightman, M.</dc:creator>
<dc:creator>Gourlay, R.</dc:creator>
<dc:creator>Morrice, N.</dc:creator>
<dc:creator>Mathea, S.</dc:creator>
<dc:creator>Knapp, S.</dc:creator>
<dc:creator>Alessi, D. R.</dc:creator>
<dc:date>2022-06-09</dc:date>
<dc:identifier>doi:10.1101/2022.06.09.495448</dc:identifier>
<dc:title><![CDATA[PKC isoforms activate LRRK1 kinase by phosphorylating conserved residues (Ser1064, Ser1074 and Thr1075) within the CORB GTPase domain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.06.07.494932v1?rss=1">
<title>
<![CDATA[
Structural conversion of α-synuclein at the mitochondria induces neuronal toxicity 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.06.07.494932v1?rss=1"
</link>
<description><![CDATA[
Aggregation of -Synuclein (-Syn) drives Parkinsons disease, although the initial stages of self-assembly and structural conversion have not been captured inside neurons. We track the intracellular conformational states of -Syn utilizing a single-molecule FRET biosensor, and show that -Syn converts from its monomeric state to form two distinct oligomeric states in neurons in a concentration dependent, and sequence specific manner. 3D FRET-CLEM reveals the structural organization, and location of aggregation hotspots inside the cell. Notably multiple intracellular seeding events occur preferentially on membrane surfaces, especially mitochondrial membranes. The mitochondrial lipid, cardiolipin triggers rapid oligomerization of A53T -Syn, and cardiolipin is sequestered within aggregating lipid-protein complexes. Mitochondrial aggregates impair complex I activity and increase mitochondrial ROS generation, which accelerates the oligomerization of A53T -Syn, and ultimately causes permeabilization of mitochondrial membranes, and cell death. Patient iPSC derived neurons harboring A53T mutations exhibit accelerated oligomerization that is dependent on mitochondrial ROS, early mitochondrial permeabilization and neuronal death. Our study highlights a mechanism of de novo oligomerization at the mitochondria and its induction of neuronal toxicity.
]]></description>
<dc:creator>Choi, M. L.</dc:creator>
<dc:creator>Chappard, A.</dc:creator>
<dc:creator>Singh, B. P.</dc:creator>
<dc:creator>Maclachlan, C.</dc:creator>
<dc:creator>Rodrigues, M.</dc:creator>
<dc:creator>Fedotova, E.</dc:creator>
<dc:creator>Berezhnov, A.</dc:creator>
<dc:creator>De, S.</dc:creator>
<dc:creator>Peddie, C.</dc:creator>
<dc:creator>Athauda, D.</dc:creator>
<dc:creator>Virdi, G. S.</dc:creator>
<dc:creator>Zhang, W.</dc:creator>
<dc:creator>Evans, J. R.</dc:creator>
<dc:creator>Wernick, A.</dc:creator>
<dc:creator>Zanjani, Z.</dc:creator>
<dc:creator>Angelova, P. R.</dc:creator>
<dc:creator>Esteras, N.</dc:creator>
<dc:creator>Andrey, A.</dc:creator>
<dc:creator>Morris, K.</dc:creator>
<dc:creator>Jeacock, K.</dc:creator>
<dc:creator>Tosatto, L.</dc:creator>
<dc:creator>Little, D.</dc:creator>
<dc:creator>Gissen, P.</dc:creator>
<dc:creator>Collinson, L.</dc:creator>
<dc:creator>Clarke, D. J.</dc:creator>
<dc:creator>Kunath, T.</dc:creator>
<dc:creator>Klenerman, D.</dc:creator>
<dc:creator>Abramov, A. Y.</dc:creator>
<dc:creator>Horrocks, M. H.</dc:creator>
<dc:creator>Gandhi, S.</dc:creator>
<dc:date>2022-06-09</dc:date>
<dc:identifier>doi:10.1101/2022.06.07.494932</dc:identifier>
<dc:title><![CDATA[Structural conversion of α-synuclein at the mitochondria induces neuronal toxicity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.06.08.495316v1?rss=1">
<title>
<![CDATA[
Metagenomics of Parkinson's disease implicates the gut microbiome in multiple disease mechanisms 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.06.08.495316v1?rss=1"
</link>
<description><![CDATA[
Parkinsons disease (PD) may start in the gut and spread to the brain. To investigate the role of gut microbiome, we enrolled 490 PD and 234 control individuals, conducted deep shotgun sequencing of fecal DNA, followed by metagenome-wide association studies requiring significance by two methods (ANCOM-BC and MaAsLin2) to declare disease association. Thirty-percent of species and pathways tested had altered abundances in PD, depicting a widespread dysbiosis. Network analysis showed PD-associated species form polymicrobial clusters that grow or shrink together, and some compete. Metagenomic profile of PD indicates a disease permissive microbiome, evidenced by overabundance of pathogens and immunogenic components, dysregulated neuroactive signaling, preponderance of molecules that induce alpha-synuclein pathology, and over-production of toxicants; with the reduction in anti-inflammatory and neuroprotective factors limiting the capacity to recover. These data provide a broad foundation with a wealth of concrete testable hypotheses to discern the role of the gut microbiome in PD.
]]></description>
<dc:creator>Wallen, Z. D.</dc:creator>
<dc:creator>Demirkan, A.</dc:creator>
<dc:creator>Twa, G.</dc:creator>
<dc:creator>Cohen, G.</dc:creator>
<dc:creator>Dean, M. N.</dc:creator>
<dc:creator>Standaert, D. G.</dc:creator>
<dc:creator>Sampson, T.</dc:creator>
<dc:creator>Payami, H.</dc:creator>
<dc:date>2022-06-12</dc:date>
<dc:identifier>doi:10.1101/2022.06.08.495316</dc:identifier>
<dc:title><![CDATA[Metagenomics of Parkinson's disease implicates the gut microbiome in multiple disease mechanisms]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.05.25.493476v1?rss=1">
<title>
<![CDATA[
Basal Ganglia Neurons in Healthy and Parkinsonian Primates Generate Recurring Sequences of Spikes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.05.25.493476v1?rss=1"
</link>
<description><![CDATA[
The spiking activity of basal ganglia neurons can be characterized by summary statistics such as the average firing rate, or by measures of firing patterns, such as burst discharges, or oscillatory fluctuations of firing rates. Many of these features are altered by the presence of parkinsonism. This study examined another distinct attribute of firing activity, i.e., the occurrence of repeating sequences of inter-spike intervals. We studied this feature in extracellular electrophysiologic recordings that were made in the basal ganglia of Rhesus monkeys, before and after they had been rendered parkinsonian by treatment with the neurotoxin 1-methyl-4-phyl-1,2,3,6-tetrahydropyridine (MPTP). Neurons in both pallidal segments and in the subthalamic nucleus tended to fire in repeating sequences, typically 2 ISIs long (i.e., involving three spikes). In recordings that were 5000 inter-spike intervals long, 20-40% of spikes participated in one of many sequences with each ISI replicating the sequence pattern with a timing error of [&le;]1%. Compared to similar analyses in shuffled representations of the same data, sequences were more common in the original representation of ISIs in the subthalamic nucleus and the external pallidal segment. Induction of parkinsonism reduced the proportion of sequence spikes in the external pallidum but increased it in the subthalamic nucleus. We found no relation between the sequence generation and the firing rate of neurons, and, at most, a weak correlation between sequence generation and the incidence of bursts. We conclude that basal ganglia neurons fire in recognizable sequences of ISIs, whose incidence is influenced by the induction of parkinsonism.
]]></description>
<dc:creator>Galvan, A.</dc:creator>
<dc:creator>Wichmann, T.</dc:creator>
<dc:date>2022-05-26</dc:date>
<dc:identifier>doi:10.1101/2022.05.25.493476</dc:identifier>
<dc:title><![CDATA[Basal Ganglia Neurons in Healthy and Parkinsonian Primates Generate Recurring Sequences of Spikes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-05-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.02.20.481207v1?rss=1">
<title>
<![CDATA[
Synaptic Location Is a Determinant of the Detrimental Effects of α-Synuclein Pathology to Glutamatergic Transmission in the Basolateral Amygdala 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.02.20.481207v1?rss=1"
</link>
<description><![CDATA[
The presynaptic protein -synuclein (Syn) has been suggested to be involved in the pathogenesis of Parkinsons disease (PD). In PD, the amygdala is prone to develop insoluble Syn aggregates, and it has been suggested that circuit dysfunction involving the amygdala contributes to the psychiatric symptoms. Yet, how Syn aggregates affect amygdala function is unknown. In this study, we examined Syn in glutamatergic axon terminals and the impact of its aggregation on glutamatergic transmission in the basolateral amygdala (BLA). We found that Syn is primarily present in the vesicular glutamate transporter 1-expressing (vGluT1+) terminals in mouse BLA, which is consistent with higher levels of Syn expression in vGluT1+ glutamatergic neurons in the cerebral cortex relative to the vGluT2+ glutamatergic neurons in the thalamus. We found that Syn aggregation selectively decreased the cortico-BLA, but not the thalamo-BLA, transmission; and that cortico-BLA synapses displayed enhanced short-term depression upon repetitive stimulation. In addition, using confocal microscopy, we found that vGluT1+ axon terminals exhibited decreased levels of soluble Syn, which suggests that lower levels of soluble Syn might underlie the enhanced short-term depression of cortico-BLA synapses. In agreement with this idea, we found that cortico-BLA synaptic depression was also enhanced in Syn knockout mice.

In conclusion, both basal and dynamic cortico-BLA transmission were disrupted by abnormal aggregation of Syn and these changes might be relevant to the perturbed cortical control of the amygdala that has been suggested to play a role in psychiatric symptoms in PD.
]]></description>
<dc:creator>Nagaraja, C.</dc:creator>
<dc:creator>Chen, L.</dc:creator>
<dc:creator>Daniels, S.</dc:creator>
<dc:creator>Meyerdirk, L.</dc:creator>
<dc:creator>Steiner, J. A.</dc:creator>
<dc:creator>Galvis, M. L. E.</dc:creator>
<dc:creator>Henderson, M.</dc:creator>
<dc:creator>Brundin, P.</dc:creator>
<dc:creator>Chu, H.-y.</dc:creator>
<dc:date>2022-02-20</dc:date>
<dc:identifier>doi:10.1101/2022.02.20.481207</dc:identifier>
<dc:title><![CDATA[Synaptic Location Is a Determinant of the Detrimental Effects of α-Synuclein Pathology to Glutamatergic Transmission in the Basolateral Amygdala]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.06.21.496850v1?rss=1">
<title>
<![CDATA[
Damaged mitochondria recruit the effector NEMO to activate NF-κB signaling 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.06.21.496850v1?rss=1"
</link>
<description><![CDATA[
Failure to clear damaged mitochondria via mitophagy disrupts physiological function and may initiate damage signaling via inflammatory cascades. However, signaling mechanisms leading from impaired mitophagy to neuro-inflammation are unclear. We discovered that NF-{kappa}B essential regulator NEMO is recruited to damaged mitochondria in a Parkin- and p62/SQSTM1-dependent manner in a time-course similar to recruitment of the structurally-related mitophagy receptor, OPTN. NEMO and p62 colocalize, partitioning into distinct domains from OPTN. Either depletion of p62 or mutation of NEMOs ubiquitin-binding domain abolishes NEMO recruitment, indicating multifactorial interactions. The active catalytic IKK component phospho-IKK{beta} colocalizes with NEMO on damaged mitochondria, initiating NF-{kappa}B signaling and the upregulation of inflammatory cytokines. These findings suggest that damaged mitochondria serve as an intracellular platform for innate immune signaling by promoting the formation of activated IKK complexes in a Parkin-dependent manner. We propose that mitophagy and NF-{kappa}B signaling are competing pathways regulating the response to cellular stress.
]]></description>
<dc:creator>Harding, O.</dc:creator>
<dc:creator>Holzbaur, E. L. F.</dc:creator>
<dc:date>2022-06-22</dc:date>
<dc:identifier>doi:10.1101/2022.06.21.496850</dc:identifier>
<dc:title><![CDATA[Damaged mitochondria recruit the effector NEMO to activate NF-κB signaling]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.06.20.496872v1?rss=1">
<title>
<![CDATA[
Dopaminergic axons track somatic signaling in behaving mice 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.06.20.496872v1?rss=1"
</link>
<description><![CDATA[
Striatal dopamine released from the axons of midbrain dopamine neurons has been linked to a wide range of functions, including movement control and reward-based learning. Recent studies have reported functional signaling differences between axons and somas of dopamine neurons, suggesting that local modulation controls dopamine release and calling into question the classical view of somatic control. However, these experiments are technically challenging, making it difficult to ensure that axonal and somatic recordings come from the same neurons, particularly given the heterogeneity of dopaminergic cell types. Here we used genetic strategies to isolate key dopaminergic neuron subtypes and monitor their axonal and somatic signaling patterns in behaving mice. Contrary to the inferences drawn from previous studies, these experiments revealed a robust correlation between somatic and axonal signaling. Thus, by exploiting a previously unknown connection between genetic and functional diversity in dopamine neurons, we establish that subtypes must be considered to understand the mechanisms of dopamine release in striatum during behavior.
]]></description>
<dc:creator>Azcorra, M.</dc:creator>
<dc:creator>Gaertner, Z.</dc:creator>
<dc:creator>Davidson, C.</dc:creator>
<dc:creator>Ramakrishnan, C.</dc:creator>
<dc:creator>Fenno, L.</dc:creator>
<dc:creator>Kim, Y. S.</dc:creator>
<dc:creator>Deisseroth, K.</dc:creator>
<dc:creator>Awatramani, R.</dc:creator>
<dc:creator>Dombeck, D. A.</dc:creator>
<dc:date>2022-06-22</dc:date>
<dc:identifier>doi:10.1101/2022.06.20.496872</dc:identifier>
<dc:title><![CDATA[Dopaminergic axons track somatic signaling in behaving mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.03.16.484460v1?rss=1">
<title>
<![CDATA[
Sst+ GPi output neurons provide direct feedback to key nodes of the basal ganglia and drive behavioral flexibility. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.03.16.484460v1?rss=1"
</link>
<description><![CDATA[
The internal globus pallidus (GPi) is a basal ganglia output nucleus with separate projections to the thalamus and the lateral habenula (LHb). Here, we show a GPi subtype with projections to LHb (GPi-LHb), genetically defined based on glutamate/GABA co-transmission and somatostatin (Sst) expression, also projects back to key nodes in the basal ganglia: the external globus pallidus (GPe), the striatal striosomes, and dopamine neurons in the substantia nigra. We found that the Sst+ GPi population showed strong movement and direction-specific selectivity in a goal-directed choice task, but not during self-paced exploration, and did not signal prediction errors or outcome modulation. During goal-directed behavior, the Sst+ GPi activity slowly evolved with learning of correct choice actions and genetic silencing disrupted the ability to update choice behavior following a task rule reversal. In summary, we have found that Sst+ GPi neurons establish a wide feedback network in the basal ganglia and drive behavioral flexibility.
]]></description>
<dc:creator>Weglage, M.</dc:creator>
<dc:creator>Aehrlund Richter, S.</dc:creator>
<dc:creator>Skara, V.</dc:creator>
<dc:creator>Lazaridis, I.</dc:creator>
<dc:creator>Meletis, K.</dc:creator>
<dc:date>2022-03-18</dc:date>
<dc:identifier>doi:10.1101/2022.03.16.484460</dc:identifier>
<dc:title><![CDATA[Sst+ GPi output neurons provide direct feedback to key nodes of the basal ganglia and drive behavioral flexibility.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.07.12.499634v1?rss=1">
<title>
<![CDATA[
Structural basis for ATG9A recruitment to the ULK1 complex in mitophagy initiation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.07.12.499634v1?rss=1"
</link>
<description><![CDATA[
The assembly of the autophagy initiation machinery nucleates autophagosome biogenesis, including in the PINK1- and Parkin-dependent mitophagy pathway implicated in Parkinsons disease. The structural interaction between the sole transmembrane autophagy protein, ATG9A, and components of the ULK1 complex is one of the major missing links needed to complete a structural map of autophagy initiation. We determined the 2.4 [A] x-ray crystallographic structure of the ternary structure of ATG9A C-terminal tail bound to the ATG13:ATG101 HORMA dimer, which is part of the ULK1 complex. We term the interacting portion of the extreme C-terminal part of the ATG9A tail the "HORMA dimer interacting region" (HDIR). This structure shows that the HDIR binds to the HORMA domain of ATG101 by {beta}-sheet complementation such that the ATG9A tail resides in a deep cleft at the ATG13:ATG101 interface. Disruption of this complex in cells impairs damage induced PINK1/Parkin mitophagy mediated by the cargo receptor NDP52.
]]></description>
<dc:creator>Ren, X.</dc:creator>
<dc:creator>Nguyen, T. N.</dc:creator>
<dc:creator>Lam, W. K.</dc:creator>
<dc:creator>Buffalo, C. Z.</dc:creator>
<dc:creator>Lazarou, M.</dc:creator>
<dc:creator>Yokom, A. L.</dc:creator>
<dc:creator>Hurley, J. H.</dc:creator>
<dc:date>2022-07-13</dc:date>
<dc:identifier>doi:10.1101/2022.07.12.499634</dc:identifier>
<dc:title><![CDATA[Structural basis for ATG9A recruitment to the ULK1 complex in mitophagy initiation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.07.20.500694v1?rss=1">
<title>
<![CDATA[
Integrated multi-cohort analysis of the Parkinson's disease gut metagenome 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.07.20.500694v1?rss=1"
</link>
<description><![CDATA[
BackgroundThe gut microbiome is altered in several neurologic disorders including Parkinsons disease (PD).

ObjectivesProfile the fecal gut metagenome in PD for alterations in microbial composition, taxon abundance, metabolic pathways, and microbial gene products, and their relationship with disease progression.

MethodsShotgun metagenomic sequencing was conducted on 244 stool donors from two independent cohorts in the United States, including individuals with PD (n=48, n=47, respectively), environmental Household Controls (HC, n=29, n=30), and community Population Controls (PC, n=41, n=49). Microbial features consistently altered in PD compared to HC and PC subjects were identified. Data were cross-referenced to public metagenomic datasets from two previous studies in Germany and China to determine generalizable microbiome features.

ResultsThe gut microbiome in PD shows significant alterations in community composition. Robust taxonomic alterations include depletion of putative "beneficial" gut commensals Faecalibacterium prausnitzii and Eubacterium and Roseburia species, and increased abundance of Akkermansia muciniphila and Bifidobacterium species. Pathway enrichment analysis and metabolic potential, constructed from microbial gene abundance, revealed disruptions in microbial carbohydrate and lipid metabolism and increased amino acid and nucleotide metabolism. These global gene-level signatures indicate an increased response to oxidative stress, decreased cellular growth and microbial motility, and disrupted inter-community signaling.

ConclusionsA metagenomic meta-analysis of PD shows consistent and novel alterations in taxonomic representation, functional metabolic potential, and microbial gene abundance across four independent studies from three continents. These data reveal stereotypic changes in the gut microbiome are a consistent feature of PD, highlighting potential diagnostic and therapeutic avenues for future research.
]]></description>
<dc:creator>Boktor, J. C.</dc:creator>
<dc:creator>Sharon, G.</dc:creator>
<dc:creator>Verhagen Metman, L. A.</dc:creator>
<dc:creator>Hall, D. A.</dc:creator>
<dc:creator>Engen, P. A.</dc:creator>
<dc:creator>Zreloff, Z.</dc:creator>
<dc:creator>Hakim, D. J.</dc:creator>
<dc:creator>Bostick, J. W.</dc:creator>
<dc:creator>Ousey, J.</dc:creator>
<dc:creator>Lange, D.</dc:creator>
<dc:creator>Humphrey, G.</dc:creator>
<dc:creator>Ackermann, G.</dc:creator>
<dc:creator>Carlin, M.</dc:creator>
<dc:creator>Knight, R.</dc:creator>
<dc:creator>Keshavarzian, A.</dc:creator>
<dc:creator>Mazmanian, S. K.</dc:creator>
<dc:date>2022-07-21</dc:date>
<dc:identifier>doi:10.1101/2022.07.20.500694</dc:identifier>
<dc:title><![CDATA[Integrated multi-cohort analysis of the Parkinson's disease gut metagenome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.06.27.497828v1?rss=1">
<title>
<![CDATA[
A prebiotic diet modulates microglial states and motor deficits in α-synuclein overexpressing mice 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.06.27.497828v1?rss=1"
</link>
<description><![CDATA[
Parkinsons disease (PD) is a movement disorder characterized by neuroinflammation, -synuclein pathology, and neurodegeneration. Most cases of PD are non-hereditary, suggesting a strong role for environmental factors, and it has been speculated that disease may originate in peripheral tissues such as the gastrointestinal (GI) tract before affecting the brain. The gut microbiome is altered in PD and may impact motor and GI symptoms as indicated by animal studies, though mechanisms of gut-brain interactions remain incompletely defined. Intestinal bacteria ferment dietary fibers into short-chain fatty acids, with fecal levels of these molecules differing between PD and healthy controls and in mouse models. Among other effects, dietary microbial metabolites can modulate activation of microglia, brain-resident immune cells implicated in PD. We therefore investigated whether a fiber-rich diet influences microglial function in -synuclein overexpressing (ASO) mice, a preclinical model with PD-like symptoms and pathology. Feeding a prebiotic high-fiber diet attenuates motor deficits and reduces -synuclein aggregation in the substantia nigra of mice. Concomitantly, the gut microbiome of ASO mice adopts a profile correlated with health upon prebiotic treatment, which also reduces microglial activation. Single-cell RNA-seq analysis of microglia from the substantia nigra and striatum uncovers increased pro-inflammatory signaling and reduced homeostatic responses in ASO mice compared to wild-type counterparts on standard diets. However, prebiotic feeding reverses pathogenic microglial states in ASO mice and promotes expansion of protective disease-associated macrophage (DAM) subsets of microglia. Notably, depletion of microglia using a CSF1R inhibitor eliminates the beneficial effects of prebiotics by restoring motor deficits to ASO mice despite feeding a prebiotic diet. These studies uncover a novel microglia-dependent interaction between diet and motor symptoms in mice, findings that may have implications for neuroinflammation and PD.
]]></description>
<dc:creator>Abdel-Haq, R.</dc:creator>
<dc:creator>Schlachetzki, J. C. M.</dc:creator>
<dc:creator>Boktor, J. C.</dc:creator>
<dc:creator>Cantu-Jungles, T. M.</dc:creator>
<dc:creator>Thron, T.</dc:creator>
<dc:creator>Zhang, M.</dc:creator>
<dc:creator>Bostick, J. W.</dc:creator>
<dc:creator>Khazaei, T.</dc:creator>
<dc:creator>Chilakala, S.</dc:creator>
<dc:creator>Morais, L. H.</dc:creator>
<dc:creator>Humphrey, G.</dc:creator>
<dc:creator>Keshavarzian, A.</dc:creator>
<dc:creator>Katz, J. E.</dc:creator>
<dc:creator>Thomson, M.</dc:creator>
<dc:creator>Knight, R.</dc:creator>
<dc:creator>Gradinaru, V.</dc:creator>
<dc:creator>Hamaker, B. R.</dc:creator>
<dc:creator>Glass, C. K.</dc:creator>
<dc:creator>Mazmanian, S. K.</dc:creator>
<dc:date>2022-06-30</dc:date>
<dc:identifier>doi:10.1101/2022.06.27.497828</dc:identifier>
<dc:title><![CDATA[A prebiotic diet modulates microglial states and motor deficits in α-synuclein overexpressing mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.07.13.499896v1?rss=1">
<title>
<![CDATA[
Structural and functional landscape of α-synuclein fibril conformations amplified from cerebrospinal fluid 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.07.13.499896v1?rss=1"
</link>
<description><![CDATA[
Lewy body dementias are pathologically defined by the deposition of -synuclein fibrils into inclusions throughout the brain. Cerebrospinal fluid(CSF) in disease harbors circulating -synuclein-fibril seeds, and parental -synuclein fibrils can template core structure into amplified fibrils. Using cryo-electron microscopy, we identify six novel -synuclein fibril assemblies amplified from ten CSF samples (3.8[A] to 2.9[A] nominal resolutions). Fibrils are classified based on two types of filament interaction, two types of {beta}-sheet stacking, and two types of hydrophobic pocket. CSF-amplified fibril products have one, two, or three distinct assemblies each. Six of ten samples share a common fibril assembly. Within this classification, the fibrils have distinct profiles in amyloid dye binding, and dramatically different potencies in both seeding new inclusions in neurons and evoked microglial pro-inflammatory responses. However, no single structural feature predicts functional phenotypes. Our results highlight CSF as a valuable resource to identify novel -synuclein assemblies potentially important in disease.
]]></description>
<dc:creator>Sokratian, A.</dc:creator>
<dc:creator>Zhou, Y.</dc:creator>
<dc:creator>Xu, E.</dc:creator>
<dc:creator>Viverette, E.</dc:creator>
<dc:creator>Dillard, L.</dc:creator>
<dc:creator>Yuan, Y.</dc:creator>
<dc:creator>Li, J.</dc:creator>
<dc:creator>Matarangas, A.</dc:creator>
<dc:creator>Bouvette, J.</dc:creator>
<dc:creator>Borgnia, M. J.</dc:creator>
<dc:creator>Bartesaghi, A.</dc:creator>
<dc:creator>West, A.</dc:creator>
<dc:date>2022-07-13</dc:date>
<dc:identifier>doi:10.1101/2022.07.13.499896</dc:identifier>
<dc:title><![CDATA[Structural and functional landscape of α-synuclein fibril conformations amplified from cerebrospinal fluid]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.07.26.501592v1?rss=1">
<title>
<![CDATA[
Nicotine-mediated rescue of α-synuclein toxicity requires synaptic vesicle glycoprotein 2 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.07.26.501592v1?rss=1"
</link>
<description><![CDATA[
BackgroundParkinsons disease (PD) is characterized by -synuclein aggregation and loss of dopamine (DA) neurons in the substantia nigra. Risk of PD arises due to a combination of genetic and environmental factors, which may interact, termed gene-environment (GxE) interactions. An inverse association between smoking and risk of PD is well-established, and a previous genome-wide GxE interaction study identified genetic variation in the synaptic-vesicle glycoprotein 2C (SV2C) locus as an important mediator of the degree to which smoking is inversely associated with PD.

ObjectiveWe sought to determine the mechanism of the smoking-SV2C interaction in a Drosophila model of PD.

MethodsFlies expressing human -synuclein in all neurons develop the hallmarks of PD, including motor dysfunction, loss of DA neurons, and formation of -synuclein inclusions. We assessed the effects of increasing doses of nicotine on these parameters of neurodegeneration, in the presence or absence of SV2 knockdown.

Results-synuclein-expressing flies treated with nicotine had improvement in locomotion, DA neuron counts, and in -synuclein aggregation. However, in -synuclein-expressing flies in which Drosophila orthologs of SV2 were knocked down, nicotine failed to rescue neurodegeneration.

ConclusionsThis work confirms a GxE interaction between nicotine and SV2, defines a role for this interaction in -synuclein proteostasis, and suggests that future clinical trials on nicotine should consider genetic variation in SV2C. Further, this provides proof of concept that our model can be used for mechanistic study of GxE, paving the way for investigation of additional GxE interactions or identification of novel GxE.
]]></description>
<dc:creator>Olsen, A. L.</dc:creator>
<dc:creator>Clemens, S. G.</dc:creator>
<dc:creator>Feany, M. B.</dc:creator>
<dc:date>2022-07-28</dc:date>
<dc:identifier>doi:10.1101/2022.07.26.501592</dc:identifier>
<dc:title><![CDATA[Nicotine-mediated rescue of α-synuclein toxicity requires synaptic vesicle glycoprotein 2]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.08.08.503142v1?rss=1">
<title>
<![CDATA[
In vivo reduction of age-dependent neuromelanin accumulation mitigates features of Parkinson's disease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.08.08.503142v1?rss=1"
</link>
<description><![CDATA[
Humans accumulate with age the dark-brown pigment neuromelanin inside specific neuronal groups. Neurons with the highest neuromelanin levels are particularly susceptible to degeneration in Parkinsons disease, especially dopaminergic neurons of the substantia nigra (SN), the loss of which leads to characteristic motor Parkinsons disease symptoms. In contrast to humans, neuromelanin does not appear spontaneously in most animals, including rodents, and Parkinsons disease is an exclusively human condition. Using humanized neuromelanin-producing rodents, we recently found that neuromelanin can trigger Parkinsons disease pathology when accumulated above a specific pathogenic threshold.

Here, by taking advantage of this newly developed animal model, we assessed whether the intracellular buildup of neuromelanin that occurs with age can be slowed down in vivo to prevent or attenuate Parkinsons disease. Because neuromelanin derives from the oxidation of free cytosolic dopamine, we enhanced dopamine vesicular encapsulation in the SN of neuromelanin-producing rats by viral vector-mediated overexpression of vesicular monoamine transporter 2 (VMAT2). This strategy reduced the formation of potentially toxic oxidized dopamine species that can convert into neuromelanin and maintained intracellular neuromelanin levels below their pathogenic threshold. Decreased neuromelanin production was associated with an attenuation of Lewy body-like inclusion formation and a long-term preservation of dopamine homeostasis, nigrostriatal neuronal integrity and motor function in these animals.

Our results demonstrate the feasibility and therapeutic potential of modulating age-dependent intracellular neuromelanin production in vivo, thereby opening an unexplored path for the treatment of Parkinsons disease and, in a broader sense, brain aging.
]]></description>
<dc:creator>Gonzalez-Sepulveda, M.</dc:creator>
<dc:creator>Compte, J.</dc:creator>
<dc:creator>Cuadros, T.</dc:creator>
<dc:creator>Nicolau, A.</dc:creator>
<dc:creator>Guillard-Sirieix, C.</dc:creator>
<dc:creator>Penuelas, N.</dc:creator>
<dc:creator>Lorente-Picon, M.</dc:creator>
<dc:creator>Parent, A.</dc:creator>
<dc:creator>Romero-Gimenez, J.</dc:creator>
<dc:creator>Cladera-Sastre, J. M.</dc:creator>
<dc:creator>Laguna, A.</dc:creator>
<dc:creator>Vila, M.</dc:creator>
<dc:date>2022-08-11</dc:date>
<dc:identifier>doi:10.1101/2022.08.08.503142</dc:identifier>
<dc:title><![CDATA[In vivo reduction of age-dependent neuromelanin accumulation mitigates features of Parkinson's disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.08.14.503930v1?rss=1">
<title>
<![CDATA[
Unconventional Initiation of PINK1/Parkin Mitophagy by Optineurin 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.08.14.503930v1?rss=1"
</link>
<description><![CDATA[
Cargo sequestration is a fundamental step of selective autophagy in which cells generate a double membrane structure termed an autophagosome on the surface of cargoes. NDP52, TAX1BP1 and p62 bind FIP200 which recruits the ULK1/2 complex to initiate autophagosome formation on cargoes. How OPTN initiates autophagosome formation during selective autophagy remains unknown despite its importance in neurodegeneration. Here, we uncover an unconventional path of PINK1/Parkin mitophagy initiation by OPTN that does not begin with FIP200 binding nor require the ULK1/2 kinases. Using gene-edited cell lines and in vitro reconstitutions, we show that OPTN utilizes the kinase TBK1 which binds directly to the class III phosphatidylinositol 3-kinase complex I to initiate mitophagy. During NDP52 mitophagy initiation, TBK1 is functionally redundant with ULK1/2, classifying TBK1s role as a selective autophagy initiating kinase. Overall, this work reveals that OPTN mitophagy initiation is mechanistically distinct and highlights the mechanistic plasticity of selective autophagy pathways.
]]></description>
<dc:creator>Nguyen, T. N.</dc:creator>
<dc:creator>Sawa-makarska, J.</dc:creator>
<dc:creator>Khuu, G.</dc:creator>
<dc:creator>Lam, W. K.</dc:creator>
<dc:creator>Adriaenssens, E.</dc:creator>
<dc:creator>Fracchiolla, D.</dc:creator>
<dc:creator>Shoebridge, S.</dc:creator>
<dc:creator>Padman, B. S.</dc:creator>
<dc:creator>Skulsuppaisarn, M.</dc:creator>
<dc:creator>Lindblom, R. S. J.</dc:creator>
<dc:creator>Martens, S.</dc:creator>
<dc:creator>Lazarou, M.</dc:creator>
<dc:date>2022-08-15</dc:date>
<dc:identifier>doi:10.1101/2022.08.14.503930</dc:identifier>
<dc:title><![CDATA[Unconventional Initiation of PINK1/Parkin Mitophagy by Optineurin]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.08.04.502831v1?rss=1">
<title>
<![CDATA[
Is Tau the Initial Pathology in Dopaminergic Nigrostriatal Degeneration? Studies in Parkinsonism and Parkinson ' Disease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.08.04.502831v1?rss=1"
</link>
<description><![CDATA[
While Parkinsons disease (PD) remains clinically defined by cardinal motor symptoms resulting from nigrostriatal degeneration, it is now appreciated that PD consists of multiple pathologies, but it is unclear which occurs first and which are responsible for the nigrostriatal degeneration. For the past number of years, we have been studying a well-characterized cohort of subjects with motor impairment that we have termed mild motor deficits (MMD). Motor deficits were determined on a modified and validated Unified Parkinsons Disease Rating Scale III (UPDRS III), but they occur to a degree insufficient to diagnose PD. We consider this population to have prodromal PD. However, in past studies, cases in this cohort had a selection bias as both a clinical syndrome in between no motor deficits and PD, plus nigral Lewy pathology as defined post-mortem, were required for inclusion. Therefore, in this study, we only based inclusion on a clinical phenotype intermediate between no motor impairment and PD. Then, we divided this group further based upon whether or not they had a synucleinopathy. Here we demonstrate that loss of nigral dopaminergic neurons, loss of putamenal dopaminergic innervation, loss of TH-phenotype in the substantia nigra and putamen, and changes in axonal transport occur equally in groups with and without nigral alpha-synuclein aggregates. Indeed, the common feature of these two groups is that both have similar degrees of AT8-expressing phospho-tau, a pathology not seen in the nigrostriatal system of aged-matched controls. These finding were confirmed with early (CP13) and late (PHF1) tau markers. This suggests that the initiation of nigrostriatal dopaminergic neurodegeneration occurs independently of alpha-synuclein aggregation and is likely tau mediated.
]]></description>
<dc:creator>Chu, Y.</dc:creator>
<dc:creator>Hirst, W. D.</dc:creator>
<dc:creator>Federoff, H. J.</dc:creator>
<dc:creator>Harms, A. S.</dc:creator>
<dc:creator>Stoessl, A. J.</dc:creator>
<dc:creator>Kordower, J. H.</dc:creator>
<dc:date>2022-08-06</dc:date>
<dc:identifier>doi:10.1101/2022.08.04.502831</dc:identifier>
<dc:title><![CDATA[Is Tau the Initial Pathology in Dopaminergic Nigrostriatal Degeneration? Studies in Parkinsonism and Parkinson ' Disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.08.10.503505v1?rss=1">
<title>
<![CDATA[
Membrane remodeling properties of the Parkinson's disease protein LRRK2 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.08.10.503505v1?rss=1"
</link>
<description><![CDATA[
Mutations in Leucine-rich repeat kinase 2 (LRRK2) are responsible for late-onset autosomal dominant Parkinsons disease (PD). LRRK2 has been implicated in a wide range of physiological processes including membrane repair in the endolysosomal system. Here, using cell free systems, we report that purified LRRK2 directly binds acidic lipid bilayers with a preference for highly curved bilayers. While this binding is nucleotide independent, LRRK2 can also deform low curvature liposomes into narrow tubules in a guanylnucleotide-dependent but ATP-independent way. Moreover, assembly of LRRK2 into scaffolds at the surface of lipid tubules can constrict them. We suggest that an interplay between the membrane remodeling and signaling properties of LRRK2 may be key to its physiological function. LRRK2, via its kinase activity, may achieve its signaling role at sites where membrane remodeling occurs.

Significance StatementLRRK2 is one of the most frequently mutated gene in familial Parkinsons disease. While much has been learned about its molecular properties, upstream regulators and protein substrates of its kinase activity, its precise function remains unclear. Recent evidence has pointed to a role of LRRK2 in membrane repair in the endo/lysosomal system. Here we show that purified LRRK2 has membrane remodeling properties. We suggest that its ability to sense and induce membrane curvature may be key to its function in membrane dynamics. These properties may help coordinate a direct role of LRRK2 at the membrane interface with its the signaling role of its kinase domain.
]]></description>
<dc:creator>Wang, X.</dc:creator>
<dc:creator>Wu, Y.</dc:creator>
<dc:creator>Cai, S.</dc:creator>
<dc:creator>Ge, J.</dc:creator>
<dc:creator>Shao, L.</dc:creator>
<dc:creator>De Camilli, P.</dc:creator>
<dc:date>2022-08-10</dc:date>
<dc:identifier>doi:10.1101/2022.08.10.503505</dc:identifier>
<dc:title><![CDATA[Membrane remodeling properties of the Parkinson's disease protein LRRK2]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.04.04.487055v1?rss=1">
<title>
<![CDATA[
Comprehensive Structural Variant Detection: From Mosaic to Population-Level 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.04.04.487055v1?rss=1"
</link>
<description><![CDATA[
Long-read Structural Variation (SV) calling remains a challenging but highly accurate way to identify complex genomic alterations. Here, we present Sniffles2, which is faster and more accurate than state-of-the-art SV caller across different coverages, sequencing technologies, and SV types. Furthermore, Sniffles2 solves the problem of family- to population-level SV calling to produce fully genotyped VCF files by introducing a gVCF file concept. Across 11 probands, we accurately identified causative SVs around MECP2, including highly complex alleles with three overlapping SVs. Sniffles2 also enables the detection of mosaic SVs in bulk long-read data. As a result, we successfully identified multiple mosaic SVs across a multiple system atrophy patient brain. The identified SV showed a remarkable diversity within the cingulate cortex, impacting both genes involved in neuron function and repetitive elements. In summary, we demonstrate the utility and versatility of Sniffles2 to identify SVs from the mosaic to population levels.
]]></description>
<dc:creator>Smolka, M.</dc:creator>
<dc:creator>Paulin, L. F.</dc:creator>
<dc:creator>Grochowski, C. M.</dc:creator>
<dc:creator>Mahmoud, M.</dc:creator>
<dc:creator>Behera, S.</dc:creator>
<dc:creator>Gandhi, M.</dc:creator>
<dc:creator>Hong, K.</dc:creator>
<dc:creator>Pehlivan, D.</dc:creator>
<dc:creator>Scholz, S. W.</dc:creator>
<dc:creator>Carvalho, C. M.</dc:creator>
<dc:creator>Proukakis, C.</dc:creator>
<dc:creator>Sedlazeck, F. J.</dc:creator>
<dc:date>2022-04-05</dc:date>
<dc:identifier>doi:10.1101/2022.04.04.487055</dc:identifier>
<dc:title><![CDATA[Comprehensive Structural Variant Detection: From Mosaic to Population-Level]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-04-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.10.21.513169v1?rss=1">
<title>
<![CDATA[
Pseudogenes limit the identification of novel common transcripts generated by their parent genes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.10.21.513169v1?rss=1"
</link>
<description><![CDATA[
The human genome contains numerous duplicated regions, such as parent-pseudogene pairs, causing sequencing reads to align equally well to either gene. The extent to which this ambiguity complicates transcriptomic analyses is currently unknown. This is concerning as many parent genes have been linked to disease, including GBA1, causally linked to both Parkinsons and Gaucher disease. We find that most of the short sequencing reads that map to GBA1, also map to its pseudogene, GBAP1. Using long-read RNA-sequencing in human brain, where all reads mapped uniquely, we demonstrate significant differences in expression compared to short-read data. We identify novel transcripts from both GBA1 and GBAP1, including protein-coding transcripts that are translated in vitro and detected in proteomic data, but that lack GCase activity. By combining long-read with single-nuclear RNA-sequencing to analyse brain-relevant cell types we demonstrate that transcript expression varies by brain region with cell-type-selectivity. Taken together, these results suggest a non-lysosomal function for both GBA1 and GBAP1 in brain. Finally, we demonstrate that inaccuracies in annotation are widespread among parent genes, with implications for many human diseases.
]]></description>
<dc:creator>Gustavsson, E. K.</dc:creator>
<dc:creator>Sethi, S.</dc:creator>
<dc:creator>Gao, Y.</dc:creator>
<dc:creator>Brenton, J.</dc:creator>
<dc:creator>Garcia Ruiz, S.</dc:creator>
<dc:creator>Zhang, D.</dc:creator>
<dc:creator>Garza, R.</dc:creator>
<dc:creator>Reynolds, R. H.</dc:creator>
<dc:creator>Evans, J. R.</dc:creator>
<dc:creator>Chen, Z.</dc:creator>
<dc:creator>Grant-Peters, M.</dc:creator>
<dc:creator>MacPherson, H.</dc:creator>
<dc:creator>Montgomery, K.</dc:creator>
<dc:creator>Dore, R.</dc:creator>
<dc:creator>Wernick, A. I.</dc:creator>
<dc:creator>Arber, C.</dc:creator>
<dc:creator>Wray, S.</dc:creator>
<dc:creator>Gandhi, S.</dc:creator>
<dc:creator>Esselborn, J.</dc:creator>
<dc:creator>Blauwendraat, C.</dc:creator>
<dc:creator>Douse, C. H.</dc:creator>
<dc:creator>Adami, A.</dc:creator>
<dc:creator>Atacho, D. A. M.</dc:creator>
<dc:creator>Kouli, A.</dc:creator>
<dc:creator>Quaegebeur, A.</dc:creator>
<dc:creator>Barker, R. A.</dc:creator>
<dc:creator>Englund, E.</dc:creator>
<dc:creator>Platt, F. M.</dc:creator>
<dc:creator>Jakobsson, J.</dc:creator>
<dc:creator>Wood, N. W.</dc:creator>
<dc:creator>Houlden, H.</dc:creator>
<dc:creator>Saini, H.</dc:creator>
<dc:creator>Bento, C. F.</dc:creator>
<dc:creator>Hardy, J.</dc:creator>
<dc:creator>Ryten, M.</dc:creator>
<dc:date>2022-10-21</dc:date>
<dc:identifier>doi:10.1101/2022.10.21.513169</dc:identifier>
<dc:title><![CDATA[Pseudogenes limit the identification of novel common transcripts generated by their parent genes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.10.03.510717v1?rss=1">
<title>
<![CDATA[
Distinct adaptations revealed by unbiased proteomic analysis of autophagy cargos in the brain in PINK1 and LRRK2 models of Parkinson's disease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.10.03.510717v1?rss=1"
</link>
<description><![CDATA[
Autophagy is essential for neuronal homeostasis, while defects in autophagy are implicated in Parkinson disease (PD), a prevalent and progressive neurodegenerative disorder. We used unbiased proteomics to compare cargos degraded by basal autophagy in the brain from two mouse models of PD, PINK1-/- and LRRK2G2019S mice. We find evidence for the upregulation of adaptive pathways to support homeostasis in both PD models. In PINK1-/- mice, we observed increased expression of the selective receptor BNIP3 along with evidence of engagement of other alternative pathways for mitophagy. Despite these changes, we find the rate of autophagic flux in PINK1-/- neurons is decreased. In LRRK2G2019S mice, hyperactive kinase activity known to impair autophagosomal and lysosomal function results in increased secretion of extracellular vesicles and autophagy cargo. In support of this observation, we find reduced levels of PIKFYVE, a negative regulator of extracellular vesicle secretion, in both brain and cortical neurons from LRRK2G2019S mice. Thus, distinct adaptive pathways are activated to compensate for perturbations induced by either loss of PINK1 or hyperactivation of LRRK2. Our findings highlight the engagement of compensatory pathways to maintain homeostasis in the brain, and provide insights into the vulnerabilities these compensatory changes may introduce that may further contribute to PD progression.
]]></description>
<dc:creator>Goldsmith, J.</dc:creator>
<dc:creator>Ordureau, A.</dc:creator>
<dc:creator>Stavoe, A. K.</dc:creator>
<dc:creator>Boecker, C. A.</dc:creator>
<dc:creator>Arany, M.</dc:creator>
<dc:creator>Harper, J. W.</dc:creator>
<dc:creator>Holzbaur, E. L.</dc:creator>
<dc:date>2022-10-04</dc:date>
<dc:identifier>doi:10.1101/2022.10.03.510717</dc:identifier>
<dc:title><![CDATA[Distinct adaptations revealed by unbiased proteomic analysis of autophagy cargos in the brain in PINK1 and LRRK2 models of Parkinson's disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.11.02.514817v1?rss=1">
<title>
<![CDATA[
PARK15/FBXO7 is dispensable for PINK1/Parkin-dependent mitophagy in iNeuron and HeLa cell systems 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.11.02.514817v1?rss=1"
</link>
<description><![CDATA[
The protein kinase PINK1 and ubiquitin ligase Parkin promote removal of damaged mitochondria via a feed-forward mechanism involving ubiquitin (Ub) phosphorylation (pUb), Parkin activation, and ubiquitylation of mitochondrial outer membrane proteins to support recruitment of mitophagy receptors. The ubiquitin ligase substrate receptor FBXO7/PARK15 is mutated in an early-onset parkinsonian-pyramidal syndrome. Previous studies have proposed a role for FBXO7 in promoting Parkin-dependent mitophagy. Here, we systematically examine the involvement of FBXO7 in depolarization and mtUPR-dependent mitophagy in the well-established HeLa and induced-neurons cell systems. We find that FBXO7-/- cells have no demonstrable defect in: 1) kinetics of pUb accumulation, 2) pUb puncta on mitochondria by super-resolution imaging, 3) recruitment of Parkin and autophagy machinery to damaged mitochondria, 4) mitophagic flux, and 5) mitochondrial clearance as quantified by global proteomics. Moreover, global proteomics of neurogenesis in the absence of FBXO7 reveals no obvious alterations in mitochondria or other organelles. These results argue against a general role for FBXO7 in Parkin-dependent mitophagy and point to the need for additional studies to define how FBXO7 mutations promote parkinsonian-pyramidal syndrome.
]]></description>
<dc:creator>Kraus, F.</dc:creator>
<dc:creator>Goodall, E. A.</dc:creator>
<dc:creator>Smith, I. R.</dc:creator>
<dc:creator>Jiang, Y.</dc:creator>
<dc:creator>Paoli, J. C.</dc:creator>
<dc:creator>Zhang, J.</dc:creator>
<dc:creator>Paulo, J. A.</dc:creator>
<dc:creator>Harper, J. W.</dc:creator>
<dc:date>2022-11-02</dc:date>
<dc:identifier>doi:10.1101/2022.11.02.514817</dc:identifier>
<dc:title><![CDATA[PARK15/FBXO7 is dispensable for PINK1/Parkin-dependent mitophagy in iNeuron and HeLa cell systems]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.10.28.514238v1?rss=1">
<title>
<![CDATA[
Protein aggregation and calcium dysregulation are the earliest hallmarks of synucleinopathy in human midbrain dopaminergic neurons 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.10.28.514238v1?rss=1"
</link>
<description><![CDATA[
Mutations in the SNCA gene cause autosomal dominant Parkinsons disease (PD), with progressive loss of dopaminergic neurons in the substantia nigra, and accumulation of aggregates of -synuclein. However, the sequence of molecular events that proceed from the SNCA mutation during development, to its end stage pathology is unknown. Utilising human induced pluripotent stem cells (hiPSCs) with SNCA mutations, we resolved the temporal sequence of pathophysiological events that occur during neuronal differentiation in order to discover the early, and likely causative, events in synucleinopathies. We adapted a small molecule-based protocol that generates highly enriched midbrain dopaminergic (mDA) neurons (>80%). We characterised their molecular identity using single-cell RNA sequencing and their functional identity through the synthesis and secretion of dopamine, the ability to generate action potentials, and form functional synapses and networks. RNA velocity analyses confirmed the developmental transcriptomic trajectory of midbrain neural precursors into mDA neurons using our approach, and identified key driver genes in mDA neuronal development. To characterise the synucleinopathy, we adopted super-resolution methods to determine the number, size and structure of aggregates in SNCA-mutant mDA neurons. At one week of differentiation, prior to maturation to mDA neurons of molecular and functional identity, we demonstrate the formation of small aggregates; specifically, {beta}-sheet rich oligomeric aggregates, in SNCA-mutant midbrain immature neurons. The aggregation progresses over time to accumulate phosphorylated aggregates, and later fibrillar aggregates. When the midbrain neurons were functional, we observed evidence of impaired physiological calcium signalling, with raised basal calcium, and impairments in cytosolic and mitochondrial calcium efflux. Once midbrain identity fully developed, SNCA-mutant neurons exhibited bioenergetic impairments, mitochondrial dysfunction and oxidative stress. During the maturation of mDA neurons, upregulation of mitophagy and autophagy occured, and ultimately these multiple cellular stresses lead to an increase in cell death by six weeks post-differentiation. Our differentiation paradigm generates an efficient model for studying disease mechanisms in PD, and highlights that protein misfolding to generate intraneuronal oligomers is one of the earliest critical events driving disease in human neurons, rather than a late-stage hallmark of the disease.
]]></description>
<dc:creator>Virdi, G. S.</dc:creator>
<dc:creator>Choi, M. L.</dc:creator>
<dc:creator>Evans, J. R.</dc:creator>
<dc:creator>Yao, Z.</dc:creator>
<dc:creator>Athauda, D.</dc:creator>
<dc:creator>Strohbuecker, S.</dc:creator>
<dc:creator>Wernick, A. I.</dc:creator>
<dc:creator>Alrashidi, H.</dc:creator>
<dc:creator>Melandri, D.</dc:creator>
<dc:creator>Perez-lloret, J.</dc:creator>
<dc:creator>Stroh, P. R.</dc:creator>
<dc:creator>Sylantyev, S.</dc:creator>
<dc:creator>Eaton, S.</dc:creator>
<dc:creator>Heales, S.</dc:creator>
<dc:creator>Kunath, T.</dc:creator>
<dc:creator>Horrocks, M. H.</dc:creator>
<dc:creator>Abramov, A. Y.</dc:creator>
<dc:creator>Patani, R.</dc:creator>
<dc:creator>Gandhi, S.</dc:creator>
<dc:date>2022-10-28</dc:date>
<dc:identifier>doi:10.1101/2022.10.28.514238</dc:identifier>
<dc:title><![CDATA[Protein aggregation and calcium dysregulation are the earliest hallmarks of synucleinopathy in human midbrain dopaminergic neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.10.12.511960v1?rss=1">
<title>
<![CDATA[
Border-associated macrophages mediate the neuroinflammatory response in an alpha-synuclein model of Parkinson disease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.10.12.511960v1?rss=1"
</link>
<description><![CDATA[
Dopaminergic cell loss due to the accumulation of -syn is a core feature of PD pathogenesis. Neuroinflammation specifically induced by -syn has been shown to exacerbate neurodegeneration, yet the role of CNS resident macrophages in this process remains unclear. We found that a specific subset of CNS resident macrophages, border-associated macrophages (BAMs), play an essential role in mediating -syn related neuroinflammation due to their unique role as the antigen presenting cells necessary to initiate a CD4 T cell response. Surprisingly, the loss of MHCII antigen presentation on microglia had no effect on neuroinflammation. Furthermore, -syn expression led to an expansion in BAM numbers and a unique damage-associated activation state. Through a combinatorial approach of single-cell RNA sequencing and depletion experiments, we found that BAMs played an essential role in immune cell recruitment, infiltration, and antigen presentation. Furthermore, BAMs were identified in post-mortem PD brain in close proximity to T cells. These results point to a critical role for BAMs in mediating PD pathogenesis through their essential role in the orchestration of the -syn-mediated neuroinflammatory response.
]]></description>
<dc:creator>Schonhoff, A. M.</dc:creator>
<dc:creator>Figge, D. A.</dc:creator>
<dc:creator>Jurkuvenaite, A. J.</dc:creator>
<dc:creator>Gallups, N. J.</dc:creator>
<dc:creator>Childers, G. M.</dc:creator>
<dc:creator>Webster, J. A.</dc:creator>
<dc:creator>Standaert, D. G.</dc:creator>
<dc:creator>Goldman, J. E.</dc:creator>
<dc:creator>Harms, A. S.</dc:creator>
<dc:date>2022-10-16</dc:date>
<dc:identifier>doi:10.1101/2022.10.12.511960</dc:identifier>
<dc:title><![CDATA[Border-associated macrophages mediate the neuroinflammatory response in an alpha-synuclein model of Parkinson disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-10-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.11.14.516471v1?rss=1">
<title>
<![CDATA[
Regulatory imbalance between LRRK2 kinase, PPM1H phosphatase, and ARF6 GTPase disrupts the axonal transport of autophagosomes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.11.14.516471v1?rss=1"
</link>
<description><![CDATA[
Gain-of-function mutations in the LRRK2 gene cause Parkinsons disease (PD), increasing phosphorylation of RAB GTPases through hyperactive kinase activity. We found that LRRK2-hyperphosphorylated RABs disrupt the axonal transport of autophagosomes by perturbing the coordinated regulation of cytoplasmic dynein and kinesin motors. In iPSC-derived human neurons, knock-in of the strongly-hyperactive LRRK2-p.R1441H mutation caused striking impairments in autophagosome transport, inducing frequent directional reversals and pauses. Knock-out of the opposing Protein Phosphatase 1H (PPM1H) phenocopied the effect of hyperactive LRRK2. Overexpression of ADP-ribosylation factor 6 (ARF6), a GTPase that acts as a switch for selective activation of dynein or kinesin, attenuated transport defects in both p.R1441H knock-in and PPM1H knock-out neurons. Together, these findings support a model where a regulatory imbalance between LRRK2-hyperphosphorylated RABs and ARF6 induces an unproductive "tug-of-war" between dynein and kinesin, disrupting processive autophagosome transport. This disruption may contribute to PD pathogenesis by impairing the essential homeostatic functions of axonal autophagy.
]]></description>
<dc:creator>Dou, D.</dc:creator>
<dc:creator>Smith, E. M.</dc:creator>
<dc:creator>Evans, C. S.</dc:creator>
<dc:creator>Boecker, C. A.</dc:creator>
<dc:creator>Holzbaur, E. L. F.</dc:creator>
<dc:date>2022-11-14</dc:date>
<dc:identifier>doi:10.1101/2022.11.14.516471</dc:identifier>
<dc:title><![CDATA[Regulatory imbalance between LRRK2 kinase, PPM1H phosphatase, and ARF6 GTPase disrupts the axonal transport of autophagosomes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.11.22.517583v1?rss=1">
<title>
<![CDATA[
Golgi-IP, a novel tool for multimodal analysis of Golgi molecular content 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.11.22.517583v1?rss=1"
</link>
<description><![CDATA[
The Golgi is a membrane-bound organelle that is essential for protein and lipid biosynthesis. It represents a central trafficking hub that sorts proteins and lipids to various destinations or for secretion from the cell. The Golgi has emerged as a docking platform for cellular signalling pathways including LRRK2 kinase whose deregulation leads to Parkinson disease. Golgi dysfunction is associated with a broad spectrum of diseases including cancer, neurodegeneration, and cardiovascular diseases. To allow the study of the Golgi at high resolution, we report a rapid immunoprecipitation technique (Golgi-IP) to isolate intact Golgi mini-stacks for subsequent analysis of their content. By fusing the Golgi resident protein TMEM115 to three tandem HA epitopes (GolgiTAG), we purified the Golgi using Golgi-IP with minimal contamination from other compartments. We then established an analysis pipeline using liquid chromatography coupled with mass spectrometry to characterize the human Golgi proteome, metabolome and lipidome. Subcellular proteomics confirmed known Golgi proteins and identified novel ones. Metabolite profiling established the first known human Golgi metabolome and revealed the selective enrichment of uridine-diphosphate (UDP) sugars and their derivatives, which is consistent with their roles in protein and lipid glycosylation. Furthermore, targeted metabolomics validated SLC35A2 as the subcellular transporter for UDP-hexose. Finally, lipidomics analysis showed that phospholipids including phosphatidylcholine, phosphatidylinositol and phosphatidylserine are the most abundant Golgi lipids and that glycosphingolipids are enriched in this compartment. Altogether, our work establishes a comprehensive molecular map of the human Golgi and provides a powerful method to study the Golgi with high precision in health and disease states.

SignificanceThe Golgi is central to protein and lipid processing. It senses and responds to diverse cell states to allow trafficking of macromolecules based on cellular demands. Traditional techniques for purifying the Golgi shaped our understanding of its functions, however such methods are too slow to preserve the labile Golgi metabolome and transient protein interactions. Here, we overcome this issue through the development of a method for the rapid capture of intact Golgi from human cells using organelle-specific immunoprecipitation (Golgi-IP). Using high resolution mass spectrometry, we demonstrate that our approach allows the unbiased characterization of the Golgi proteome, metabolome and lipidome. Thus, we believe that the Golgi-IP will be useful for the study of the Golgi in health and disease states.
]]></description>
<dc:creator>Fasimoye, R.</dc:creator>
<dc:creator>Dong, W.</dc:creator>
<dc:creator>Nirujogi, R. S.</dc:creator>
<dc:creator>Rawat, E. S.</dc:creator>
<dc:creator>Iguchi, M.</dc:creator>
<dc:creator>Nyame, K.</dc:creator>
<dc:creator>Phung, T. K.</dc:creator>
<dc:creator>Bagnoli, E.</dc:creator>
<dc:creator>Prescott, A. R.</dc:creator>
<dc:creator>Alessi, D. R.</dc:creator>
<dc:creator>Abu-remaileh, M.</dc:creator>
<dc:date>2022-11-23</dc:date>
<dc:identifier>doi:10.1101/2022.11.22.517583</dc:identifier>
<dc:title><![CDATA[Golgi-IP, a novel tool for multimodal analysis of Golgi molecular content]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.11.30.518286v1?rss=1">
<title>
<![CDATA[
Temporal landscape of mitochondrial proteostasis governed by the UPRmt 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.11.30.518286v1?rss=1"
</link>
<description><![CDATA[
Breakdown of mitochondrial proteostasis activates quality control pathways including the mitochondrial unfolded protein response (UPRmt) and PINK1/Parkin mitophagy. However, beyond the upregulation of chaperones and proteases, we have a limited understanding of how the UPRmt remodels and restores damaged mito-proteomes. Here, we have developed a functional proteomics framework, termed MitoPQ (Mitochondrial Proteostasis Quantification), to dissect the UPRmts role in maintaining proteostasis during stress. We discover essential roles for the UPRmt in both protecting and repairing proteostasis, with oxidative phosphorylation metabolism being a central target of the UPRmt. Transcriptome analyses together with MitoPQ reveal that UPRmt transcription factors drive independent signaling arms that act in concert to maintain proteostasis. Unidirectional interplay between the UPRmt and PINK1/Parkin mitophagy was found to promote oxidative phosphorylation recovery when the UPRmt failed. Collectively, this study defines the network of proteostasis mediated by the UPRmt and highlights the value of functional proteomics in decoding stressed proteomes.
]]></description>
<dc:creator>Uoselis, L.</dc:creator>
<dc:creator>Lindblom, R.</dc:creator>
<dc:creator>Skulsuppaisarn, M.</dc:creator>
<dc:creator>Khuu, G.</dc:creator>
<dc:creator>Nguyen, T. N.</dc:creator>
<dc:creator>Rudler, D. L.</dc:creator>
<dc:creator>Filipovska, A.</dc:creator>
<dc:creator>Schittenhelm, R. B.</dc:creator>
<dc:creator>Lazarou, M.</dc:creator>
<dc:date>2022-11-30</dc:date>
<dc:identifier>doi:10.1101/2022.11.30.518286</dc:identifier>
<dc:title><![CDATA[Temporal landscape of mitochondrial proteostasis governed by the UPRmt]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.12.05.519143v1?rss=1">
<title>
<![CDATA[
Sex-specific microglial responses to glucocerebrosidase inhibition: relevance to GBA1-linked Parkinson disease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.12.05.519143v1?rss=1"
</link>
<description><![CDATA[
Microglia are heterogenous cells characterized by distinct populations each contributing to specific biological processes in the nervous system, including neuroprotection. To elucidate the impact of sex-specific microglia heterogenicity to the susceptibility of neuronal stress, we analysed the dynamic changes in shape and motility occurring in primary mouse microglia following pro-inflammatory or neurotoxic insults, thus finding sex-specific responses of microglial subpopulations. Male microglia exhibited a pro-inflammatory phenotype, whereas female microglia showed enhanced neuroprotective capabilities associated with the activation of Nrf2 detoxification pathway in neurons. The sex difference in neuroprotective functions is lost by inhibition of glucocerebrosidase, the product of the GBA1 gene, mutations of which are the major risk factor for Parkinsons disease (PD). This finding is consistent with the increased risk of PD observed in female carriers of GBA1 mutation, when compared with wild type population, suggesting a role for microglial functionality in the etiopathogenesis of PD-GBA1.
]]></description>
<dc:creator>Ciana, P.</dc:creator>
<dc:creator>Brunialti, E.</dc:creator>
<dc:creator>Villa, A.</dc:creator>
<dc:creator>Toffoli, M.</dc:creator>
<dc:creator>Lucas Del Pozo, S.</dc:creator>
<dc:creator>Rizzi, N.</dc:creator>
<dc:creator>Meda, C.</dc:creator>
<dc:creator>Maggi, A. C.</dc:creator>
<dc:creator>Schapira, A. H. V.</dc:creator>
<dc:date>2022-12-05</dc:date>
<dc:identifier>doi:10.1101/2022.12.05.519143</dc:identifier>
<dc:title><![CDATA[Sex-specific microglial responses to glucocerebrosidase inhibition: relevance to GBA1-linked Parkinson disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.11.08.515615v1?rss=1">
<title>
<![CDATA[
Rapid iPSC inclusionopathy models shed light on formation, consequence and molecular subtype of alpha-synuclein inclusions 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.11.08.515615v1?rss=1"
</link>
<description><![CDATA[
Intracellular inclusions accompanying neurodegeneration are histopathologically and ultrastructurally heterogeneous but the significance of this heterogeneity is unclear. iPSC models, while promising for disease modeling, do not form inclusions in a reasonable timeframe and suffer from limited tractability. Here, we developed an iPSC toolbox utilizing piggyBac-based or targeted transgenes to rapidly induce CNS cells with concomitant expression of aggregation-prone proteins. This system is amenable to screening and longitudinal tracking at single-cell and single-inclusion resolution. For proof-of-principle, cortical neuron -synuclein "inclusionopathy" models were engineered to form inclusions through exogenous seeding or -synuclein mutation. These models recapitulated known fibril- and lipid-rich inclusion subtypes, uncovering dynamic interactions between them, and refined the classification of inclusions in postmortem brain. Genetic-modifier and protein-interaction screens pinpointed proteins like RhoA whose sequestration into specific inclusion subtypes is likely to be toxic. This iPSC platform should enhance our understanding of proteinaceous pathologies in neurodegeneration and facilitate therapeutics development.
]]></description>
<dc:creator>Lam, I.</dc:creator>
<dc:creator>Ndayisaba, A.</dc:creator>
<dc:creator>Lewis, A. J.</dc:creator>
<dc:creator>Fu, Y.</dc:creator>
<dc:creator>Sagredo, G. T.</dc:creator>
<dc:creator>Zaccagnini, L.</dc:creator>
<dc:creator>Sandoe, J.</dc:creator>
<dc:creator>Sanz, R. L.</dc:creator>
<dc:creator>Vahdatshoar, A.</dc:creator>
<dc:creator>Martin, T. D.</dc:creator>
<dc:creator>Morshed, N.</dc:creator>
<dc:creator>Ichihashi, T.</dc:creator>
<dc:creator>Tripathi, A.</dc:creator>
<dc:creator>Ramalingam, N.</dc:creator>
<dc:creator>Oettgen-Suazo, C.</dc:creator>
<dc:creator>Bartels, T.</dc:creator>
<dc:creator>Schabinger, M.</dc:creator>
<dc:creator>Hallacli, E.</dc:creator>
<dc:creator>Jiang, X.</dc:creator>
<dc:creator>Verma, A.</dc:creator>
<dc:creator>Tea, C.</dc:creator>
<dc:creator>Wang, Z.</dc:creator>
<dc:creator>Hakozaki, H.</dc:creator>
<dc:creator>Yu, X.</dc:creator>
<dc:creator>Hyles, K.</dc:creator>
<dc:creator>Park, C.</dc:creator>
<dc:creator>Theunissen, T. W.</dc:creator>
<dc:creator>Wang, H.</dc:creator>
<dc:creator>Jaenisch, R.</dc:creator>
<dc:creator>Lindquist, S.</dc:creator>
<dc:creator>Stevens, B.</dc:creator>
<dc:creator>Stefanova, N.</dc:creator>
<dc:creator>Wenning, G.</dc:creator>
<dc:creator>Luk, K. C.</dc:creator>
<dc:creator>Sanchez Pernaute, R.</dc:creator>
<dc:creator>Gomez-Esteban, J. C.</dc:creator>
<dc:creator>Felsky, D.</dc:creator>
<dc:creator>Kiyota, Y.</dc:creator>
<dc:creator>Sahni, N.</dc:creator>
<dc:creator>Yi, S. S.</dc:creator>
<dc:creator>Chung, C.-Y.</dc:creator>
<dc:creator>Stahlberg, H.</dc:creator>
<dc:creator>Ferrer, I.</dc:creator>
<dc:creator>Schoneberg, J.</dc:creator>
<dc:creator>Ell</dc:creator>
<dc:date>2022-11-09</dc:date>
<dc:identifier>doi:10.1101/2022.11.08.515615</dc:identifier>
<dc:title><![CDATA[Rapid iPSC inclusionopathy models shed light on formation, consequence and molecular subtype of alpha-synuclein inclusions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.12.15.520427v1?rss=1">
<title>
<![CDATA[
Novel green fluorescent polyamines to analyze ATP13A2 and ATP13A3 activity in the mammalian polyamine transport system 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.12.15.520427v1?rss=1"
</link>
<description><![CDATA[
Cells acquire the polyamines putrescine (PUT), spermidine (SPD) and spermine (SPM) via the complementary action of polyamine uptake and synthesis pathways. The endosomal P5B-type ATPases ATP13A2 and ATP13A3 emerge as major determinants of mammalian polyamine uptake. Our biochemical evidence shows that fluorescently labeled polyamines are genuine substrates of ATP13A2. They can be used to measure polyamine uptake in ATP13A2 and ATP13A3-dependent cell models resembling radiolabeled polyamine uptake. We further report that ATP13A3 enables faster and stronger cellular polyamine uptake than ATP13A2. We also compared the uptake of new green-fluorescent PUT, SPD and SPM analogs using different coupling strategies (amide, triazole or isothiocyanate) and fluorophores (symmetrical BODIPY, BODIPY-FL and FITC). ATP13A2 promotes the uptake of various SPD and SPM analogs, whereas ATP13A3 mainly stimulates the uptake of PUT and SPD conjugates. However, the polyamine linker and coupling position on the fluorophore impacts the transport capacity, whereas replacing the fluorophore affects polyamine selectivity. The highest uptake in ATP13A2 or ATP13A3 cells is observed with BODIPY-FL-amide conjugated to SPD, whereas BODIPY-PUT analogs are specifically taken up via ATP13A3. We found that P5B-type ATPase isoforms transport fluorescently labeled polyamine analogs with a distinct structure-activity relationship (SAR) suggesting that isoform-specific polyamine probes can be designed.
]]></description>
<dc:creator>Houdou, M. C.</dc:creator>
<dc:creator>Jacobs, N.</dc:creator>
<dc:creator>Coene, J.</dc:creator>
<dc:creator>Azfar, M.</dc:creator>
<dc:creator>Vanhoutte, R.</dc:creator>
<dc:creator>Van den Haute, C.</dc:creator>
<dc:creator>Eggermont, J.</dc:creator>
<dc:creator>Daniels, V.</dc:creator>
<dc:creator>Verhelst, S. H. L.</dc:creator>
<dc:creator>Vangheluwe, P.</dc:creator>
<dc:date>2022-12-15</dc:date>
<dc:identifier>doi:10.1101/2022.12.15.520427</dc:identifier>
<dc:title><![CDATA[Novel green fluorescent polyamines to analyze ATP13A2 and ATP13A3 activity in the mammalian polyamine transport system]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.12.06.519342v1?rss=1">
<title>
<![CDATA[
Quantitative mapping of autophagic cargo during nutrient stress reveals YIPF3-YIPF4 as membrane receptors for Golgiphagy 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.12.06.519342v1?rss=1"
</link>
<description><![CDATA[
During nutrient stress, macroautophagy is employed to degrade cellular macromolecules, thereby providing biosynthetic building blocks while simultaneously remodeling the proteome. While the machinery responsible for initiation of macroautophagy is well characterized, our understanding of the extent to which individual proteins, protein complexes and organelles are selected for autophagic degradation, and the underlying targeting mechanisms is limited. Here, we use orthogonal proteomic strategies to provide a global molecular inventory of autophagic cargo during nutrient stress in mammalian cell lines. Through prioritization of autophagic cargo, we identify a heterodimeric pair of membrane-embedded proteins, YIPF3 and YIPF4, as receptors for Golgiphagy. During nutrient stress, YIPF4 is mobilized into ATG8-positive vesicles that traffic to lysosomes as measured via Golgiphagy flux reporters in a process that requires the VPS34 and ULK1-FIP200 arms of the autophagy system. Cells lacking YIPF3 or YIPF4 are selectively defective in elimination of Golgi membrane proteins during nutrient stress. By merging absolute protein abundance with autophagic turnover, we create a global protein census describing how autophagic degradation maps onto protein abundance and subcellular localization. Our results, available via an interactive web tool, reveal that autophagic turnover prioritizes membrane-bound organelles (principally Golgi and ER) for proteome remodeling during nutrient stress.

One-Sentence SummaryDuring nutrient stress, macroautophagy uses organelle-phagy receptors to prioritize recycling of Golgi and ER membrane proteins.
]]></description>
<dc:creator>Hickey, K. L.</dc:creator>
<dc:creator>Swarup, S.</dc:creator>
<dc:creator>Smith, I. R.</dc:creator>
<dc:creator>Paoli, J. C.</dc:creator>
<dc:creator>Paulo, J. A.</dc:creator>
<dc:creator>Harper, J. W.</dc:creator>
<dc:date>2022-12-07</dc:date>
<dc:identifier>doi:10.1101/2022.12.06.519342</dc:identifier>
<dc:title><![CDATA[Quantitative mapping of autophagic cargo during nutrient stress reveals YIPF3-YIPF4 as membrane receptors for Golgiphagy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.12.22.521485v1?rss=1">
<title>
<![CDATA[
Serine-129 phosphorylation of α-synuclein is a trigger for physiologic protein-protein interactions and synaptic function 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.12.22.521485v1?rss=1"
</link>
<description><![CDATA[
Phosphorylation of -synuclein at the Serine-129 site (-syn Ser129P) is an established pathologic hallmark of synucleinopathies, and also a therapeutic target. In physiologic states, only a small fraction of total -syn is phosphorylated at this site, and consequently, almost all studies to date have focused on putative pathologic roles of this post-translational modification. We noticed that unlike native (total) -syn that is widely expressed throughout the brain, the overall pattern of -syn Ser129P is restricted, suggesting intrinsic regulation and putative physiologic roles. Surprisingly, preventing phosphorylation at the Ser-129 site blocked the ability of -syn to attenuate activity-dependent synaptic vesicle (SV) recycling - widely thought to reflect its normal function. Exploring mechanisms, we found that neuronal activity augments -syn Ser-129P, and this phosphorylation is required for -syn binding to VAMP2 and synapsin - two functional binding-partners that are necessary for -syn function. AlphaFold2-driven modeling suggests a scenario where Ser129P induces conformational changes in the C-terminus that stabilizes this region and facilitates protein-protein interactions. Our experiments indicate that the pathology-associated Ser129P is an unexpected physiologic trigger of -syn function, which has broad implications for pathophysiology and drug-development.
]]></description>
<dc:creator>Parra-Rivas, L. A.</dc:creator>
<dc:creator>Madhivanan, K.</dc:creator>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Boyer, N. P.</dc:creator>
<dc:creator>Prakashchand, D. D.</dc:creator>
<dc:creator>Aulston, B. D.</dc:creator>
<dc:creator>Pizzo, D. P.</dc:creator>
<dc:creator>Branes-Guerrero, K.</dc:creator>
<dc:creator>Tang, Y.</dc:creator>
<dc:creator>Das, U.</dc:creator>
<dc:creator>Scott, D. A.</dc:creator>
<dc:creator>Rangamani, P.</dc:creator>
<dc:creator>Roy, S.</dc:creator>
<dc:date>2022-12-23</dc:date>
<dc:identifier>doi:10.1101/2022.12.22.521485</dc:identifier>
<dc:title><![CDATA[Serine-129 phosphorylation of α-synuclein is a trigger for physiologic protein-protein interactions and synaptic function]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.02.27.530238v1?rss=1">
<title>
<![CDATA[
From Policy to Practice: Tracking an Open Science Funding Initiative 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.02.27.530238v1?rss=1"
</link>
<description><![CDATA[
This is a critical moment in the open science landscape. Over the past few years there has been growing momentum to improve open research policies and require grantees to share all research outputs, from datasets to code to protocols, in FAIR (findable, accessible, interoperable and reusable [FAIR]) repositories with persistent identifiers attached. The Aligning Science Across Parkinsons (ASAP) initiative has made substantial investments in improving open science compliance monitoring for its grantees, requiring grantees to update their manuscripts if not all research outputs have been linked in the initial manuscript version. Here, we evaluate ASAPs effectiveness in improving research output sharing for all articles processed through the ASAP compliance workflow between March 1, 2022, and October 1, 2022. Our ultimate goal in sharing our findings is to assist other funders and institutions as they consider open science implementation. By normalizing the open science and compliance process across funding bodies, we hope to simplify and streamline researcher, institutional, and funder workflows, allowing researchers to focus on science by easily leveraging resources and building upon the work of others.
]]></description>
<dc:creator>Ratan, K.</dc:creator>
<dc:creator>McIntosh, S.</dc:creator>
<dc:creator>Shah, H. V.</dc:creator>
<dc:creator>Lewis, M.</dc:creator>
<dc:creator>Dumanis, S. B.</dc:creator>
<dc:creator>Vines, T. H.</dc:creator>
<dc:creator>Schekman, R.</dc:creator>
<dc:creator>Riley, E. A. U.</dc:creator>
<dc:date>2023-02-28</dc:date>
<dc:identifier>doi:10.1101/2023.02.27.530238</dc:identifier>
<dc:title><![CDATA[From Policy to Practice: Tracking an Open Science Funding Initiative]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.01.27.524249v1?rss=1">
<title>
<![CDATA[
Protein network analysis links the NSL complex to Parkinson's disease and mitochondrial biology 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.01.27.524249v1?rss=1"
</link>
<description><![CDATA[
Whilst the majority of PD cases are sporadic, much of our understanding of the pathophysiological basis of disease can be traced back to the study of rare, monogenic forms of disease. In the past decade, the availability of Genome-Wide Association Studies (GWAS) has facilitated a shift in focus, toward identifying common risk variants conferring increased risk of developing PD across the population. A recent mitophagy screening assay of GWAS candidates has functionally implicated the non-specific lethal (NSL) complex in the regulation of PINK1-mitophagy. Here, a bioinformatics approach has been taken to investigate the proteome of the NSL complex, to unpick its relevance to PD progression. The mitochondrial NSL interactome has been built, mining 3 separate repositories: PINOT, HIPPIE and MIST, for curated, literature-derived protein-protein interaction (PPI) data. We built; i) the  mitochondrial interactome, applying gene-set enrichment analysis (GSEA) to explore the relevance of the NSL mitochondrial interactome to PD and, ii) the PD-oriented interactome to uncover biological pathways underpinning the NSL /PD association. In this study, we find the mitochondrial NSL interactome to be significantly enriched for the protein products of PD associated genes, including the Mendelian PD genes LRRK2 and VPS35. Additionally, the PD associated interactome is enriched for mitochondrial processes; "mitochondrial cell death", "mitochondrial protein localisation", "membrane protein localisation" and "mitochondrial transport". Our data points to NSL complex members OGT and WDR5 as key drivers of this increased PD association. These findings strengthen a role for mitochondrial quality control in both familial and sporadic disease.
]]></description>
<dc:creator>Kelly, K.</dc:creator>
<dc:creator>Lewis, P.</dc:creator>
<dc:creator>Plun-Favreau, H.</dc:creator>
<dc:creator>Manzoni, C.</dc:creator>
<dc:date>2023-01-27</dc:date>
<dc:identifier>doi:10.1101/2023.01.27.524249</dc:identifier>
<dc:title><![CDATA[Protein network analysis links the NSL complex to Parkinson's disease and mitochondrial biology]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.02.17.529028v1?rss=1">
<title>
<![CDATA[
Genome-wide screen reveals Rab12 GTPase as a critical activator of pathogenic LRRK2 kinase 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.02.17.529028v1?rss=1"
</link>
<description><![CDATA[
Activating mutations in the Leucine Rich Repeat Kinase 2 (LRRK2) cause Parkinsons disease. LRRK2 phosphorylates a subset of Rab GTPases, particularly Rab10 and Rab8A, and we showed previously that phosphoRabs play an important role in LRRK2 membrane recruitment and activation (Vides et al., 2022). To learn more about LRRK2 pathway regulation, we carried out an unbiased, CRISPR-based genome-wide screen to identify modifiers of cellular phosphoRab10 levels. A flow cytometry assay was developed to detect changes in phosphoRab10 levels in pools of mouse NIH-3T3 cells harboring unique CRISPR guide sequences. Multiple negative and positive regulators were identified; surprisingly, knockout of the Rab12 gene was especially effective in decreasing phosphoRab10 levels in multiple cell types and knockout mouse tissues. Rab-driven increases in phosphoRab10 were specific for Rab12, LRRK2 dependent and PPM1H phosphatase reversible; they were seen with wild type and pathogenic G2019S and R1441C LRRK2. AlphaFold modeling revealed a novel Rab12 binding site in the LRRK2 Armadillo domain and we show that residues predicted to be essential for Rab12 interaction at this site influence overall phosphoRab levels in a manner distinct from Rab29 activation of LRRK2. Our data support a model in which Rab12 binding to a new site in the LRRK2 Armadillo domain activates LRRK2 kinase for Rab phosphorylation and could serve as a new therapeutic target for a novel class of LRRK2 inhibitors that do not target the kinase domain.
]]></description>
<dc:creator>Dhekne, H. S.</dc:creator>
<dc:creator>Tonelli, F.</dc:creator>
<dc:creator>Yeshaw, W. M.</dc:creator>
<dc:creator>Chiang, C. Y.</dc:creator>
<dc:creator>Limouse, C.</dc:creator>
<dc:creator>Jaimon, E.</dc:creator>
<dc:creator>Purlyte, E.</dc:creator>
<dc:creator>Alessi, D. R.</dc:creator>
<dc:creator>Pfeffer, S. R.</dc:creator>
<dc:date>2023-02-18</dc:date>
<dc:identifier>doi:10.1101/2023.02.17.529028</dc:identifier>
<dc:title><![CDATA[Genome-wide screen reveals Rab12 GTPase as a critical activator of pathogenic LRRK2 kinase]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.03.04.531072v1?rss=1">
<title>
<![CDATA[
L1 retrotransposons drive human neuronal transcriptome complexity and functional diversification 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.03.04.531072v1?rss=1"
</link>
<description><![CDATA[
The genetic mechanisms underlying the expansion in size and complexity of the human brain remains poorly understood. L1 retrotransposons are a source of divergent genetic information in hominoid genomes, but their importance in physiological functions and their contribution to human brain evolution is largely unknown. Using multi-omic profiling we here demonstrate that L1-promoters are dynamically active in the developing and adult human brain. L1s generate hundreds of developmentally regulated and cell-type specific transcripts, many which are co-opted as chimeric transcripts or regulatory RNAs. One L1-derived lncRNA, LINC01876, is a human-specific transcript expressed exclusively during brain development. CRISPRi-silencing of LINC01876 results in reduced size of cerebral organoids and premature differentiation of neural progenitors, implicating L1s in human-specific developmental processes. In summary, our results demonstrate that L1-derived transcripts provide a previously undescribed layer of primate- and human-specific transcriptome complexity that contributes to the functional diversification of the human brain.
]]></description>
<dc:creator>Garza, R.</dc:creator>
<dc:creator>Atacho, D.</dc:creator>
<dc:creator>Adami, A.</dc:creator>
<dc:creator>Gerdes, P.</dc:creator>
<dc:creator>Vinod, M.</dc:creator>
<dc:creator>Hsieh, P.</dc:creator>
<dc:creator>Karlsson, O.</dc:creator>
<dc:creator>Horvath, V.</dc:creator>
<dc:creator>Johansson, P. A.</dc:creator>
<dc:creator>Pandiloski, N.</dc:creator>
<dc:creator>Matas, J.</dc:creator>
<dc:creator>Quaegebeur, A.</dc:creator>
<dc:creator>Kouli, A.</dc:creator>
<dc:creator>Sharma, Y.</dc:creator>
<dc:creator>Jonsson, M. E.</dc:creator>
<dc:creator>Monni, E.</dc:creator>
<dc:creator>Englund, E.</dc:creator>
<dc:creator>Eichler, E. E.</dc:creator>
<dc:creator>Gale Hammell, M.</dc:creator>
<dc:creator>Barker, R. A.</dc:creator>
<dc:creator>Kokaia, Z.</dc:creator>
<dc:creator>Douse, C. H.</dc:creator>
<dc:creator>Jakobsson, J.</dc:creator>
<dc:date>2023-03-06</dc:date>
<dc:identifier>doi:10.1101/2023.03.04.531072</dc:identifier>
<dc:title><![CDATA[L1 retrotransposons drive human neuronal transcriptome complexity and functional diversification]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.04.20.537553v1?rss=1">
<title>
<![CDATA[
Single cell spatial transcriptomic and translatomic profiling of dopaminergic neurons in health, ageing and disease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.04.20.537553v1?rss=1"
</link>
<description><![CDATA[
The brain is spatially organized and contains unique cell types, each performing diverse functions, and exhibiting differential susceptibility to neurodegeneration. This is exemplified in Parkinsons disease with the preferential loss of dopaminergic neurons of the substantia nigra pars compacta. Using a Parkinsons transgenic model, we conducted a single-cell spatial transcriptomic and dopaminergic neuron translatomic analysis of young and old mouse brains. Through the high resolving capacity of single-cell spatial transcriptomics, we provide a deep characterization of the expression features of dopaminergic neurons and 27 other cell types within their spatial context, identifying markers of healthy and aging cells, spanning Parkinsons-relevant pathways. We integrate gene enrichment and GWAS data to prioritize putative causative genes for disease investigation, identifying CASR as a novel regulator of dopaminergic calcium handling. These datasets (see: spatialbrain.org) represent the largest public resource for the investigation of spatial gene expression in brain cells in health, aging and disease.
]]></description>
<dc:creator>Kilfeather, P.</dc:creator>
<dc:creator>Khoo, J. H.</dc:creator>
<dc:creator>Wagner, K.</dc:creator>
<dc:creator>Liang, H.</dc:creator>
<dc:creator>Caiazza, M.-C.</dc:creator>
<dc:creator>An, Y.</dc:creator>
<dc:creator>Zhang, X.</dc:creator>
<dc:creator>Chen, X.</dc:creator>
<dc:creator>Connor-Robson, N.</dc:creator>
<dc:creator>Shang, Z.</dc:creator>
<dc:creator>Wade-Martins, R.</dc:creator>
<dc:date>2023-04-20</dc:date>
<dc:identifier>doi:10.1101/2023.04.20.537553</dc:identifier>
<dc:title><![CDATA[Single cell spatial transcriptomic and translatomic profiling of dopaminergic neurons in health, ageing and disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.03.01.530566v1?rss=1">
<title>
<![CDATA[
Early striatal hyperexcitability in an in vitro human striatal microcircuit model carrying the Parkinson's GBA-N370S mutation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.03.01.530566v1?rss=1"
</link>
<description><![CDATA[
Understanding medium spiny neuron (MSN) physiology is essential to understand motor impairments in Parkinsons disease (PD) given the architecture of the basal ganglia. Here, we developed a custom three-chamber microfluidic platform and established a cortico-striato-nigral microcircuit recapitulating the striatal presynaptic triad in vitro using induced pluripotent stem cell (iPSC)-derived neurons. We found that, although cortical glutamatergic projections facilitated MSN synaptic activity, dopaminergic transmission was essential for excitability maturation of MSNs in vitro. Replacement of wild-type iPSC-dopamine neurons (iPSC-DaNs) in the striatal microcircuit with those carrying the PD-related GBA-N370S mutation induced early hyperexcitability in iPSC-MSNs through reduction of voltage-gated sodium and potassium intrinsic currents. Such deficits were resolved in aged cultures or with antagonism of protein kinase A activity in nigrostriatal iPSC-DaNs. Hence, our results highlight the unique utility of modelling striatal neurons in a modular and highly physiological circuit which is essential to reveal mechanistic insights of the loss of electrical functional integrity in the striata of GBA1 PD patients.
]]></description>
<dc:creator>Do, Q. B.</dc:creator>
<dc:creator>Ng, B.</dc:creator>
<dc:creator>Marquez-Gomez, R.</dc:creator>
<dc:creator>Beccano-Kelly, D.</dc:creator>
<dc:creator>Ibarra-Aizpurua, N.</dc:creator>
<dc:creator>Caiazza, M.-C.</dc:creator>
<dc:creator>Lang, C.</dc:creator>
<dc:creator>Baleriola, J.</dc:creator>
<dc:creator>Bengoa-Vergniory, N.</dc:creator>
<dc:creator>Wade-Martins, R.</dc:creator>
<dc:date>2023-03-02</dc:date>
<dc:identifier>doi:10.1101/2023.03.01.530566</dc:identifier>
<dc:title><![CDATA[Early striatal hyperexcitability in an in vitro human striatal microcircuit model carrying the Parkinson's GBA-N370S mutation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.04.15.537033v1?rss=1">
<title>
<![CDATA[
Scalable, flexible carbon fiber electrode thread arrays for three-dimensional spatial profiling of neurochemical activity in deep brain structures of rodents 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.04.15.537033v1?rss=1"
</link>
<description><![CDATA[
We developed a flexible "electrode-thread" array for recording dopamine neurochemical activity from a lateral distribution of subcortical targets (up to 16) transverse to the axis of insertion. Ultrathin ([~] 10 {micro}m diameter) carbon fiber (CF) electrode-threads (CFETs) are clustered into a tight bundle to introduce them into the brain from a single entry point. The individual CFETs splay laterally in deep brain tissue during insertion due to their innate flexibility. This spatial redistribution allows navigation of the CFETs towards deep brain targets spreading horizontally from the axis of insertion. Commercial "linear" arrays provide single entry insertion but only allow measurements along the axis of insertion. Horizontally configured neurochemical recording arrays inflict separate penetrations for each individual channel (i.e., electrode). We tested functional performance of our CFET arrays in vivo for recording dopamine neurochemical dynamics and for providing lateral spread to multiple distributed sites in the striatum of rats. Spatial spread was further characterized using agar brain phantoms to measure electrode deflection as a function of insertion depth. We also developed protocols to slice the embedded CFETs within fixed brain tissue using standard histology techniques. This method allowed extraction of the precise spatial coordinates of the implanted CFETs and their recording sites as integrated with immunohistochemical staining for surrounding anatomical, cytological, and protein expression labels. Neurochemical recording operations tested here can be integrated with already widely established capabilities of CF-based electrodes to record single neuron activity and local field potentials, to enable multi-modal recording functions. Our CFET array has the potential to unlock a wide range of applications, from uncovering the role of neuromodulators in synaptic plasticity, to addressing critical safety barriers in clinical translation towards diagnostic and adaptive treatment in Parkinsons disease and major mood disorders.
]]></description>
<dc:creator>Xia, M.</dc:creator>
<dc:creator>Agca, B. N.</dc:creator>
<dc:creator>Yoshida, T.</dc:creator>
<dc:creator>Choi, J.</dc:creator>
<dc:creator>Amjad, U.</dc:creator>
<dc:creator>Bose, K.</dc:creator>
<dc:creator>Keren, N.</dc:creator>
<dc:creator>Zukerman, S.</dc:creator>
<dc:creator>Cima, M. J.</dc:creator>
<dc:creator>Graybiel, A. M.</dc:creator>
<dc:creator>Schwerdt, H. N.</dc:creator>
<dc:date>2023-04-17</dc:date>
<dc:identifier>doi:10.1101/2023.04.15.537033</dc:identifier>
<dc:title><![CDATA[Scalable, flexible carbon fiber electrode thread arrays for three-dimensional spatial profiling of neurochemical activity in deep brain structures of rodents]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.07.08.499339v1?rss=1">
<title>
<![CDATA[
Mitoguardin-2 is a lipid transporter and its lipid transfer ability is required for function 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.07.08.499339v1?rss=1"
</link>
<description><![CDATA[
Lipid transport proteins at membrane contact sites, where organelles are closely apposed, are critical in redistributing lipids from the endoplasmic reticulum (ER), where they are made, to other cellular membranes. Such protein mediated transfer is especially important for maintaining organelles disconnected from secretory pathways, like mitochondria. Here we identify mitoguardin-2, a mitochondrial protein at contacts with the ER and/or lipid droplets (LDs), as a lipid transporter. An X-ray structure shows that the C-terminal domain of mitoguardin-2 has a hydrophobic cavity that binds lipids. Mass spectrometry analysis reveals that both glycerophospholipids and free-fatty acids co-purify with mitoguardin-2 from cells, and that each mitoguardin-2 can accommodate up to two lipids. Mitoguardin-2 transfers glycerophospholipids between membranes in vitro, and this transport ability is required for roles both in mitochondrial and LD biology. While it is not established that protein-mediated transfer at contacts plays a role in LD metabolism, our findings raise the possibility that mitoguardin-2 functions in transporting fatty acids and glycerophospholipids at mitochondria-LD contacts.
]]></description>
<dc:creator>Hong, Z.</dc:creator>
<dc:creator>Adlakha, J.</dc:creator>
<dc:creator>Guinn, E.</dc:creator>
<dc:creator>Giska, F.</dc:creator>
<dc:creator>Gupta, K.</dc:creator>
<dc:creator>Melia, T.</dc:creator>
<dc:creator>Reinisch, K. M.</dc:creator>
<dc:date>2022-07-09</dc:date>
<dc:identifier>doi:10.1101/2022.07.08.499339</dc:identifier>
<dc:title><![CDATA[Mitoguardin-2 is a lipid transporter and its lipid transfer ability is required for function]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.01.16.523926v1?rss=1">
<title>
<![CDATA[
The chromatin modulating NSL complex regulates genes and pathways genetically linked to Parkinsons disease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.01.16.523926v1?rss=1"
</link>
<description><![CDATA[
Genetic variants conferring risk for Parkinsons disease have been highlighted through genome-wide association studies, yet exploration of their specific disease mechanisms is lacking. Two Parkinsons disease candidate genes, KAT8 and KANSL1, identified through genome-wide studies and a PINK1-mitophagy screen, encode part of the histone acetylating non-specific lethal complex. This complex localises to the nucleus, where it has a role in transcriptional activation, and to mitochondria, where it has been suggested to have a role in mitochondrial transcription. In this study, we sought to identify whether the non-specific lethal complex has potential regulatory relationships with other genes associated with Parkinsons disease in human brain.

Correlation in the expression of non-specific lethal genes and Parkinsons disease-associated genes was investigated in primary gene co-expression networks utilising publicly available transcriptomic data from multiple brain regions (provided by the Genotype-Tissue Expression Consortium and UK Brain Expression Consortium), whilst secondary networks were used to examine cell-type specificity. Reverse engineering of gene regulatory networks generated regulons of the complex, which were tested for heritability using stratified linkage disequilibrium score regression and then validated in vitro using the QuantiGene multiplex assay.

Significant clustering of non-specific lethal genes was revealed alongside Parkinsons disease-associated genes in frontal cortex primary co-expression modules. Both primary and secondary co-expression modules containing these genes were enriched for mainly neuronal cell types. Regulons of the complex contained Parkinsons disease-associated genes and were enriched for biological pathways genetically linked to disease. When examined in a neuroblastoma cell line, 41% of prioritised gene targets showed significant changes in mRNA expression following KANSL1 or KAT8 perturbation.

In conclusion, genes encoding the non-specific lethal complex are highly correlated with and regulate genes associated with Parkinsons disease. Overall, these findings reveal a potentially wider role for this protein complex in regulating genes and pathways implicated in Parkinsons disease.
]]></description>
<dc:creator>Hicks, A. R.</dc:creator>
<dc:creator>Reynolds, R. H.</dc:creator>
<dc:creator>O'Callaghan, B.</dc:creator>
<dc:creator>Garcia Ruiz, S.</dc:creator>
<dc:creator>Gil Martinez, A. L.</dc:creator>
<dc:creator>Botia, J. A.</dc:creator>
<dc:creator>Plun-Favreau, H.</dc:creator>
<dc:creator>Ryten, M.</dc:creator>
<dc:date>2023-01-18</dc:date>
<dc:identifier>doi:10.1101/2023.01.16.523926</dc:identifier>
<dc:title><![CDATA[The chromatin modulating NSL complex regulates genes and pathways genetically linked to Parkinsons disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.06.01.543278v1?rss=1">
<title>
<![CDATA[
Structure and activation of the human autophagy-initiating ULK1C:PI3KC3-C1 supercomplex 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.06.01.543278v1?rss=1"
</link>
<description><![CDATA[
The unc-51-like kinase protein kinase complex (ULK1C) is the most upstream and central player in the initiation of macroautophagy in mammals. Here, the cryo-EM structure of the human ULK1C core was determined at amino acid residue-level resolution. A moderate resolution structure of the ULK1C core in complex with another autophagy core complex, the class III phosphatidylinositol 3-kinase complex I (PI3KC3-C1) was also determined. The two complexes co-assemble through extensive contacts between the FIP200 scaffold subunit of ULK1C and the VPS15, ATG14, and BECN1 subunits of PI3KC3-C1.

The FIP200:ATG13:ULK1 core of ULK1C undergoes a rearrangement from 2:1:1 to 2:2:2 stoichiometry in the presence of PI3KC3-C1. This suggests a structural mechanism for the initiation of autophagy through formation of a ULK1C:PI3KC3-C1 supercomplex and dimerization of ULK1 on the FIP200 scaffold.
]]></description>
<dc:creator>Chen, M. H.</dc:creator>
<dc:creator>Ren, X.</dc:creator>
<dc:creator>Cook, A. S. I.</dc:creator>
<dc:creator>Hurley, J. H.</dc:creator>
<dc:date>2023-06-01</dc:date>
<dc:identifier>doi:10.1101/2023.06.01.543278</dc:identifier>
<dc:title><![CDATA[Structure and activation of the human autophagy-initiating ULK1C:PI3KC3-C1 supercomplex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.03.29.534819v1?rss=1">
<title>
<![CDATA[
TNF-NFkB-p53 axis restricts in vivo survival of hPSC-derived dopamine neuron 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.03.29.534819v1?rss=1"
</link>
<description><![CDATA[
Ongoing, first-in-human clinical trials illustrate the feasibility and translational potential of human pluripotent stem cell (hPSC)-based cell therapies in Parkinsons disease (PD). However, a major unresolved challenge in the field is the extensive cell death following transplantation with <10% of grafted dopamine neurons surviving. Here, we performed a pooled CRISPR/Cas9 screen to enhance survival of postmitotic dopamine neurons in vivo. We identified p53-mediated apoptotic cell death as major contributor to dopamine neuron loss and uncovered a causal link of TNFa-NF{kappa}B signaling in limiting cell survival. As a translationally applicable strategy to purify postmitotic dopamine neurons, we performed a cell surface marker screen that enabled purification without the need for genetic reporters. Combining cell sorting with adalimumab pretreatment, a clinically approved and widely used TNFa inhibitor, enabled efficient engraftment of postmitotic dopamine neurons leading to extensive re-innervation and functional recovery in a preclinical PD mouse model. Thus, transient TNFa inhibition presents a clinically relevant strategy to enhance survival and enable engraftment of postmitotic human PSC-derived dopamine neurons in PD.

HighlightsO_LIIn vivo CRISPR-Cas9 screen identifies p53 limiting survival of grafted human dopamine neurons.
C_LIO_LITNF-NF{kappa}B pathway mediates p53-dependent human dopamine neuron death
C_LIO_LICell surface marker screen to enrich human dopamine neurons for translational use.
C_LIO_LIFDA approved TNF-alpha inhibitor rescues in vivo dopamine neuron survival with in vivo function.
C_LI
]]></description>
<dc:creator>Kim, T. W.</dc:creator>
<dc:creator>Koo, S. Y.</dc:creator>
<dc:creator>Riessland, M.</dc:creator>
<dc:creator>Cho, H.</dc:creator>
<dc:creator>Chaudhry, F.</dc:creator>
<dc:creator>Kolisnyk, B.</dc:creator>
<dc:creator>Russo, M. V.</dc:creator>
<dc:creator>Saurat, N.</dc:creator>
<dc:creator>Mehta, S.</dc:creator>
<dc:creator>Garippa, R.</dc:creator>
<dc:creator>Betel, D.</dc:creator>
<dc:creator>Studer, L.</dc:creator>
<dc:date>2023-03-31</dc:date>
<dc:identifier>doi:10.1101/2023.03.29.534819</dc:identifier>
<dc:title><![CDATA[TNF-NFkB-p53 axis restricts in vivo survival of hPSC-derived dopamine neuron]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.09.24.509340v1?rss=1">
<title>
<![CDATA[
Reduced Thalamic Excitation to Motor Cortical Pyramidal Tract Neurons in a Mouse Model of Parkinsonism 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.09.24.509340v1?rss=1"
</link>
<description><![CDATA[
Degeneration of midbrain dopaminergic (DA) neurons causes a reduced motor output from the primary motor cortex (M1), underlying the motor symptoms of Parkinsons disease (PD). However, cellular and circuitry mechanisms of M1 dysfunction in PD remain undefined. Using multidisciplinary approaches, we found that DA degeneration induces cell-subtype- and inputs-specific reduction of thalamic excitation to M1 pyramidal tract (PT) neurons. Physiological and anatomical analyses suggest that DA degeneration induces a loss of thalamocortical synapses to M1 PT neurons, resulting in an impaired thalamic driving of their activities. Moreover, we showed that the decreased thalamocortical connectivity are mediated by an excessive activation of NMDA receptors of M1 PT neurons. Further, the decreased thalamocortical transmission in parkinsonism can be rescued by chemogenetically suppressing basal ganglia outputs. Together, our data suggest that the reduced motor cortical outputs in parkinsonism are not only an immediate consequence of basal ganglia inhibition but also involves specific local circuitry adaptations within M1. This study reveals novel insight in the pathophysiology of parkinsonian motor deficits.
]]></description>
<dc:creator>Chen, L.</dc:creator>
<dc:creator>Daniels, S.</dc:creator>
<dc:creator>Dvorak, R.</dc:creator>
<dc:creator>Chu, H.-y.</dc:creator>
<dc:date>2022-09-26</dc:date>
<dc:identifier>doi:10.1101/2022.09.24.509340</dc:identifier>
<dc:title><![CDATA[Reduced Thalamic Excitation to Motor Cortical Pyramidal Tract Neurons in a Mouse Model of Parkinsonism]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.04.02.535274v1?rss=1">
<title>
<![CDATA[
α-synuclein expression in response to bacterial ligands and metabolites in gut enteroendocrine cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.04.02.535274v1?rss=1"
</link>
<description><![CDATA[
BackgroundCaudo-rostral migration of pathological forms of -synuclein from the gut to the brain is proposed as an early feature in Parkinson disease (PD) pathogenesis, but the underlying mechanisms remain unknown. Intestinal enteroendocrine cells sense and respond to numerous luminal signals, including bacterial factors, and transmit this information to the brain via the enteric nervous system and vagus nerve. There is evidence that gut bacteria composition and their metabolites change in PD patients and these alterations can trigger -synuclein pathology in animal models.

ObjectiveHere we investigated the effect of toll-like receptor (TLR) and free fatty acid receptor (FFA2/3) agonists on -synuclein levels in mouse STC-1 enteroendocrine cells.

MethodsSTC-1 cells were treated with TLR and FFA2/3 agonists alone and in combination with selective antagonists. The level of -synuclein protein was measured in cell lysates and cell culture media by western blot and ELISA. And the level of -synuclein and tumour necrosis factor (TNF) mRNA was measured by quantitative RT-PCR.

ResultsTLR and FFA receptor agonists significantly increased intracellular and extracellular -synuclein levels and antagonists significantly reduced these effects. TLR and FFA receptor agonists also significantly increased TNF transcription and this was inhibited by corresponding antagonists.

ConclusionsElevated intracellular -synuclein increases the likelihood of aggregation and conversion to toxic forms. Factors derived from bacteria induce -synuclein accumulation in STC-1 cells. Here we provide support for a mechanism by which exposure of enteroendocrine cells to specific bacterial factors found in PD gut dysbiosis might facilitate accumulation and transmission of -synuclein pathology from the gut to the brain.
]]></description>
<dc:creator>Hurley, M. J.</dc:creator>
<dc:creator>Menozzi, E.</dc:creator>
<dc:creator>Koletsi, S.</dc:creator>
<dc:creator>Bates, R.</dc:creator>
<dc:creator>Gegg, M. E.</dc:creator>
<dc:creator>Chau, K.-Y.</dc:creator>
<dc:creator>Blottiere, H. M.</dc:creator>
<dc:creator>Macnaughtan, J.</dc:creator>
<dc:creator>Schapira, A. H. V.</dc:creator>
<dc:date>2023-04-02</dc:date>
<dc:identifier>doi:10.1101/2023.04.02.535274</dc:identifier>
<dc:title><![CDATA[α-synuclein expression in response to bacterial ligands and metabolites in gut enteroendocrine cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.09.01.505977v1?rss=1">
<title>
<![CDATA[
LRRK2-G2019S SYNERGIZES WITH AGEING AND LOW-GRADE INFLAMMATION TO PROMOTE GUT AND PERIPHERAL IMMUNE CELL ACTIVATION THAT PRECEDE NIGROSTRIATAL DEGENERATION 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.09.01.505977v1?rss=1"
</link>
<description><![CDATA[
BackgroundMutations in the leucine-rich repeat kinase 2 (LRRK2) gene are the most frequent cause of familial Parkinsons disease (PD). The incomplete penetrance of LRRK2 mutations suggest that additional hits are required for disease onset. We hypothesized that chronic low-grade inflammation interacts with LRRK2 G2019S, the most frequent PD-associated mutation, to activate peripheral and central immune reactions and drive age-dependent neurodegeneration.

Methods and ResultsWe exposed wild-type and LRRK2 G2019S mice to a low chronic dose of lipopolysaccharide, and we performed a longitudinal analysis of central and peripheral immune reactions and neurodegeneration. Low-dose inflammation triggered nigrostriatal degeneration, macrophage/monocyte brain infiltration, and astro-/microgliosis. LRRK2 G2019S mice showed an early dysregulation of peripheral cytokines, increased CD4+ T-cell infiltration and -synuclein aggregation in the colon. Interestingly, peripheral immune activation and colonic -synuclein aggregation precede astro-/microgliosis and neurodegeneration.

ConclusionsOur study suggests an early role of the peripheral immune system and the gut in LRRK2 PD and provides a novel model to study early therapeutic immune targets and biomarkers.
]]></description>
<dc:creator>Giachino, C.</dc:creator>
<dc:creator>Tirolo, C.</dc:creator>
<dc:creator>Caniglia, S.</dc:creator>
<dc:creator>Serapide, M. F.</dc:creator>
<dc:creator>L'Episcopo, F.</dc:creator>
<dc:creator>Bertoli, F.</dc:creator>
<dc:creator>Giuliano, C.</dc:creator>
<dc:creator>Mearelli, M.</dc:creator>
<dc:creator>Jakobi, M.</dc:creator>
<dc:creator>Schneiderhan-Marra, N.</dc:creator>
<dc:creator>Deleidi, M.</dc:creator>
<dc:creator>Marchetti, B.</dc:creator>
<dc:date>2022-09-03</dc:date>
<dc:identifier>doi:10.1101/2022.09.01.505977</dc:identifier>
<dc:title><![CDATA[LRRK2-G2019S SYNERGIZES WITH AGEING AND LOW-GRADE INFLAMMATION TO PROMOTE GUT AND PERIPHERAL IMMUNE CELL ACTIVATION THAT PRECEDE NIGROSTRIATAL DEGENERATION]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.03.24.534149v1?rss=1">
<title>
<![CDATA[
Post-fibrillization nitration of alpha-synuclein abolishes its seeding activity and pathology formation in primary neurons and in vivo 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.03.24.534149v1?rss=1"
</link>
<description><![CDATA[
Increasing evidence points to post-translational modifications (PTMs) as key regulators of alpha-synuclein (-Syn) function in health and disease. However, whether these PTMs occur before or after -Syn pathology formation and their role in regulating -Syn toxicity remain unclear. In this study, we demonstrate that post-fibrillization nitration of -Syn fibrils induced their fragmentation, modified their surface and dynamic properties but not their structure, and nearly abolished their seeding activity in primary neurons and in vivo. Furthermore, we show that the dynamic and surface properties of the fibrils, rather than simply their length, are important determinants of -Syn fibril seeding activity. Altogether, our work demonstrates that post-aggregation modifications of -Syn may provide novel approaches to target a central process that contributes to pathology formation and disease progression. Finally, our results suggest that the pattern of PTMs on pathological aggregates, rather than simply their presence, could be a key determinant of their toxicity and neurodegeneration. This calls for reconsidering current approaches relying solely on quantifying and correlating the level of pathology to assess the efficacy of novel therapies, as not all -Syn aggregates in the brain are pathogenic.
]]></description>
<dc:creator>Donzelli, S.</dc:creator>
<dc:creator>OSullivan, S.</dc:creator>
<dc:creator>Mahul-Mellier, A.-L.</dc:creator>
<dc:creator>Ulusoy, A.</dc:creator>
<dc:creator>Fusco, G.</dc:creator>
<dc:creator>Kumar, S. T.</dc:creator>
<dc:creator>Chiki, A.</dc:creator>
<dc:creator>Burtscher, J.</dc:creator>
<dc:creator>Boussouf, M. L. D.</dc:creator>
<dc:creator>Rostami, I.</dc:creator>
<dc:creator>De Simone, A. D. S.</dc:creator>
<dc:creator>Di Monte, D. A.</dc:creator>
<dc:creator>Lashuel, H. A.</dc:creator>
<dc:date>2023-03-25</dc:date>
<dc:identifier>doi:10.1101/2023.03.24.534149</dc:identifier>
<dc:title><![CDATA[Post-fibrillization nitration of alpha-synuclein abolishes its seeding activity and pathology formation in primary neurons and in vivo]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.08.05.552108v1?rss=1">
<title>
<![CDATA[
Feed-forward metabotropic signaling by Cav1 Ca2+ channels supports pacemaking in pedunculopontine cholinergic neurons 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.08.05.552108v1?rss=1"
</link>
<description><![CDATA[
Like a handful of other neuronal types in the brain, cholinergic neurons (CNs) in the pedunculopontine nucleus (PPN) are lost in the course of Parkinsons disease (PD). Why this is the case is unknown. One neuronal trait implicated in PD selective neuronal vulnerability is the engagement of feed-forward stimulation of mitochondrial oxidative phosphorylation (OXPHOS) to meet high bioenergetic demand, leading to sustained oxidant stress and ultimately degeneration. The extent to which this trait is shared by PPN CNs is unresolved. To address this question, a combination of molecular and physiological approaches were used. These studies revealed that PPN CNs are autonomous pacemakers with modest spike-associated cytosolic Ca2+ transients. These Ca2+ transients were attributable in part to the opening of high-threshold Cav1.2 Ca2+ channels, but not Cav1.3 channels. Nevertheless, Cav1.2 channel signaling through endoplasmic reticulum ryanodine receptors stimulated mitochondrial OXPHOS to help maintain cytosolic adenosine triphosphate (ATP) levels necessary for pacemaking. Inhibition of Cav1.2 channels led to recruitment of ATP-sensitive K+ channels and slowing of pacemaking. Cav1.2 channel-mediated stimulation of mitochondria increased oxidant stress. Thus, PPN CNs have a distinctive physiological phenotype that shares some, but not all, of the features of other neurons that are selectively vulnerable in PD.
]]></description>
<dc:creator>Tubert, C.</dc:creator>
<dc:creator>Zampese, E.</dc:creator>
<dc:creator>Pancani, T.</dc:creator>
<dc:creator>Tkatch, T.</dc:creator>
<dc:creator>Surmeier, D. J.</dc:creator>
<dc:date>2023-08-07</dc:date>
<dc:identifier>doi:10.1101/2023.08.05.552108</dc:identifier>
<dc:title><![CDATA[Feed-forward metabotropic signaling by Cav1 Ca2+ channels supports pacemaking in pedunculopontine cholinergic neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.08.04.551615v1?rss=1">
<title>
<![CDATA[
Neuromelanin accumulation drives endogenous synucleinopathy in non-human primates 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.08.04.551615v1?rss=1"
</link>
<description><![CDATA[
Although neuromelanin (NMel) is a dark pigment characteristic of dopaminergic neurons in the human substantia nigra pars compacta (SNpc), its potential role in the pathogenesis of Parkinsons disease (PD) has often been neglected since most commonly used laboratory animals lack NMel. Here we took advantage of adeno-associated viral vectors encoding the human tyrosinase gene for triggering a time-dependent NMel accumulation within SNpc dopaminergic neurons in macaques up to similar levels of pigmentation as observed in elderly humans. Furthermore, NMel accumulation induced an endogenous synucleinopathy mimicking intracellular inclusions typically observed in PD together with a progressive degeneration of NMel-expressing dopaminergic neurons. Moreover, Lewy body-like intracellular inclusions were observed in cortical areas of the frontal lobe receiving dopaminergic innervation, supporting a circuit-specific anterograde spread of endogenous synucleinopathy by permissive trans-synaptic templating. In summary, the conducted strategy resulted in the development and characterization of a new macaque model of PD matching the known neuropathology of this disorder with unprecedented accuracy. Most importantly, evidence is provided showing that intracellular aggregation of endogenous alpha-synuclein is triggered by NMel accumulation, therefore any therapeutic approach intended to decrease NMel levels may provide appealing choices for the successful implementation of novel PD therapeutics.
]]></description>
<dc:creator>Chocarro, J.</dc:creator>
<dc:creator>Rico, A. J.</dc:creator>
<dc:creator>Ariznabarreta, G.</dc:creator>
<dc:creator>Roda, E.</dc:creator>
<dc:creator>Honrubia, A.</dc:creator>
<dc:creator>Collantes, M.</dc:creator>
<dc:creator>Penuelas, I.</dc:creator>
<dc:creator>Vazquez, A.</dc:creator>
<dc:creator>Rodriguez-Perez, A. I.</dc:creator>
<dc:creator>Labandeira-Garcia, J. L.</dc:creator>
<dc:creator>Vila Bover, M.</dc:creator>
<dc:creator>Lanciego, J. L.</dc:creator>
<dc:date>2023-08-06</dc:date>
<dc:identifier>doi:10.1101/2023.08.04.551615</dc:identifier>
<dc:title><![CDATA[Neuromelanin accumulation drives endogenous synucleinopathy in non-human primates]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.08.08.552400v1?rss=1">
<title>
<![CDATA[
Modelling human brain-wide pigmentation in rodents recapitulates age-related multisystem neurodegenerative deficits 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.08.08.552400v1?rss=1"
</link>
<description><![CDATA[
One key limitation in developing effective treatments for neurodegenerative diseases is the lack of models accurately mimicking the complex physiopathology of the human disease. Humans accumulate with age the pigment neuromelanin inside neurons that synthesize catecholamines. Neurons reaching the highest neuromelanin levels preferentially degenerate in Parkinsons, Alzheimers and apparently healthy aging individuals. However, this brain pigment is not taken into consideration in current animal models because common laboratory species, such as rodents, do not produce neuromelanin. Here we generate a tissue-specific transgenic mouse, termed tgNM, that mimics the human age-dependent brain-wide distribution of neuromelanin within catecholaminergic regions, based on the constitutive catecholamine-specific expression of human melanin-producing enzyme tyrosinase. We show that, in parallel to progressive human-like neuromelanin pigmentation, these animals display age-related neuronal dysfunction and degeneration affecting numerous brain circuits and body tissues, linked to motor and non-motor deficits, reminiscent of early neurodegenerative stages. This model may open new research avenues in brain aging and neurodegeneration.
]]></description>
<dc:creator>Laguna, A.</dc:creator>
<dc:creator>Penuelas, N.</dc:creator>
<dc:creator>Gonzalez-Sepulveda, M.</dc:creator>
<dc:creator>Nicolau, A.</dc:creator>
<dc:creator>Arthaud, S.</dc:creator>
<dc:creator>Guilliard.Sirieix, C.</dc:creator>
<dc:creator>Lorente-Picon, M.</dc:creator>
<dc:creator>Compte, J.</dc:creator>
<dc:creator>Miquel-Rio, L.</dc:creator>
<dc:creator>Xicoy, H.</dc:creator>
<dc:creator>Liu, J.</dc:creator>
<dc:creator>Parent, A.</dc:creator>
<dc:creator>Cuadros, T.</dc:creator>
<dc:creator>Romero-Gimenez, J.</dc:creator>
<dc:creator>Pujol, G.</dc:creator>
<dc:creator>Gimenez-Llort, L.</dc:creator>
<dc:creator>Fort, P.</dc:creator>
<dc:creator>Bortolozzi, A.</dc:creator>
<dc:creator>Carballo-Carbajal, I.</dc:creator>
<dc:creator>Vila, M.</dc:creator>
<dc:date>2023-08-08</dc:date>
<dc:identifier>doi:10.1101/2023.08.08.552400</dc:identifier>
<dc:title><![CDATA[Modelling human brain-wide pigmentation in rodents recapitulates age-related multisystem neurodegenerative deficits]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.06.01.543059v1?rss=1">
<title>
<![CDATA[
The lipid flippase ATP10B enables cellular lipid uptake under stress conditions 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.06.01.543059v1?rss=1"
</link>
<description><![CDATA[
Pathogenic ATP10B variants have been described in patients with Parkinsons disease and dementia with Lewy body disease, and we previously established ATP10B as a late endo-/lysosomal lipid flippase transporting both phosphatidylcholine (PC) and glucosylceramide (GluCer) from the lysosomal exoplasmic to cytoplasmic membrane leaflet. Since several other lipid flippases regulate cellular lipid uptake, we here examined whether also ATP10B impacts cellular lipid uptake. Transient co-expression of ATP10B with its obligatory subunit CDC50A stimulated the uptake of fluorescently (NBD-) labeled PC in HeLa cells. This uptake is dependent on the transport function of ATP10B, is impaired by disease-associated variants and appears specific for NBD-PC. Uptake of non-ATP10B substrates, such as NBD-sphingomyelin or NBD-phosphatidylethanolamine is not increased. Remarkably, in stable cell lines co-expressing ATP10B/CDC50A we only observed increased NBD-PC uptake following treatment with rotenone, a mitochondrial complex I inhibitor that induces transport-dependent ATP10B phenotypes. Conversely, Im95m and WM-115 cells with endogenous ATP10B expression, present a decreased NBD-PC uptake following ATP10B knockdown, an effect that is exacerbated under rotenone stress. Our data show that the endo-/lysosomal lipid flippase ATP10B contributes to cellular PC uptake under specific cell stress conditions.
]]></description>
<dc:creator>Wouters, R.</dc:creator>
<dc:creator>Beletchi, I.</dc:creator>
<dc:creator>Van den Haute, C.</dc:creator>
<dc:creator>Baekelandt, V.</dc:creator>
<dc:creator>Martin, S.</dc:creator>
<dc:creator>Eggermont, J.</dc:creator>
<dc:creator>Vangheluwe, P.</dc:creator>
<dc:date>2023-06-01</dc:date>
<dc:identifier>doi:10.1101/2023.06.01.543059</dc:identifier>
<dc:title><![CDATA[The lipid flippase ATP10B enables cellular lipid uptake under stress conditions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.08.08.552291v1?rss=1">
<title>
<![CDATA[
Autophagy preferentially degrades non-fibrillar polyQ aggregates 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.08.08.552291v1?rss=1"
</link>
<description><![CDATA[
Aggregation of proteins containing expanded polyglutamine (polyQ) repeats is the cytopathologic hallmark of a group of dominantly inherited neurodegenerative diseases, including Huntingtons disease (HD). Huntingtin (Htt), the disease protein of HD, forms amyloid-like fibrils by liquid-to-solid phase transition. Macroautophagy has been proposed to clear polyQ aggregates, but the efficiency of aggrephagy is limited. Here, we used cryo-electron tomography to visualize the interactions of autophagosomes with polyQ aggregates in cultured cells in situ. We found that an amorphous aggregate phase exists next to the radially organized polyQ fibrils. Autophagosomes preferentially engulfed this amorphous material, mediated by interactions between the autophagy receptor p62/SQSTM1 and the non-fibrillar aggregate surface. In contrast, amyloid fibrils excluded p62 and evaded clearance, resulting in trapping of autophagic structures. These results suggest that the limited efficiency of autophagy in clearing polyQ aggregates is due to the inability of autophagosomes to interact productively with the non-deformable, fibrillar disease aggregates.
]]></description>
<dc:creator>Zhao, D. Y.</dc:creator>
<dc:creator>Bauerlein, F. J.</dc:creator>
<dc:creator>Saha, I.</dc:creator>
<dc:creator>Hartl, F.-U.</dc:creator>
<dc:creator>Baumeister, W.</dc:creator>
<dc:creator>Wilfling, F.</dc:creator>
<dc:date>2023-08-09</dc:date>
<dc:identifier>doi:10.1101/2023.08.08.552291</dc:identifier>
<dc:title><![CDATA[Autophagy preferentially degrades non-fibrillar polyQ aggregates]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.08.09.552634v1?rss=1">
<title>
<![CDATA[
Erythroid differentiation dependent interaction of VPS13A with XK at the plasma membrane of K562 cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.08.09.552634v1?rss=1"
</link>
<description><![CDATA[
Mutations of the bridge-like lipid transport protein VPS13A and of the lipid scramblase XK result in Chorea Acanthocytosis (ChAc) and McLeod syndrome (MLS) respectively, two similar conditions involving neurodegeneration and deformed erythrocytes (acanthocytes). VPS13A binds XK, suggesting a model in which VPS13A forms a lipid transport bridge between the ER and the plasma membrane (PM) where XK resides. However, studies of VPS13A in HeLa and COS7 cells showed that this protein localizes primarily at contacts of the ER with mitochondria. Overexpression of XK in these cells redistributed VPS13A to the biosynthetic XK pool in the ER but not to PM localized XK. Colocalization of VPS13A with XK at the PM was only observed if overexpressed XK harbored mutations that disengage its VPS13A binding site from an intramolecular interaction. As the acanthocytosis phenotype of ChAc and MLS suggests a role of the two proteins in cells of the erythroid lineage, we explored their localization in K562 cells, which differentiate into erythroblasts upon hemin addition. When tagged VPS13A was overexpressed in hemin treated K562 cells, robust formation of ER-PM contacts positive for VPS13A were observed and their formation was abolished in XK KO cells. ER-PM contacts positive for VPS13A were seldomly observed in undifferentiated K562 cells, in spite of the presence of XK in these cells at concentrations similar to those observed after differentiation. These findings reveal that the interaction of VPS13A with XK at ER-PM contacts requires a permissive state which depends upon cell type and/or functional state of the cell.
]]></description>
<dc:creator>Amos, C.</dc:creator>
<dc:creator>Xu, P.</dc:creator>
<dc:creator>De Camilli, P.</dc:creator>
<dc:date>2023-08-10</dc:date>
<dc:identifier>doi:10.1101/2023.08.09.552634</dc:identifier>
<dc:title><![CDATA[Erythroid differentiation dependent interaction of VPS13A with XK at the plasma membrane of K562 cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.12.17.520834v1?rss=1">
<title>
<![CDATA[
LRRK2 Suppresses Lysosome Degradative Activity in Macrophages and Microglia via Transcription Factor E3 Inhibition 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.12.17.520834v1?rss=1"
</link>
<description><![CDATA[
Cells maintain optimal levels of lysosome degradative activity to protect against pathogens, clear waste and generate nutrients. Here we show that LRRK2, a protein that is tightly linked to Parkinsons disease, negatively regulates lysosome degradative activity in macrophages and microglia via a transcriptional mechanism. Depletion of LRRK2 and inhibition of LRRK2 kinase activity enhanced lysosomal proteolytic activity and increased the expression of multiple lysosomal hydrolases. Conversely, the kinase hyperactive LRRK2 G2019S Parkinsons disease mutant suppressed lysosomal degradative activity and gene expression. We identified MiT-TFE transcription factors (TFE3, TFEB and MITF) as mediators of LRRK2-dependent control of lysosomal gene expression. LRRK2 negatively regulated the abundance and nuclear localization of these transcription factors and their depletion prevented LRRK2-dependent changes in lysosome protein levels. These discoveries define a role for LRRK2 in controlling lysosome degradative activity and support a model wherein LRRK2 hyperactivity may increase Parkinsons disease risk by suppressing lysosome degradative activity.

Significance StatementThis study defines a homeostatic mechanism that allows macrophages and microglia to match the degradative activity of their lysosomes to ongoing changes in cellular demand. It shows that the leucine rich repeat kinase 2 (LRRK2) protein suppresses lysosome degradative activity by inhibiting the expression and nuclear localization of the MiT-TFE family of transcription factors that control the expression of multiple genes that encode lysosome proteins. It further demonstrates that a Parkinsons disease mutation that hyperactivates LRRK2 kinase activity limits the degradative activity of lysosomes more strongly. These findings support a model wherein LRRK2 protects cells from excessive lysosome degradative activity and suggest that overactivation of this pathway may increase Parkinsons disease risk by limiting the degradative activity of lysosomes.
]]></description>
<dc:creator>Yadavalli, N.</dc:creator>
<dc:creator>Ferguson, S.</dc:creator>
<dc:date>2022-12-18</dc:date>
<dc:identifier>doi:10.1101/2022.12.17.520834</dc:identifier>
<dc:title><![CDATA[LRRK2 Suppresses Lysosome Degradative Activity in Macrophages and Microglia via Transcription Factor E3 Inhibition]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.08.08.552410v1?rss=1">
<title>
<![CDATA[
Non-ablative disease-modifying effects of magnetic resonance-guided focused ultrasound in neuromelanin-producing parkinsonian rodents 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.08.08.552410v1?rss=1"
</link>
<description><![CDATA[
Age-dependent accumulation of the brain pigment neuromelanin has been implicated in the pathogenesis of Parkinsons disease (PD). In humans, intracellular and extracellular neuromelanin levels are increased in PD postmortem brains and boosting neuromelanin production in rodents compromises neuronal function and viability and triggers a PD-like phenotype. Focused ultrasound has been shown to reduce ultraviolet light-induced skin hyperpigmentation in guinea pig and to remove brain extracellular {beta}-amyloid plaques in Alzheimers mouse models. Here we show that repeated application of transcranial focused ultrasound (tFUS) is able to decrease intracellular and extracellular neuromelanin levels in neuromelanin-producing parkinsonian rats, compared to sham-treated animals, without the need for any additional therapeutic agent or intervention. Reduced neuromelanin levels in tFUS-treated animals were associated with decreased Lewy-like pathology, preserved dopaminergic phenotype, attenuated nigrostriatal degeneration, reduced glial activation, and long-term recovery of motor function. Our findings indicate that tFUS treatment applied at prodromal/early disease stages provides by itself extended structural and functional preservation of the nigrostriatal pathway in neuromelanin-producing parkinsonian rats without causing overt neuronal damage. This FDA-approved technology should thus be explored further as a noninvasive method with neuroprotective potential in PD and to maintain neuromelanin to levels below its pathogenic threshold within the aging population.

One Sentence SummaryTranscranial focused ultrasound reduces age-dependent neuromelanin accumulation and provides therapeutic benefit in parkinsonian rats
]]></description>
<dc:creator>Compte, J.</dc:creator>
<dc:creator>Tible, M.</dc:creator>
<dc:creator>Cuadros, T.</dc:creator>
<dc:creator>Romero-Jimenez, J.</dc:creator>
<dc:creator>Laguna, A.</dc:creator>
<dc:creator>Aubry, J.-F.</dc:creator>
<dc:creator>Dumont, E.</dc:creator>
<dc:creator>Constans, C.</dc:creator>
<dc:creator>Tiennot, T.</dc:creator>
<dc:creator>Santin, M. D.</dc:creator>
<dc:creator>Lehericy, S.</dc:creator>
<dc:creator>Vila, M.</dc:creator>
<dc:date>2023-08-11</dc:date>
<dc:identifier>doi:10.1101/2023.08.08.552410</dc:identifier>
<dc:title><![CDATA[Non-ablative disease-modifying effects of magnetic resonance-guided focused ultrasound in neuromelanin-producing parkinsonian rodents]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.08.07.552338v1?rss=1">
<title>
<![CDATA[
Adaptor Protein-3 Produces Synaptic Vesicles that Release Phasic Dopamine 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.08.07.552338v1?rss=1"
</link>
<description><![CDATA[
The burst firing of midbrain dopamine neurons releases a phasic dopamine signal that mediates reinforcement learning. At many synapses, however, high firing rates deplete synaptic vesicles (SVs), resulting in synaptic depression that limits release. What accounts for the increased release of dopamine by stimulation at high frequency? We find that adaptor protein-3 (AP-3) and its coat protein VPS41 promote axonal dopamine release by targeting vesicular monoamine transporter VMAT2 to the axon rather than dendrites. AP-3 and VPS41 also produce SVs that respond preferentially to high frequency stimulation, independent of their role in axonal polarity. In addition, conditional inactivation of VPS41 in dopamine neurons impairs reinforcement learning, and this involves a defect in the frequency dependence of release rather than the amount of dopamine released. Thus, AP-3 and VPS41 promote the axonal polarity of dopamine release but enable learning by producing a novel population of SVs tuned specifically to high firing frequency that confers the phasic release of dopamine.

Significance statementReinforcement learning requires the phasic dopamine produced by burst firing but synaptic vesicle depletion limits the ability to convey information at high firing rates. We now find that AP-3 has two independent roles in dopamine release. First, AP-3 confers the axonal polarity of dopamine release by targeting vesicular monoamine transporter 2 (VMAT2) to the axon. Second, AP-3 acting locally at the nerve terminal produces synaptic vesicles that respond specifically to high frequency stimulation. Consistent with this, loss of AP-3 impairs reinforcement learning and this reflects the defect in release at high frequency, not the reduction in axonal dopamine.
]]></description>
<dc:creator>Jain, S.</dc:creator>
<dc:creator>Yee, A. G.</dc:creator>
<dc:creator>Maas, J.</dc:creator>
<dc:creator>Gierok, S.</dc:creator>
<dc:creator>Xu, H.</dc:creator>
<dc:creator>Stansil, J.</dc:creator>
<dc:creator>Eriksen, J.</dc:creator>
<dc:creator>Nelson, A.</dc:creator>
<dc:creator>Silm, K.</dc:creator>
<dc:creator>Ford, C. P.</dc:creator>
<dc:creator>Edwards, R. H.</dc:creator>
<dc:date>2023-08-08</dc:date>
<dc:identifier>doi:10.1101/2023.08.07.552338</dc:identifier>
<dc:title><![CDATA[Adaptor Protein-3 Produces Synaptic Vesicles that Release Phasic Dopamine]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.07.03.186411v1?rss=1">
<title>
<![CDATA[
L-type calcium channel contribution to striatal dopamine release is governed by calbindin-D28K, the dopamine transporter, D2-receptors, α2δ-subunits and sex differences 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.07.03.186411v1?rss=1"
</link>
<description><![CDATA[
Ca2+ entry into nigrostriatal dopamine (DA) neurons and axons via L-type voltage-gated Ca2+ channels (LTCCs) contributes respectively to pacemaker activity and DA release, and has long been thought to contribute to vulnerability to degeneration in Parkinsons disease. LTCC function is greater in DA axons and neurons from substantia nigra pars compacta than from ventral tegmental area, but this is not explained by channel expression level. We tested the hypothesis that LTCC-control of DA release is governed rather by local mechanisms, focussing on candidate biological factors known to operate differently between types of DA neurons and/or be associated with their differing vulnerability to parkinsonism, including biological sex, -synuclein, DA transporters (DATs), and calbindin-D28k (Calb1). We detected evoked DA release ex vivo in mouse striatal slices using fast-scan cyclic voltammetry, and assessed LTCC support of DA release by detecting the inhibition of DA release by the LTCC inhibitors isradipine or CP8. Using genetic knockouts or pharmacological manipulations we identified that striatal LTCC support of DA release depended on multiple intersecting factors, in a regionally and sexually divergent manner. LTCC function was promoted by factors associated with Parkinsonian risk, including male sex, -synuclein, DAT, and a dorsolateral co-ordinate, but limited by factors associated with protection i.e. female sex, glucocerebrosidase activity, Calb1, and ventromedial co-ordinate. Together, these data show that LTCC function in DA axons, and isradipine effect, are locally governed and suggest they vary in a manner that in turn might impact on, or reflect, the cellular stress that leads to parkinsonian degeneration.



O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=133 SRC="FIGDIR/small/186411v2_ufig1.gif" ALT="Figure 1">
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]]></description>
<dc:creator>Brimblecombe, K. R.</dc:creator>
<dc:creator>Conor-Robson, N.</dc:creator>
<dc:creator>Roberts, B. M.</dc:creator>
<dc:creator>Gracie, C.</dc:creator>
<dc:creator>te Water Naude, R.</dc:creator>
<dc:creator>Karthik, G.</dc:creator>
<dc:creator>Wade-Martins, R.</dc:creator>
<dc:creator>Cragg, S. J.</dc:creator>
<dc:date>2020-07-04</dc:date>
<dc:identifier>doi:10.1101/2020.07.03.186411</dc:identifier>
<dc:title><![CDATA[L-type calcium channel contribution to striatal dopamine release is governed by calbindin-D28K, the dopamine transporter, D2-receptors, α2δ-subunits and sex differences]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-07-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.08.23.554510v1?rss=1">
<title>
<![CDATA[
M4-mediated cholinergic transmission is reduced in parkinsonian mice and its restoration alleviates motor deficits and levodopa-induced dyskinesia 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.08.23.554510v1?rss=1"
</link>
<description><![CDATA[
The dynamic equilibrium between dopamine (DA) and acetylcholine (ACh) in the dorsal striatum is thought to be essential for motor function, as imbalances in their levels are associated with Parkinsons disease (PD) and levodopa-induced dyskinesia (LID). While loss of DA leads to enhanced striatal ACh, whether this translates to specific alterations in transmission remains unclear. To address this, we examined how the strength of ACh release and signaling onto direct-pathway medium spiny neurons is altered in parkinsonian mice. Rather than the predicted cholinergic enhancement, we found that the strength of muscarinic M4-receptor mediated transmission was reduced following DA loss, resulting from downregulated receptors and downstream signaling. Despite M4-receptors being thought to mediate anti-kinetic effects, restoring M4-receptor function partially rescued parkinsonian balance and coordination deficits and limited the development of levodopa-induced dyskinetic behaviors, indicating that decreased M4-function contributed to circuit and motor dysfunctions in response to DA loss.
]]></description>
<dc:creator>Nielsen, B. E.</dc:creator>
<dc:creator>Ford, C. P.</dc:creator>
<dc:date>2023-08-24</dc:date>
<dc:identifier>doi:10.1101/2023.08.23.554510</dc:identifier>
<dc:title><![CDATA[M4-mediated cholinergic transmission is reduced in parkinsonian mice and its restoration alleviates motor deficits and levodopa-induced dyskinesia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.08.28.555228v1?rss=1">
<title>
<![CDATA[
A RAB7A Phosphoswitch Coordinates Rubicon Homology Protein Regulation of PINK1/Parkin-Dependent Mitophagy 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.08.28.555228v1?rss=1"
</link>
<description><![CDATA[
Activation of PINK1 and Parkin in response to mitochondrial damage initiates a cytoprotective mitophagy response that includes phosphorylation of RAB7A at Ser72. Rubicon is a RAB7A binding protein that acts as a negative regulator of autophagy. The structure of the Rubicon-RAB7A complex suggests that phosphorylation of RAB7A at Ser72 would block Rubicon binding. Indeed, in vitro phosphorylation of RAB7A by TBK1 abrogates Rubicon-RAB7A binding. Pacer, a positive regulator of autophagy, has an RH domain with a basic triad predicted to bind an introduced phosphate. Consistent with this, Pacer-RH binds to phosho-RAB7A but not to unphosphorylated RAB7A. In cells, mitochondrial depolarization reduces Rubicon:RAB7A colocalization whilst recruiting Pacer to phospho-RAB7A-positive puncta. Pacer knockout reduces Parkin mitophagy with little effect on bulk autophagy or Parkin-independent mitophagy. Rescue of Parkin-dependent mitophagy requires the intact pRAB7A phosphate-binding basic triad of Pacer. Together these structural and functional data support a model in which the TBK1-dependent phosphorylation of RAB7A serves as a switch, promoting mitophagy by relieving Rubicon inhibition and favoring Pacer activation.
]]></description>
<dc:creator>Tudorica, D. A.</dc:creator>
<dc:creator>Basak, B. L. F.</dc:creator>
<dc:creator>Cordova, A. P.</dc:creator>
<dc:creator>Khuu, G.</dc:creator>
<dc:creator>Rose, K.</dc:creator>
<dc:creator>Lazarou, M.</dc:creator>
<dc:creator>Holzbaur, E.</dc:creator>
<dc:creator>Hurley, J. H.</dc:creator>
<dc:date>2023-08-29</dc:date>
<dc:identifier>doi:10.1101/2023.08.28.555228</dc:identifier>
<dc:title><![CDATA[A RAB7A Phosphoswitch Coordinates Rubicon Homology Protein Regulation of PINK1/Parkin-Dependent Mitophagy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.03.14.532627v1?rss=1">
<title>
<![CDATA[
State-dependent GABAergic regulation of striatal spiny projection neuron excitability 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.03.14.532627v1?rss=1"
</link>
<description><![CDATA[
Synaptic transmission mediated by GABAA receptors (GABAARs) in adult, principal striatal spiny projection neurons (SPNs) can suppress ongoing spiking, but its effect on synaptic integration at sub-threshold membrane potentials is less well characterized, particularly those near the resting down-state. To fill this gap, a combination of molecular, optogenetic, optical and electrophysiological approaches were used to study SPNs in mouse ex vivo brain slices, and computational tools were used to model somatodendritic synaptic integration. Activation of GABAARs, either by uncaging of GABA or by optogenetic stimulation of GABAergic synapses, evoked currents with a reversal potential near -60 mV in perforated patch recordings from both juvenile and adult SPNs. Molecular profiling of SPNs suggested that this relatively positive reversal potential was not attributable to NKCC1 expression, but rather to a dynamic equilibrium between KCC2 and Cl-/HCO3-cotransporters. Regardless, from down-state potentials, optogenetic activation of dendritic GABAergic synapses depolarized SPNs. This GABAAR-mediated depolarization summed with trailing ionotropic glutamate receptor (iGluR) stimulation, promoting dendritic spikes and increasing somatic depolarization. Simulations revealed that a diffuse dendritic GABAergic input to SPNs effectively enhanced the response to coincident glutamatergic input. Taken together, our results demonstrate that GABAARs can work in concert with iGluRs to excite adult SPNs when they are in the resting down-state, suggesting that their inhibitory role is limited to brief periods near spike threshold. This state-dependence calls for a reformulation of the role intrastriatal GABAergic circuits.
]]></description>
<dc:creator>Day, M.</dc:creator>
<dc:creator>Belal, M.</dc:creator>
<dc:creator>Surmeier, C.</dc:creator>
<dc:creator>Melendez-Zaidi, A.</dc:creator>
<dc:creator>Tkatch, T.</dc:creator>
<dc:creator>Clarke, V. R. J.</dc:creator>
<dc:creator>Surmeier, D. J.</dc:creator>
<dc:date>2023-03-14</dc:date>
<dc:identifier>doi:10.1101/2023.03.14.532627</dc:identifier>
<dc:title><![CDATA[State-dependent GABAergic regulation of striatal spiny projection neuron excitability]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.08.24.554624v1?rss=1">
<title>
<![CDATA[
iATPSnFR2: a high dynamic range fluorescent sensor for monitoring intracellular ATP 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.08.24.554624v1?rss=1"
</link>
<description><![CDATA[
We developed a significantly improved genetically encoded quantitative adenosine triphosphate (ATP) sensor to provide real-time dynamics of ATP levels in subcellular compartments. iATPSnFR2 is a variant of iATPSnFR1, a previously developed sensor that has circularly permuted super-folder GFP inserted between the ATP-binding helices of the{varepsilon} -subunit of a bacterial F0-F1 ATPase. Optimizing the linkers joining the two domains resulted in a [~] 5-6 fold improvement in the dynamic range compared to the previous generation sensor, with excellent discrimination against other analytes and affinity variants varying from 4 M to 500 M. A chimeric version of this sensor fused to either the HaloTag protein or a suitably spectrally separated fluorescent protein, provides a ratiometric readout allowing comparisons of ATP across cellular regions. Subcellular targeting of the sensor to nerve terminals reveals previously uncharacterized single synapse metabolic signatures, while targeting to the mitochondrial matrix allowed direct quantitative probing of oxidative phosphorylation dynamics.

Significance StatementAdenosine triphosphate (ATP) is a key metabolite necessary for cellular life. Here we develop a next-generation genetically encoded ratiometric fluorescent ATP sensor that allows subcellular tracking of ATP levels in living cells. The large dynamic range makes it possible to follow the dynamics of this metabolite across cells and subcellular regions under different metabolic stressors. We expect that iATPSnFR2 will provide researchers with exciting new opportunities to study ATP dynamics with temporal and spatial resolution that has, until now, been unavailable.
]]></description>
<dc:creator>Marvin, J. S.</dc:creator>
<dc:creator>Kokotos, A. C.</dc:creator>
<dc:creator>Kumar, M.</dc:creator>
<dc:creator>Pulido, C.</dc:creator>
<dc:creator>Tkachuk, A. N.</dc:creator>
<dc:creator>Yao, J. S.</dc:creator>
<dc:creator>Brown, T. A.</dc:creator>
<dc:creator>Ryan, T. A.</dc:creator>
<dc:date>2023-08-24</dc:date>
<dc:identifier>doi:10.1101/2023.08.24.554624</dc:identifier>
<dc:title><![CDATA[iATPSnFR2: a high dynamic range fluorescent sensor for monitoring intracellular ATP]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.09.28.559894v1?rss=1">
<title>
<![CDATA[
Structural pathway for class III PI 3-kinase activation by the myristoylated GTP-binding pseudokinase VPS15 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.09.28.559894v1?rss=1"
</link>
<description><![CDATA[
The class III phosphatidylinositol (PI) 3-kinase complexes I and II (PI3KC3-C1 and -C2) are central to the initiation of macroautophagy and endosomal maturation, respectively. Through three-dimensional classification of a large cryo-EM dataset of human PI3KC3-C1 bound to the small GTPase RAB1A, we were able to map the structural pathway of enzyme activation. The inactive conformation is stabilized by an N-myristoyl modification of the pseudokinase (PK) subunit VPS15. The N-myristate is sequestered in the N-lobe of the VPS15 PK domain, which stabilizes a series of interactions whereby VPS15 sequesters and blocks the catalytic and membrane binding units of the VPS34 lipid kinase. In the activated conformation, the N-myristate and the VPS34 lipid kinase domain are liberated to interact with membranes and catalyze PI3P formation. The VPS15 PK domain contains a unique Arg at the gatekeeper position and binds tightly to GTP. GTP binding structurally stabilizes the N-myristate "in" conformation, which promotes the inactive conformation. This pathway provides a general mechanism for PI3KC3 activation in autophagy and endosome biogenesis and a roadmap for their pharmacological upregulation.
]]></description>
<dc:creator>Cook, A. S. I.</dc:creator>
<dc:creator>Chen, M. H.</dc:creator>
<dc:creator>Ren, X.</dc:creator>
<dc:creator>Rao, S.</dc:creator>
<dc:creator>Garcia, S. N.</dc:creator>
<dc:creator>Claveras Cabezudo, A.</dc:creator>
<dc:creator>Iavarone, A. T.</dc:creator>
<dc:creator>Hummer, G.</dc:creator>
<dc:creator>Hurley, J. H.</dc:creator>
<dc:date>2023-09-28</dc:date>
<dc:identifier>doi:10.1101/2023.09.28.559894</dc:identifier>
<dc:title><![CDATA[Structural pathway for class III PI 3-kinase activation by the myristoylated GTP-binding pseudokinase VPS15]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.07.17.549391v1?rss=1">
<title>
<![CDATA[
Three-step docking by WIPI2, ATG16L1 and ATG3 delivers LC3 to the phagophore 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.07.17.549391v1?rss=1"
</link>
<description><![CDATA[
The covalent attachment of ubiquitin-like LC3 proteins prepares the autophagic membrane for cargo recruitment. We resolve key steps in LC3 lipidation by combining molecular dynamics simulations and experiments in vitro and in cellulo. We show how the E3-like ligase ATG12- ATG5-ATG16L1 in complex with the E2-like conjugase ATG3 docks LC3 onto the membrane in three steps by (1) the PI(3)P effector protein WIPI2, (2) helix 2 of ATG16L1, and (3) a membrane-interacting surface of ATG3. Phosphatidylethanolamine (PE) lipids concentrate in a region around the thioester bond between ATG3 and LC3, highlighting residues with a possible role in the catalytic transfer of LC3 to PE, including two conserved histidines. In a near-complete pathway from the initial membrane recruitment to the LC3 lipidation reaction, the three-step targeting of the ATG12-ATG5-ATG16L1 machinery establishes a high level of regulatory control.
]]></description>
<dc:creator>Rao, S.</dc:creator>
<dc:creator>Strong, L. M.</dc:creator>
<dc:creator>Ren, X.</dc:creator>
<dc:creator>Skulsuppaisarn, M.</dc:creator>
<dc:creator>Lazarou, M.</dc:creator>
<dc:creator>Hurley, J. H.</dc:creator>
<dc:creator>Hummer, G.</dc:creator>
<dc:date>2023-07-17</dc:date>
<dc:identifier>doi:10.1101/2023.07.17.549391</dc:identifier>
<dc:title><![CDATA[Three-step docking by WIPI2, ATG16L1 and ATG3 delivers LC3 to the phagophore]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.09.11.557213v1?rss=1">
<title>
<![CDATA[
Parkinson's genes orchestrate pyroptosis through selective trafficking of mtDNA to leaky lysosomes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.09.11.557213v1?rss=1"
</link>
<description><![CDATA[
Inflammation is an age-related factor that underlies numerous human disorders. A key driver of inflammation is the release of mitochondrial DNA (mtDNA), which binds and activates cytosolic sensors. This induces transcriptional responses and, ultimately, pyroptotic cell death. The main challenge has been to understand how mtDNA can cross the two mitochondrial membranes to access the cytosol. Through a genome-wide CRISPR knockout screen we identified a new pyroptotic pathway defined by mtDNA exit within mitochondrial-derived vesicles that are delivered to lysosomes. Critically, breach of lysosomes allows mtDNA to access cytosol, requiring multiple Parkinsons Disease-related proteins and Gasdermin pores, identified in the screen. These data place mitochondria-to-lysosome transport as a driver of pyroptosis and link multiple PD proteins along a common pathway.

One sentence summaryParkinsons disease-related proteins regulate pyroptosis
]]></description>
<dc:creator>Nguyen, M.</dc:creator>
<dc:creator>Collier, J. J.</dc:creator>
<dc:creator>Ignatenko, O.</dc:creator>
<dc:creator>Morin, G.</dc:creator>
<dc:creator>Huang, S.</dc:creator>
<dc:creator>Desjardins, M.</dc:creator>
<dc:creator>McBride, H. M.</dc:creator>
<dc:date>2023-09-12</dc:date>
<dc:identifier>doi:10.1101/2023.09.11.557213</dc:identifier>
<dc:title><![CDATA[Parkinson's genes orchestrate pyroptosis through selective trafficking of mtDNA to leaky lysosomes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.10.06.561190v1?rss=1">
<title>
<![CDATA[
LRRK2 kinase inhibition reverses G2019S mutation-dependent effects on tau pathology spread 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.10.06.561190v1?rss=1"
</link>
<description><![CDATA[
Mutations in leucine-rich repeat kinase 2 (LRRK2) are the most common cause of familial Parkinsons disease (PD). These mutations elevate LRRK2 kinase activity, making LRRK2 kinase inhibitors an attractive therapeutic target. LRRK2 kinase activity has been consistently linked to specific cell signaling pathways, mostly related to organelle trafficking and homeostasis, but its relationship to PD pathogenesis has been more difficult to define. LRRK2-PD patients consistently present with loss of dopaminergic neurons in the substantia nigra but show variable development of Lewy body or tau tangle pathology. Animal models carrying LRRK2 mutations do not develop robust PD-related phenotypes spontaneously, hampering the assessment of LRRK2 inhibitors efficacy against disease processes. We hypothesized that mutations in LRRK2 may not be directly related to a single disease pathway, but instead may elevate susceptibility to multiple disease processes, depending on the disease trigger. To test this hypothesis, we have previously evaluated progression of -synuclein and tau pathologies following injection of proteopathic seeds. We demonstrated that transgenic mice overexpressing mutant LRRK2 show alterations in the brain-wide progression of pathology, especially at older ages. Here, we assess tau pathology progression in relation to long-term LRRK2 kinase inhibition. Wildtype or LRRK2G2019S knock-in mice were injected with tau fibrils and treated with control diet or diet containing LRRK2 kinase inhibitor MLi-2 targeting the IC50 or IC90 of LRRK2 for 3 to 6 months. Mice were evaluated for tau pathology by brain-wide quantitative pathology in 665 brain regions and subsequent linear diffusion modeling of progression. Consistent with our previous work, we found systemic alterations in the progression of tau pathology in LRRK2G2019S mice that were most pronounced at 6 months. Importantly, LRRK2 kinase inhibition reversed these effects in LRRK2G2019S mice, but had minimal effect in wildtype mice, suggesting that LRRK2 kinase inhibition is likely to reverse specific disease processes in G2019S mutation carriers, but additional work may be necessary to determine the potential effect in non-carriers. This work supports a protective role of LRRK2 kinase inhibition in G2019S carriers and provides a rational workflow for systematic evaluation of brain-wide phenotypes in therapeutic development.
]]></description>
<dc:creator>Lubben, N.</dc:creator>
<dc:creator>Brynildsen, J. K.</dc:creator>
<dc:creator>Leyns, C. E. G.</dc:creator>
<dc:creator>Webb, C. M.</dc:creator>
<dc:creator>Li, H. L.</dc:creator>
<dc:creator>Changolkar, L.</dc:creator>
<dc:creator>Zhang, B.</dc:creator>
<dc:creator>Meymand, E.</dc:creator>
<dc:creator>O'Reilly, M.</dc:creator>
<dc:creator>Madaj, Z.</dc:creator>
<dc:creator>DeWeerd, D.</dc:creator>
<dc:creator>Fell, M. J.</dc:creator>
<dc:creator>Lee, V. M. Y.</dc:creator>
<dc:creator>Bassett, D. S.</dc:creator>
<dc:creator>Henderson, M. X.</dc:creator>
<dc:date>2023-10-06</dc:date>
<dc:identifier>doi:10.1101/2023.10.06.561190</dc:identifier>
<dc:title><![CDATA[LRRK2 kinase inhibition reverses G2019S mutation-dependent effects on tau pathology spread]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.10.05.559770v1?rss=1">
<title>
<![CDATA[
A topographical atlas of alpha-Synuclein dosage and cell-type expression in the mouse brain and periphery 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.10.05.559770v1?rss=1"
</link>
<description><![CDATA[
Parkinsons disease (PD) is the second most common neurodegenerative disease worldwide and presents pathologically with Lewy pathology and dopaminergic neuron loss. Lewy pathology contains aggregated Synuclein (Syn), a protein encoded by the SNCA gene which is also mutated or duplicated in a subset of familial PD cases. Due to its predominant presynaptic localization, immunostaining for the protein results in diffuse signal, providing little insight into the types of cells expressing Syn. As a result, insight into Syn expression-driven cellular vulnerability has been difficult to ascertain. Using a combination of knock-in mice that target Syn to the nucleus of cells (SncaNLS) and in situ hybridization of Snca in wild-type mice, we systematically map the topography and cell types expressing Syn in the mouse brain, spinal cord, retina, and gut. We find a high degree of correlation between Syn protein and RNA levels across multiple brain regions and further identify cell types with low and high Syn. We found that Syn is highly expressed in neurons, particularly those involved in PD and to a lower extent in non-neuronal cell types, notably those of oligodendrocyte lineage. We also find that Syn is devoid in certain neuron types (e.g. ChAT-positive motor neurons), and that all enteric neurons express Syn to a certain degree. Taken together, this atlas provides much-needed insight into the cellular topography of Syn, and provides a quantitative map to test assumptions about the role of Syn in network vulnerability in PD and other Synucleinopathies.
]]></description>
<dc:creator>Geertsma, H. M.</dc:creator>
<dc:creator>Fisk, Z. A.</dc:creator>
<dc:creator>Sauline, L.</dc:creator>
<dc:creator>Prigent, A.</dc:creator>
<dc:creator>Kurgat, K.</dc:creator>
<dc:creator>Callaghan, S. M.</dc:creator>
<dc:creator>aSCENT PD Consortium,</dc:creator>
<dc:creator>Henderson, M.</dc:creator>
<dc:creator>Rousseaux, M. W. C.</dc:creator>
<dc:date>2023-10-07</dc:date>
<dc:identifier>doi:10.1101/2023.10.05.559770</dc:identifier>
<dc:title><![CDATA[A topographical atlas of alpha-Synuclein dosage and cell-type expression in the mouse brain and periphery]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.10.06.560646v1?rss=1">
<title>
<![CDATA[
Integrative analysis reveals a conserved role for the amyloid precursor protein in proteostasis during aging 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.10.06.560646v1?rss=1"
</link>
<description><![CDATA[
A{beta} peptides derived from the amyloid precursor protein (APP) have been strongly implicated in the pathogenesis of Alzheimers disease. However, the normal function of APP and the importance of that role in neurodegenerative disease is less clear. We recovered the Drosophila ortholog of APP, Appl, in an unbiased forward genetic screen for neurodegeneration mutants. We performed comprehensive single cell transcriptional and proteomic studies of Appl mutant flies to investigate Appl function in the aging brain. We found an unexpected role for Appl in control of multiple cellular pathways, including translation, mitochondrial function, nucleic acid and lipid metabolism, cellular signaling and proteostasis. We mechanistically defined a role for Appl in regulating autophagy through TGF{beta} signaling and documented the broader relevance of our findings using mouse genetic, human iPSC and in vivo tauopathy models. Our results demonstrate a conserved role for APP in controlling age-dependent proteostasis with plausible relevance to Alzheimers disease.
]]></description>
<dc:creator>Nithianandam, V.</dc:creator>
<dc:creator>Bukhari, H.</dc:creator>
<dc:creator>Leventhal, M. J.</dc:creator>
<dc:creator>Battaglia, R. A.</dc:creator>
<dc:creator>Dong, X.</dc:creator>
<dc:creator>Fraenkel, E.</dc:creator>
<dc:creator>Feany, M. B.</dc:creator>
<dc:date>2023-10-07</dc:date>
<dc:identifier>doi:10.1101/2023.10.06.560646</dc:identifier>
<dc:title><![CDATA[Integrative analysis reveals a conserved role for the amyloid precursor protein in proteostasis during aging]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.06.15.545139v1?rss=1">
<title>
<![CDATA[
Localization of PPM1H phosphatase tunes Parkinsons disease-linked LRRK2 kinase-mediated Rab GTPase phosphorylation and ciliogenesis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.06.15.545139v1?rss=1"
</link>
<description><![CDATA[
PPM1H phosphatase reverses Parkinsons disease-associated, LRRK2-mediated Rab GTPase phosphorylation. We show here that PPM1H relies on an N-terminal amphipathic helix for Golgi localization. The amphipathic helix enables PPM1H to bind to liposomes in vitro, and small, highly curved liposomes stimulate PPM1H activity. We artificially anchored PPM1H to the Golgi, mitochondria, or mother centriole. Our data show that regulation of Rab10 GTPase phosphorylation requires PPM1H access to Rab10 at or near the mother centriole. Moreover, poor co-localization of Rab12 explains in part why it is a poor substrate for PPM1H in cells but not in vitro. These data support a model in which localization drives PPM1H substrate selection and centriolar PPM1H is critical for regulation of Rab GTPase-regulated ciliogenesis. Moreover, Golgi localized PPM1H maintains active Rab GTPases on the Golgi to carry out their non-ciliogenesis-related functions in membrane trafficking.

Significance StatementPathogenic, hyperactive LRRK2 kinase is strongly linked to Parkinsons disease and LRRK2 phosphorylates a subset of Rab GTPases that are master regulators of membrane trafficking. PPM1H phosphatase specifically dephosphorylates Rab8A and Rab10, the major LRRK2 substrates. Here we provide novel cell biological and biochemical insight related to the localization and activation of PPM1H phosphatase. Understanding how PPM1H modulates LRRK2 activity is of fundamental interest and also important, as activators of PPM1H may eventually benefit Parkinsons disease patients.
]]></description>
<dc:creator>Yeshaw, W. M.</dc:creator>
<dc:creator>Adhikari, A.</dc:creator>
<dc:creator>Chiang, C. Y.</dc:creator>
<dc:creator>Dhekne, H. S.</dc:creator>
<dc:creator>Wawro, P. S.</dc:creator>
<dc:creator>Pfeffer, S. R.</dc:creator>
<dc:date>2023-06-15</dc:date>
<dc:identifier>doi:10.1101/2023.06.15.545139</dc:identifier>
<dc:title><![CDATA[Localization of PPM1H phosphatase tunes Parkinsons disease-linked LRRK2 kinase-mediated Rab GTPase phosphorylation and ciliogenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.07.25.550405v1?rss=1">
<title>
<![CDATA[
CURTAIN - A Unique Web-based tool for exploration and sharing of MS-based proteomics data 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.07.25.550405v1?rss=1"
</link>
<description><![CDATA[
To facilitate analysis and sharing of mass spectrometry (MS)-based proteomics data we created tools called CURTAIN (https://curtain.proteo.info) and CURTAIN-PTM (https://curtainptm.proteo.info). These enable the non-MS expert to interactively peruse volcano plots; deconvolute primary experimental data to individual replicates that can be visualized in bar charts or violin plots allowing statistical analysis; and export of plots in SVG format. They also permit assessment of experimental quality by correlation matrix and profile plot. Within CURTAIN, the user can analyze domain structure, AlphaFold predicted structure, reported interactors, relative expression, disease and pharmaceutical links, and mutagenesis information on all selected hits. Moreover, CURTAIN-PTM permits the comparison of all identified PTM sites on protein(s) of interest with PTM information contained within selected databases. For phosphorylation site analysis CURTAIN-PTM links with the kinase library to predict upstream kinases that phosphorylate sites of interest. We provide examples of the utility of CURTAIN and CURTAIN-PTM in analyzing how targeted degradation of the PPM1H Rab phosphatase that counteracts the Parkinsons LRRK2 kinase impacts cellular protein levels and phosphorylation sites. We reanalyzed a ubiquitylation dataset, characterizing the PINK1-Parkin pathway activation in primary neurons, revealing new data of interest not highlighted previously. CURTAIN and CURTAIN-PTM are free to use and open-source and will enable researchers to share and maximize the analysis and impact of their proteomics data. We advocate that differential expression proteomic data should be published containing a shareable CURTAIN web-link, allowing readers to better explore their data.

Significance StatementTo enable non-experts to better share and explore mass spectrometry data, we have generated using open-source software, interactive tools termed CURTAIN and CURTAIN-PTM. These tools enable users to save their analysis sessions with a sharable unique web-link, enabling other researchers to visualize and further analyze these datasets. These links can also be reported in publications allowing readers to further survey the reported data. We discuss benefits for the research community of publishing proteomic data containing a shareable web-link.
]]></description>
<dc:creator>Phung, T. K.</dc:creator>
<dc:creator>Berndsen, K.</dc:creator>
<dc:creator>Phan, T. L.</dc:creator>
<dc:creator>Muqit, M. M.</dc:creator>
<dc:creator>Alessi, D. R.</dc:creator>
<dc:creator>Nirujogi, R. S.</dc:creator>
<dc:date>2023-07-25</dc:date>
<dc:identifier>doi:10.1101/2023.07.25.550405</dc:identifier>
<dc:title><![CDATA[CURTAIN - A Unique Web-based tool for exploration and sharing of MS-based proteomics data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.06.07.544051v1?rss=1">
<title>
<![CDATA[
Parkinson's VPS35 mutation induces LRRK2 mediated lysosomal association of RILPL1 and TMEM55B 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.06.07.544051v1?rss=1"
</link>
<description><![CDATA[
The Parkinsons VPS35[D620N] mutation causes lysosome dysfunction enhancing LRRK2 kinase activity. We find the VPS35[D620N] mutation alters expression of [~]350 lysosomal proteins and stimulates LRRK2 phosphorylation of Rab proteins at the lysosome. This recruits the phosphoRab effector protein RILPL1 to the lysosome where it binds to the lysosomal integral membrane protein TMEM55B. We identify highly conserved regions of RILPL1 and TMEM55B that interact and design mutations that block binding. In mouse fibroblasts, brain, and lung, we demonstrate that the VPS35 [D620N] mutation reduces RILPL1 levels, in a manner reversed by LRRK2 inhibition. Knock-out of RILPL1 enhances phosphorylation of Rab substrates and knock-out of TMEM55B increases RILPL1 levels. The lysosomotropic agent LLOMe, also induced LRRK2 kinase mediated association of RILPL1 to the lysosome, but to a lower extent than the D620N mutation. Our study uncovers a pathway through which dysfunctional lysosomes resulting from the VPS35[D620N] mutation recruit and activate LRRK2 on the lysosomal surface, driving assembly of the RILPL1-TMEM55B complex.
]]></description>
<dc:creator>Pal, P.</dc:creator>
<dc:creator>Taylor, M.</dc:creator>
<dc:creator>Lam, P. Y.</dc:creator>
<dc:creator>Tonelli, F.</dc:creator>
<dc:creator>Hecht, C. A.</dc:creator>
<dc:creator>Lis, P.</dc:creator>
<dc:creator>Nirujogi, R. S.</dc:creator>
<dc:creator>Phung, T. K.</dc:creator>
<dc:creator>Dickie, E. A.</dc:creator>
<dc:creator>Wightman, M.</dc:creator>
<dc:creator>Macartney, T.</dc:creator>
<dc:creator>Pfeffer, S. R.</dc:creator>
<dc:creator>Alessi, D. R.</dc:creator>
<dc:date>2023-06-07</dc:date>
<dc:identifier>doi:10.1101/2023.06.07.544051</dc:identifier>
<dc:title><![CDATA[Parkinson's VPS35 mutation induces LRRK2 mediated lysosomal association of RILPL1 and TMEM55B]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.12.19.521076v1?rss=1">
<title>
<![CDATA[
Unique functional responses differentially map onto genetic subtypes of dopamine neurons 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.12.19.521076v1?rss=1"
</link>
<description><![CDATA[
Dopamine neurons are characterized by their response to unexpected rewards, but they also fire during movement and aversive stimuli. Dopamine neuron diversity has been observed based on molecular expression profiles; however, whether different functions map onto such genetic subtypes remains unclear. Here, we establish that three genetic dopamine subtypes within the substantia nigra pars compacta each have a unique set of responses to rewards, aversive stimuli, accelerations and decelerations, and these signaling patterns are highly-correlated between somas and axons within subtypes. Remarkably, reward responses were not detected in one subtype, which instead displayed acceleration-correlated signaling. Our findings establish a connection between functional and genetic dopamine subtypes and demonstrate that molecular expression patterns can serve as a common framework to dissect dopaminergic functions.
]]></description>
<dc:creator>Azcorra, M.</dc:creator>
<dc:creator>Gaertner, Z.</dc:creator>
<dc:creator>Davidson, C.</dc:creator>
<dc:creator>Hayes, C. K.</dc:creator>
<dc:creator>Ramakrishnan, C.</dc:creator>
<dc:creator>Fenno, L.</dc:creator>
<dc:creator>Kim, Y. S.</dc:creator>
<dc:creator>Deisseroth, K.</dc:creator>
<dc:creator>Longnecker, R.</dc:creator>
<dc:creator>Awatramani, R.</dc:creator>
<dc:creator>Dombeck, D. A.</dc:creator>
<dc:date>2022-12-19</dc:date>
<dc:identifier>doi:10.1101/2022.12.19.521076</dc:identifier>
<dc:title><![CDATA[Unique functional responses differentially map onto genetic subtypes of dopamine neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.05.17.541144v1?rss=1">
<title>
<![CDATA[
Spatial transcriptomics reveals molecular dysfunction associated with Lewy pathology 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.05.17.541144v1?rss=1"
</link>
<description><![CDATA[
Lewy pathology composed of -synuclein is the key pathological hallmark of Parkinsons disease (PD), found both in dopaminergic neurons that control motor function, and throughout cortical regions that control cognitive function. Recent work has investigated which dopaminergic neurons are most susceptible to death, but little is known about which neurons are vulnerable to developing Lewy pathology and what molecular changes an aggregate induces. In the current study, we use spatial transcriptomics to selectively capture whole transcriptome signatures from cortical neurons with Lewy pathology compared to those without pathology in the same brains. We find, both in PD and in a mouse model of PD, that there are specific classes of excitatory neurons that are vulnerable to developing Lewy pathology in the cortex. Further, we identify conserved gene expression changes in aggregate-bearing neurons that we designate the Lewy-associated molecular dysfunction from aggregates (LAMDA) signature. This gene signature indicates that neurons with aggregates downregulate synaptic, mitochondrial, ubiquitin-proteasome, endo-lysosomal, and cytoskeletal genes and upregulate DNA repair and complement/cytokine genes. However, beyond DNA repair gene upregulation, we find that neurons also activate apoptotic pathways, suggesting that if DNA repair fails, neurons undergo programmed cell death. Our results identify neurons vulnerable to Lewy pathology in the PD cortex and identify a conserved signature of molecular dysfunction in both mice and humans.
]]></description>
<dc:creator>Goralski, T.</dc:creator>
<dc:creator>Meyerdirk, L.</dc:creator>
<dc:creator>Breton, L.</dc:creator>
<dc:creator>Brasseur, L.</dc:creator>
<dc:creator>Kurgat, K.</dc:creator>
<dc:creator>DeWeerd, D.</dc:creator>
<dc:creator>Turner, L.</dc:creator>
<dc:creator>Becker, K.</dc:creator>
<dc:creator>Adams, M.</dc:creator>
<dc:creator>Newhouse, D.</dc:creator>
<dc:creator>Henderson, M. X.</dc:creator>
<dc:date>2023-05-17</dc:date>
<dc:identifier>doi:10.1101/2023.05.17.541144</dc:identifier>
<dc:title><![CDATA[Spatial transcriptomics reveals molecular dysfunction associated with Lewy pathology]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-05-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.04.09.536178v1?rss=1">
<title>
<![CDATA[
Astrocytic LRRK2 Controls Synaptic Connectivity through ERM Phosphorylation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.04.09.536178v1?rss=1"
</link>
<description><![CDATA[
Astrocytes are highly complex cells that mediate critical roles in synapse formation and maintenance by establishing thousands of direct contacts with synapses through their perisynaptic processes. Here, we found that the most common Parkinsonism gene mutation, LRRK2 G2019S, enhances the phosphorylation of the ERM proteins (Ezrin, Radixin, and Moesin), components of the perisynaptic astrocyte processes in a subset of cortical astrocytes. The ERM hyperphosphorylation was accompanied by decreased astrocyte morphological complexity and reduced excitatory synapse density and function. Dampening ERM phosphorylation levels in LRRK2 G2019S mouse astrocytes restored both their morphology and the excitatory synapse density in the anterior cingulate cortex. To determine how LRRK2 mutation impacts Ezrin interactome, we used an in vivo BioID proteomic approach, and we found that astrocytic Ezrin interacts with Atg7, a master regulator of autophagy. The Ezrin/Atg7 interaction is inhibited by Ezrin phosphorylation, thus diminished in LRRK2 G2019S astrocytes. Importantly, the Atg7 function is required to maintain proper astrocyte morphology. Our data provide a molecular pathway through which the LRRK2 G2019S mutation alters astrocyte morphology and synaptic density in a brain-region-specific manner.
]]></description>
<dc:creator>Wang, S.</dc:creator>
<dc:creator>Sivadasan Bindu, D.</dc:creator>
<dc:creator>Tan, C. X.</dc:creator>
<dc:creator>Sakers, K.</dc:creator>
<dc:creator>Takano, T.</dc:creator>
<dc:creator>Rodriguez Salazar, M. P.</dc:creator>
<dc:creator>Dimond, K.</dc:creator>
<dc:creator>Soderling, S. H.</dc:creator>
<dc:creator>La Spada, A. R.</dc:creator>
<dc:creator>Eroglu, C.</dc:creator>
<dc:date>2023-04-10</dc:date>
<dc:identifier>doi:10.1101/2023.04.09.536178</dc:identifier>
<dc:title><![CDATA[Astrocytic LRRK2 Controls Synaptic Connectivity through ERM Phosphorylation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.01.27.477991v1?rss=1">
<title>
<![CDATA[
Unconventional secretion of α-synuclein mediated by palmitoylated DNAJC5 oligomers 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.01.27.477991v1?rss=1"
</link>
<description><![CDATA[
Alpha-synuclein (-syn), a major component of Lewy bodies found in Parkinsons disease (PD) patients, has been found exported outside of cells and may mediate its toxicity via cell-to-cell transmission. Here, we reconstituted soluble, monomeric -syn secretion by the expression of DnaJ homolog subfamily C member 5 (DNAJC5) in HEK293T cells. DNAJC5 undergoes palmitoylation and anchors on the membrane. Palmitoylation is essential for DNAJC5-induced -syn secretion, and the secretion is not limited by substrate size or unfolding. Cytosolic -syn is actively translocated and sequestered in an endosomal membrane compartment in a DNAJC5-dependent manner. Reduction of -syn secretion caused by a palmitoylation-deficient mutation in DNAJC5 can be reversed by a membrane-targeting peptide fusion-induced oligomerization of DNAJC5. The secretion of endogenous -syn mediated by DNAJC5 is also found in a human neuroblastoma cell line, SH-SY5Y, differentiated into neurons in the presence of retinoic acid, and in human induced pluripotent stem cell-derived midbrain dopamine neurons. We propose that DNAJC5 forms a palmitoylated oligomer to accommodate and export -syn.
]]></description>
<dc:creator>Wu, S.</dc:creator>
<dc:creator>Sirkis, D. W.</dc:creator>
<dc:creator>Schekman, R.</dc:creator>
<dc:date>2022-01-27</dc:date>
<dc:identifier>doi:10.1101/2022.01.27.477991</dc:identifier>
<dc:title><![CDATA[Unconventional secretion of α-synuclein mediated by palmitoylated DNAJC5 oligomers]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-01-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.10.14.562004v1?rss=1">
<title>
<![CDATA[
A fluid-walled microfluidic platform for human neuron microcircuits and directed axotomy 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.10.14.562004v1?rss=1"
</link>
<description><![CDATA[
In our brains, different neurons make appropriate connections; however, there remain few in vitro models of such circuits. We use an open microfluidic approach to build and study neuronal circuits in vitro in ways that fit easily into existing bio-medical workflows. Dumbbell-shaped circuits are built in minutes in standard Petri dishes; the aqueous phase is confined by fluid walls - interfaces between cell-growth medium and an immiscible fluorocarbon, FC40. Conditions are established that ensure post-mitotic neurons derived from human induced pluripotent stem cells (iPSCs) plated in one chamber of a dumbbell remain where deposited. After seeding cortical neurons on one side, axons grow through the connecting conduit to ramify amongst striatal neurons on the other - an arrangement mimicking unidirectional cortico-striatal connectivity. We also develop a moderate-throughput non-contact axotomy assay. Cortical axons in conduits are severed by a media jet; then, brain-derived neurotrophic factor and striatal neurons in distal chambers promote axon regeneration. As additional conduits and chambers are easily added, this opens up the possibility of mimicking complex neuronal networks, and screening drugs for their effects on connectivity.
]]></description>
<dc:creator>Nebuloni, F.</dc:creator>
<dc:creator>Do, Q.</dc:creator>
<dc:creator>Cook, P. R.</dc:creator>
<dc:creator>Walsh, E.</dc:creator>
<dc:creator>Wade-Martins, R.</dc:creator>
<dc:date>2023-10-17</dc:date>
<dc:identifier>doi:10.1101/2023.10.14.562004</dc:identifier>
<dc:title><![CDATA[A fluid-walled microfluidic platform for human neuron microcircuits and directed axotomy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.10.10.561760v1?rss=1">
<title>
<![CDATA[
Phosphoglycerate kinase 1 is a central leverage point in Parkinson's disease driven neuronal metabolic deficits 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.10.10.561760v1?rss=1"
</link>
<description><![CDATA[
Phosphoglycerate kinase 1 (PGK1), the first ATP producing glycolytic enzyme, has emerged as a therapeutic target for Parkinsons Disease (PD), since a potential enhancer of its activity was reported to significantly lower PD risk. We carried out a suppressor screen of hypometabolic synaptic deficits and demonstrated that PGK1 is a rate limiting enzyme in nerve terminal ATP production. Increasing PGK1 expression in mid-brain dopamine neurons protected against hydroxy-dopamine driven striatal dopamine nerve terminal dysfunction in-vivo and modest changes in PGK1 activity dramatically suppressed hypometabolic synapse dysfunction in vitro. Furthermore, PGK1 is cross-regulated by PARK7 (DJ-1), a PD associated molecular chaperone, and synaptic deficits driven by PARK20 (Synaptojanin-1) can be reversed by increasing local synaptic PGK1 activity. These data indicate that nerve terminal bioenergetic deficits may underly a spectrum of PD susceptibilities and the identification of PGK1 as the limiting enzyme in axonal glycolysis provides a mechanistic underpinning for therapeutic protection.

O_LISuppressor screen identifies PGK1 as the rate limiting enzyme in axon metabolism
C_LIO_LIPGK1 expression in-vivo protects against striatal dopaminergic axon degeneration
C_LIO_LILoss of DJ-1 impairs axonal ATP production
C_LIO_LIPGK1 function is regulated by PARK7/DJ-1, and can reverse PARK20 synaptic deficits
C_LI
]]></description>
<dc:creator>Kokotos, A.</dc:creator>
<dc:creator>Antoniazzi, A.</dc:creator>
<dc:creator>Unda, S.</dc:creator>
<dc:creator>Kokotos, M. S.</dc:creator>
<dc:creator>Park, D.</dc:creator>
<dc:creator>Eliezer, D.</dc:creator>
<dc:creator>Kaplitt, M.</dc:creator>
<dc:creator>De Camilli, P.</dc:creator>
<dc:creator>Ryan, T.</dc:creator>
<dc:date>2023-10-10</dc:date>
<dc:identifier>doi:10.1101/2023.10.10.561760</dc:identifier>
<dc:title><![CDATA[Phosphoglycerate kinase 1 is a central leverage point in Parkinson's disease driven neuronal metabolic deficits]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.09.15.460511v1?rss=1">
<title>
<![CDATA[
The calcium sensor synaptotagmin-1 is critical for phasic axonal dopamine release in the striatum and mesencephalon, but is dispensable for basic motor behaviors in mice 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.09.15.460511v1?rss=1"
</link>
<description><![CDATA[
In Parkinsons disease (PD), motor dysfunctions only become apparent after extensive loss of DA innervation. This resilience has been hypothesized to be due to the ability of many motor behaviors to be sustained through a diffuse basal tone of DA; but experimental evidence for this is limited. Here we show that conditional deletion of the calcium sensor synaptotagmin-1 (Syt1) in DA neurons (Syt1 cKODA mice) abrogates most activity-dependent axonal DA release in the striatum and mesencephalon, leaving somatodendritic (STD) DA release intact. Strikingly, Syt1 cKODA mice showed intact performance in multiple unconditioned DA-dependent motor tasks and even in a task evaluating conditioned motivation for food. Considering that basal extracellular DA levels in the striatum were unchanged, our findings suggest that activity-dependent DA release is dispensable for such tasks and that they can be sustained by a basal tone of extracellular DA. Taken together, our findings reveal the striking resilience of DA-dependent motor functions in the context of a near-abolition of phasic DA release, shedding new light on why extensive loss of DA innervation is required to reveal motor dysfunctions in PD.
]]></description>
<dc:creator>Delignat-Lavaud, B.</dc:creator>
<dc:creator>Kano, J.</dc:creator>
<dc:creator>Ducrot, C.</dc:creator>
<dc:creator>Masse, I.</dc:creator>
<dc:creator>Mukherjee, S.</dc:creator>
<dc:creator>Giguere, N.</dc:creator>
<dc:creator>Moquin, L.</dc:creator>
<dc:creator>Levesque, C.</dc:creator>
<dc:creator>Nanni, S. B.</dc:creator>
<dc:creator>Bourque, M.-J.</dc:creator>
<dc:creator>Rosa-Neto, P.</dc:creator>
<dc:creator>Levesque, D.</dc:creator>
<dc:creator>De Beaumont, L.</dc:creator>
<dc:creator>Trudeau, L.-E.</dc:creator>
<dc:date>2021-09-16</dc:date>
<dc:identifier>doi:10.1101/2021.09.15.460511</dc:identifier>
<dc:title><![CDATA[The calcium sensor synaptotagmin-1 is critical for phasic axonal dopamine release in the striatum and mesencephalon, but is dispensable for basic motor behaviors in mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.06.14.545012v1?rss=1">
<title>
<![CDATA[
LRRK2 regulates the activation of the unfolded protein response and antigen presentation in macrophages during inflammation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.06.14.545012v1?rss=1"
</link>
<description><![CDATA[
While the contribution of inflammation in the pathological process leading to Parkinsons disease (PD) is well established, a growing body of evidence supports a role for the long-lasting adaptive immune system in the disease. We showed that, in inflammatory conditions, the PD-proteins PINK1 and Parkin negatively regulate the presentation of mitochondrial antigens on MHC I molecules, a process referred to as MitAP (Mitochondrial Antigen Presentation). In the absence of PINK1, over-activation of this pathway in antigen-presenting cells (APCs) engages autoimmune mechanisms leading to the establishment of cytotoxic CD8+ T cells. Co-culture of dopaminergic neurons (DNs) with these T cells led to neuronal cell death, suggesting that the MitAP in DNs made them susceptible to T cell-mediated destruction. In the present study, we used a pharmacological and genetic approach to characterize the MitAP pathway at the molecular level. We showed that this antigen presentation pathway is induced in APCs in response to inflammatory signals through the sequential activation of TLR4, cGAS-STING and the Unfolded Protein Response (UPR). A "UPR motif" present on a STING cytoplasmic domain was shown to specifically activate the UPR sensor IRE1. Remarkably, the PD-related protein LRRK2, acted with STING upstream of the UPR to regulate the transition from innate to adaptive immunity, thereby identifying this PD-related protein as a key player in the immune response during inflammation.
]]></description>
<dc:creator>Fahmy, A. M.</dc:creator>
<dc:creator>Cannon, T.</dc:creator>
<dc:creator>Paredes, C. H.</dc:creator>
<dc:creator>Ahmadi, A.</dc:creator>
<dc:creator>Xu, Y. Z.</dc:creator>
<dc:creator>Barette, B.</dc:creator>
<dc:creator>Ibrahim, M.</dc:creator>
<dc:creator>Lanoix, J.</dc:creator>
<dc:creator>Arango-Duque, G.</dc:creator>
<dc:creator>Schurr, E.</dc:creator>
<dc:creator>Chevet, E.</dc:creator>
<dc:creator>Thibault, P.</dc:creator>
<dc:creator>Gruenheid, S.</dc:creator>
<dc:creator>McBride, H.</dc:creator>
<dc:creator>Desjardins, M.</dc:creator>
<dc:date>2023-06-15</dc:date>
<dc:identifier>doi:10.1101/2023.06.14.545012</dc:identifier>
<dc:title><![CDATA[LRRK2 regulates the activation of the unfolded protein response and antigen presentation in macrophages during inflammation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.06.16.545271v1?rss=1">
<title>
<![CDATA[
Characterizing the diversity of enteric neurons using Dopamine Transporter (DAT)-Cre reporter mice 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.06.16.545271v1?rss=1"
</link>
<description><![CDATA[
The enteric nervous system (ENS) comprises a complex network of neurons whereby a subset appears to be dopaminergic, although the characteristics, roles, and implications in disease are less understood. Most investigations relating to enteric dopamine (DA) neurons rely on immunoreactivity to tyrosine hydroxylase (TH) - a rate-limiting enzyme in the production of DA. However, TH immunoreactivity is likely to provide an incomplete picture given previous work has showed that some DA neurons contain little if any TH and its levels tend to be decreased in response to cellular stress. This study herein provides a comprehensive characterization of DA neurons in the gut using a well-accepted reporter mouse line, expressing a fluorescent protein (tdTomato) under control of the DA transporter (DAT) promoter. Our findings confirm a unique localization of DA neurons in the gut and unveil the discrete subtypes of DA neurons in this organ, which we characterized using both immunofluorescence and single-cell transcriptomics, as well as validated using in situ hybridization. We observed distinct subtypes of DAT-tdTomato neurons expressing co-transmitters and modulators across both plexuses; some of them likely co-releasing acetylcholine, while others were positive for a slew of canonical DA markers (TH, VMAT2 and GIRK2). Interestingly, we uncovered a seemingly novel population of DA neurons unique to the ENS which were ChAT/DAT-tdTomato-immunoreactive neurons and were characterised by the expression of Grp, Calcb and Sst. Given the clear heterogeneity of DAergic gut neurons, further investigation is warranted to define their functional signatures and discover any inherent vulnerabilities in disease.



O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=81 SRC="FIGDIR/small/545271v3_ufig1.gif" ALT="Figure 1">
View larger version (24K):
org.highwire.dtl.DTLVardef@1e1830borg.highwire.dtl.DTLVardef@625579org.highwire.dtl.DTLVardef@1d9b59aorg.highwire.dtl.DTLVardef@219cab_HPS_FORMAT_FIGEXP  M_FIG C_FIG Using a reporter mouse line, expressing a fluorescent protein under control of the dopamine transporter (DAT) promoter, discrete subtypes of dopaminergic neurons were unveiled across the ganglionated plexuses of the gut. A novel subpopulations of enteric DA neurons, expressing genes previously reported involved in dopamine signaling in the brain, exhibit a cholinergic phenotype.
]]></description>
<dc:creator>Recinto, S.</dc:creator>
<dc:creator>Premachandran, S.</dc:creator>
<dc:creator>Mukherjee, S.</dc:creator>
<dc:creator>McDonald, A.</dc:creator>
<dc:creator>Gruenheid, S.</dc:creator>
<dc:creator>Trudeau, L. E.</dc:creator>
<dc:creator>Stratton, J. A.</dc:creator>
<dc:date>2023-06-19</dc:date>
<dc:identifier>doi:10.1101/2023.06.16.545271</dc:identifier>
<dc:title><![CDATA[Characterizing the diversity of enteric neurons using Dopamine Transporter (DAT)-Cre reporter mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.07.31.549036v1?rss=1">
<title>
<![CDATA[
Comparative Analysis of Methods to Reduce Activation Signature Gene Expression in PBMCs 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.07.31.549036v1?rss=1"
</link>
<description><![CDATA[
Preserving the in vivo cell transcriptome is essential for accurate profiling, yet factors during cell isolation including time ex vivo and temperature induce artifactual gene expression, particularly in stress-responsive immune cells. In this study, we investigated two methods to mitigate ex vivo activation signature gene (ASG) expression in peripheral blood mononuclear cells (PBMCs): transcription and translation inhibitors (TTis) and cold temperatures during isolation. Comparative analysis of PBMCs isolated with TTis revealed reduced ASG expression. However, TTi treatment impaired responsiveness to LPS stimulation in subsequent in vitro experiments. In contrast, cold isolation methods also prevented ASG expression; up to a point where the addition of TTis during cold isolation offered minimal additional advantage. These findings highlight the importance of considering the advantages and drawbacks of different isolation methods to ensure accurate interpretation of PBMC transcriptomic profiles.

HighlightsO_LITraditional room temperature isolation methods trigger activation signature gene expression in PBMCs, even when rapidly isolated, whereas 4{degrees}C isolation methods do not.
C_LIO_LITranscription and translation inhibitors and cold processing techniques reduce activation signature gene expression via shared mechanisms.
C_LIO_LIPBMCs treated with transcription and translation inhibitors lose responsiveness to external stimuli.
C_LIO_LICold isolation methods offer a suitable and inexpensive alternative to mitigate activation signature gene expression in PBMCs.
C_LI
]]></description>
<dc:creator>Andriamboavonjy, L.</dc:creator>
<dc:creator>MacDonald, A.</dc:creator>
<dc:creator>Hamilton, L. K.</dc:creator>
<dc:creator>Labrecque, M.</dc:creator>
<dc:creator>Boivin, M.-N.</dc:creator>
<dc:creator>Karamchandani, J.</dc:creator>
<dc:creator>Stratton, J. A.</dc:creator>
<dc:creator>Tetreault, M.</dc:creator>
<dc:date>2023-08-02</dc:date>
<dc:identifier>doi:10.1101/2023.07.31.549036</dc:identifier>
<dc:title><![CDATA[Comparative Analysis of Methods to Reduce Activation Signature Gene Expression in PBMCs]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.10.14.562276v1?rss=1">
<title>
<![CDATA[
Systems-level analyses dissociate genetic regulators of reactive oxygen species and energy production 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.10.14.562276v1?rss=1"
</link>
<description><![CDATA[
Respiratory chain dysfunction can decrease ATP and increase reactive oxygen species (ROS) levels. Despite the importance of these metabolic parameters to a wide range of cellular functions and disease, we lack an integrated understanding of how they are differentially regulated. To address this question, we adapted a CRISPRi- and FACS-based platform to compare the effects of respiratory gene knockdown on ROS to their effects on ATP. Focusing on genes whose knockdown is known to decrease mitochondria-derived ATP, we showed that knockdown of genes in specific respiratory chain complexes (I, III and CoQ10 biosynthesis) increased ROS, whereas knockdown of other low ATP hits either had no impact (mitochondrial ribosomal proteins) or actually decreased ROS (complex IV). Moreover, although shifting metabolic conditions profoundly altered mitochondria-derived ATP levels, it had little impact on mitochondrial or cytosolic ROS. In addition, knockdown of a subset of complex I subunits--including NDUFA8, NDUFB4, and NDUFS8--decreased complex I activity, mitochondria-derived ATP and supercomplex level, but knockdown of these genes had differential effects on ROS. Conversely, we found an essential role for ether lipids in the dynamic regulation of mitochondrial ROS levels independent of ATP. Thus, our results identify specific metabolic regulators of cellular ATP and ROS balance that may help dissect the roles of these processes in disease and identify therapeutic strategies to independently target energy failure and oxidative stress.

SignificanceMitochondrial respiration generates both energy (ATP) and reactive oxygen species (ROS). Insufficient energy and increased ROS from respiratory chain dysfunction may be central to the pathophysiology of neurodegenerative diseases and aging. We established a screening platform using CRISPR and fluorescent-cell sorting to compare the impact of decreasing respiratory chain proteins on ROS and ATP levels. The results provide the first systems-level analysis of how ROS and ATP are differentially regulated, and identify genes and respiratory chain complexes that can manipulate each independently. These findings advance our understanding of the relative contributions of ATP and ROS to disease pathophysiology, and guide the development of therapies to preserve energy while minimizing ROS.
]]></description>
<dc:creator>Bennett, N. K.</dc:creator>
<dc:creator>Lee, M.</dc:creator>
<dc:creator>Orr, A. L.</dc:creator>
<dc:creator>Nakamura, K.</dc:creator>
<dc:date>2023-10-18</dc:date>
<dc:identifier>doi:10.1101/2023.10.14.562276</dc:identifier>
<dc:title><![CDATA[Systems-level analyses dissociate genetic regulators of reactive oxygen species and energy production]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.06.24.546170v1?rss=1">
<title>
<![CDATA[
Synapsin E-domain is essential for α-synuclein function 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.06.24.546170v1?rss=1"
</link>
<description><![CDATA[
The cytosolic proteins synucleins and synapsins are thought to play cooperative roles in regulating synaptic vesicle (SV) recycling, but mechanistic insight is lacking. Here we identify the synapsin E-domain as an essential functional binding-partner of -synuclein (-syn). Synapsin E-domain allows -syn functionality, binds to -syn, and is necessary and sufficient for enabling effects of -syn at the synapse. Together with previous studies implicating the E-domain in clustering SVs, our experiments advocate a cooperative role for these two proteins in maintaining physiologic SV clusters.
]]></description>
<dc:creator>Stavsky, A.</dc:creator>
<dc:creator>Parra-Rivas, L. A.</dc:creator>
<dc:creator>Tal, S.</dc:creator>
<dc:creator>Madhivanan, K.</dc:creator>
<dc:creator>Roy, S.</dc:creator>
<dc:creator>Gitler, D.</dc:creator>
<dc:date>2023-06-26</dc:date>
<dc:identifier>doi:10.1101/2023.06.24.546170</dc:identifier>
<dc:title><![CDATA[Synapsin E-domain is essential for α-synuclein function]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.11.11.378976v1?rss=1">
<title>
<![CDATA[
A Markov Random Field Model for Network-based Differential Expression Analysis of Single-cell RNA-seq Data 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.11.11.378976v1?rss=1"
</link>
<description><![CDATA[
BackgroundRecent development of single cell sequencing technologies has made it possible to identify genes with different expression (DE) levels at the cell type level between different groups of samples. In this article, we propose to borrow information through known biological networks to increase statistical power to identify differentially expressed genes (DEGs).

ResultsWe develop MRFscRNAseq, which is based on a Markov Random Field (MRF) model to appropriately accommodate gene network information as well as dependencies among cell types to identify cell-type specific DEGs. We implement an Expectation-Maximization (EM) algorithm with mean field-like approximation to estimate model parameters and a Gibbs sampler to infer DE status. Simulation study shows that our method has better power to detect cell-type specific DEGs than conventional methods while appropriately controlling type I error rate. The usefulness of our method is demonstrated through its application to study the pathogenesis and biological processes of idiopathic pulmonary fibrosis (IPF) using a single-cell RNA-sequencing (scRNA-seq) data set, which contains 18,150 protein-coding genes across 38 cell types on lung tissues from 32 IPF patients and 28 normal controls.

ConclusionsThe proposed MRF model is implemented in the R package MRFscRNAseq available on GitHub. By utilizing gene-gene and cell-cell networks, our method increases statistical power to detect differentially expressed genes from scRNA-seq data.
]]></description>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Xu, Z.</dc:creator>
<dc:creator>Adams, T.</dc:creator>
<dc:creator>Kaminski, N.</dc:creator>
<dc:creator>Zhao, H.</dc:creator>
<dc:date>2020-11-12</dc:date>
<dc:identifier>doi:10.1101/2020.11.11.378976</dc:identifier>
<dc:title><![CDATA[A Markov Random Field Model for Network-based Differential Expression Analysis of Single-cell RNA-seq Data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.03.29.534370v1?rss=1">
<title>
<![CDATA[
Splicing accuracy varies across human introns, tissues and age 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.03.29.534370v1?rss=1"
</link>
<description><![CDATA[
Alternative splicing impacts most multi-exonic human genes. Inaccuracies during this process may have an important role in ageing and disease. Here, we investigated mis-splicing using RNA-sequencing data from ~14K control samples and 42 human body sites, focusing on split reads partially mapping to known transcripts in annotation. We show that mis-splicing occurs at different rates across introns and tissues and that these splicing inaccuracies are primarily affected by the abundance of core components of the spliceosome assembly and its regulators. Using publicly available data on short-hairpin RNA-knockdowns of numerous spliceosomal components and related regulators, we found support for the importance of RNA-binding proteins in mis-splicing. We also demonstrated that age is positively correlated with mis-splicing, and it affects genes implicated in neurodegenerative diseases. This in-depth characterisation of mis-splicing can have important implications for our understanding of the role of splicing inaccuracies in human disease and the interpretation of long-read RNA-sequencing data.
]]></description>
<dc:creator>Garcia-Ruiz, S.</dc:creator>
<dc:creator>Zhang, D.</dc:creator>
<dc:creator>Gustavsson, E. K.</dc:creator>
<dc:creator>Rocamora-Perez, G.</dc:creator>
<dc:creator>Grant-Peters, M.</dc:creator>
<dc:creator>Fairbrother-Browne, A.</dc:creator>
<dc:creator>Reynolds, R. H.</dc:creator>
<dc:creator>Brenton, J. W.</dc:creator>
<dc:creator>Gil-Martinez, A. L.</dc:creator>
<dc:creator>Chen, Z.</dc:creator>
<dc:creator>Rio, D. C.</dc:creator>
<dc:creator>Botia, J. A.</dc:creator>
<dc:creator>Guelfi, S.</dc:creator>
<dc:creator>Collado-Torres, L.</dc:creator>
<dc:creator>Ryten, M.</dc:creator>
<dc:date>2023-03-30</dc:date>
<dc:identifier>doi:10.1101/2023.03.29.534370</dc:identifier>
<dc:title><![CDATA[Splicing accuracy varies across human introns, tissues and age]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.08.03.551835v1?rss=1">
<title>
<![CDATA[
KAT8 compound inhibition inhibits PINK1/Parkin-dependent mitophagy and initiates mitochondrial delivery to lysosomes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.08.03.551835v1?rss=1"
</link>
<description><![CDATA[
It has recently been shown that KAT8, a genome-wide association study (GWAS) candidate risk gene for Parkinsons Disease, is involved in PINK1/Parkin-dependant mitophagy. The KAT8 gene encodes a lysine acetyltransferase and represents the catalytically active subunit of the non-specific lethal (NSL) epigenetic remodelling complex. In the current study, we show that contrary to KAT5 inhibition, dual inhibition of KAT5 and KAT8 via the MG149 compound inhibits the initial steps of the PINK1-dependant mitophagy process. More specifically, our study shows that following mitochondrial depolarisation induced by mitochondrial toxins, MG149 treatment inhibits PINK1-dependant mitophagy initiation by impairing PINK1 activation, and subsequent phosphorylation of Parkin and ubiquitin. While this inhibitory effect of MG149 on PINK1-activation is potent, MG149 treatment in the absence of mitochondrial toxins is sufficient to depolarise the mitochondrial membrane, recruit PINK1 and promote partial downstream recruitment of the autophagy receptor p62, leading to an increase in mitochondrial delivery to the lysosomes. Altogether, our study provides additional support for KAT8 as a regulator of mitophagy and autophagy processes.
]]></description>
<dc:creator>de Talhouet, C.</dc:creator>
<dc:creator>O'Callaghan, B.</dc:creator>
<dc:creator>Esteras Gallego, N.</dc:creator>
<dc:creator>Plun-Favreau, H.</dc:creator>
<dc:date>2023-08-05</dc:date>
<dc:identifier>doi:10.1101/2023.08.03.551835</dc:identifier>
<dc:title><![CDATA[KAT8 compound inhibition inhibits PINK1/Parkin-dependent mitophagy and initiates mitochondrial delivery to lysosomes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.11.02.365825v1?rss=1">
<title>
<![CDATA[
The Hsc70 Disaggregation Machinery RemovesMonomer Units Directly from α-Synuclein FibrilEnds 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.11.02.365825v1?rss=1"
</link>
<description><![CDATA[
Molecular chaperones contribute to the maintenance of cellular protein homeostasis through a wide range of mechanisms, including the assistance of de novo protein folding, the rescue of misfolded proteins, and the prevention of amyloid formation. Chaperones of the Hsp70 family have a striking capability of disaggregating otherwise irreversible aggregate structures such as amyloid fibrils that accumulate during the development of neurodegenerative diseases. However, the mechanisms of this key emerging functionality remain largely unknown. Here, we bring together microfluidic measurements with kinetic analysis and show that that the Hsp70 protein heat chock complement Hsc70 together with its two co-chaperones DnaJB1 and the nucleotide exchange factor Apg2 is able to completely reverse the aggregation process of alpha-synuclein, associated with Parkinsons disease, back to its soluble monomeric state. Moreover, we show that this reaction proceeds with first order kinetics in a process where monomer units are taken off directly from the fibril ends. Our results demonstrate that all components of the chaperone triad are essential for fibril disaggregation. Lastly, we quantify the interactions between the three chaperones as well as between the chaperones and the fibrils in solution, yielding both binding stoichiometries and dissociation constants. Crucially, we find that the stoichiometry of Hsc70 binding to fibrils suggests Hsc70 clustering at the fibril ends. Taken together, our results show that the mechanism of action of the Hsc70-DnaJB1-Apg2 chaperone system in disaggregating -synuclein fibrils involves the removal of monomer units without any intermediate fragmentation steps. These findings are fundamental to our understanding of the suppression of amyloid proliferation early in life and the natural clearance mechanisms of fibrillar deposits in Parkinsons disease, and inform on the possibilities and limitations of this strategy in the development of therapeutics against synucleinopathies and related neurodegenerative diseases.
]]></description>
<dc:creator>Schneider, M. M.</dc:creator>
<dc:creator>Gautam, S.</dc:creator>
<dc:creator>Herling, T. W.</dc:creator>
<dc:creator>Andrzejwska, E.</dc:creator>
<dc:creator>Krainer, G.</dc:creator>
<dc:creator>Miller, A. M.</dc:creator>
<dc:creator>Peter, Q. A. E.</dc:creator>
<dc:creator>Ruggeri, F. S.</dc:creator>
<dc:creator>Vendruscolo, M.</dc:creator>
<dc:creator>Bracher, A.</dc:creator>
<dc:creator>Dobson, C. M.</dc:creator>
<dc:creator>Hartl, F.-U.</dc:creator>
<dc:creator>Knowles, T.</dc:creator>
<dc:date>2020-11-03</dc:date>
<dc:identifier>doi:10.1101/2020.11.02.365825</dc:identifier>
<dc:title><![CDATA[The Hsc70 Disaggregation Machinery RemovesMonomer Units Directly from α-Synuclein FibrilEnds]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.09.30.461806v1?rss=1">
<title>
<![CDATA[
Progranulin deficiency results in reduced bis(monoacylglycero)phosphate (BMP) levels and gangliosidosis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.09.30.461806v1?rss=1"
</link>
<description><![CDATA[
Homozygous mutations of granulin precursor (GRN) lead to neuronal ceroid lipofuscinosis1, a severe neurodevelopmental disease, in humans and neuroinflammation in mice2. Haploinsufficiency of GRN almost invariably causes frontotemporal dementia (FTD)3,4. The GRN locus produces progranulin (PGRN), a lysosomal precursor protein that is cleaved to granulin peptides5,6. Despite intensive investigation, the function of granulins and the reason why their absence causes neurodegeneration remain unclear. Here, we investigated PGRN function in lipid degradation, a major function of lysosomes. We show that PGRN-knockout human cells, PGRN-deficient murine brain, and frontal lobes of human brains from patients with GRN mutation-related FTD have increased levels of gangliosides, highly abundant sialic acid-containing glycosphingolipids (GSL) that are degraded in lysosomes. Probing how PGRN deficiency causes these changes, we found normal levels and activities of enzymes that catabolize gangliosides. However, levels of bis(monoacylglycero)phosphate (BMP), a lysosomal lipid required for ganglioside catabolism7, were markedly reduced in PGRN-deficient cells and patient brain tissues. These data indicate that granulins are required to maintain BMP levels, which regulate ganglioside catabolism, and that PGRN deficiency in lysosomes leads to gangliosidosis. This aberrant accumulation of gangliosides may contribute to neuroinflammation and neurodegeneration susceptibility.
]]></description>
<dc:creator>Boland, S.</dc:creator>
<dc:creator>Swarup, S.</dc:creator>
<dc:creator>Ambaw, Y. A.</dc:creator>
<dc:creator>Richards, R. C.</dc:creator>
<dc:creator>Fischer, A. W.</dc:creator>
<dc:creator>Singh, S.</dc:creator>
<dc:creator>Aggarwal, G.</dc:creator>
<dc:creator>Spina, S.</dc:creator>
<dc:creator>Nana, A. L.</dc:creator>
<dc:creator>Grinberg, L. T.</dc:creator>
<dc:creator>Seeley, W. W.</dc:creator>
<dc:creator>Surma, M. A.</dc:creator>
<dc:creator>Klose, C.</dc:creator>
<dc:creator>Paulo, J. A.</dc:creator>
<dc:creator>Nguyen, A. D.</dc:creator>
<dc:creator>Harper, J. W.</dc:creator>
<dc:creator>Walther, T. C.</dc:creator>
<dc:creator>Farese, R. V.</dc:creator>
<dc:date>2021-09-30</dc:date>
<dc:identifier>doi:10.1101/2021.09.30.461806</dc:identifier>
<dc:title><![CDATA[Progranulin deficiency results in reduced bis(monoacylglycero)phosphate (BMP) levels and gangliosidosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-09-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.06.26.546565v1?rss=1">
<title>
<![CDATA[
Combinatorial selective ER-phagy remodels the ER during neurogenesis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.06.26.546565v1?rss=1"
</link>
<description><![CDATA[
The endoplasmic reticulum (ER) employs a diverse proteome landscape to orchestrate many cellular functions ranging from protein and lipid synthesis to calcium ion flux and inter-organelle communication. A case in point concerns the process of neurogenesis: a refined tubular ER network is assembled via ER shaping proteins into the newly formed neuronal projections to create highly polarized dendrites and axons. Previous studies have suggested a role for autophagy in ER remodeling, as autophagy-deficient neurons in vivo display axonal ER accumulation within synaptic boutons, and the membrane-embedded ER-phagy receptor FAM134B has been genetically linked with human sensory and autonomic neuropathy. However, our understanding of the mechanisms underlying selective removal of ER and the role of individual ER-phagy receptors is limited. Here, we combine a genetically tractable induced neuron (iNeuron) system for monitoring ER remodeling during in vitro differentiation with proteomic and computational tools to create a quantitative landscape of ER proteome remodeling via selective autophagy. Through analysis of single and combinatorial ER-phagy receptor mutants, we delineate the extent to which each receptor contributes to both magnitude and selectivity of ER protein clearance. We define specific subsets of ER membrane or lumenal proteins as preferred clients for distinct receptors. Using spatial sensors and flux reporters, we demonstrate receptor-specific autophagic capture of ER in axons, and directly visualize tubular ER membranes within autophagosomes in neuronal projections by cryo-electron tomography. This molecular inventory of ER proteome remodeling and versatile genetic toolkit provides a quantitative framework for understanding contributions of individual ER-phagy receptors for reshaping ER during cell state transitions.
]]></description>
<dc:creator>Hoyer, M. J.</dc:creator>
<dc:creator>Smith, I. R.</dc:creator>
<dc:creator>Paoli, J. C.</dc:creator>
<dc:creator>Jiang, Y.</dc:creator>
<dc:creator>Paulo, J. A.</dc:creator>
<dc:creator>Harper, W.</dc:creator>
<dc:date>2023-06-26</dc:date>
<dc:identifier>doi:10.1101/2023.06.26.546565</dc:identifier>
<dc:title><![CDATA[Combinatorial selective ER-phagy remodels the ER during neurogenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.03.27.534444v1?rss=1">
<title>
<![CDATA[
Proteostasis and lysosomal quality control deficits in Alzheimer's disease neurons 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.03.27.534444v1?rss=1"
</link>
<description><![CDATA[
Aging is the most prominent risk factor for Alzheimers disease (AD). However, the cellular mechanisms linking neuronal proteostasis decline to the characteristic aberrant protein deposits in AD brains remain elusive. Here, we develop transdifferentiated neurons (tNeurons) from human dermal fibroblasts as a neuronal model that retains aging hallmarks and exhibits AD-linked vulnerabilities. Remarkably, AD tNeurons accumulate proteotoxic deposits, including phospho-Tau and A{beta}, resembling those in AD patient and APP mouse brains. Quantitative tNeuron proteomics identify aging and AD-linked deficits in proteostasis and organelle homeostasis, most notably in endosome-lysosomal components. Lysosomal deficits in aged tNeurons, including constitutive lysosomal damage and ESCRT-mediated lysosomal repair defects, are exacerbated in AD tNeurons and linked to inflammatory cytokine secretion and cell death. Supporting lysosomal deficits centrality in AD, compounds ameliorating lysosomal function reduce A{beta} deposits and cytokine secretion. Thus, the tNeuron model system reveals impaired lysosomal homeostasis as an early event of aging and AD.
]]></description>
<dc:creator>Chou, C.-C.</dc:creator>
<dc:creator>Vest, R.</dc:creator>
<dc:creator>Prado, M. A.</dc:creator>
<dc:creator>Wilson-Grady, J.</dc:creator>
<dc:creator>Paulo, J. A.</dc:creator>
<dc:creator>Shibuya, Y.</dc:creator>
<dc:creator>Moran-Losada, P.</dc:creator>
<dc:creator>Lee, T.-T.</dc:creator>
<dc:creator>Luo, J.</dc:creator>
<dc:creator>Gygi, S. P.</dc:creator>
<dc:creator>Kelly, J. W.</dc:creator>
<dc:creator>Finley, D.</dc:creator>
<dc:creator>Wernig, M.</dc:creator>
<dc:creator>Wyss-Coray, T.</dc:creator>
<dc:creator>Frydman, J.</dc:creator>
<dc:date>2023-03-27</dc:date>
<dc:identifier>doi:10.1101/2023.03.27.534444</dc:identifier>
<dc:title><![CDATA[Proteostasis and lysosomal quality control deficits in Alzheimer's disease neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.07.25.550521v1?rss=1">
<title>
<![CDATA[
RAB3 phosphorylation by pathogenic LRRK2 impairs trafficking of synaptic vesicle precursors 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.07.25.550521v1?rss=1"
</link>
<description><![CDATA[
Gain-of-function mutations in the LRRK2 gene cause Parkinsons disease (PD), characterized by debilitating motor and non-motor symptoms. Increased phosphorylation of a subset of RAB GTPases by LRRK2 is implicated in PD pathogenesis. We find that increased phosphorylation of RAB3A, a cardinal synaptic vesicle precursor (SVP) protein, disrupts anterograde axonal transport of SVPs in iPSC-derived human neurons (iNeurons) expressing hyperactive LRRK2-p.R1441H. Knockout of the opposing protein phosphatase 1H (PPM1H) in iNeurons phenocopies this effect. In these models, the compartmental distribution of synaptic proteins is altered; synaptophysin and synaptobrevin-2 become sequestered in the neuronal soma with decreased delivery to presynaptic sites along the axon. We find that RAB3A phosphorylation disrupts binding to the motor adapter MADD, potentially preventing formation of the RAB3A-MADD-KIF1A/1B{beta} complex driving anterograde SVP transport. RAB3A hyperphosphorylation also disrupts interactions with RAB3GAP and RAB-GDI1. Our results reveal a mechanism by which pathogenic hyperactive LRRK2 may contribute to the altered synaptic homeostasis associated with characteristic non-motor and cognitive manifestations of PD.

SUMMARYDou et al. demonstrate that Parkinsons disease-associated hyperactive LRRK2 decreases trafficking of synaptic vesicle proteins within neurons by disrupting regulation of the synaptic vesicle precursor protein RAB3A. Impaired delivery of synaptic proteins to presynaptic sites could contribute to progression of debilitating non-motor PD symptoms.
]]></description>
<dc:creator>Dou, D.</dc:creator>
<dc:creator>Aiken, J.</dc:creator>
<dc:creator>Holzbaur, E. L. F.</dc:creator>
<dc:date>2023-07-25</dc:date>
<dc:identifier>doi:10.1101/2023.07.25.550521</dc:identifier>
<dc:title><![CDATA[RAB3 phosphorylation by pathogenic LRRK2 impairs trafficking of synaptic vesicle precursors]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.09.25.559255v1?rss=1">
<title>
<![CDATA[
Control of mitophagy initiation and progression by the TBK1 adaptors NAP1 and SINTBAD 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.09.25.559255v1?rss=1"
</link>
<description><![CDATA[
Mitophagy preserves overall mitochondrial fitness by selectively targeting damaged mitochondria for degradation. The regulatory mechanisms that prevent PINK1/Parkin-dependent mitophagy and other selective autophagy pathways from overreacting while ensuring swift progression once initiated are largely elusive. Here, we demonstrate how the TBK1 adaptors NAP1 and SINTBAD restrict the initiation of OPTN-driven mitophagy by competing with OPTN for TBK1. Conversely, they promote the progression of NDP52-driven mitophagy by recruiting TBK1 to NDP52 and stabilizing its interaction with FIP200. Notably, OPTN emerges as the primary recruiter of TBK1 during mitophagy initiation, which in return boosts NDP52-mediated mitophagy. Our results thus define NAP1 and SINTBAD as cargo receptor rheostats, elevating the threshold for mitophagy initiation by OPTN while promoting the progression of the pathway once set in motion by supporting NDP52. These findings shed light on the cellular strategy to prevent pathway hyperactivity while still ensuring efficient progression.
]]></description>
<dc:creator>Adriaenssens, E.</dc:creator>
<dc:creator>Nguyen, T. N.</dc:creator>
<dc:creator>Sawa-Makarska, J.</dc:creator>
<dc:creator>Khuu, G.</dc:creator>
<dc:creator>Schuschnig, M.</dc:creator>
<dc:creator>Shoebridge, S.</dc:creator>
<dc:creator>Watts, E. M.</dc:creator>
<dc:creator>Csalyi, K. D.</dc:creator>
<dc:creator>Padman, B. S.</dc:creator>
<dc:creator>Lazarou, M.</dc:creator>
<dc:creator>Martens, S.</dc:creator>
<dc:date>2023-09-25</dc:date>
<dc:identifier>doi:10.1101/2023.09.25.559255</dc:identifier>
<dc:title><![CDATA[Control of mitophagy initiation and progression by the TBK1 adaptors NAP1 and SINTBAD]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.08.10.552488v1?rss=1">
<title>
<![CDATA[
A leaky gut dysregulates gene networks in the brain associated with immune activation, oxidative stress, and myelination in a mouse model of colitis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.08.10.552488v1?rss=1"
</link>
<description><![CDATA[
The gut and brain are increasingly linked in human disease, with neuropsychiatric conditions classically attributed to the brain showing an involvement of the intestine and inflammatory bowel diseases (IBDs) displaying an ever-expanding list of neurological comorbidities. To identify molecular systems that underpin this gut-brain connection and thus discover therapeutic targets, experimental models of gut dysfunction must be evaluated for brain effects. In the present study, we examine disturbances along the gut-brain axis in a widely used murine model of colitis, the dextran sodium sulfate (DSS) model, using high-throughput transcriptomics and an unbiased network analysis strategy coupled with standard biochemical outcome measures to achieve a comprehensive approach to identify key disease processes in both colon and brain. We examine the reproducibility of colitis induction with this model and its resulting genetic programs during different phases of disease, finding that DSS-induced colitis is largely reproducible with a few site-specific molecular features. We focus on the circulating immune system as the intermediary between the gut and brain, which exhibits an activation of pro-inflammatory innate immunity during colitis. Our unbiased transcriptomics analysis provides supporting evidence for immune activation in the brain during colitis, suggests that myelination may be a process vulnerable to increased intestinal permeability, and identifies a possible role for oxidative stress and brain oxygenation. Overall, we provide a comprehensive evaluation of multiple systems in a prevalent experimental model of intestinal permeability, which will inform future studies using this model and others, assist in the identification of druggable targets in the gut-brain axis, and contribute to our understanding of the concomitance of intestinal and neuropsychiatric dysfunction.
]]></description>
<dc:creator>Boles, J. S.</dc:creator>
<dc:creator>Krueger, M.</dc:creator>
<dc:creator>Jernigan, J.</dc:creator>
<dc:creator>Cole, C.</dc:creator>
<dc:creator>Neighbarger, N.</dc:creator>
<dc:creator>Huarte, O. U.</dc:creator>
<dc:creator>Tansey, M. G.</dc:creator>
<dc:date>2023-08-13</dc:date>
<dc:identifier>doi:10.1101/2023.08.10.552488</dc:identifier>
<dc:title><![CDATA[A leaky gut dysregulates gene networks in the brain associated with immune activation, oxidative stress, and myelination in a mouse model of colitis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.08.14.553305v1?rss=1">
<title>
<![CDATA[
Gut mucosal cells transfer α-synuclein to the vagus nerve 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.08.14.553305v1?rss=1"
</link>
<description><![CDATA[
Epidemiological and histopathological findings have raised the possibility that misfolded -synuclein protein might spread from the gut to the brain and increase the risk of Parkinsons disease (PD). While past experimental studies in mouse models have relied on gut injections of exogenous recombinant -synuclein fibrils to study gut to brain -synuclein transfer, the possible origins of misfolded -synuclein within the gut have remained elusive. We recently demonstrated that sensory cells of the gut mucosa express -synuclein. In this study, we employed mouse intestinal organoids expressing human -synuclein to observe the transfer of -synuclein protein from gut epithelial cells in organoids co-cultured with vagal nodose neurons that are otherwise devoid of -synuclein expression. In intact mice that express pathological human -synuclein, but no mouse -synuclein, -synuclein fibril templating activity emerges in -synuclein seeded fibril aggregation assays in tissues from the gut, vagus nerve, and dorsal motor nucleus. In newly engineered transgenic mice that restrict pathological human -synuclein expression to intestinal epithelial cells, -synuclein fibril-templating activity transfers to the vagus nerve and to the dorsal motor nucleus. Subdiaphragmatic vagotomy prior to the induction of -synuclein expression in the gut epithelial cells effectively protects the hindbrain from the emergence of -synuclein fibril templating activity. Overall, these findings highlight a novel potential non-neuronal source of fibrillar -synuclein protein that might arise in gut mucosal cells.
]]></description>
<dc:creator>Chandra, R.</dc:creator>
<dc:creator>Sokratian, A.</dc:creator>
<dc:creator>Chavez, K. R.</dc:creator>
<dc:creator>King, S.</dc:creator>
<dc:creator>Swain, S. M.</dc:creator>
<dc:creator>Snyder, J. C.</dc:creator>
<dc:creator>West, A. B.</dc:creator>
<dc:creator>Liddle, R. A.</dc:creator>
<dc:date>2023-08-14</dc:date>
<dc:identifier>doi:10.1101/2023.08.14.553305</dc:identifier>
<dc:title><![CDATA[Gut mucosal cells transfer α-synuclein to the vagus nerve]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.11.22.517582v1?rss=1">
<title>
<![CDATA[
Structure of LRRK1 and mechanisms of autoinhibition and activation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.11.22.517582v1?rss=1"
</link>
<description><![CDATA[
Leucine Rich Repeat Kinase 1 and 2 (LRRK1 and LRRK2) are homologs in the ROCO family of proteins in humans. Despite their shared domain architecture and involvement in intracellular trafficking, their disease associations are strikingly different: LRRK2 is involved in familial Parkinsons Disease (PD) while LRRK1 is linked to bone diseases. Furthermore, PD-linked mutations in LRRK2 are typically autosomal dominant gain-of-function while those in LRRK1 are autosomal recessive loss-of-function. To understand these differences, we solved cryo-EM structures of LRRK1 in its monomeric and dimeric forms. Both differ from the corresponding LRRK2 structures. Unlike LRRK2, which is sterically autoinhibited as a monomer, LRRK1 is sterically autoinhibited in a dimer-dependent manner. LRRK1 has an additional level of autoinhibition that prevents activation of the kinase and is absent in LRRK2. Finally, we place the structural signatures of LRRK1 and LRRK2 in the context of the evolution of the LRRK family of proteins.
]]></description>
<dc:creator>Reimer, J. M.</dc:creator>
<dc:creator>Dickey, A. M.</dc:creator>
<dc:creator>Lin, Y. X.</dc:creator>
<dc:creator>Abrisch, R. G.</dc:creator>
<dc:creator>Mathea, S.</dc:creator>
<dc:creator>Chatterjee, D.</dc:creator>
<dc:creator>Fay, E. J.</dc:creator>
<dc:creator>Daugherty, M. D.</dc:creator>
<dc:creator>Reck-Peterson, S. L.</dc:creator>
<dc:creator>Leschziner, A. E.</dc:creator>
<dc:date>2022-11-23</dc:date>
<dc:identifier>doi:10.1101/2022.11.22.517582</dc:identifier>
<dc:title><![CDATA[Structure of LRRK1 and mechanisms of autoinhibition and activation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.02.03.478943v1?rss=1">
<title>
<![CDATA[
A mono- and intralink filter (mi-filter) to improve false-discovery rates in cross-linking mass spectrometry data 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.02.03.478943v1?rss=1"
</link>
<description><![CDATA[
Cross-Linking Mass Spectrometry (XL-MS) has become an indispensable tool for the emerging field of systems structural biology over the recent years. However, the confidence in individual protein-protein interactions (PPIs) depends on the correct assessment of individual inter protein cross-links. This can be challenging, in particularly in samples where relatively few PPIs are detected, as is often the case in complex samples containing low abundant proteins or in in-vivo settings. In this manuscript we are describing a novel mono- and intralink filter (mi-filter) that is applicable to any kind of crosslinking data and workflow. It stipulates that only proteins for which at least one monolink or intra-protein crosslink has been identified within a given dataset are considered for an inter-protein cross-link and therefore participate in a PPI. We show that this simple and intuitive filter has a dramatic effect on different types of crosslinking-data ranging from single protein complexes, over medium-complexity affinity enrichments to proteome-wide cell lysates and significantly lowers the number of false-positive identifications resulting in improved false-discovery rates for inter-protein links in all these types of XL-MS data.
]]></description>
<dc:creator>Chen, X.</dc:creator>
<dc:creator>Sailer, C.</dc:creator>
<dc:creator>Kammer, K.-M.</dc:creator>
<dc:creator>Fuersch, J.</dc:creator>
<dc:creator>Eisele, M. R.</dc:creator>
<dc:creator>Sakata, E.</dc:creator>
<dc:creator>Stengel, F.</dc:creator>
<dc:date>2022-02-03</dc:date>
<dc:identifier>doi:10.1101/2022.02.03.478943</dc:identifier>
<dc:title><![CDATA[A mono- and intralink filter (mi-filter) to improve false-discovery rates in cross-linking mass spectrometry data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.09.07.556689v1?rss=1">
<title>
<![CDATA[
Inhibition of Parkinsons Disease-related LRRK2 by type-I and type-II kinase inhibitors: activity and structures 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.09.07.556689v1?rss=1"
</link>
<description><![CDATA[
Mutations in Leucine Rich Repeat Kinase 2 (LRRK2) are a common cause of familial Parkinsons Disease (PD), and a risk factor for the sporadic form. Increased kinase activity has been shown in both familial and sporadic PD patients, making LRRK2 kinase inhibitors a major focus of drug development efforts in PD. Although significant progress has been made in understanding the structural biology of LRRK2, there are no available structures of LRRK2 inhibitor complexes. To this end, we solved cryo-EM structures of LRRK2, wild-type and PD-linked mutants, bound to the LRRK2-specific type-I inhibitor MLi-2 and the broad-spectrum type-II inhibitor GZD-824. Our structures revealed LRRK2s kinase in the active-like state, stabilized by type-I inhibitor interactions, and an inactive DYG-out type-II inhibitor complex. Our structural analysis also showed how inhibitor-induced conformational changes in LRRK2 are affected by its autoinhibitory N-terminal repeats. The structural models provide a template for the rational development of LRRK2 kinase inhibitors covering both canonical inhibitor binding modes.
]]></description>
<dc:creator>Leschziner, A. E.</dc:creator>
<dc:creator>Sanz Murillo, M.</dc:creator>
<dc:creator>Suarez, A. V.</dc:creator>
<dc:creator>Dederer, V.</dc:creator>
<dc:creator>Chatterjee, D.</dc:creator>
<dc:creator>Louro, J. A.</dc:creator>
<dc:creator>Knapp, S.</dc:creator>
<dc:creator>Mathea, S.</dc:creator>
<dc:date>2023-09-09</dc:date>
<dc:identifier>doi:10.1101/2023.09.07.556689</dc:identifier>
<dc:title><![CDATA[Inhibition of Parkinsons Disease-related LRRK2 by type-I and type-II kinase inhibitors: activity and structures]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.11.09.375048v1?rss=1">
<title>
<![CDATA[
Sphingolipid changes in Parkinson L444P GBA mutation fibroblasts promote α-synuclein aggregation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.11.09.375048v1?rss=1"
</link>
<description><![CDATA[
Intraneuronal accumulation of aggregated -synuclein is a pathological hallmark of Parkinsons disease. Therefore, mechanisms capable of promoting -synuclein deposition bear important pathogenetic implications. Mutations of the glucocerebrosidase 1 (GBA) gene represent a prevalent Parkinsons disease risk factor. They are associated with loss of activity of a key enzyme involved in lipid metabolism, glucocerebrosidase, supporting a mechanistic relationship between abnormal -synuclein-lipid interactions and the development of Parkinson pathology. In this study, the lipid membrane composition of fibroblasts isolated from control subjects, patients with idiopathic Parkinsons disease (iPD) and Parkinson patients carrying the L444P GBA mutation (PD-GBA) was assayed using shotgun lipidomics. The lipid profile of PD-GBA fibroblasts differed significantly from that of control and iPD cells. It was characterized by an overall increase in sphingolipid levels. It also featured a significant change in the proportion of ceramide, sphingomyelin and hexosylceramide molecules with shorter and longer hydrocarbon chain length; levels of shorter-chain molecules were increased while the percent of longer-chain sphingolipids was decreased in PD-GBA lipid extracts. The extent of this shift was correlated to the degree of reduction of fibroblast glucocerebrosidase activity. In a second set of experiments, lipid extracts from control and PD-GBA fibroblasts were added to incubations of recombinant -synuclein. The kinetics of -synuclein aggregation, as assessed by the binding of thioflavin T to amyloid structures, was significantly accelerated after addition of PD-GBA extracts as compared to control samples. Amyloid fibrils collected at the end of these incubations contained lipids, indicating -synuclein-lipid co-assembly. Lipids extracted from -synuclein fibrils were also analysed by shotgun lipidomics. Data revealed that the lipid content of these fibrils was significantly enriched of shorter-chain sphingolipids. Taken together, findings of this study indicate that the L444P GBA mutation and consequent enzymatic loss are associated with a distinctly altered membrane lipid profile that provides a biological fingerprint of this mutation in Parkinson fibroblasts. This altered lipid profile, which includes an increased content of shorter-chain sphingolipids, could also be an indicator of increased risk for -synuclein aggregate pathology. Shorter-chain molecules may act as preferred reactants during lipid-induced -synuclein fibrillation.
]]></description>
<dc:creator>Galvagnion, C.</dc:creator>
<dc:creator>Cerri, S.</dc:creator>
<dc:creator>Schapira, A. H. V.</dc:creator>
<dc:creator>Blandini, F.</dc:creator>
<dc:creator>Di Monte, D. A.</dc:creator>
<dc:date>2020-11-09</dc:date>
<dc:identifier>doi:10.1101/2020.11.09.375048</dc:identifier>
<dc:title><![CDATA[Sphingolipid changes in Parkinson L444P GBA mutation fibroblasts promote α-synuclein aggregation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-11-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.06.01.494130v1?rss=1">
<title>
<![CDATA[
The GBA variant E326K is associated with alpha-synuclein aggregation and lipid droplet accumulation in human cell lines. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.06.01.494130v1?rss=1"
</link>
<description><![CDATA[
Sequence variants or mutations in the GBA gene are numerically the most important risk factor for Parkinson disease (PD). The GBA gene encodes for the lysosomal hydrolase enzyme, glucocerebrosidase (GCase). GBA mutations often reduce GCase activity and lead to impairment of the autophagy-lysosomal pathway, which is important in the turnover of alpha-synuclein, accumulation of which is a key pathological hallmark of PD. Although the E326K variant is one of the most common GBA variants associated with PD, there is limited understanding of its biochemical effects. We have characterised homozygous and heterozygous E326K variants in human fibroblasts. We found that E326K variants did not cause significant loss of GCase protein or activity, endoplasmic reticulum (ER) retention or ER stress, in contrast to the L444P GBA mutation. This was confirmed in human dopaminergic SH-SY5Y neuroblastoma cell lines over-expressing GCase with either E326K or L444P protein. Despite no loss of GCase activity, a significant increase of insoluble alpha-synuclein aggregates in E326K and L444P mutants was observed. Notably, SH-SY5Y over-expressing E326K demonstrated a significant increase in lipid droplet number under basal conditions, which was exacerbated following treatment with the fatty acid oleic acid. Similarly, a significant increase in lipid droplet formation following lipid loading was observed in heterozygous and homozygous E326K fibroblasts. In conclusion, the work presented here demonstrates that the E326K mutation behaves differently to common loss of function GBA mutations, however lipid dyshomeostasis and alpha-synuclein pathology is still evident.
]]></description>
<dc:creator>Smith, L. J.</dc:creator>
<dc:creator>Bolsinger, M. M.</dc:creator>
<dc:creator>Chau, K.-Y.</dc:creator>
<dc:creator>Gegg, M. E.</dc:creator>
<dc:creator>Schapira, A. H. V.</dc:creator>
<dc:date>2022-06-01</dc:date>
<dc:identifier>doi:10.1101/2022.06.01.494130</dc:identifier>
<dc:title><![CDATA[The GBA variant E326K is associated with alpha-synuclein aggregation and lipid droplet accumulation in human cell lines.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-06-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.10.23.563557v1?rss=1">
<title>
<![CDATA[
Reduction of a-synuclein aggregates by PIKfyve inhibition via TFEB-mediated lysosomal biogenesis in a Parkinson disease model 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.10.23.563557v1?rss=1"
</link>
<description><![CDATA[
Parkinson disease is a neurodegenerative disorder characterised by impairment of motor function, and is associated with a progressive accumulation of insoluble aggregates of misfolded alpha-synuclein. In the present study, we exploited the SH-SY5Y cell model overexpressing a pro-aggregation form of alpha-synuclein to investigate the efficacy of PIKfyve-mediated lysosomal biogenesis, through TFEB, as potential target for Parkinson therapy.

To investigate this, we exploited high-content imaging along with enzymatic assays to follow the progression of lysosomal biogenesis, lysosomal function and alpha-synuclein accumulation. The cellular model exploited in this study recapitulated important elements of the biochemical phenotype observed in Parkinson patient-derived neurons, including synuclein aggregates and impaired glucocerebrosidase (GCase) function. PIKfyve inhibition by YM201636 resulted in a lysosomal-dependant reduction of alpha-synuclein aggregates as early as 24 hours post-treatment. The mechanism of action of YM201636 was shown to be TFEB-mediated, with an increase in TFEB in the nuclei which subsequently resulted in increased lysosomal markers LAMP1 and GCase. PIKfyve inhibtion efficacy was also tested in differentiated SH-SY5Y cells, exhibiting a neuron-like morphology. In these cells, YM201636 also significantly reduced alpha-synuclein aggregates and increased TFEB nuclear presence.

These findings suggest that PIKfyve inhibition could be exploited as therapeutic target for Parkinson disease.
]]></description>
<dc:creator>Lucas-Del-Pozo, S.</dc:creator>
<dc:creator>Uras, G.</dc:creator>
<dc:creator>Fierli, F.</dc:creator>
<dc:creator>Lentini, V.</dc:creator>
<dc:creator>Koletsi, S.</dc:creator>
<dc:creator>Chau, K.-Y.</dc:creator>
<dc:creator>Hughes, D.</dc:creator>
<dc:creator>Schapira, A. H.</dc:creator>
<dc:date>2023-10-24</dc:date>
<dc:identifier>doi:10.1101/2023.10.23.563557</dc:identifier>
<dc:title><![CDATA[Reduction of a-synuclein aggregates by PIKfyve inhibition via TFEB-mediated lysosomal biogenesis in a Parkinson disease model]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.07.11.451956v1?rss=1">
<title>
<![CDATA[
Chemically Induced Senescence in Human Stem Cell-Derived Neurons Promotes Phenotypic Presentation of Neurodegeneration 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.07.11.451956v1?rss=1"
</link>
<description><![CDATA[
Modeling age-related neurodegenerative disorders with human stem cells is difficult due to the embryonic nature of stem cell derived neurons. We developed a chemical cocktail to induce senescence of iPSC-derived neurons to address this challenge. We first screened small molecules that induce embryonic fibroblasts to exhibit features characteristic of aged fibroblasts. We then optimized a cocktail of small molecules that induced senescence in fibroblasts and cortical neurons without causing DNA damage. The utility of the "senescence cocktail" was validated in motor neurons derived from ALS patient iPSCs which exhibited protein aggregation and axonal degeneration substantially earlier than those without cocktail treatment. Our "senescence cocktail" will likely enhance the manifestation of disease-related phenotypes in neurons derived from iPSCs, enabling the generation of reliable drug discovery platforms.
]]></description>
<dc:creator>Fathi, A.</dc:creator>
<dc:creator>Mathivanan, S.</dc:creator>
<dc:creator>Kong, L.</dc:creator>
<dc:creator>Petersen, A. J.</dc:creator>
<dc:creator>Harder, C. R. K.</dc:creator>
<dc:creator>Block, J.</dc:creator>
<dc:creator>Miller, J. M.</dc:creator>
<dc:creator>Bhattacharyya, A.</dc:creator>
<dc:creator>Wang, D.</dc:creator>
<dc:creator>Zhang, S.-C.</dc:creator>
<dc:date>2021-07-12</dc:date>
<dc:identifier>doi:10.1101/2021.07.11.451956</dc:identifier>
<dc:title><![CDATA[Chemically Induced Senescence in Human Stem Cell-Derived Neurons Promotes Phenotypic Presentation of Neurodegeneration]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-07-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.04.01.535194v1?rss=1">
<title>
<![CDATA[
Circular RNAs in the human brain are tailored to neuron identity and neuropsychiatric disease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.04.01.535194v1?rss=1"
</link>
<description><![CDATA[
Little is known about circular RNAs (circRNAs) in specific brain cells and human neuropsychiatric disease. Here, we systematically identified over 11,039 circRNAs expressed in vulnerable dopamine and pyramidal neurons laser-captured from 190 human brains and non-neuronal cells using ultra-deep, total RNA sequencing. 1,526 and 3,308 circRNAs were custom-tailored to the cell identity of dopamine and pyramidal neurons and enriched in synapse pathways. 88% of Parkinsons and 80% of Alzheimers disease-associated genes produced circRNAs. circDNAJC6, produced from a juvenile-onset Parkinsons gene, was already dysregulated during prodromal, onset stages of common Parkinsons disease neuropathology. Globally, addiction-associated genes preferentially produced circRNAs in dopamine neurons, autism-associated genes in pyramidal neurons, and cancers in non-neuronal cells. This study shows that circular RNAs in the human brain are tailored to neuron identity and implicate circRNA- regulated synaptic specialization in neuropsychiatric diseases.
]]></description>
<dc:creator>Dong, X.</dc:creator>
<dc:creator>Bai, Y.</dc:creator>
<dc:creator>Liao, Z.</dc:creator>
<dc:creator>Gritsch, D.</dc:creator>
<dc:creator>Liu, X.</dc:creator>
<dc:creator>Wang, T.</dc:creator>
<dc:creator>Borges-Monroy, R.</dc:creator>
<dc:creator>Ehrlich, A.</dc:creator>
<dc:creator>Serano, G. E.</dc:creator>
<dc:creator>Feany, M. B.</dc:creator>
<dc:creator>Beach, T. G.</dc:creator>
<dc:creator>Scherzer, C. R.</dc:creator>
<dc:date>2023-04-03</dc:date>
<dc:identifier>doi:10.1101/2023.04.01.535194</dc:identifier>
<dc:title><![CDATA[Circular RNAs in the human brain are tailored to neuron identity and neuropsychiatric disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-04-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.06.02.543481v1?rss=1">
<title>
<![CDATA[
α-synuclein promotes neuronal dysfunction and death by disrupting the binding of ankyrin to β-spectrin 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.06.02.543481v1?rss=1"
</link>
<description><![CDATA[
-synuclein plays a key role in the pathogenesis of Parkinsons disease and related disorders, but critical interacting partners and molecular mechanisms mediating neurotoxicity are incompletely understood. We show that -synuclein binds directly to {beta}-spectrin. Using males and females in a Drosophila model of -synuclein-related disorders we demonstrate that {beta}-spectrin is critical for -synuclein neurotoxicity. Further, the ankyrin binding domain of {beta}-spectrin is required for -synuclein binding and neurotoxicity. A key plasma membrane target of ankyrin, Na+/K+ ATPase, is mislocalized when human -synuclein is expressed in Drosophila. Accordingly, membrane potential is depolarized in -synuclein transgenic fly brains. We examine the same pathway in human neurons and find that Parkinsons disease patient-derived neurons with a triplication of the -synuclein locus show disruption of the spectrin cytoskeleton, mislocalization of ankyrin and Na+/K+ ATPase, and membrane potential depolarization. Our findings define a specific molecular mechanism by which elevated levels of -synuclein in Parkinsons disease and related -synucleinopathies leads to neuronal dysfunction and death.

Significance StatementThe small synaptic vesicle associate protein -synuclein plays a critical role in the pathogenesis of Parkinsons disease and related disorders, but the disease-relevant binding partners of -synuclein and proximate pathways critical for neurotoxicity require further definition. We show that -synuclein binds directly to {beta}-spectrin, a key cytoskeletal protein required for localization of plasma membrane proteins and maintenance of neuronal viability. Binding of -synuclein to {beta}-spectrin alters the organization of the spectrin-ankyrin complex, which is critical for localization and function of integral membrane proteins, including Na+/K+ ATPase. These finding outline a previously undescribed mechanism of -synuclein neurotoxicity and thus suggest potential new therapeutic approaches in Parkinsons disease and related disorders.
]]></description>
<dc:creator>Maor, G.</dc:creator>
<dc:creator>Dubreuil, R. R.</dc:creator>
<dc:creator>Feany, M. B.</dc:creator>
<dc:date>2023-06-05</dc:date>
<dc:identifier>doi:10.1101/2023.06.02.543481</dc:identifier>
<dc:title><![CDATA[α-synuclein promotes neuronal dysfunction and death by disrupting the binding of ankyrin to β-spectrin]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.06.21.545840v1?rss=1">
<title>
<![CDATA[
Detecting Full-Length EccDNA with FLED and long-reads sequencing 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.06.21.545840v1?rss=1"
</link>
<description><![CDATA[
Reconstructing the full-length sequence of extrachromosomal circular DNA (eccDNA) from short sequencing reads has proved challenging given the similarity of eccDNAs and their corresponding linear DNAs. Previous sequencing methods were unable to achieve high-throughput detection of full-length eccDNAs. Here we describe a new strategy that combined rolling circle amplification (RCA) and nanopore long-reads sequencing technology to generate full-length eccDNAs. We further developed a novel algorithm, called Full-Length eccDNA Detection (FLED), to reconstruct the sequence of eccDNAs. We used FLED to analyze seven human epithelial and cancer cell line samples and identified over 5,000 full-length eccDNAs per sample. The structures of identified eccDNAs were validated by both PCR and Sanger sequencing. Compared to other published nanopore-based eccDNA detectors, FLED exhibited higher sensitivity. In cancer cell lines, the genes overlapped with eccDNA regions were enriched in cancer-related pathways and cis-regulatory elements can be predicted in the up-stream or downstream of intact genes on eccDNA molecules, and the expressions of these cancer-related genes were dysregulated in tumor cell lines, indicating the regulatory potency of eccDNAs in biological processes. Our method takes advantage of nanopore long reads and enables unbiased reconstruction of full-length eccDNA sequences. FLED is imple-mented using Python3 which is freely available on GitHub (https://github.com/FuyuLi/FLED).
]]></description>
<dc:creator>Li, F.</dc:creator>
<dc:creator>Ming, W.</dc:creator>
<dc:creator>Lu, W.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Dong, X.</dc:creator>
<dc:creator>Bai, Y.</dc:creator>
<dc:date>2023-06-24</dc:date>
<dc:identifier>doi:10.1101/2023.06.21.545840</dc:identifier>
<dc:title><![CDATA[Detecting Full-Length EccDNA with FLED and long-reads sequencing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.10.13.464123v1?rss=1">
<title>
<![CDATA[
Constitutive nuclear accumulation of endogenous alpha-synuclein in mice causes motor dysfunction and cortical atrophy, independent of protein aggregation. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.10.13.464123v1?rss=1"
</link>
<description><![CDATA[
BackgroundA growing body of evidence suggests that nuclear alpha-synuclein (Syn) plays a role in the pathogenesis of Parkinsons disease (PD). However, this question has been difficult to address as controlling the localization of Syn in experimental systems often requires protein overexpression, which affects its aggregation propensity.

MethodsWe engineered SncaNLS mice which localize endogenous Syn to the nucleus. We characterized these mice on a behavioral, histological, and biochemical level to determine whether the increase of nuclear Syn is sufficient to elicit PD-like phenotypes.

ResultsSncaNLS mice exhibit age-dependent motor deficits and altered gastrointestinal function. We found that these phenotypes were not linked to Syn aggregation or phosphorylation. Through histological analyses, we observed motor cortex atrophy in the absence of midbrain dopaminergic neurodegeneration. We sampled cortical proteomes of SncaNLS mice and controls to determine the molecular underpinnings of these pathologies. Interestingly, we found several dysregulated proteins involved in dopaminergic signaling, namely Darpp-32, which we further confirmed was decreased in cortical samples of the SncaNLS mice compared to controls via immunoblotting.

ConclusionsThese results suggest that chronic endogenous nuclear Syn can elicit toxic phenotypes in mice, independent of its aggregation. This model raises key questions related to the mechanism of Syn toxicity in PD and provides a new model to study an underappreciated aspect of PD pathogenesis.
]]></description>
<dc:creator>Geertsma, H. M.</dc:creator>
<dc:creator>Suk, T. R.</dc:creator>
<dc:creator>Ricke, K. M.</dc:creator>
<dc:creator>Horsthuis, K.</dc:creator>
<dc:creator>Parmasad, J.-L. A.</dc:creator>
<dc:creator>Fisk, Z.</dc:creator>
<dc:creator>Callaghan, S. M.</dc:creator>
<dc:creator>Rousseaux, M. W. C.</dc:creator>
<dc:date>2021-10-14</dc:date>
<dc:identifier>doi:10.1101/2021.10.13.464123</dc:identifier>
<dc:title><![CDATA[Constitutive nuclear accumulation of endogenous alpha-synuclein in mice causes motor dysfunction and cortical atrophy, independent of protein aggregation.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.09.08.556120v1?rss=1">
<title>
<![CDATA[
Detecting rhythmic spiking through the power spectra of point process model residuals 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.09.08.556120v1?rss=1"
</link>
<description><![CDATA[
ObjectiveOscillations figure prominently as neurological disease hallmarks and neuromodulation targets. To detect oscillations in a neurons spiking, one might attempt to seek peaks in the spike trains power spectral density (PSD) which exceed a flat baseline. Yet for a non-oscillating neuron, the PSD is not flat: The recovery period ("RP", the post-spike drop in spike probability, starting with the refractory period) introduces global spectral distortion. An established "shuffling" procedure corrects for RP distortion by removing the spectral component explained by the inter-spike interval (ISI) distribution. However, this procedure sacrifices oscillation-related information present in the ISIs, and therefore in the PSD. We asked whether point process models (PPMs) might achieve more selective RP distortion removal, thereby enabling improved oscillation detection.

ApproachIn a novel "residuals" method, we first estimate the RP duration (nr) from the ISI distribution. We then fit the spike train with a PPM that predicts spike likelihood based on the time elapsed since the most recent of any spikes falling within the preceding nr milliseconds. Finally, we compute the PSD of the models residuals.

Main resultsWe compared the residuals and shuffling methods ability to enable accurate oscillation detection with flat baseline-assuming tests. Over synthetic data, the residuals method generally outperformed the shuffling method in classification of true-versus false-positive oscillatory power, principally due to enhanced sensitivity in sparse spike trains. In single-unit data from the internal globus pallidus (GPi) and ventrolateral anterior thalamus (VLa) of a parkinsonian monkey -- in which alpha-beta oscillations (8-30 Hz) were anticipated -- the residuals method reported the greatest incidence of significant alpha-beta power, with low firing rates predicting residuals-selective oscillation detection.

SignificanceThese results encourage continued development of the residuals approach, to support more accurate oscillation detection. Improved identification of oscillations could promote improved disease models and therapeutic technologies.
]]></description>
<dc:creator>Cox, K. M.</dc:creator>
<dc:creator>Kase, D.</dc:creator>
<dc:creator>Znati, T.</dc:creator>
<dc:creator>Turner, R. S.</dc:creator>
<dc:date>2023-09-08</dc:date>
<dc:identifier>doi:10.1101/2023.09.08.556120</dc:identifier>
<dc:title><![CDATA[Detecting rhythmic spiking through the power spectra of point process model residuals]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.07.19.549710v1?rss=1">
<title>
<![CDATA[
The SATB1-MIR22-GBA axis mediates glucocerebroside accumulation inducing a cellular senescence-like phenotype in dopaminergic neurons 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.07.19.549710v1?rss=1"
</link>
<description><![CDATA[
Idiopathic Parkinsons Disease (PD) is characterized by the loss of dopaminergic neurons in the substantia nigra pars compacta, which is associated with neuroinflammation and reactive gliosis. The underlying cause of PD and the concurrent neuroinflammation are not well understood. In this study, we utilized human and murine neuronal lines, stem cell-derived dopaminergic neurons, and mice to demonstrate that three previously identified genetic risk factors for PD, namely SATB1, MIR22HG, and GBA, are components of a single gene regulatory pathway. Our findings indicate that dysregulation of this pathway leads to the upregulation of glucocerebrosides (GluCer), which triggers a cellular senescence-like phenotype in dopaminergic neurons. Specifically, we discovered that downregulation of the transcriptional repressor SATB1 results in the derepression of the microRNA miR-22-3p, leading to decreased GBA expression and subsequent accumulation of GluCer. Furthermore, our results demonstrate that an increase in GluCer alone is sufficient to impair lysosomal and mitochondrial function, thereby inducing cellular senescence dependent on S100A9 and stress factors. Dysregulation of the SATB1-MIR22-GBA pathway, observed in both PD patients and normal aging, leads to lysosomal and mitochondrial dysfunction due to the GluCer accumulation, ultimately resulting in a cellular senescence-like phenotype in dopaminergic neurons. Therefore, our study highlights a novel pathway involving three genetic risk factors for PD and provides a potential mechanism for the senescence-induced neuroinflammation and reactive gliosis observed in both PD and normal aging.
]]></description>
<dc:creator>Russo, T.</dc:creator>
<dc:creator>Kolisnyk, B.</dc:creator>
<dc:creator>BS, A.</dc:creator>
<dc:creator>Kim, T. W.</dc:creator>
<dc:creator>Martin, J.</dc:creator>
<dc:creator>Plessis-Belair, J.</dc:creator>
<dc:creator>Ni, J.</dc:creator>
<dc:creator>Pearson, J. A.</dc:creator>
<dc:creator>Park, E. J.</dc:creator>
<dc:creator>Sher, R. B.</dc:creator>
<dc:creator>Studer, L.</dc:creator>
<dc:creator>Riessland, M.</dc:creator>
<dc:date>2023-07-21</dc:date>
<dc:identifier>doi:10.1101/2023.07.19.549710</dc:identifier>
<dc:title><![CDATA[The SATB1-MIR22-GBA axis mediates glucocerebroside accumulation inducing a cellular senescence-like phenotype in dopaminergic neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.02.03.479014v1?rss=1">
<title>
<![CDATA[
Interaction of an α-synuclein epitope with HLA-DRB1*15:01 initiates early enteric features of Parkinson's disease in humanized mice 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.02.03.479014v1?rss=1"
</link>
<description><![CDATA[
Enteric symptoms, including constipation, are hallmarks of prodromal Parkinsons disease (PD) that can appear decades before the onset of motor symptoms and diagnosis. PD patients possess circulating T cells that recognize specific -synuclein-(-syn)-derived epitopes. One epitope, -syn32-46, binds with strong affinity to the HLA-DRB1*15:01 allele implicated in autoimmune diseases. We report that -syn32-46 immunization in a mouse expressing HLA-DRB1*15:01 triggers intestinal inflammation leading to loss of enteric neurons, damage of enteric dopaminergic neurons, constipation and weight loss. -Syn32-46 immunization activates innate and adaptive immune gene signatures in the gut and induces changes in CD4+ TH1/ TH17 transcriptome that resemble tissue resident memory cells found in mucosal barriers during inflammation. Depletion of CD4+, but not CD8+, T cells partially rescues enteric neurodegeneration. Therefore, interaction of -syn32-46 and HLA-DRB1*15:0 is critical for gut inflammation and CD4+ T cell-mediated loss of enteric neurons in humanized mice, suggesting potential mechanisms of prodromal enteric PD.

HIGHLIGHTS AND eTOC BlurbO_LI-syn32-46 immunization of an HLA-DRB1*15:01 mouse triggers weight loss and constipation.
C_LIO_LI-syn32-46 immunizations induce gut inflammation, loss of enteric neurons and damage to dopaminergic neurons.
C_LIO_LI-syn32-46 immunization induces innate and adaptive immune responses in the gut.
C_LIO_LIDepletion of CD4+, but not CD8+, T cells partially rescues enteric neural loss.
C_LIO_LIAn interaction between -syn32-46 and HLA-DRB1*15:01 is critical for this model of prodromal PD.
C_LI

Parkinsons disease (PD) patients exhibit elevated number of circulating T cells that recognize -synuclein-(-syn)- epitopes, particularly during early disease stages. One epitope, -syn32-46, interacts with the HLA-DRB1*15:01; however, its role in PD pathogenesis remains unknown. Garretti et al. show that -syn32-46 immunization of a mouse expressing HLA-DRB1*15:01 triggers intestinal inflammation, a loss of enteric neurons, constipation and weight loss, suggesting a critical role for -syn autoimmunity in HLA-DRB1*15:01 carriers in prodromal PD.
]]></description>
<dc:creator>Garretti, F.</dc:creator>
<dc:creator>Monahan, C.</dc:creator>
<dc:creator>Sloan, N.</dc:creator>
<dc:creator>Shariar, S.</dc:creator>
<dc:creator>Kim, S. W.</dc:creator>
<dc:creator>Sette, A.</dc:creator>
<dc:creator>Cutforth, T.</dc:creator>
<dc:creator>Kanter, E.</dc:creator>
<dc:creator>Sulzer, D.</dc:creator>
<dc:creator>Agalliu, D.</dc:creator>
<dc:date>2022-02-05</dc:date>
<dc:identifier>doi:10.1101/2022.02.03.479014</dc:identifier>
<dc:title><![CDATA[Interaction of an α-synuclein epitope with HLA-DRB1*15:01 initiates early enteric features of Parkinson's disease in humanized mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-02-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.12.14.520294v1?rss=1">
<title>
<![CDATA[
Conserved and cell type-specific transcriptional responses to IFN-γ in the ventral midbrain 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.12.14.520294v1?rss=1"
</link>
<description><![CDATA[
Dysregulated inflammation within the central nervous system (CNS) contributes to neuropathology in infectious, autoimmune, and neurodegenerative disease. With the exception of microglia, major histocompatibility complex (MHC) proteins are virtually undetectable in the mature, healthy central nervous system (CNS). Neurons have generally been considered incapable of antigen presentation, and although interferon gamma (IFN-{gamma}) can elicit neuronal MHC class I (MHC-I) expression and antigen presentation in vitro, it remains unclear whether similar responses occur in vivo. Here we directly injected IFN-{gamma} into the ventral midbrain of mature mice and analyzed gene expression profiles of specific CNS cell types. We find that IFN-{gamma} induces cellular proliferation and expression of MHC-II and associated genes only in microglia. However, IFN-{gamma} upregulated MHC-I and associated mRNAs in ventral midbrain microglia, astrocytes, oligodendrocytes, and GABAergic, glutamatergic, and dopaminergic neurons. The core set of IFN-{gamma}-induced genes and their response kinetics were conserved across neurons and glia, with a lower amplitude of expression in neurons. A diverse repertoire of genes was upregulated in glia, particularly microglia, while no neuron-specific responses to IFN-{gamma} were observed. Using mutant mice to selectively delete the IFN-{gamma}-binding domain of IFNGR1 in dopaminergic neurons, we demonstrate that dopaminergic neurons respond directly to IFN-{gamma}. Our results suggest that most neurons are capable of responding directly to IFN-{gamma} and upregulating MHC-I and related genes in vivo, but their expression amplitude and repertoire is limited compared to oligodendrocytes, astrocytes, and microglia.

One-sentence summaryWe find that IFN-{gamma} induces transcription of MHC class I antigen processing and presentation machinery in all major parenchymal cell types in the ventral midbrain; however, neuronal responses are low amplitude and limited to a small set of genes, MHC class II expression and cellular proliferation are restricted to microglia, and dopamine neuronal responses require cell autonomous expression of IFNGR1.
]]></description>
<dc:creator>Sulzer, D.</dc:creator>
<dc:creator>Hobson, B.</dc:creator>
<dc:creator>Stanley, A.</dc:creator>
<dc:creator>De Los Santos, M.</dc:creator>
<dc:creator>Culbertson, B.</dc:creator>
<dc:creator>Mosharov, E.</dc:creator>
<dc:creator>Sims, P.</dc:creator>
<dc:date>2022-12-15</dc:date>
<dc:identifier>doi:10.1101/2022.12.14.520294</dc:identifier>
<dc:title><![CDATA[Conserved and cell type-specific transcriptional responses to IFN-γ in the ventral midbrain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-12-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.11.09.566376v1?rss=1">
<title>
<![CDATA[
Disrupted sleep-wake regulation in the MCI-Park mouse model of Parkinson's Disease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.11.09.566376v1?rss=1"
</link>
<description><![CDATA[
Disrupted sleep has a profound adverse impact on lives of Parkinsons disease (PD) patients and their caregivers. Sleep disturbances are exceedingly common in PD, with substantial heterogeneity in type, timing, and severity. Among the most common sleep-related symptoms reported by PD patients are insomnia, excessive daytime sleepiness, and sleep fragmentation, characterized by interruptions and decreased continuity of sleep. Alterations in brain wave activity, as measured on the electroencephalogram (EEG), also occur in PD, with changes in the pattern and relative contributions of different frequency bands of the EEG spectrum to overall EEG activity in different vigilance states consistently observed. The mechanisms underlying these PD-associated sleep-wake abnormalities are poorly understood, and they are ineffectively treated by conventional PD therapies. To help fill this gap in knowledge, a new progressive model of PD - the MCI-Park mouse - was studied. Near the transition to the parkinsonian state, these mice exhibited significantly altered sleep-wake regulation, including increased wakefulness, decreased non-rapid eye movement (NREM) sleep, increased sleep fragmentation, reduced rapid eye movement (REM) sleep, and altered EEG activity patterns. These sleep-wake abnormalities mirror those identified in PD patients. Thus, this model may help elucidate the circuit mechanisms underlying sleep disruption in PD and identify targets for novel therapeutic approaches.
]]></description>
<dc:creator>Summa, K. C.</dc:creator>
<dc:creator>Jiang, P.</dc:creator>
<dc:creator>Gonzalez-Rodriguez, P.</dc:creator>
<dc:creator>Huang, X.</dc:creator>
<dc:creator>Lin, X.</dc:creator>
<dc:creator>Vitaterna, M. H.</dc:creator>
<dc:creator>Dan, Y.</dc:creator>
<dc:creator>Surmeier, D. J.</dc:creator>
<dc:creator>Turek, F. W.</dc:creator>
<dc:date>2023-11-13</dc:date>
<dc:identifier>doi:10.1101/2023.11.09.566376</dc:identifier>
<dc:title><![CDATA[Disrupted sleep-wake regulation in the MCI-Park mouse model of Parkinson's Disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.08.26.505472v1?rss=1">
<title>
<![CDATA[
Disruption of lysosomal proteolysis in astrocytes facilitates midbrain proteostasis failure in an early-onset PD model. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.08.26.505472v1?rss=1"
</link>
<description><![CDATA[
Accumulation of advanced glycation end products (AGEs) on biopolymers accompany cellular aging and drives poorly understood disease processes. Here, we studied how AGEs contribute to development of early on-set Parkinsons Disease (PD) caused by loss-of-function of DJ1, a protein deglycase. In induced pluripotent stem cell (iPSC)-derived midbrain organoid models deficient for DJ1 activity, we find that lysosomal proteolysis is impaired, causing AGEs to accumulate, -synuclein (-syn) phosphorylation to increase, and proteins to aggregate. These processes are at least partly driven by astrocytes, as DJ1 loss reduces their capacity to provide metabolic support and triggers acquisition of a pro-inflammatory phenotype. Consistently, in co-cultures, we find that DJ1-expressing astrocytes are able to reverse the proteolysis deficits of DJ1 knockout midbrain neurons. In conclusion, astrocytes capacity to clear toxic damaged proteins is critical to preserve neuronal function and their dysfunction contributes to the neurodegeneration observed in PD.
]]></description>
<dc:creator>Morrone Parfitt, G.</dc:creator>
<dc:creator>Coccia, E.</dc:creator>
<dc:creator>Goldman, C.</dc:creator>
<dc:creator>Whitney, K.</dc:creator>
<dc:creator>Reyes, R.</dc:creator>
<dc:creator>Sarrafha, L.</dc:creator>
<dc:creator>Nam, K. H.</dc:creator>
<dc:creator>Jones, D.</dc:creator>
<dc:creator>crary, J. F.</dc:creator>
<dc:creator>Ordureau, A.</dc:creator>
<dc:creator>Blanchard, J.</dc:creator>
<dc:creator>Ahfeldt, T. D.</dc:creator>
<dc:date>2022-08-27</dc:date>
<dc:identifier>doi:10.1101/2022.08.26.505472</dc:identifier>
<dc:title><![CDATA[Disruption of lysosomal proteolysis in astrocytes facilitates midbrain proteostasis failure in an early-onset PD model.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-08-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.08.21.552829v1?rss=1">
<title>
<![CDATA[
Functional characterization of ATP13A2 variants associated with distinct neurodegenerative disorders 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.08.21.552829v1?rss=1"
</link>
<description><![CDATA[
ATP13A2 is a late endolysosomal transporter that exports the polyamines spermine and spermidine from the organellar lumen to the cytosol. Loss-of-function variants in ATP13A2 are causative for Kufor-Rakeb syndrome (KRS, a recessive juvenile-onset parkinsonism with dementia) and have also been identified in early-onset PD (EOPD) and hereditary spastic paraplegia (HSP). Furthermore, candidate pathogenic ATP13A2 variants have been identified in neuronal ceroid lipofuscinosis (NCL; M854R), multiple system atrophy (MSA; Y1020C) and amyotrophic lateral sclerosis (ALS; I411M) suggesting that ATP13A2 may be implicated in a broader range of neurodegenerative disorders. Since the functional consequences of the NCL, MSA, and ALS variants have not yet been examined, we here characterized these ATP13A2 variants in terms of subcellular localization, cellular polyamine uptake, and transport activity. We found that the homozygous NCL-associated M854R variant results in an instable protein with low expression levels, leading to complete loss of ATPase and cellular polyamine uptake activity. The heterozygous MSA-linked Y1020C variant is properly localized and presents only partially decreased ATPase activity without affecting cellular polyamine uptake. The ALS-associated I411M variant is also correctly localized and exhibits a minor effect on cellular polyamine uptake, however, without a significant impact on ATPase activity. Taken together, only the homozygous NCL variant of ATP13A2 causes a complete loss-of-function, validating that ATP13A2 dysfunction is implicated in NCL. The ALS and MSA variants only presented a subtle functional defect, questioning whether these heterozygous variants are pathogenic and whether ATP13A2 dysfunction may cause MSA or ALS.
]]></description>
<dc:creator>Vrijsen, S.</dc:creator>
<dc:creator>El Asrar, R. A.</dc:creator>
<dc:creator>Houdou, M. C.</dc:creator>
<dc:creator>Van den Haute, C.</dc:creator>
<dc:creator>Baekelandt, V.</dc:creator>
<dc:creator>Lyons, J. A.</dc:creator>
<dc:creator>Eggermont, J.</dc:creator>
<dc:creator>Vangheluwe, P.</dc:creator>
<dc:date>2023-08-21</dc:date>
<dc:identifier>doi:10.1101/2023.08.21.552829</dc:identifier>
<dc:title><![CDATA[Functional characterization of ATP13A2 variants associated with distinct neurodegenerative disorders]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.01.21.525025v1?rss=1">
<title>
<![CDATA[
Adult-specific Reelin expression alters striatal neuronal organization. Implications for neuropsychiatric disorders. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.01.21.525025v1?rss=1"
</link>
<description><![CDATA[
In addition to neuronal migration, brain development and adult plasticity, the extracellular matrix protein Reelin has been extensively implicated in human psychiatric disorders such as schizophrenia, bipolar disorder and autistic spectrum disorder. Moreover, heterozygous reeler mice exhibit features reminiscent of these disorders, while overexpression of Reelin protects against its manifestation. However, how Reelin influences the structure and circuits of the striatal complex, a key region for the above-mentioned disorders, is far from being understood, especially when altered Reelin expression levels are found at adult stages. In the present study, we took advantage of complementary conditional gain- and loss-of-function mouse models to investigate how Reelin levels may modify adult brains striatal structure and neuronal composition. Using immunohistochemical techniques, we determined that Reelin does not seem to influence the striatal patch and matrix organization (studied by -opioid receptor immunohistochemistry) nor the density of medium spiny neurons (MSNs, studied with DARPP-32). We show that overexpression of Reelin leads to increased numbers of striatal Parvalbumin- and Cholinergic-interneurons, and to a slight increase in the tyrosine hydroxylase-positive projections. We conclude that increased Reelin levels might modulate the numbers of striatal interneurons and the density of the nigrostriatal dopaminergic projections, suggesting that these changes may be involved in the protection of Reelin against neuropsychiatric disorders.
]]></description>
<dc:creator>Pardo, M.</dc:creator>
<dc:creator>Gregorio, S.</dc:creator>
<dc:creator>Montalban, E.</dc:creator>
<dc:creator>Pujadas, L.</dc:creator>
<dc:creator>Elias-Tersa, A.</dc:creator>
<dc:creator>Masachs, N.</dc:creator>
<dc:creator>Vilchez-Acosta, A.</dc:creator>
<dc:creator>Parent, A.</dc:creator>
<dc:creator>Auladell, C.</dc:creator>
<dc:creator>Girault, J. A.</dc:creator>
<dc:creator>Vila, M.</dc:creator>
<dc:creator>Nairn, A. C.</dc:creator>
<dc:creator>Manso, Y.</dc:creator>
<dc:creator>Soriano, E.</dc:creator>
<dc:date>2023-01-22</dc:date>
<dc:identifier>doi:10.1101/2023.01.21.525025</dc:identifier>
<dc:title><![CDATA[Adult-specific Reelin expression alters striatal neuronal organization. Implications for neuropsychiatric disorders.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.08.07.552289v1?rss=1">
<title>
<![CDATA[
Single-cell somatic copy number variants in brain using different amplification methods and reference genomes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.08.07.552289v1?rss=1"
</link>
<description><![CDATA[
The presence of somatic mutations, including copy number variants (CNVs), in the brain is well recognized. Comprehensive study requires single-cell whole genome amplification, with several methods available, prior to sequencing. We compared PicoPLEX with two recent adaptations of multiple displacement amplification (MDA): primary template-directed amplification (PTA) and droplet MDA, across 93 human brain cortical nuclei. We demonstrated different properties for each, with PTA providing the broadest amplification, PicoPLEX the most even, and distinct chimeric profiles. Furthermore, we performed CNV calling on two brains with multiple system atrophy and one control brain using different reference genomes. We found that 38% of brain cells have at least one Mb-scale CNV, with some supported by bulk sequencing or single-cells from other brain regions. Our study highlights the importance of selecting whole genome amplification method and reference genome for CNV calling, while supporting the existence of somatic CNVs in healthy and diseased human brain.
]]></description>
<dc:creator>Kalef-Ezra, E.</dc:creator>
<dc:creator>Turan, Z. G.</dc:creator>
<dc:creator>Perez-Rodriguez, D.</dc:creator>
<dc:creator>Bomann, I.</dc:creator>
<dc:creator>Behera, S.</dc:creator>
<dc:creator>Morley, C.</dc:creator>
<dc:creator>Scholz, S. W.</dc:creator>
<dc:creator>Jaunmuktane, Z.</dc:creator>
<dc:creator>Demeulemeester, J.</dc:creator>
<dc:creator>Sedlazeck, F.</dc:creator>
<dc:creator>Proukakis, C.</dc:creator>
<dc:date>2023-08-08</dc:date>
<dc:identifier>doi:10.1101/2023.08.07.552289</dc:identifier>
<dc:title><![CDATA[Single-cell somatic copy number variants in brain using different amplification methods and reference genomes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.03.20.533519v1?rss=1">
<title>
<![CDATA[
Neuronal phase shifts differ for excitation vs. inhibition: a computer modeling study 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.03.20.533519v1?rss=1"
</link>
<description><![CDATA[
Rhythmic activity is ubiquitous in neural systems, and impedance analysis has been widely used to examine frequency-dependent responses of neuronal membranes to rhythmic inputs. Impedance analysis assumes the neuronal membrane is a linear system, requiring the use of small signals to stay in a near-linear regime. However, postsynaptic potentials are often large and trigger nonlinear mechanisms. We therefore augmented impedance analysis to evaluate membrane responses in this nonlinear domain, analyzing responses to injected current for subthreshold membrane voltage (Vmemb), suprathreshold spike-blocked Vmemb, and spiking in a validated neocortical pyramidal neuron computer model. Responses in these output regimes were asymmetrical, with different phase shifts during hyperpolarizing and depolarizing half-cycles. Suprathreshold chirp stimulation gave equivocal results due to nonstationarity of response, requiring us to use fixed-frequency sinusoids. Sinusoidal inputs produced phase retreat: action potentials occurred progressively later in cycles of the input stimulus, resulting from adaptation. Conversely, sinusoidal current with increasing amplitude over cycles produced a pattern of phase advance: action potentials occurred progressively earlier. Phase retreat was dependent on Ih and IAHP currents; phase advance was modulated by these currents. Our results suggest differential responses of cortical neurons depending on the frequency of oscillatory input in the delta - beta range, which will play a role in neuronal responses to shifts in network state. We hypothesize that intrinsic cellular properties complement network properties and contribute to in vivo phase-shift phenomena such as phase precession, seen in place and grid cells, and phase roll, observed in hippocampal CA1 neurons.

New & NoteworthyWe augmented electrical impedance analysis to characterize phase shifts between large amplitude current stimuli and nonlinear, asymmetric membrane potential responses. We predict different frequency-dependent phase shifts in response excitation versus inhibition, as well as shifts in spike timing over multiple input cycles, in resonant pyramidal neurons. We hypothesize that these effects contribute to navigation-related phenomena like phase precession and phase roll. Our neuron-level hypothesis complements, rather than falsifies, prior network-level explanations of these phenomena.
]]></description>
<dc:creator>Kelley, C.</dc:creator>
<dc:creator>Antic, S. D.</dc:creator>
<dc:creator>Carnevale, N. T.</dc:creator>
<dc:creator>Kubie, J. L.</dc:creator>
<dc:creator>Lytton, W.</dc:creator>
<dc:date>2023-03-21</dc:date>
<dc:identifier>doi:10.1101/2023.03.20.533519</dc:identifier>
<dc:title><![CDATA[Neuronal phase shifts differ for excitation vs. inhibition: a computer modeling study]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.02.15.528722v1?rss=1">
<title>
<![CDATA[
Integrated single cell and unsupervised spatial transcriptomic analysis defines molecular anatomy of the human dorsolateral prefrontal cortex 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.02.15.528722v1?rss=1"
</link>
<description><![CDATA[
The molecular organization of the human neocortex has been historically studied in the context of its histological layers. However, emerging spatial transcriptomic technologies have enabled unbiased identification of transcriptionally-defined spatial domains that move beyond classic cytoarchitecture. Here we used the Visium spatial gene expression platform to generate a data-driven molecular neuroanatomical atlas across the anterior-posterior axis of the human dorsolateral prefrontal cortex (DLPFC). Integration with paired single nucleus RNA-sequencing data revealed distinct cell type compositions and cell-cell interactions across spatial domains. Using PsychENCODE and publicly available data, we map the enrichment of cell types and genes associated with neuropsychiatric disorders to discrete spatial domains. Finally, we provide resources for the scientific community to explore these integrated spatial and single cell datasets at research.libd.org/spatialDLPFC/.

SummaryGeneration of a molecular neuroanatomical map of the human prefrontal cortex reveals novel spatial domains and cell-cell interactions relevant for psychiatric disease.
]]></description>
<dc:creator>Huuki-Myers, L. A.</dc:creator>
<dc:creator>Spangler, A.</dc:creator>
<dc:creator>Eagles, N. J.</dc:creator>
<dc:creator>Montgomery, K. D.</dc:creator>
<dc:creator>Kwon, S. H.</dc:creator>
<dc:creator>Guo, B.</dc:creator>
<dc:creator>Grant-Peters, M.</dc:creator>
<dc:creator>Divecha, H. R.</dc:creator>
<dc:creator>Tippani, M.</dc:creator>
<dc:creator>Sriworarat, C.</dc:creator>
<dc:creator>Nguyen, A. B.</dc:creator>
<dc:creator>Ravichandran, P.</dc:creator>
<dc:creator>Tran, M. N.</dc:creator>
<dc:creator>Seyedian, A.</dc:creator>
<dc:creator>PsychENCODE Consortium,</dc:creator>
<dc:creator>Hyde, T. M.</dc:creator>
<dc:creator>Kleinman, J. E.</dc:creator>
<dc:creator>Battle, A.</dc:creator>
<dc:creator>Page, S. C.</dc:creator>
<dc:creator>Ryten, M.</dc:creator>
<dc:creator>Hicks, S. C.</dc:creator>
<dc:creator>Martinowich, K.</dc:creator>
<dc:creator>Collado-Torres, L.</dc:creator>
<dc:creator>Maynard, K. R.</dc:creator>
<dc:date>2023-02-15</dc:date>
<dc:identifier>doi:10.1101/2023.02.15.528722</dc:identifier>
<dc:title><![CDATA[Integrated single cell and unsupervised spatial transcriptomic analysis defines molecular anatomy of the human dorsolateral prefrontal cortex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.12.11.571045v1?rss=1">
<title>
<![CDATA[
Alpha-Synuclein pathology disrupts mitochondrial function in dopaminergic and cholinergic neurons at-risk in Parkinsons disease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.12.11.571045v1?rss=1"
</link>
<description><![CDATA[
BackgroundPathological accumulation of aggregated -synuclein (aSYN) is a common feature of Parkinsons disease (PD). However, the mechanisms by which intracellular aSYN pathology contributes to dysfunction and degeneration of neurons in the brain are still unclear. A potentially relevant target of aSYN is the mitochondrion. To test this hypothesis, genetic and physiological methods were used to monitor mitochondrial function in substantia nigra pars compacta (SNc) dopaminergic and pedunculopontine nucleus (PPN) cholinergic neurons after stereotaxic injection of aSYN pre-formed fibrils (PFFs) into the mouse brain.

MethodsaSYN PPFs were stereotaxically injected into the SNc or PPN of mice. Twelve weeks later, mice were studied using a combination of approaches, including immunocytochemical analysis, cell- type specific transcriptomic profiling, electron microscopy, electrophysiology and two-photon-laser- scanning microscopy of genetically encoded sensors for bioenergetic and redox status.

ResultsIn addition to inducing a significant neuronal loss, SNc injection of PFFs induced the formation of intracellular, phosphorylated aSYN aggregates selectively in dopaminergic neurons. In these neurons, PFF-exposure decreased mitochondrial gene expression, reduced the number of mitochondria, increased oxidant stress, and profoundly disrupted mitochondrial adenosine triphosphate production. Consistent with an aSYN-induced bioenergetic deficit, the autonomous spiking of dopaminergic neurons slowed or stopped. PFFs also up-regulated lysosomal gene expression and increased lysosomal abundance, leading to the formation of Lewy-like inclusions. Similar changes were observed in PPN cholinergic neurons following aSYN PFF exposure.

ConclusionsTaken together, our findings suggest that disruption of mitochondrial function, and the subsequent bioenergetic deficit, is a proximal step in the cascade of events induced by aSYN pathology leading to dysfunction and degeneration of neurons at-risk in PD.
]]></description>
<dc:creator>Geibl, F. F.</dc:creator>
<dc:creator>Henrich, M. T.</dc:creator>
<dc:creator>Xie, Z.</dc:creator>
<dc:creator>Zampese, E.</dc:creator>
<dc:creator>Tkatch, T.</dc:creator>
<dc:creator>Wokosin, D. L.</dc:creator>
<dc:creator>Nasiri, E.</dc:creator>
<dc:creator>Grotmann, C. A.</dc:creator>
<dc:creator>Dawson, V. L.</dc:creator>
<dc:creator>Dawson, T. M.</dc:creator>
<dc:creator>Chandel, N. S.</dc:creator>
<dc:creator>Oertel, W. H.</dc:creator>
<dc:creator>Surmeier, D. J.</dc:creator>
<dc:date>2023-12-11</dc:date>
<dc:identifier>doi:10.1101/2023.12.11.571045</dc:identifier>
<dc:title><![CDATA[Alpha-Synuclein pathology disrupts mitochondrial function in dopaminergic and cholinergic neurons at-risk in Parkinsons disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.11.01.563607v1?rss=1">
<title>
<![CDATA[
FASlpr gene dosage tunes the extent of lymphoproliferation and T cell differentiation in lupus 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.11.01.563607v1?rss=1"
</link>
<description><![CDATA[
Sle1 and Faslpr are two lupus susceptibility loci that lead to manifestations of systemic lupus erythematosus. To evaluate dosage effects of FASlpr in determining cellular and serological phenotypes associated with lupus, we developed a new C57BL/6 (B6) congenic lupus strain, B6.Sle1/Sle1.Faslpr/+ (sle1homo.lprhet) and compared it with B6.Faslpr/lpr(lprhomo), B6.Sle1/Sle1 (sle1homo), and B6.Sle1/Sle1.Faslpr/lpr (sle1homo.lprhomo) strains. Whereas Sle1homo.lprhomo mice exhibited profound lymphoproliferation and early mortality, sle1homo.lprhet mice had a lifespan comparable to B6 mice, with no evidence of splenomegaly or lymphadenopathy. Compared to B6 monogenic lupus strains, sle1homo.lprhet mice exhibited significantly elevated serum anti-dsDNA antibodies and increased proteinuria. Additionally, Sle1homo.lprhet T cells had an increased propensity to differentiate into Th1 cells. Gene dose effects of Faslpr were noted in upregulating serum IL-1, IL-2, and IL-27. Taken together, sle1homo.lprhet mice emerge as a more faithful model of human SLE, ideal for genetic studies, autoantibody repertoire investigation, and for exploring Th1 effector cell skewing.
]]></description>
<dc:creator>Bohat, R.</dc:creator>
<dc:creator>Liang, X.</dc:creator>
<dc:creator>Chen, Y.</dc:creator>
<dc:creator>Xu, C.</dc:creator>
<dc:creator>Zheng, N.</dc:creator>
<dc:creator>Guerrero, A.</dc:creator>
<dc:creator>Jaffery, R.</dc:creator>
<dc:creator>Egan, N. A.</dc:creator>
<dc:creator>Robles, A.</dc:creator>
<dc:creator>Hicks, M. J.</dc:creator>
<dc:creator>Du, Y.</dc:creator>
<dc:creator>Chen, X.</dc:creator>
<dc:creator>Mohan, C.</dc:creator>
<dc:creator>Peng, W.</dc:creator>
<dc:date>2023-11-02</dc:date>
<dc:identifier>doi:10.1101/2023.11.01.563607</dc:identifier>
<dc:title><![CDATA[FASlpr gene dosage tunes the extent of lymphoproliferation and T cell differentiation in lupus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.12.15.571818v1?rss=1">
<title>
<![CDATA[
Development And Characterization Of A Non-Human Primate Model Of Disseminated Synucleinopathy 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.12.15.571818v1?rss=1"
</link>
<description><![CDATA[
The presence of a widespread cortical synucleinopathy is the main neuropathological hallmark underlying clinical entities such as Parkinsons disease with dementia (PDD) and dementia with Lewy bodies (DLB). There currently is a pressing need for the development of non-human primate (NHPs) models of PDD and DLB to further overcome existing limitations in drug discovery. Here we took advantage of a retrogradely-spreading adeno-associated viral vector serotype 9 coding for the alpha-synuclein A53T mutated gene to induce a widespread synucleinopathy of cortical and subcortical territories innervating the putamen. Four weeks post-AAV deliveries animals were sacrificed and a comprehensive biodistribution study was conducted, comprising the quantification of neurons expressing alpha-synuclein, rostrocaudal distribution and their specific location. In brief, cortical afferent systems were found to be the main contributors to putaminal afferents (superior frontal and precentral gyrus in particular), together with neurons located in the caudal intralaminar nuclei and in the substantia nigra pars compacta (leading to thalamostriatal and nigrostriatal projections, respectively). Obtained data extends current models of synucleinopathies in NHPs, providing a reproducible platform enabling the adequate implementation of end-stage preclinical screening of new drugs targeting alpha-synuclein.
]]></description>
<dc:creator>Rico, A. J.</dc:creator>
<dc:creator>Corcho, A.</dc:creator>
<dc:creator>Chocarro, J.</dc:creator>
<dc:creator>Ariznabarreta, G.</dc:creator>
<dc:creator>Roda, E.</dc:creator>
<dc:creator>Honrubia, A.</dc:creator>
<dc:creator>Arnaiz Esteban, P.</dc:creator>
<dc:creator>Lanciego, J. L.</dc:creator>
<dc:date>2023-12-15</dc:date>
<dc:identifier>doi:10.1101/2023.12.15.571818</dc:identifier>
<dc:title><![CDATA[Development And Characterization Of A Non-Human Primate Model Of Disseminated Synucleinopathy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.12.18.572180v1?rss=1">
<title>
<![CDATA[
Neuropathological assessment of the olfactory bulb and tract in individuals with COVID-19 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.12.18.572180v1?rss=1"
</link>
<description><![CDATA[
The majority of patients with Parkinson disease (PD) experience a loss in their sense of smell and accumulate insoluble -synuclein aggregates in their olfactory bulbs (OB). Subjects affected by a SARS-CoV-2-linked illness (COVID-19) frequently experience hyposmia. We previously hypothesized that -synuclein and tau misprocessing could occur following host responses to microbial triggers. Using semiquantitative measurements of immunohistochemical signals, we examined OB and olfactory tract specimens collected serially at autopsies between 2020 and 2023. Deceased subjects comprised 50 adults, which included COVID19+ patients (n=22), individuals with Lewy body disease (e.g., PD and dementia with Lewy bodies (DLB; n=6)), Alzheimer disease (AD; n=3), other non-synucleinopathy-linked degenerative diseases (e.g., progressive supranuclear palsy (PSP; n=2) and multisystem atrophy (MSA; n=1)). Further, we included neurologically healthy controls (HCO; n=9) and those with an inflammation-rich brain disorder as neurological controls (NCO; n=7). When probing for inflammatory changes focusing on anterior olfactory nuclei (AON) using anti-CD68 immunostaining, scores were consistently elevated in NCO and AD cases. In contrast, inflammation on average was not significantly altered in COVID19+ patients relative to controls, although anti-CD68 reactivity in their OB and tracts declined with progression in age. Mild-to-moderate increases in phospho-Syn and phospho-tau signals were detected in the AON of tauopathy-and synucleinopathy-afflicted brains, respectively, consistent with mixed pathology, as described by others. Lastly, when both sides were available for comparison in our case series, we saw no asymmetry in the degree of pathology of the left versus right OB and tracts. We concluded from our autopsy series that after a fatal course of COVID-19, microscopic changes -when present-in the rostral, intracranial portion of the olfactory circuitry generally reflected neurodegenerative processes seen elsewhere in the brain. In general, inflammation correlated best with the degree of Alzheimers-linked tauopathy and declined with progression of age in COVID19+ patients.
]]></description>
<dc:creator>Lengacher, N. A.</dc:creator>
<dc:creator>Tomlinson, J. J.</dc:creator>
<dc:creator>Jochum, A.-K.</dc:creator>
<dc:creator>Franz, J.</dc:creator>
<dc:creator>Hasan Ali, O.</dc:creator>
<dc:creator>Flatz, L.</dc:creator>
<dc:creator>Jochum, W.</dc:creator>
<dc:creator>Penninger, J.</dc:creator>
<dc:creator>aSCENT-PD Investigators,</dc:creator>
<dc:creator>Stadelmann-Nessler, C.</dc:creator>
<dc:creator>Woulfe, J. M.</dc:creator>
<dc:creator>Schlossmacher, M. G.</dc:creator>
<dc:date>2023-12-19</dc:date>
<dc:identifier>doi:10.1101/2023.12.18.572180</dc:identifier>
<dc:title><![CDATA[Neuropathological assessment of the olfactory bulb and tract in individuals with COVID-19]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.12.23.573130v1?rss=1">
<title>
<![CDATA[
Synchronous Measurements of Extracellular Action Potentials and Neurochemical Activity with Carbon Fiber Electrodes in Nonhuman Primates 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.12.23.573130v1?rss=1"
</link>
<description><![CDATA[
Measuring the dynamic relationship between neuromodulators, such as dopamine, and neuronal action potentials is imperative to understand how these fundamental modes of neural signaling interact to mediate behavior. Here, we developed methods to measure concurrently dopamine and extracellular action potentials (i.e., spikes) and applied these in a monkey performing a behavioral task. Standard fast-scan cyclic voltammetric (FSCV) electrochemical (EChem) and electrophysiological (EPhys) recording systems are combined and used to collect spike and dopamine signals, respectively, from an array of carbon fiber (CF) sensors implanted in the monkey striatum. FSCV requires the application of small voltages at the implanted sensors to measure redox currents generated from target molecules, such as dopamine. These applied voltages create artifacts at neighboring EPhys-measurement sensors, producing signals that may falsely be classified as physiological spikes. Therefore, simple automated temporal interpolation algorithms were designed to remove these artifacts and enable accurate spike extraction. We validated these methods using simulated artifacts and demonstrated an average spike recovery rate of 84.5%. This spike extraction was performed on data collected from concurrent EChem and EPhys recordings made in a task-performing monkey to discriminate cell-type specific striatal units. These identified units were shown to correlate to specific behavioral task parameters related to reward size and eye-movement direction. Synchronous measures of spike and dopamine signals displayed contrasting relations to the behavioral task parameters, as taken from our small set of representative data, suggesting a complex relationship between these two modes of neural signaling. Future application of our methods will help advance our understanding of the interactions between neuromodulator signaling and neuronal activity, to elucidate more detailed mechanisms of neural circuitry and plasticity mediating behaviors in health and in disease.

Significance statementWe present a simple method for recording synchronous molecular and neuronal spike signals. Conventional electrophysiological and electrochemical instruments are combined without the need for additional hardware. A custom-designed algorithm was made and validated for extracting neuronal action potential signals with high fidelity. We were able to compute cell-type specific spike activity along with molecular dopamine signals related to reward and movement behaviors from measurements made in the monkey striatum. Such combined measurements of neurochemical and extracellular action potentials may help pave the way to elucidating mechanisms of plasticity, and how neuromodulators and neurons are orchestrated to mediate behavior.
]]></description>
<dc:creator>Amjad, U.</dc:creator>
<dc:creator>Choi, J.</dc:creator>
<dc:creator>Gibson, D. J.</dc:creator>
<dc:creator>Murray, R.</dc:creator>
<dc:creator>Graybiel, A. M.</dc:creator>
<dc:creator>Schwerdt, H. N.</dc:creator>
<dc:date>2023-12-24</dc:date>
<dc:identifier>doi:10.1101/2023.12.23.573130</dc:identifier>
<dc:title><![CDATA[Synchronous Measurements of Extracellular Action Potentials and Neurochemical Activity with Carbon Fiber Electrodes in Nonhuman Primates]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.12.19.572409v1?rss=1">
<title>
<![CDATA[
Mutations in GPNMB associated with Amyloid cutis dyschromica alter intracellular trafficking and processing of GPNMB 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.12.19.572409v1?rss=1"
</link>
<description><![CDATA[
Amyloid cutis dyschromica (ACD) is a rare skin condition characterized by focal areas of hyperpigmentation with hypopigmented macules and distinct regions of amyloid deposition. Until recently, the genetic cause of ACD remained unknown. Several studies have since named GPNMB truncation mutations as causal, with protein loss-of-function underlying ACD pathogenesis. GPNMB missense mutations have also been observed in patients, but these are less well characterized; especially on a cellular level. Here, we observed that GPNMB missense mutations implicated in familial ACD show distinct cellular phenotypes that result in impaired protein maturation and processing from the endoplasmic reticulum (ER) to the trans-Golgi network (TGN), prompting failed trafficking to lysosomes. These missense mutations also show failed secretion of the extracellular fragment of GPNMB, a well-characterized property of the protein. Overall, this work highlights previously undescribed cellular characteristics of GPNMB missense mutations implicated in ACD and helps to better inform the clinically observed phenotypes, as well as underscore GPNMBs role at melanosomes.
]]></description>
<dc:creator>Bogacki, E. C.</dc:creator>
<dc:creator>Lewis, P. A.</dc:creator>
<dc:creator>Herbst, S.</dc:creator>
<dc:date>2023-12-20</dc:date>
<dc:identifier>doi:10.1101/2023.12.19.572409</dc:identifier>
<dc:title><![CDATA[Mutations in GPNMB associated with Amyloid cutis dyschromica alter intracellular trafficking and processing of GPNMB]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.12.16.571979v1?rss=1">
<title>
<![CDATA[
Lysosomal TBK1 Responds to Amino Acid Availability to Relieve Rab7-Dependent mTORC1 Inhibition 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.12.16.571979v1?rss=1"
</link>
<description><![CDATA[
Lysosomes play a pivotal role in coordinating macromolecule degradation and regulating cell growth and metabolism. Despite substantial progress in identifying lysosomal signaling proteins, understanding the pathways that synchronize lysosome functions with changing cellular demands remains incomplete. This study uncovers a role for TANK-binding kinase 1 (TBK1), well known for its role in innate immunity and organelle quality control, in modulating lysosomal responsiveness to nutrients. Specifically, we identify a pool of TBK1 that is recruited to lysosomes in response to elevated amino acid levels. At lysosomes, this TBK1 phosphorylates Rab7 on serine 72. This is critical for alleviating Rab7-mediated inhibition of amino acid-dependent mTORC1 activation. Furthermore, a TBK1 mutant (E696K) associated with amyotrophic lateral sclerosis and frontotemporal dementia constitutively accumulates at lysosomes, resulting in elevated Rab7 phosphorylation and increased mTORC1 activation. This data establishes the lysosome as a site of amino acid regulated TBK1 signaling that is crucial for efficient mTORC1 activation. This lysosomal pool of TBK1 has broader implications for lysosome homeostasis, and its dysregulation could contribute to the pathogenesis of ALS-FTD.
]]></description>
<dc:creator>Talaia, G.</dc:creator>
<dc:creator>Bentley-DeSousa, A.</dc:creator>
<dc:creator>Ferguson, S.</dc:creator>
<dc:date>2023-12-17</dc:date>
<dc:identifier>doi:10.1101/2023.12.16.571979</dc:identifier>
<dc:title><![CDATA[Lysosomal TBK1 Responds to Amino Acid Availability to Relieve Rab7-Dependent mTORC1 Inhibition]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.12.18.572081v1?rss=1">
<title>
<![CDATA[
VPS13B is localized at the cis-trans Golgi complex interface and is a functional partner of FAM177A1 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.12.18.572081v1?rss=1"
</link>
<description><![CDATA[
Mutations in VPS13B, a member of a protein family implicated in bulk lipid transport between adjacent membranes, cause Cohen syndrome. VPS13B is known to be concentrated in the Golgi complex, but its precise location within this organelle and thus the site(s) where it achieves lipid transport remains unclear. Here we show that VPS13B is localized at the interface between cis and trans Golgi sub-compartments and that Golgi complex re-formation after Brefeldin A (BFA) induced disruption is delayed in VPS13B KO cells. This delay is phenocopied by loss of FAM177A1, a Golgi complex protein of unknown function reported to be a VPS13B interactor and whose mutations also result in a developmental disorder. In zebrafish, the vps13b orthologue, not previously annotated in this organism, genetically interacts with fam177a1. Collectively, these findings raise the possibility that bulk lipid transport by VPS13B may play a role in expanding Golgi membranes and that VPS13B may be assisted in this function by FAM177A1.
]]></description>
<dc:creator>Ugur, B.</dc:creator>
<dc:creator>Schueder, F.</dc:creator>
<dc:creator>Shin, J.</dc:creator>
<dc:creator>Hanna, M. G.</dc:creator>
<dc:creator>Wu, Y.</dc:creator>
<dc:creator>Leonzino, M.</dc:creator>
<dc:creator>Su, M.</dc:creator>
<dc:creator>McAdow, A. R.</dc:creator>
<dc:creator>Wilson, C. A.</dc:creator>
<dc:creator>Postlethwait, J.</dc:creator>
<dc:creator>Solnica-Krezel, L.</dc:creator>
<dc:creator>Bewersdorf, J.</dc:creator>
<dc:creator>De Camilli, P.</dc:creator>
<dc:date>2023-12-18</dc:date>
<dc:identifier>doi:10.1101/2023.12.18.572081</dc:identifier>
<dc:title><![CDATA[VPS13B is localized at the cis-trans Golgi complex interface and is a functional partner of FAM177A1]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.12.18.572148v1?rss=1">
<title>
<![CDATA[
RASP: Optimal single fluorescent puncta detection in complex cellular backgrounds 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.12.18.572148v1?rss=1"
</link>
<description><![CDATA[
Super-resolution and single-molecule microscopy are increasingly applied to complex biological systems. A major challenge of this approach is that fluorescent puncta must be detected in the low signal, high noise, heterogeneous background environments of cells and tissue. We present RASP, Radiality Analysis of Single Puncta, a bioimaging-segmentation method that solves this problem. RASP removes false positive puncta that other analysis methods detect, and detects features over a broad range of spatial scales: from single proteins to complex cell phenotypes. RASP outperforms the state-of-the-art in precision and speed, using image gradients to separate Gaussian-shaped objects from background. We demonstrate RASPs power by showing it can extract spatial correlations between microglia, neurons, and -synuclein oligomers in the human brain. This sensitive, computationally efficient approach enables fluorescent puncta and cellular features to be distinguished in cellular and tissue environments with a sensitivity down to the level of the single protein.
]]></description>
<dc:creator>Fu, B.</dc:creator>
<dc:creator>Brock, E. E.</dc:creator>
<dc:creator>Breiter, J. C.</dc:creator>
<dc:creator>Tian, R.</dc:creator>
<dc:creator>Andrews, R.</dc:creator>
<dc:creator>Toomey, C. E.</dc:creator>
<dc:creator>Lachica, J.</dc:creator>
<dc:creator>Lashley, T.</dc:creator>
<dc:creator>Ryten, M.</dc:creator>
<dc:creator>Wood, N. W.</dc:creator>
<dc:creator>Vendruscolo, M.</dc:creator>
<dc:creator>Gandhi, S.</dc:creator>
<dc:creator>Weiss, L. E.</dc:creator>
<dc:creator>Beckwith, J. S.</dc:creator>
<dc:creator>Lee, S. F.</dc:creator>
<dc:date>2023-12-18</dc:date>
<dc:identifier>doi:10.1101/2023.12.18.572148</dc:identifier>
<dc:title><![CDATA[RASP: Optimal single fluorescent puncta detection in complex cellular backgrounds]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2020.03.11.986455v1?rss=1">
<title>
<![CDATA[
Pathogenic LRRK2 R1441C mutation is associated with striatal alterations 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2020.03.11.986455v1?rss=1"
</link>
<description><![CDATA[
LRRK2 mutations are associated with both familial and sporadic forms of Parkinsons disease (PD). Convergent evidence suggests that LRRK2 plays critical roles in regulating striatal function. Here, by using knock-in mouse lines that express the two most common LRRK2 pathogenic mutations--G2019S and R1441C--we investigated how pathogenic LRRK2 mutations altered striatal physiology. We found that R1441C mice displayed a reduced nigrostriatal dopamine release and hypoexcitability in indirect-pathway striatal projection neurons. These alterations were associated with an impaired striatal-dependent motor learning. This deficit in motor learning was rescued following the subchronic administration of the LRRK2 kinase inhibitor Mli-2. In contrast, though a decreased release of dopamine was observed in the G2019S knock-in mice no concomitant cellular and behavioral alterations were found. In summary, our data argue that the impact of LRRK2 mutations cannot be simply generalized. Our findings offer mechanistic insights for devising treatment strategies for PD patients.
]]></description>
<dc:creator>Xenias, H.</dc:creator>
<dc:creator>Chen, C.</dc:creator>
<dc:creator>Kang, S.</dc:creator>
<dc:creator>Shanmugasundaram, B.</dc:creator>
<dc:creator>Cherian, S.</dc:creator>
<dc:creator>Situ, X.</dc:creator>
<dc:creator>Scesa, G.</dc:creator>
<dc:creator>Chan, C. S.</dc:creator>
<dc:creator>Parisiadou, L.</dc:creator>
<dc:date>2020-03-12</dc:date>
<dc:identifier>doi:10.1101/2020.03.11.986455</dc:identifier>
<dc:title><![CDATA[Pathogenic LRRK2 R1441C mutation is associated with striatal alterations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2020-03-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.10.28.466357v1?rss=1">
<title>
<![CDATA[
Motor learning selectively strengthens cortical and striatal synapses of motor engram neurons 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.10.28.466357v1?rss=1"
</link>
<description><![CDATA[
Learning and consolidation of new motor skills require adaptations of neuronal activity and connectivity in the motor cortex and striatum, two key motor regions of the brain. Yet, how neurons undergo synaptic changes and become recruited during motor learning to form a memory engram remains an open question. Here, we train mice on a single-pellet reaching motor learning task and use a genetic approach to identify and manipulate behavior-relevant neurons selectively in the primary motor cortex (M1). We find that the degree of reactivation of M1 engram neurons correlates strongly with motor performance. We further demonstrate that learning-induced dendritic spine reorganization specifically occurs in these M1 engram neurons. In addition, we find that motor learning leads to an increase in the number and strength of outputs from M1 engram neurons onto striatal spiny projection neurons (SPNs) and that these synapses form local clusters along SPN dendrites. These results identify a highly specific synaptic plasticity during the formation of long-lasting motor memory traces in the corticostriatal circuit.

HIGHLIGHTS- Motor performance is correlated with the reactivation of motor engram neurons
- Motor learning increases spine density and new spine survival selectively on M1 engram neurons
- Motor learning strengthens motor engram outputs to the striatum
- M1 engram outputs converge onto clusters of dendritic spines on striatal spiny projection neurons
]]></description>
<dc:creator>Hwang, F.-J.</dc:creator>
<dc:creator>Roth, R. H.</dc:creator>
<dc:creator>Wu, Y.-W.</dc:creator>
<dc:creator>Sun, Y.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Ding, J. B.</dc:creator>
<dc:date>2021-10-28</dc:date>
<dc:identifier>doi:10.1101/2021.10.28.466357</dc:identifier>
<dc:title><![CDATA[Motor learning selectively strengthens cortical and striatal synapses of motor engram neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-10-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.03.08.482579v1?rss=1">
<title>
<![CDATA[
In situ architecture of the lipid transport protein VPS13C at ER-lysosomes membrane contacts 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.03.08.482579v1?rss=1"
</link>
<description><![CDATA[
VPS13 is a eukaryotic lipid transport protein localized at membrane contact sites. Previous studies suggested that it may transfer lipids between adjacent bilayers by a bridge-like mechanism. Direct evidence for this hypothesis from a full-length structure and from EM studies in situ, however, is still missing. Here we have capitalized on AlphaFold predictions to complement the structural information already available about VPS13 and to generate a full-length model of human VPS13C, the Parkinsons disease-linked VPS13 paralog localized at contacts between the ER and endo/lysosomes. Such model predicts a ~30-nm rod with a hydrophobic groove that extends throughout its length. We further investigated whether such a structure can be observed in situ at ER-endo/lysosome contacts. To this aim, we combined genetic approaches with cryo-focused-ion-beam (cryo-FIB) milling and cryo-electron tomography (cryo-ET) to examine HeLa cells overexpressing this protein (either full length or with an internal truncation) along with VAP, its anchoring binding partner at the ER. Using these methods we identified rod-like densities that span the space separating the two adjacent membranes and that match the predicted structures of either full length VPS13C or its shorter truncated mutant, thus providing the first in-situ evidence for a bridge-model of VPS13 in lipid transport. Intriguingly, the majority of the VPS13C rods were separated from the ER membranes by a narrow gap, suggesting that while VAP anchors the protein to the ER, direct contact of the VPS13C rod with the ER bilayer to allow lipid transport may be independently regulated.
]]></description>
<dc:creator>Cai, S.</dc:creator>
<dc:creator>Wu, Y.</dc:creator>
<dc:creator>Guillen-Samander, A.</dc:creator>
<dc:creator>Hancock-Cerutti, W. F.</dc:creator>
<dc:creator>Liu, J.</dc:creator>
<dc:creator>De Camilli, P.</dc:creator>
<dc:date>2022-03-08</dc:date>
<dc:identifier>doi:10.1101/2022.03.08.482579</dc:identifier>
<dc:title><![CDATA[In situ architecture of the lipid transport protein VPS13C at ER-lysosomes membrane contacts]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-03-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.09.22.507905v1?rss=1">
<title>
<![CDATA[
Dynamic refinement of behavioral structure mediates dopamine-dependent credit assignment 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.09.22.507905v1?rss=1"
</link>
<description><![CDATA[
Animals exhibit a diverse behavioral repertoire when exploring new environments and can learn which actions or action sequences produce positive outcomes. Dopamine release upon encountering reward is critical for reinforcing reward-producing actions1-3. However, it has been challenging to understand how credit is assigned to the exact action that produced dopamine release during continuous behavior. We investigated this problem with a novel self-stimulation paradigm in which specific spontaneous movements triggered optogenetic stimulation of dopaminergic neurons. Dopamine self-stimulation rapidly and dynamically changes the structure of the entire behavioral repertoire. Initial stimulations reinforced not only the stimulation-producing target action, but also actions similar to target and actions that occurred a few seconds before stimulation. Repeated pairings led to gradual refinement of the behavioral repertoire to home in on the target. Reinforcement of action sequences revealed further temporal dependencies of refinement. Action pairs spontaneously separated by long time intervals promoted a stepwise credit assignment, with early refinement of actions most proximal to stimulation and subsequent refinement of more distal actions. Thus, a retrospective reinforcement mechanism promotes not only reinforcement, but gradual refinement of the entire behavioral repertoire to assign credit to specific actions and action sequences that lead to dopamine release.
]]></description>
<dc:creator>Tang, J. C. Y.</dc:creator>
<dc:creator>Paixao, V.</dc:creator>
<dc:creator>Carvalho, F.</dc:creator>
<dc:creator>Silva, A.</dc:creator>
<dc:creator>Klaus, A.</dc:creator>
<dc:creator>Alves da Silva, J.</dc:creator>
<dc:creator>Costa, R. M.</dc:creator>
<dc:date>2022-09-22</dc:date>
<dc:identifier>doi:10.1101/2022.09.22.507905</dc:identifier>
<dc:title><![CDATA[Dynamic refinement of behavioral structure mediates dopamine-dependent credit assignment]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-09-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.02.07.527479v1?rss=1">
<title>
<![CDATA[
POLCAM: Instant molecular orientation microscopy for the life sciences 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.02.07.527479v1?rss=1"
</link>
<description><![CDATA[
Current methods for single-molecule orientation localization microscopy (SMOLM) require optical setups and algorithms that can be prohibitively slow and complex, limiting the widespread adoption for biological applications. We present POLCAM, a simplified SMOLM method based on polarized detection using a polarization camera, that can be easily implemented on any wide-field fluorescence microscope. To make polarization cameras compatible with single-molecule detection, we developed theory to minimize field of view errors, used simulations to optimize experimental design, and developed a fast algorithm based on Stokes parameter estimation which can operate over 1000 fold faster than the state of the art, enabling near instant determination of molecular anisotropy. To aid in the adoption of POLCAM, we developed open-source image analysis software, and a website detailing hardware installation and software use. To illustrate the potential of POLCAM in the life sciences, we applied our method to study alpha-synuclein fibrils, the actin cytoskeleton of mammalian cells, fibroblast-like cells and the plasma membrane of live human T cells.
]]></description>
<dc:creator>Bruggeman, E.</dc:creator>
<dc:creator>Zhang, O.</dc:creator>
<dc:creator>Needham, L.-M.</dc:creator>
<dc:creator>Koerbel, M.</dc:creator>
<dc:creator>Daly, S.</dc:creator>
<dc:creator>Cheetham, M.</dc:creator>
<dc:creator>Peters, R.</dc:creator>
<dc:creator>Wu, T.</dc:creator>
<dc:creator>Klymchenko, A. S.</dc:creator>
<dc:creator>Davis, S. J.</dc:creator>
<dc:creator>Paluch, E. K.</dc:creator>
<dc:creator>Klenerman, D.</dc:creator>
<dc:creator>Lew, M. D.</dc:creator>
<dc:creator>O'Holleran, K.</dc:creator>
<dc:creator>Lee, S. F.</dc:creator>
<dc:date>2023-02-08</dc:date>
<dc:identifier>doi:10.1101/2023.02.07.527479</dc:identifier>
<dc:title><![CDATA[POLCAM: Instant molecular orientation microscopy for the life sciences]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-02-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.07.18.549513v1?rss=1">
<title>
<![CDATA[
Into the deep: The subthalamic and para-subthalamic nuclei in behavioral avoidance 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.07.18.549513v1?rss=1"
</link>
<description><![CDATA[
The subthalamic nucleus (STN) is a key component of the brain network for movement control. However, the STN is strikingly heterogeneous and also intricately engaged in limbic and cognitive functions. The STN shows aberrant firing activity in several neurological and neuropsychiatric disorders, including Parkinso[n]s disease (PD). Deep brain stimulation (DBS) in the STN alleviates motor impairment in PD, but patients have reported altered mood as adverse side-effect. Recent observations suggest that optogenetic STN activation in mice induces flight behavior. We hypothesized that STN activation stand at risk of causing an aversive response with behavioral avoidance as consequence. The STN is directly adjoined with the para-STN (pSTN), a hypothalamic area correlated with appetitive and aversive behavior. STN-DBS aiming to correct STN might thereby also modulate pSTN. To dissociate the impact of STN and pSTN, we took advantage of selective promoters in mice, identified in our recent RNA- sequencing of the subthalamic area, to selectively direct optogenetic excitation. Acute photostimulation resulted in aversion via both the STN and pSTN, but only STN- stimulation-paired cues resulted in conditioned avoidance. Viral-genetic tracing coupled with electrophysiological recordings identified a polysynaptic pathway from the STN to the lateral habenula, a critical hub for aversion and associated with clinical depression. This study demonstrates that STN activation is directly correlated with aversion, and thereby contributes neurobiological underpinnings to emotional affect upon STN manipulation with implications for STN-targeted treatment outcome.
]]></description>
<dc:creator>Serra, G. P.</dc:creator>
<dc:creator>Guillaumin, A.</dc:creator>
<dc:creator>Vlcek, B.</dc:creator>
<dc:creator>Delagado-Zabalza, L.</dc:creator>
<dc:creator>Ricci, A.</dc:creator>
<dc:creator>Rubino, E.</dc:creator>
<dc:creator>Dumas, S.</dc:creator>
<dc:creator>Baufreton, J. M.</dc:creator>
<dc:creator>Georges, F.</dc:creator>
<dc:creator>Wallen-Mackenzie, A.</dc:creator>
<dc:date>2023-07-19</dc:date>
<dc:identifier>doi:10.1101/2023.07.18.549513</dc:identifier>
<dc:title><![CDATA[Into the deep: The subthalamic and para-subthalamic nuclei in behavioral avoidance]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-07-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.10.12.562063v1?rss=1">
<title>
<![CDATA[
The R1441C-LRRK2 mutation induces myeloid immune cell exhaustion in an age- and sex-dependent manner 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.10.12.562063v1?rss=1"
</link>
<description><![CDATA[
Considering age is the greatest risk factor for many neurodegenerative diseases, aging, in particular aging of the immune system, is the most underappreciated and understudied contributing factor in the neurodegeneration field. Genetic variation around the LRRK2 gene affects risk of both familial and sporadic Parkinsons disease (PD). The leucine-rich repeat kinase 2 (LRRK2) protein has been implicated in peripheral immune signaling, however, the effects of an aging immune system on LRRK2 function have been neglected to be considered. We demonstrate here that the R1441C mutation induces a hyper-responsive phenotype in macrophages from young female mice, characterized by increased effector functions, including stimulation-dependent antigen presentation, cytokine release, phagocytosis, and lysosomal function. This is followed by age-acquired immune cell exhaustion in a Lrrk2-kinase-dependent manner. Immune-exhausted macrophages exhibit suppressed antigen presentation and hypophagocytosis, which is also demonstrated in myeloid cells from R1441C and Y1699C-PD patients. Our novel findings that LRRK2 mutations confer immunological advantage at a young age but may predispose the carrier to age-acquired immune cell exhaustion have significant implications for LRRK2 biology and therapeutic development. Indeed, LRRK2 has become an appealing target in PD, but our findings suggest that more research is required to understand the cell-type specific consequences and optimal timing of LRRK2-targeting therapeutics.

One Sentence SummaryThe R1441C-LRRK2 mutation causes an age-acquired immune cell exhaustion in macrophages in a sex-dependent manner
]]></description>
<dc:creator>Wallings, R.</dc:creator>
<dc:creator>McFarland, K.</dc:creator>
<dc:creator>Staley, H.</dc:creator>
<dc:creator>Neighbarger, N.</dc:creator>
<dc:creator>Schaake, S.</dc:creator>
<dc:creator>Brueggemann, N.</dc:creator>
<dc:creator>Zittel, S.</dc:creator>
<dc:creator>Usnich, T.</dc:creator>
<dc:creator>Klein, C.</dc:creator>
<dc:creator>Sammler, E.</dc:creator>
<dc:creator>Tansey, M. G.</dc:creator>
<dc:date>2023-10-17</dc:date>
<dc:identifier>doi:10.1101/2023.10.12.562063</dc:identifier>
<dc:title><![CDATA[The R1441C-LRRK2 mutation induces myeloid immune cell exhaustion in an age- and sex-dependent manner]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.10.31.564602v1?rss=1">
<title>
<![CDATA[
A STING-CASM-GABARAP Pathway Activates LRRK2 at Lysosomes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.10.31.564602v1?rss=1"
</link>
<description><![CDATA[
Mutations that increase LRRK2 kinase activity have been linked to Parkinsons disease and Crohns disease. LRRK2 is also activated by lysosome damage. However, the endogenous cellular mechanisms that control LRRK2 kinase activity are not well understood. In this study, we identify signaling through Stimulator of Interferon Genes (STING) as an activator of LRRK2 via the Conjugation of ATG8 to Single Membranes (CASM) pathway. We furthermore establish that multiple chemical stimuli that perturb lysosomal homeostasis also converge on CASM to activate LRRK2. Although CASM results in the lipidation of multiple ATG8 protein family members, we establish that LRRK2 lysosome recruitment and kinase activation is highly dependent on interactions with the GABARAP member of this family. Collectively these results define a pathway that integrates multiple stimuli at lysosomes to control the kinase activity of LRRK2. Aberrant activation of LRRK2 via this pathway may be of relevance in both Parkinsons and Crohns diseases.
]]></description>
<dc:creator>Bentley-DeSousa, A.</dc:creator>
<dc:creator>Ferguson, S.</dc:creator>
<dc:date>2023-11-01</dc:date>
<dc:identifier>doi:10.1101/2023.10.31.564602</dc:identifier>
<dc:title><![CDATA[A STING-CASM-GABARAP Pathway Activates LRRK2 at Lysosomes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.12.07.570617v1?rss=1">
<title>
<![CDATA[
Movement-related increases in subthalamic activity optimize locomotion 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.12.07.570617v1?rss=1"
</link>
<description><![CDATA[
The subthalamic nucleus (STN) is traditionally thought to restrict movement. Lesion or prolonged STN inhibition increases movement vigor and propensity, while optogenetic excitation typically has opposing effects. Subthalamic and motor activity are also inversely correlated in movement disorders. However, most STN neurons exhibit movement-related increases in firing. To address this paradox, STN activity was recorded and manipulated in head-fixed mice at rest and during self-initiated and -paced treadmill locomotion. The majority of STN neurons (type 1) exhibited locomotion-dependent increases in activity, with half encoding the locomotor cycle. A minority of neurons exhibited dips in activity or were uncorrelated with movement. Brief optogenetic inhibition of the dorsolateral STN (where type 1 neurons are concentrated) slowed and prematurely terminated locomotion. In Q175 Huntingtons disease mice abnormally brief, low-velocity locomotion was specifically associated with type 1 hypoactivity. Together these data argue that movement-related increases in STN activity contribute to optimal locomotor performance.
]]></description>
<dc:creator>Callahan, J. W.</dc:creator>
<dc:creator>Morales, J. C.</dc:creator>
<dc:creator>Atherton, J. F.</dc:creator>
<dc:creator>Wang, D.</dc:creator>
<dc:creator>Kostic, S.</dc:creator>
<dc:creator>Bevan, M. D.</dc:creator>
<dc:date>2023-12-08</dc:date>
<dc:identifier>doi:10.1101/2023.12.07.570617</dc:identifier>
<dc:title><![CDATA[Movement-related increases in subthalamic activity optimize locomotion]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.12.23.573181v1?rss=1">
<title>
<![CDATA[
Mechanism of human PINK1 activation at the TOM complex in a reconstituted system 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.12.23.573181v1?rss=1"
</link>
<description><![CDATA[
Loss of function mutations in PTEN-induced kinase 1 (PINK1) are a frequent cause of early-onset Parkinsons disease (PD). Stabilisation of PINK1 at the Translocase of Outer Membrane (TOM) complex of damaged mitochondria is a critical step for its activation. To date the mechanism of how PINK1 is activated in the TOM complex is unclear. Herein we report co-expression of human PINK1 and all seven TOM subunits in Saccharomyces cerevisiae is sufficient for PINK1 activation. We use this reconstitution system to systematically assess the role of each TOM subunit towards PINK1 activation. We unambiguously demonstrate that the TOM20 and TOM70 receptor subunits are required for optimal PINK1 activation and map their sites of interaction with PINK1 using AlphaFold structural modelling and mutagenesis. We also demonstrate an essential role of the pore-containing subunit TOM40 and its structurally associated subunits TOM7 and TOM22 for PINK1 activation. These molecular findings will aid in the development of small molecule activators of PINK1 as a therapeutic strategy for PD.
]]></description>
<dc:creator>Raimi, O.</dc:creator>
<dc:creator>Ojha, H.</dc:creator>
<dc:creator>Ehses, K.</dc:creator>
<dc:creator>Dederer, V.</dc:creator>
<dc:creator>Lange, S.</dc:creator>
<dc:creator>Rivera, C. P.</dc:creator>
<dc:creator>Deegan, T.</dc:creator>
<dc:creator>Chen, Y.</dc:creator>
<dc:creator>Wightman, M.</dc:creator>
<dc:creator>Toth, R.</dc:creator>
<dc:creator>Labib, K.</dc:creator>
<dc:creator>Mathea, S.</dc:creator>
<dc:creator>Ranson, N.</dc:creator>
<dc:creator>Fernandez-Busnadiego, R. F.</dc:creator>
<dc:creator>Muqit, M.</dc:creator>
<dc:date>2023-12-23</dc:date>
<dc:identifier>doi:10.1101/2023.12.23.573181</dc:identifier>
<dc:title><![CDATA[Mechanism of human PINK1 activation at the TOM complex in a reconstituted system]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.01.08.574634v1?rss=1">
<title>
<![CDATA[
Pathways controlling neurotoxicity and proteostasis in mitochondrial complex I deficiency 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.01.08.574634v1?rss=1"
</link>
<description><![CDATA[
Neuromuscular disorders caused by dysfunction of the mitochondrial respiratory chain are common, severe and untreatable. We recovered a number of mitochondrial genes, including electron transport chain components, in a large forward genetic screen for mutations causing age-related neurodegeneration in the context of proteostasis dysfunction. We created a model of complex I deficiency in the Drosophila retina to probe the role of protein degradation abnormalities in mitochondrial encephalomyopathies. Using our genetic model, we found that complex I deficiency regulates both the ubiquitin/proteasome and autophagy/lysosome arms of the proteostasis machinery. We further performed an in vivo kinome screen to uncover new and potentially druggable mechanisms contributing to complex I related neurodegeneration and proteostasis failure. Reduction of RIOK kinases and the innate immune signaling kinase pelle prevented neurodegeneration in complex I deficiency animals. Genetically targeting oxidative stress, but not RIOK1 or pelle knockdown, normalized proteostasis markers. Our findings outline distinct pathways controlling neurodegeneration and protein degradation in complex I deficiency and introduce an experimentally facile model in which to study these debilitating and currently treatment-refractory disorders.
]]></description>
<dc:creator>Nithianadam, V.</dc:creator>
<dc:creator>Sarkar, S.</dc:creator>
<dc:creator>Feany, M. B.</dc:creator>
<dc:date>2024-01-08</dc:date>
<dc:identifier>doi:10.1101/2024.01.08.574634</dc:identifier>
<dc:title><![CDATA[Pathways controlling neurotoxicity and proteostasis in mitochondrial complex I deficiency]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-01-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.01.15.575737v1?rss=1">
<title>
<![CDATA[
Loss of primary cilia and dopaminergic neuroprotection in pathogenic LRRK2-driven and idiopathic Parkinsons disease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.01.15.575737v1?rss=1"
</link>
<description><![CDATA[
Activating LRRK2 mutations cause Parkinsons disease. Previously, we showed that cholinergic interneurons and astrocytes but not medium spiny neurons of the dorsal striatum lose primary cilia in LRRK2 mutant mice. Single nucleus RNA sequencing shows that cilia loss in cholinergic interneurons correlates with higher LRRK2 expression and decreased glial derived neurotrophic factor transcription. Nevertheless, much higher LRRK2 expression is seen in medium spiny neurons that have normal cilia in mice and humans. In parallel with decreased striatal dopaminergic neurite density, LRRK2 G2019S neurons show increased autism-linked CNTN5 adhesion protein expression; glial cells show significant loss of ferritin heavy chain. Human striatal tissue from LRRK2 pathway mutation carriers and idiopathic Parkinsons disease show similar cilia loss in cholinergic interneurons and astrocytes and overall loss of such neurons. These data strongly suggest that loss of cilia in specific striatal cell types decreases neuroprotection for dopamine neurons in mice and human Parkinsons disease.

TeaserCilia loss in Parkinsons disease decreases dopaminergic neuroprotection due to inability to sense Hedgehog signals
]]></description>
<dc:creator>Khan, S. S.</dc:creator>
<dc:creator>Jaimon, E.</dc:creator>
<dc:creator>Lin, Y.-E.</dc:creator>
<dc:creator>Nikoloff, J.</dc:creator>
<dc:creator>Tonelli, F.</dc:creator>
<dc:creator>Alessi, D. R.</dc:creator>
<dc:creator>Pfeffer, S. R.</dc:creator>
<dc:date>2024-01-16</dc:date>
<dc:identifier>doi:10.1101/2024.01.15.575737</dc:identifier>
<dc:title><![CDATA[Loss of primary cilia and dopaminergic neuroprotection in pathogenic LRRK2-driven and idiopathic Parkinsons disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-01-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.01.18.576289v1?rss=1">
<title>
<![CDATA[
3D Bioprinting of Human Neural Tissues with Functional Connectivity 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.01.18.576289v1?rss=1"
</link>
<description><![CDATA[
Probing how the human neural networks operate is hindered by the lack of reliable human neural tissues amenable for dynamic functional assessment of neural circuits. We developed a 3D bioprinting platform to assemble tissues with defined human neural cell types in a desired dimension using a commercial bioprinter. The printed neuronal progenitors differentiate to neurons and form functional neural circuits in and between tissue layers with specificity within weeks, evidenced by the cortical-to-striatal projection, spontaneous synaptic currents and synaptic response to neuronal excitation. Printed astrocyte progenitors develop into mature astrocytes with elaborated processes and form functional neuron-astrocyte networks, indicated by calcium flux and glutamate uptake in response to neuronal excitation under physiological and pathological conditions. These designed human neural tissues will likely be useful for understanding the wiring of human neural networks, modeling pathological processes, and serving as platforms for drug testing.
]]></description>
<dc:creator>Yan, Y.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Gao, Y.</dc:creator>
<dc:creator>Mathivanan, S.</dc:creator>
<dc:creator>Kong, L.</dc:creator>
<dc:creator>Tao, Y.</dc:creator>
<dc:creator>Dong, Y.</dc:creator>
<dc:creator>Li, X.</dc:creator>
<dc:creator>Bhattacharyya, A.</dc:creator>
<dc:creator>Zhao, X.</dc:creator>
<dc:creator>Zhang, S.-C.</dc:creator>
<dc:date>2024-01-23</dc:date>
<dc:identifier>doi:10.1101/2024.01.18.576289</dc:identifier>
<dc:title><![CDATA[3D Bioprinting of Human Neural Tissues with Functional Connectivity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-01-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.02.05.579034v1?rss=1">
<title>
<![CDATA[
Biochemical fractionation of human α-Synuclein in a Drosophila model of synucleinopathies 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.02.05.579034v1?rss=1"
</link>
<description><![CDATA[
Synucleinopathies are a group of central nervous system pathologies that are characterized by neuronal accumulation of misfolded and aggregated -synuclein in proteinaceous depositions known as Lewy Bodies (LBs). The transition of -synuclein from its physiological to pathological form has been associated with several post-translational modifications such as phosphorylation and an increasing degree of insolubility, which also correlate with disease progression in post-mortem specimens from human patients. Neuronal expression of -synuclein in model organisms, including Drosophila melanogaster, has been a typical approach employed to study its physiological effects. Biochemical analysis of -synuclein solubility via high-speed ultracentrifugation with buffers of increasing detergent strength offers a potent method for identification of -synuclein biochemical properties and the associated pathology stage. Unfortunately, the development of a robust and reproducible method for evaluation of human -synuclein solubility isolated from Drosophila tissues has remained elusive. Here, we tested different detergents for their ability to solubilize human -synuclein carrying the pathological mutation A53T from brains of aged flies. We also assessed the effect of sonication on solubility of human -synuclein and optimized a protocol to discriminate relative amounts of soluble/insoluble human -synuclein from dopaminergic neurons of the Drosophila brain. Our data established that, using a 5% SDS buffer, the 3-step protocol distinguishes between cytosolic soluble proteins in fraction 1, detergent-soluble proteins in fraction 2 and insoluble proteins in fraction 3. This protocol shows that sonication breaks down -synuclein insoluble complexes from the fly brain, making them soluble in the SDS buffer and enriching fraction 2 of the protocol.
]]></description>
<dc:creator>Imomnazarov, K.</dc:creator>
<dc:creator>Lopez-Scarim, J.</dc:creator>
<dc:creator>Bagheri, I.</dc:creator>
<dc:creator>Joers, V.</dc:creator>
<dc:creator>Tansey, M. G.</dc:creator>
<dc:creator>Martin-Pena, A.</dc:creator>
<dc:date>2024-02-07</dc:date>
<dc:identifier>doi:10.1101/2024.02.05.579034</dc:identifier>
<dc:title><![CDATA[Biochemical fractionation of human α-Synuclein in a Drosophila model of synucleinopathies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.02.09.579465v1?rss=1">
<title>
<![CDATA[
PINK1 is a target of T cell responses in Parkinson's disease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.02.09.579465v1?rss=1"
</link>
<description><![CDATA[
Parkinsons disease (PD) is associated with autoimmune T cells that recognize the protein alpha-synuclein in a subset of individuals. Multiple neuroantigens are targets of autoinflammatory T cells in classical central nervous system autoimmune diseases such as multiple sclerosis (MS). Here, we explored whether additional autoantigenic targets of T cells in PD. We generated 15-mer peptide pools spanning several PD-related proteins implicated in PD pathology, including GBA, SOD1, PINK1, parkin, OGDH, and LRRK2. Cytokine production (IFN{gamma}, IL-5, IL-10) against these proteins was measured using a fluorospot assay and PBMCs from patients with PD and age-matched healthy controls. This approach identified unique epitopes and their HLA restriction from the mitochondrial-associated protein PINK1, a regulator of mitochondrial stability, as an autoantigen targeted by T cells. The T cell reactivity was predominantly found in male patients with PD, which may contribute to the heterogeneity of PD. Identifying and characterizing PINK1 and other autoinflammatory targets may lead to antigen-specific diagnostics, progression markers, and/or novel therapeutic strategies for PD.
]]></description>
<dc:creator>Williams, G. P.</dc:creator>
<dc:creator>Michaelis, T.</dc:creator>
<dc:creator>Rodrigues Lima-Junior, J.</dc:creator>
<dc:creator>Frazier, A.</dc:creator>
<dc:creator>Tran, N. K.</dc:creator>
<dc:creator>Phillips, E. J.</dc:creator>
<dc:creator>Mallal, S. A.</dc:creator>
<dc:creator>Litvan, I.</dc:creator>
<dc:creator>Goldman, J. G.</dc:creator>
<dc:creator>Alcalay, R. N.</dc:creator>
<dc:creator>Sidney, J.</dc:creator>
<dc:creator>Sulzer, D.</dc:creator>
<dc:creator>Sette, A.</dc:creator>
<dc:creator>Lindestam Arlehamn, C. S.</dc:creator>
<dc:date>2024-02-12</dc:date>
<dc:identifier>doi:10.1101/2024.02.09.579465</dc:identifier>
<dc:title><![CDATA[PINK1 is a target of T cell responses in Parkinson's disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.02.01.578464v1?rss=1">
<title>
<![CDATA[
Alpha-synuclein overexpression can drive microbiome dysbiosis in mice 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.02.01.578464v1?rss=1"
</link>
<description><![CDATA[
Growing evidence indicates that persons living with Parkinson disease (PD), have a unique composition of indigenous gut microbes. Given the long prodromal or pre-diagnosed period, longitudinal studies of the human and rodent gut microbiome prior to symptomatic onset and for the duration of the disease period are currently lacking. PD is characterized in part by accumulation of the protein -synuclein (-syn) into insoluble aggregates, in both the central and enteric nervous systems. As such, a number of experimental rodent and non-human primate models of -syn overexpression recapitulate some of hallmark pathophysiologies of PD. These animal models provide an opportunity to assess how the gut microbiome changes with age under disease relevant conditions. Here, we used a transgenic mouse strain, the Thy1-hSYN "line 61" mice which over express wild-type human -syn to test how the gut microbiome composition responds in this model of PD pathology during aging. Using shotgun metagenomics, we find significant, age and genotype dependent bacterial taxa that become altered over age. We reveal that -syn overexpression can drive alterations to the gut microbiome composition and suggest that it limits the expansion of diversity through age. Given emerging data on potential contributions of the gut microbiome to PD pathologies, our data provide an experimental foundation to understand how the PD-associated microbiome may arise as a trigger or co-pathology to disease.
]]></description>
<dc:creator>Sampson, T. R.</dc:creator>
<dc:creator>Wallen, Z. D.</dc:creator>
<dc:creator>Won, W.-J.</dc:creator>
<dc:creator>Standaert, D. G.</dc:creator>
<dc:creator>Payami, H.</dc:creator>
<dc:creator>Harms, A. S.</dc:creator>
<dc:date>2024-02-01</dc:date>
<dc:identifier>doi:10.1101/2024.02.01.578464</dc:identifier>
<dc:title><![CDATA[Alpha-synuclein overexpression can drive microbiome dysbiosis in mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.02.02.578624v1?rss=1">
<title>
<![CDATA[
Transcriptional programs mediating neuronal toxicity and altered glial-neuronal signaling in a Drosophila knock-in tauopathy model 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.02.02.578624v1?rss=1"
</link>
<description><![CDATA[
Missense mutations in the gene encoding the microtubule-associated protein tau cause autosomal dominant forms of frontotemporal dementia. Multiple models of frontotemporal dementia based on transgenic expression of human tau in experimental model organisms, including Drosophila, have been described. These models replicate key features of the human disease, but do not faithfully recreate the genetic context of the human disorder. Here we use CRISPR-Cas mediated gene editing to model frontotemporal dementia caused by the tau P301L mutation by creating the orthologous mutation, P251L, in the endogenous Drosophila tau gene. Flies heterozygous or homozygous for tau P251L display age-dependent neurodegeneration, metabolic defects and accumulate DNA damage in affected neurons. To understand the molecular events promoting neuronal dysfunction and death in knock-in flies we performed single-cell RNA sequencing on approximately 130,000 cells from brains of tau P251L mutant and control flies. We found that expression of disease-associated mutant tau altered gene expression cell autonomously in all neuronal cell types identified and non-cell autonomously in glial cells. Cell signaling pathways, including glial-neuronal signaling, were broadly dysregulated as were brain region and cell-type specific protein interaction networks and gene regulatory programs. In summary, we present here a genetic model of tauopathy, which faithfully recapitulates the genetic context and phenotypic features of the human disease and use the results of comprehensive single cell sequencing analysis to outline pathways of neurotoxicity and highlight the role of non-cell autonomous changes in glia.
]]></description>
<dc:creator>Bukhari, H.</dc:creator>
<dc:creator>Nithianandam, V.</dc:creator>
<dc:creator>Battaglia, R.</dc:creator>
<dc:creator>Cicalo, A.</dc:creator>
<dc:creator>Sarkar, S.</dc:creator>
<dc:creator>Comjean, A.</dc:creator>
<dc:creator>Hu, Y.</dc:creator>
<dc:creator>Leventhal, M.</dc:creator>
<dc:creator>Dong, X.</dc:creator>
<dc:creator>Feany, M.</dc:creator>
<dc:date>2024-02-04</dc:date>
<dc:identifier>doi:10.1101/2024.02.02.578624</dc:identifier>
<dc:title><![CDATA[Transcriptional programs mediating neuronal toxicity and altered glial-neuronal signaling in a Drosophila knock-in tauopathy model]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.01.27.577538v1?rss=1">
<title>
<![CDATA[
Visualizing chaperone-mediated multistep assembly of the human 20S proteasome 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.01.27.577538v1?rss=1"
</link>
<description><![CDATA[
Dedicated assembly factors orchestrate stepwise production of many molecular machines, including the 28-subunit proteasome core particle (CP) that mediates protein degradation. Here, we report cryo-EM reconstructions of seven recombinant human subcomplexes that visualize all five chaperones and the three active site propeptides across a wide swath of the assembly pathway. Comparison of these chaperone-bound intermediates and a matching mature CP reveals molecular mechanisms determining the order of successive subunit additions, and how proteasome subcomplexes and assembly factors structurally adapt upon progressive subunit incorporation to stabilize intermediates, facilitate the formation of subsequent intermediates, and ultimately rearrange to coordinate proteolytic activation with gated access to active sites. The structural findings reported here explain many previous biochemical and genetic observations. This work establishes a methodologic approach for structural analysis of multiprotein complex assembly intermediates, illuminates specific functions of assembly factors, and reveals conceptual principles underlying human proteasome biogenesis.
]]></description>
<dc:creator>Adolf, F.</dc:creator>
<dc:creator>Du, J.</dc:creator>
<dc:creator>Goodall, E. A.</dc:creator>
<dc:creator>Walsh, R. M.</dc:creator>
<dc:creator>rawson, S.</dc:creator>
<dc:creator>von Gronaua, S.</dc:creator>
<dc:creator>Harper, J. W.</dc:creator>
<dc:creator>Hanna, J.</dc:creator>
<dc:creator>Schulman, B. A.</dc:creator>
<dc:date>2024-01-28</dc:date>
<dc:identifier>doi:10.1101/2024.01.27.577538</dc:identifier>
<dc:title><![CDATA[Visualizing chaperone-mediated multistep assembly of the human 20S proteasome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-01-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.01.08.574736v1?rss=1">
<title>
<![CDATA[
The landscape of glial pathology and T-call response in Parkinson's substantia nigra 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.01.08.574736v1?rss=1"
</link>
<description><![CDATA[
Parkinsons Disease (PD) is a progressive neurodegenerative disease that leads to debilitating movement disorders and often dementia. Recent evidence, including identification of specific peripheral T-cell receptor sequences, indicates that the adaptive immune response is associated with disease pathogenesis. However, the properties of T-cells in the brain regions where neurons degenerate are not well characterized. We have analyzed the identities and interactions of T-cells in PD in post-mortem brain tissue using single nucleus RNA sequencing, spatial transcriptomics and T-cell receptor sequencing. We found that T-cells in the substantia nigra of PD brain donors exhibit a CD8+ resident memory phenotype, increased clonal expansion, and altered spatial relationships with astrocytes, myeloid cells, and endothelial cells. We also describe regional differences in astrocytic responses to neurodegeneration. Our findings nominate potential molecular and cellular candidates that allow a deeper understanding of the pathophysiology of neurodegeneration in PD. Together, our work represents a major single nucleus and spatial transcriptional resource for the fields of neurodegeneration and PD.
]]></description>
<dc:creator>Jakubiak, K.</dc:creator>
<dc:creator>Paryani, F.</dc:creator>
<dc:creator>Kannan, A.</dc:creator>
<dc:creator>Lee, J.</dc:creator>
<dc:creator>Madden, N.</dc:creator>
<dc:creator>Li, J.</dc:creator>
<dc:creator>Chen, D.</dc:creator>
<dc:creator>Mahajan, A.</dc:creator>
<dc:creator>Xia, S.</dc:creator>
<dc:creator>Flowers, X.</dc:creator>
<dc:creator>Menon, V.</dc:creator>
<dc:creator>Sulzer, D.</dc:creator>
<dc:creator>Goldman, J. E.</dc:creator>
<dc:creator>Sims, P. A.</dc:creator>
<dc:creator>Al-Dalahmah, O.</dc:creator>
<dc:date>2024-01-09</dc:date>
<dc:identifier>doi:10.1101/2024.01.08.574736</dc:identifier>
<dc:title><![CDATA[The landscape of glial pathology and T-call response in Parkinson's substantia nigra]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-01-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.01.30.577912v1?rss=1">
<title>
<![CDATA[
Systemic inflammation triggers long-lasting neuroinflammation and accelerates neurodegeneration in a rat model of Parkinson's disease overexpressing human alpha-synuclein 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.01.30.577912v1?rss=1"
</link>
<description><![CDATA[
Increasing efforts have been made to elucidate how genetic and environmental factors interact in Parkinsons disease (PD). In the present study, we assessed the development of symptoms on a genetic PD rat model that overexpresses human -synuclein (Snca+/+) at a presymptomatic age, exposed to a pro-inflammatory insult by intraperitoneal injection of lipopolysaccharide (LPS), using immunohistology, high-dimensional flow cytometry, constant potential amperometry, and behavioral analyses. A single injection of LPS into WT and Snca+/+ rats triggered long-lasting increase in the activation of pro-inflammatory microglial markers, monocytes, and T lymphocytes. However, only LPS Snca+/+ rats showed dopaminergic neuronal loss in the substantia nigra pars compacta (SNpc), associated with a reduction in the release of evoked dopamine in the striatum. No significant changes were observed in the behavioral domain.

We propose our double-hit animal as a reliable model to investigate the mechanisms whereby -synuclein and inflammation interact to promote neurodegeneration in PD.
]]></description>
<dc:creator>Massaro Cenere, M.</dc:creator>
<dc:creator>Tiberi, M.</dc:creator>
<dc:creator>Paldino, E.</dc:creator>
<dc:creator>D'Addario, S. L.</dc:creator>
<dc:creator>Federici, M.</dc:creator>
<dc:creator>Giacomet, C.</dc:creator>
<dc:creator>Cutuli, D.</dc:creator>
<dc:creator>Matteocci, A.</dc:creator>
<dc:creator>Cossa, F.</dc:creator>
<dc:creator>Zarrilli, B.</dc:creator>
<dc:creator>Ledonne, A.</dc:creator>
<dc:creator>Petrosini, L.</dc:creator>
<dc:creator>Berretta, N.</dc:creator>
<dc:creator>Fusco, F. R.</dc:creator>
<dc:creator>Chiurchiu, V.</dc:creator>
<dc:creator>Mercuri, N. B.</dc:creator>
<dc:date>2024-01-30</dc:date>
<dc:identifier>doi:10.1101/2024.01.30.577912</dc:identifier>
<dc:title><![CDATA[Systemic inflammation triggers long-lasting neuroinflammation and accelerates neurodegeneration in a rat model of Parkinson's disease overexpressing human alpha-synuclein]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-01-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.02.12.580010v1?rss=1">
<title>
<![CDATA[
Locus coeruleus injury modulates ventral midbrain neuroinflammation during DSS-induced colitis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.02.12.580010v1?rss=1"
</link>
<description><![CDATA[
Parkinsons disease (PD) is characterized by a decades-long prodrome, consisting of a collection of non-motor symptoms that emerges prior to the motor manifestation of the disease. Of these non-motor symptoms, gastrointestinal dysfunction and deficits attributed to central norepinephrine (NE) loss, including mood changes and sleep disturbances, are frequent in the PD population and emerge early in the disease. Evidence is mounting that injury and inflammation in the gut and locus coeruleus (LC), respectively, underlie these symptoms, and the injury of these systems is central to the progression of PD. In this study, we generate a novel two-hit mouse model that captures both features, using dextran sulfate sodium (DSS) to induce gut inflammation and N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine (DSP-4) to lesion the LC. We first confirmed the specificity of DSP-4 for central NE using neurochemical methods and fluorescence light-sheet microscopy of cleared tissue, and established that DSS-induced outcomes in the periphery, including weight loss, gross indices of gut injury and systemic inflammation, the loss of tight junction proteins in the colonic epithelium, and markers of colonic inflammation, were unaffected with DSP-4 pre-administration. We then measured alterations in neuroimmune gene expression in the ventral midbrain in response to DSS treatment alone as well as the extent to which prior LC injury modified this response. In this two-hit model we observed that DSS-induced colitis activates the expression of key cytokines and chemokines in the ventral midbrain only in the presence of LC injury and the typical DSS-associated neuroimmune is blunted by pre-LC lesioning with DSP-4. In all, this study supports the growing appreciation for the LC as neuroprotective against inflammation-induced brain injury and draws attention to the potential for NEergic interventions to exert disease-modifying effects under conditions where peripheral inflammation may compromise ventral midbrain dopaminergic neurons and increase the risk for development of PD.
]]></description>
<dc:creator>Boles, J. S.</dc:creator>
<dc:creator>Holt, J.</dc:creator>
<dc:creator>Cole, C. L.</dc:creator>
<dc:creator>Neighbarger, N. K.</dc:creator>
<dc:creator>Urs, N. M.</dc:creator>
<dc:creator>Uriarte Huarte, O.</dc:creator>
<dc:creator>Tansey, M. G.</dc:creator>
<dc:date>2024-02-13</dc:date>
<dc:identifier>doi:10.1101/2024.02.12.580010</dc:identifier>
<dc:title><![CDATA[Locus coeruleus injury modulates ventral midbrain neuroinflammation during DSS-induced colitis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.01.19.524720v1?rss=1">
<title>
<![CDATA[
AlphaFold Prediction of Structural Ensembles of Disordered Proteins 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.01.19.524720v1?rss=1"
</link>
<description><![CDATA[
Deep learning methods of predicting protein structures have reached an accuracy comparable to that of high-resolution experimental methods. It is thus possible to generate accurate models of the native states of hundreds of millions of proteins. An open question, however, concerns whether these advances can be translated to disordered proteins, which should be represented as structural ensembles because of their heterogeneous and dynamical nature. Here we show that the inter-residue distances predicted by AlphaFold for disordered proteins can be used to construct accurate structural ensembles. These results illustrate the application to disordered proteins of deep learning methods originally trained for predicting the structures of folded proteins.
]]></description>
<dc:creator>Brotzakis, Z. F.</dc:creator>
<dc:creator>Zhang, S.</dc:creator>
<dc:creator>Vendruscolo, M.</dc:creator>
<dc:date>2023-01-19</dc:date>
<dc:identifier>doi:10.1101/2023.01.19.524720</dc:identifier>
<dc:title><![CDATA[AlphaFold Prediction of Structural Ensembles of Disordered Proteins]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.11.17.567425v1?rss=1">
<title>
<![CDATA[
Targeted micro-fiber arrays for measuring and manipulating localized multi-scale neural dynamics over large, deep brain volumes during behavior 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.11.17.567425v1?rss=1"
</link>
<description><![CDATA[
Neural population dynamics relevant for behavior vary over multiple spatial and temporal scales across 3-dimensional volumes. Current optical approaches lack the spatial coverage and resolution necessary to measure and manipulate naturally occurring patterns of large-scale, distributed dynamics within and across deep brain regions such as the striatum. We designed a new micro-fiber array and imaging approach capable of chronically measuring and optogenetically manipulating local dynamics across over 100 targeted locations simultaneously in head-fixed and freely moving mice. We developed a semi-automated micro-CT based strategy to precisely localize positions of each optical fiber. This highly-customizable approach enables investigation of multi-scale spatial and temporal patterns of cell-type and neurotransmitter specific signals over arbitrary 3-D volumes at a spatial resolution and coverage previously inaccessible. We applied this method to resolve rapid dopamine release dynamics across the striatum volume which revealed distinct, modality specific spatiotemporal patterns in response to salient sensory stimuli extending over millimeters of tissue. Targeted optogenetics through our fiber arrays enabled flexible control of neural signaling on multiple spatial scales, better matching endogenous signaling patterns, and spatial localization of behavioral function across large circuits.
]]></description>
<dc:creator>Vu, M.-A. T.</dc:creator>
<dc:creator>Brown, E. H.</dc:creator>
<dc:creator>Wen, M. J.</dc:creator>
<dc:creator>Noggle, C. A.</dc:creator>
<dc:creator>Zhang, Z.</dc:creator>
<dc:creator>Monk, K. J.</dc:creator>
<dc:creator>Bouabid, S.</dc:creator>
<dc:creator>Mroz, L.</dc:creator>
<dc:creator>Graham, B. M.</dc:creator>
<dc:creator>Zhuo, Y.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Otchy, T. M.</dc:creator>
<dc:creator>Tian, L.</dc:creator>
<dc:creator>Davison, I. G.</dc:creator>
<dc:creator>Boas, D. A.</dc:creator>
<dc:creator>Howe, M. W.</dc:creator>
<dc:date>2023-11-17</dc:date>
<dc:identifier>doi:10.1101/2023.11.17.567425</dc:identifier>
<dc:title><![CDATA[Targeted micro-fiber arrays for measuring and manipulating localized multi-scale neural dynamics over large, deep brain volumes during behavior]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.11.22.567373v1?rss=1">
<title>
<![CDATA[
Polyglucosan body density in the aged mouse hippocampus is controlled by a novel modifier locus on chromosome 1 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.11.22.567373v1?rss=1"
</link>
<description><![CDATA[
In aged humans and mice, aggregates of hypobranched glycogen molecules called polyglucosan bodies (PGBs) accumulate in hippocampal astrocytes. PGBs are known to drive cognitive decline in neurological diseases but remain largely unstudied in the context of typical brain aging. Here, we show that PGBs arise in autophagy-dysregulated astrocytes of the aged C57BL/6J mouse hippocampus. To map the genetic cause of age-related PGB accumulation, we quantified PGB burden in 32 fully sequenced BXD-recombinant inbred mouse strains, which display a 400-fold variation in hippocampal PGB burden at 16-18 months of age. A major modifier locus was mapped to chromosome 1 at 72-75 Mb, which we defined as the Pgb1 locus. To evaluate candidate genes and downstream mechanisms by which Pgb1 controls the aggregation of glycogen, extensive hippocampal transcriptomic and proteomic datasets were produced for aged mice of the BXD family. We utilized these datasets to identify Smarcal1 and Usp37 as potential regulators of PGB accumulation. To assess the effect of PGB burden on age-related cognitive decline, we performed phenome-wide association scans, transcriptomic analyses as well as conditioned fear memory and Y-maze testing. Importantly, we did not find any evidence suggesting a negative impact of PGBs on cognition. Taken together, our study demonstrates that the Pgb1 locus controls glycogen aggregation in astrocytes of the aged hippocampus without affecting age-related cognitive decline.
]]></description>
<dc:creator>Gomez-Pascual, A.</dc:creator>
<dc:creator>Glikman, D. M.</dc:creator>
<dc:creator>Ng, H. X.</dc:creator>
<dc:creator>Tomkins, J. E.</dc:creator>
<dc:creator>Lu, L.</dc:creator>
<dc:creator>Xu, Y.</dc:creator>
<dc:creator>Ashbrook, D. G.</dc:creator>
<dc:creator>Kaczorowski, C.</dc:creator>
<dc:creator>Kempermann, G.</dc:creator>
<dc:creator>Killmar, J.</dc:creator>
<dc:creator>Mozhui, K.</dc:creator>
<dc:creator>Aebersold, R.</dc:creator>
<dc:creator>Williams, E. G.</dc:creator>
<dc:creator>Williams, R. W.</dc:creator>
<dc:creator>Overall, R. W.</dc:creator>
<dc:creator>Jucker, M.</dc:creator>
<dc:creator>de Bakker, D. E. M.</dc:creator>
<dc:date>2023-11-22</dc:date>
<dc:identifier>doi:10.1101/2023.11.22.567373</dc:identifier>
<dc:title><![CDATA[Polyglucosan body density in the aged mouse hippocampus is controlled by a novel modifier locus on chromosome 1]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.01.31.578239v1?rss=1">
<title>
<![CDATA[
Open-Source Platform for Kinematic Analysis of Mouse Forelimb Movement 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.01.31.578239v1?rss=1"
</link>
<description><![CDATA[
We present an open-source behavioral platform and software solution for studying fine motor skills in mice performing reach-to-grasp task. The behavioral platform uses readily available and 3D-printed components and was designed to be affordable and universally reproducible. The protocol describes how to assemble the box, train mice to perform the task and process the video with the custom software pipeline to analyze forepaw kinematics. All the schematics, 3D models, code and assembly instructions are provided in the open GitHub repository.

Graphical abstract
]]></description>
<dc:creator>Berezhnoi, D.</dc:creator>
<dc:creator>Chehade, H. D.</dc:creator>
<dc:creator>Chu, H.-y.</dc:creator>
<dc:date>2024-02-02</dc:date>
<dc:identifier>doi:10.1101/2024.01.31.578239</dc:identifier>
<dc:title><![CDATA[Open-Source Platform for Kinematic Analysis of Mouse Forelimb Movement]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.02.10.579775v1?rss=1">
<title>
<![CDATA[
A proteome-wide quantitative guide for nanoscale spatially resolved extraction of membrane proteins into native nanodiscs 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.02.10.579775v1?rss=1"
</link>
<description><![CDATA[
The intricate molecular environment of the native membrane profoundly influences every aspect of membrane protein (MP) biology. Despite this, the most prevalent method of studying MPs uses detergent-like molecules that disrupt and remove this vital local membrane context. This severely impedes our ability to quantitatively decipher the local molecular context and comprehend its regulatory role in the structure, function, and biogenesis of MPs. Using a library of membrane-active polymers we have developed a platform for the high-throughput analysis of the membrane proteome. The platform enables near-complete spatially resolved extraction of target MPs directly from their endogenous membranes into native nanodiscs that maintain the local membrane context. We accompany this advancement with an open-access database that quantifies the polymer-specific extraction variability for 2065 unique mammalian MPs and provides the most optimized condition for each of them. Our method enables rapid and near-complete extraction and purification of target MPs directly from their endogenous organellar membranes at physiological expression levels while maintaining the nanoscale local membrane environment. Going beyond the plasma membrane proteome, our platform enables extraction from any target organellar membrane including the endoplasmic reticulum, mitochondria, lysosome, Golgi, and even transient organelles such as the autophagosome. To further validate this platform, we took several independent MPs and demonstrated how our resource can enable rapid extraction and purification of target MPs from different organellar membranes with high efficiency and purity. Further, taking two synaptic vesicle MPs, we show how the database can be extended to capture multiprotein complexes between overexpressed MPs. We expect these publicly available resources to empower researchers across disciplines to efficiently capture membrane  nano-scoops containing a target MP and interface with structural, functional, and other bioanalytical approaches. We demonstrate an example of this by combining our extraction platform with single-molecule TIRF imaging to demonstrate how it can enable rapid determination of homo-oligomeric states of target MPs in native cell membranes.
]]></description>
<dc:creator>Brown, C.</dc:creator>
<dc:creator>Ghosh, S.</dc:creator>
<dc:creator>McAllister, R.</dc:creator>
<dc:creator>Kumar, M.</dc:creator>
<dc:creator>Walker, G.</dc:creator>
<dc:creator>Sun, E.</dc:creator>
<dc:creator>Aman, T.</dc:creator>
<dc:creator>Panda, A.</dc:creator>
<dc:creator>Kumar, S.</dc:creator>
<dc:creator>Li, W. V.</dc:creator>
<dc:creator>Coleman, J.</dc:creator>
<dc:creator>Liu, Y.</dc:creator>
<dc:creator>Rothman, J. E.</dc:creator>
<dc:creator>Bhattacharyya, M.</dc:creator>
<dc:creator>Gupta, K.</dc:creator>
<dc:date>2024-02-12</dc:date>
<dc:identifier>doi:10.1101/2024.02.10.579775</dc:identifier>
<dc:title><![CDATA[A proteome-wide quantitative guide for nanoscale spatially resolved extraction of membrane proteins into native nanodiscs]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.02.12.579917v1?rss=1">
<title>
<![CDATA[
iSCORE-PD: an isogenic stem cell collection to research Parkinson Disease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.02.12.579917v1?rss=1"
</link>
<description><![CDATA[
Parkinsons disease (PD) is a neurodegenerative disorder caused by complex genetic and environmental factors. Genome-edited human pluripotent stem cells (hPSCs) offer a unique experimental platform to advance our understanding of PD etiology by enabling the generation of disease-relevant cell types carrying patient mutations along with isogenic control cells. To facilitate this approach, we generated a collection of 65 human stem cell lines genetically engineered to harbor high risk or causal variants in genes associated with PD (SNCA A53T, SNCA A30P, PRKN Ex3del, PINK1 Q129X, DJ1/PARK7 Ex1-5del, LRRK2 G2019S, ATP13A2 FS, FBXO7 R498X/FS, DNAJC6 c.801 A>G/FS, SYNJ1 R258Q/FS, VPS13C A444P/FS, VPS13C W395C/FS, GBA1 IVS2+1/FS). All mutations were introduced into a fully characterized and sequenced female human embryonic stem cell (hESC) line (WIBR3; NIH approval number NIHhESC-10-0079) using different genome editing techniques. To ensure the genetic integrity of these cell lines, we implemented rigorous quality controls, including whole-genome sequencing of each line. Our analysis of the genetic variation in this cell line collection revealed that while genome editing, particularly using CRISPR/Cas9, can introduce rare off-target mutations, the predominant source of genetic variants arises from routine cell culture and are fixed in cell lines during clonal isolation. The observed genetic variation was minimal compared to that typically found in patient-derived iPSC experiments and predominantly affected non-coding regions of the genome. Importantly, our analysis outlines strategies for effectively managing genetic variation through stringent quality control measures and careful experimental design. This systematic approach ensures the high quality of our stem cell collection, highlights advantages of prime editing over conventional CRISPR/Cas9 methods and provides a roadmap for the generation of gene-edited hPSC collections at scale in an academic setting. Our iSCORE-PD collection represents an easily accessible and valuable platform to study PD, which can be used by investigators to understand the molecular pathophysiology of PD in a human cellular setting.
]]></description>
<dc:creator>Busquets, O.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Syed, K. M.</dc:creator>
<dc:creator>Jerez, P. A.</dc:creator>
<dc:creator>Dunnack, J.</dc:creator>
<dc:creator>Bu, R. L.</dc:creator>
<dc:creator>Verma, Y.</dc:creator>
<dc:creator>Pangilinan, G. R.</dc:creator>
<dc:creator>Martin, A.</dc:creator>
<dc:creator>Straub, J.</dc:creator>
<dc:creator>Du, Y.</dc:creator>
<dc:creator>Simon, V. M.</dc:creator>
<dc:creator>Poser, S.</dc:creator>
<dc:creator>Bush, Z.</dc:creator>
<dc:creator>Diaz, J.</dc:creator>
<dc:creator>Sahagun, A.</dc:creator>
<dc:creator>Gao, J.</dc:creator>
<dc:creator>Hernandez, D.</dc:creator>
<dc:creator>Levine, K. S.</dc:creator>
<dc:creator>Booth, E. O.</dc:creator>
<dc:creator>Bateup, H.</dc:creator>
<dc:creator>Rio, D.</dc:creator>
<dc:creator>Hockemeyer, D.</dc:creator>
<dc:creator>Blauwendraat, C.</dc:creator>
<dc:creator>Soldner, F.</dc:creator>
<dc:date>2024-02-13</dc:date>
<dc:identifier>doi:10.1101/2024.02.12.579917</dc:identifier>
<dc:title><![CDATA[iSCORE-PD: an isogenic stem cell collection to research Parkinson Disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.02.17.580698v1?rss=1">
<title>
<![CDATA[
Large-scale visualisation of α-synuclein oligomers in Parkinson's disease brain tissue 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.02.17.580698v1?rss=1"
</link>
<description><![CDATA[
Parkinsons disease (PD) is a common neurodegenerative condition characterised by the presence in the brain of large intraneuronal aggregates, known as Lewy bodies and Lewy neurites, containing fibrillar -synuclein. According to the amyloid hypothesis, these large end-stage species form from smaller soluble protein assemblies, often termed oligomers, which are proposed as early drivers of pathogenesis. To date, however, this hypothesis has remained controversial, at least in part because it has not been possible to directly visualise oligomeric aggregates in human brain tissue. Therefore, their presence, abundance and distributions have remained elusive. Here, we present ASA-PD (Advanced Sensing of Aggregates - Parkinsons Disease), an imaging method to generate large-scale -synuclein oligomer maps in post-mortem human brain tissue. We combined autofluorescence suppression with single-molecule fluorescence methods, which together, enable the detection of nanoscale -synuclein aggregates. To demonstrate the utility of this platform, we captured [~]1.2 million oligomers from the anterior cingulate cortex in human post-mortem brain samples from PD and healthy control patients. Our data revealed a specific subpopulation of nanoscale oligomers that represent an early hallmark of the proteinopathy that underlies PD. We anticipate that quantitative information about oligomer distributions provided by ASA-PD will enable mechanistic studies to reveal the pathological processes caused by -synuclein aggregation.
]]></description>
<dc:creator>Andrews, R.</dc:creator>
<dc:creator>Fu, B.</dc:creator>
<dc:creator>Toomey, C. E.</dc:creator>
<dc:creator>Breiter, J. C.</dc:creator>
<dc:creator>Lachica, J.</dc:creator>
<dc:creator>Tian, R.</dc:creator>
<dc:creator>Beckwith, J. S.</dc:creator>
<dc:creator>Needham, L.-M.</dc:creator>
<dc:creator>Chant, G. J.</dc:creator>
<dc:creator>Loiseau, C.</dc:creator>
<dc:creator>Deconfin, A.</dc:creator>
<dc:creator>Baspin, K.</dc:creator>
<dc:creator>Magill, P. J.</dc:creator>
<dc:creator>Jaunmuktane, Z.</dc:creator>
<dc:creator>Freeman, O. J.</dc:creator>
<dc:creator>Taylor, B. J. M.</dc:creator>
<dc:creator>Hardy, J.</dc:creator>
<dc:creator>Lashley, T.</dc:creator>
<dc:creator>Ryten, M.</dc:creator>
<dc:creator>Vendruscolo, M.</dc:creator>
<dc:creator>Wood, N. W.</dc:creator>
<dc:creator>Weiss, L. E.</dc:creator>
<dc:creator>Gandhi, S.</dc:creator>
<dc:creator>Lee, S. F.</dc:creator>
<dc:date>2024-02-19</dc:date>
<dc:identifier>doi:10.1101/2024.02.17.580698</dc:identifier>
<dc:title><![CDATA[Large-scale visualisation of α-synuclein oligomers in Parkinson's disease brain tissue]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.02.17.580767v1?rss=1">
<title>
<![CDATA[
An open-source MRI compatible frame for multimodal presurgical mapping in macaque and capuchin monkeys 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.02.17.580767v1?rss=1"
</link>
<description><![CDATA[
HighlightsO_LIWe present a compact MRI-compatible stereotaxic frame for large nonhuman primates.
C_LIO_LIThe design is 3D printable, inexpensive, and matches size of an adult human head.
C_LIO_LIEnabled real-time, accurate, MRI-guided deep-brain viral vector injection.
C_LIO_LIFacilitated multimodal alignment for deep-brain electrophysiology planning.
C_LIO_LIAll computer-aided-design files are modularized and publicly available and editable.
C_LI

BackgroundHigh-precision neurosurgical targeting in nonhuman primates (NHPs) often requires presurgical anatomy mapping with noninvasive neuroimaging techniques (MRI, CT, PET), allowing for translation of individual anatomical coordinates to surgical stereotaxic apparatus. Given the varied tissue contrasts that these imaging techniques produce, precise alignment of imaging-based coordinates to surgical apparatus can be cumbersome. MRI-compatible stereotaxis with radiopaque fiducial markers offer a straight-forward and reliable solution, but existing commercial options do not fit in conformal head coils that maximize imaging quality.

New methodWe developed a compact MRI-compatible stereotaxis suitable for a variety of NHP species (Macaca mulatta, Macaca fascicularis, and Cebus apella) that allows multimodal alignment through technique-specific fiducial markers.

Comparison with existing methodsWith the express purpose of compatibility with clinically available MRI, CT, and PET systems, the frame is no larger than a human head, while allowing for imaging NHPs in the supinated position. This design requires no marker implantation, special software, or additional knowledge other than the operation of a common large animal stereotaxis.

ResultsWe demonstrated the applicability of this 3D-printable apparatus across a diverse set of experiments requiring presurgical planning: 1) We demonstrate the accuracy of the fiducial system through a within-MRI cannula insertion and subcortical injection of viral vectors. 2) We also demonstrated accuracy of multimodal (MRI and CT) alignment and coordinate transfer to guide a surgical robot electrode implantation for deep-brain electrophysiology.

ConclusionsThe computer-aided design files and engineering drawings are publicly available, with the modular design allowing for low cost and manageable manufacturing.
]]></description>
<dc:creator>Liang, L.</dc:creator>
<dc:creator>Zimmermann Rollin, I.</dc:creator>
<dc:creator>Alikaya, A.</dc:creator>
<dc:creator>Ho, J. C.</dc:creator>
<dc:creator>Santini, T.</dc:creator>
<dc:creator>Bostan, A. C.</dc:creator>
<dc:creator>Schwerdt, H. N.</dc:creator>
<dc:creator>Stauffer, W. R.</dc:creator>
<dc:creator>Ibrahim, T. S.</dc:creator>
<dc:creator>Pirondini, E.</dc:creator>
<dc:creator>Schaeffer, D.</dc:creator>
<dc:date>2024-02-20</dc:date>
<dc:identifier>doi:10.1101/2024.02.17.580767</dc:identifier>
<dc:title><![CDATA[An open-source MRI compatible frame for multimodal presurgical mapping in macaque and capuchin monkeys]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.02.17.580796v1?rss=1">
<title>
<![CDATA[
An axonal brake on striatal dopamine output by cholinergic interneurons 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.02.17.580796v1?rss=1"
</link>
<description><![CDATA[
Depolarisation of distal axons is necessary for neurons to translate somatic action potentials into neurotransmitter release. Studies have shown that striatal cholinergic interneurons (ChIs) can directly drive ectopic action potentials in dopamine (DA) axons and trigger DA release. However, here we show that this action occurs within a broader context of axonal signal integration whereby activation of ChIs and depolarisation of DA axons by nicotinic receptors (nAChRs) limits the subsequent depolarisation and release of DA in response to ensuing activity. We demonstrate that activation of ChIs and nAChRs in ex vivo mouse striatum, even when it does not trigger DA release that is detectable by fast-scan cyclic voltammetry, limits DA release for [~]100 ms by depressing subsequent axonal depolarisation and calcium summation. This axonal brake on DA release is stronger in dorsal than ventral striatum, and is unrelated to DA depletion. In vivo, antagonism of nAChRs in dorsal striatum elevated extracellular DA levels and promoted conditioned place-preference, underscoring its physiological relevance. Our findings reveal that under physiological conditions in vivo, ChIs acting via nAChRs dynamically attenuate DA output driven by DA neuron activity, leading to a predominantly inverse relationship between ACh and DA signalling that varies continuously with ChI activity.
]]></description>
<dc:creator>Zhang, Y.-F.</dc:creator>
<dc:creator>Luan, P.</dc:creator>
<dc:creator>Qiao, Q.</dc:creator>
<dc:creator>He, Y.</dc:creator>
<dc:creator>Zatka-Haas, P.</dc:creator>
<dc:creator>Zhang, G.</dc:creator>
<dc:creator>Lin, M.</dc:creator>
<dc:creator>Lak, A.</dc:creator>
<dc:creator>Jing, M.</dc:creator>
<dc:creator>Mann, E. O.</dc:creator>
<dc:creator>Cragg, S.</dc:creator>
<dc:date>2024-02-21</dc:date>
<dc:identifier>doi:10.1101/2024.02.17.580796</dc:identifier>
<dc:title><![CDATA[An axonal brake on striatal dopamine output by cholinergic interneurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.02.22.581576v1?rss=1">
<title>
<![CDATA[
Nova-ST: Nano-Patterned Ultra-Dense platform for spatial transcriptomics 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.02.22.581576v1?rss=1"
</link>
<description><![CDATA[
Spatial transcriptomics workflows using barcoded capture arrays are commonly used for resolving gene expression in tissues. However, existing techniques are either limited by capture array density or are cost prohibitive for large scale atlasing. We present Nova-ST, a dense nano-patterned spatial transcriptomics technique derived from randomly barcoded Illumina sequencing flow cells. Nova-ST enables customized, low cost, flexible, and high-resolution spatial profiling of large tissue sections. Benchmarking on mouse brain sections demonstrates significantly higher sensitivity compared to existing methods, at reduced cost.

MotivationSpatial transcriptomics analysis is becoming exceedingly important in biomedical and clinical research. Several platforms for spatial transcriptomics are currently available, but most of these technologies are commercialized making them inflexible and cost prohibitive. The motivation for this work was to establish an open source, flexible and sensitive sequencing-based spatial transcriptomics platform that can provide a considerable cost advantage for performing large scale spatial profiling. We provide thorough and in-depth guidance and resources both for the experimental and computational components of the workflow, to facilitate easy implementation of Nova-ST.
]]></description>
<dc:creator>Poovathingal, S.</dc:creator>
<dc:creator>Davie, K.</dc:creator>
<dc:creator>Vandepoel, R.</dc:creator>
<dc:creator>Poulvellarie, N.</dc:creator>
<dc:creator>Verfaillie, A.</dc:creator>
<dc:creator>Corthout, N.</dc:creator>
<dc:creator>Aerts, S.</dc:creator>
<dc:date>2024-02-24</dc:date>
<dc:identifier>doi:10.1101/2024.02.22.581576</dc:identifier>
<dc:title><![CDATA[Nova-ST: Nano-Patterned Ultra-Dense platform for spatial transcriptomics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.02.25.580545v1?rss=1">
<title>
<![CDATA[
Modeling gene-environment interactions in Parkinson's Disease: Helicobacter pylori infection of Pink1-/- mice induces CD8 T cell-dependent motor and cognitive dysfunction. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.02.25.580545v1?rss=1"
</link>
<description><![CDATA[
Parkinsons disease (PD) is a chronic neurodegenerative disorder characterized by progressive loss of motor function. Diagnosis occurs late: after motor symptom development downstream of the irreparable loss of a large proportion of the dopaminergic neurons in the substantia nigra of the brain. Understanding PD pathophysiology in its pre-motor prodromal phase is needed for earlier diagnosis and intervention. Genetic risk factors, environmental triggers, and dysregulated immunity have all been implicated in PD development. Here, we demonstrate in a mouse model deficient in the PD-associated gene Pink, that infection with the human PD-associated gastric bacterium Helicobacter pylori leads to development of motor and cognitive signs resembling prodromal features of PD. This was also associated with proliferation and activation of primary mitochondria-reactive CD8 T cells and infiltration of CD8 T cells into the brain. Development of the motor and cognitive phenotypes in the infected Pink1-/- mice was abrogated when CD8 T cells were depleted prior to infection. We anticipate that this new model, which integrates genetic PD susceptibility, a PD-relevant environmental trigger, and specific immune changes that are required for symptom development, will be a valuable tool for increasing our understanding of this complex disease.
]]></description>
<dc:creator>Kazanova, A.</dc:creator>
<dc:creator>Sung, J.</dc:creator>
<dc:creator>Oliveira, N. L.</dc:creator>
<dc:creator>Gavino, C.</dc:creator>
<dc:creator>Bessaiah, H.</dc:creator>
<dc:creator>Pei, J.</dc:creator>
<dc:creator>Burns, L.</dc:creator>
<dc:creator>Miller, W.</dc:creator>
<dc:creator>Brouillard-Galipeau, M.</dc:creator>
<dc:creator>Zhu, L.</dc:creator>
<dc:creator>Guerra, L. M.</dc:creator>
<dc:creator>Elemeery, M. N.</dc:creator>
<dc:creator>Recinto, S. J.</dc:creator>
<dc:creator>MacDonald, A.</dc:creator>
<dc:creator>Lanoix, J.</dc:creator>
<dc:creator>Thibault, P.</dc:creator>
<dc:creator>McBride, H. M.</dc:creator>
<dc:creator>Desjardins, M.</dc:creator>
<dc:creator>Stratton, J. A.</dc:creator>
<dc:creator>Labrecque, N.</dc:creator>
<dc:creator>Gruenheid, S.</dc:creator>
<dc:date>2024-02-26</dc:date>
<dc:identifier>doi:10.1101/2024.02.25.580545</dc:identifier>
<dc:title><![CDATA[Modeling gene-environment interactions in Parkinson's Disease: Helicobacter pylori infection of Pink1-/- mice induces CD8 T cell-dependent motor and cognitive dysfunction.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.02.25.581985v1?rss=1">
<title>
<![CDATA[
Comparative study of enriched dopaminergic neurons from siblings with Gaucher disease discordant for parkinsonism 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.02.25.581985v1?rss=1"
</link>
<description><![CDATA[
Inducible pluripotent stem cells (iPSCs) derived from patient samples have significantly enhanced our ability to model neurological diseases. Comparative studies of dopaminergic (DA) neurons differentiated from iPSCs derived from siblings with Gaucher disease discordant for parkinsonism provides a valuable avenue to explore genetic modifiers contributing to GBA1-associated parkinsonism in disease-relevant cells. However, such studies are often complicated by the inherent heterogeneity in differentiation efficiency among iPSC lines derived from different individuals. To address this technical challenge, we devised a selection strategy to enrich dopaminergic (DA) neurons expressing tyrosine hydroxylase (TH). A neomycin resistance gene (neo) was inserted at the C-terminus of the TH gene following a T2A self-cleavage peptide, placing its expression under the control of the TH promoter. This allows for TH+ DA neuron enrichment through geneticin selection. This method enabled us to generate comparable, high-purity DA neuron cultures from iPSC lines derived from three sisters that we followed for over a decade: one sibling is a healthy individual, and the other two have Gaucher disease (GD) with GBA1 genotype N370S/c.203delC+R257X (p.N409S/c.203delC+p.R296X). Notably, the younger sister with GD later developed Parkinson disease (PD). A comprehensive analysis of these high-purity DA neurons revealed that although GD DA neurons exhibited decreased levels of glucocerebrosidase (GCase), there was no substantial difference in GCase protein levels or lipid substrate accumulation between DA neurons from the GD and GD/PD sisters, suggesting that the PD discordance is related to of other genetic modifiers.
]]></description>
<dc:creator>Hertz, E.</dc:creator>
<dc:creator>Perez, G.</dc:creator>
<dc:creator>Hao, Y.</dc:creator>
<dc:creator>Rytel, K.</dc:creator>
<dc:creator>Ma, C.</dc:creator>
<dc:creator>Kirby, M.</dc:creator>
<dc:creator>Anderson, S.</dc:creator>
<dc:creator>Wincovitch, S.</dc:creator>
<dc:creator>Andersh, K.</dc:creator>
<dc:creator>Ahfeldt, T.</dc:creator>
<dc:creator>Blanchard, J.</dc:creator>
<dc:creator>Qi, Y. A.</dc:creator>
<dc:creator>Lopez, G.</dc:creator>
<dc:creator>Tayebi, N.</dc:creator>
<dc:creator>Sidransky, E.</dc:creator>
<dc:creator>Chen, Y.</dc:creator>
<dc:date>2024-02-28</dc:date>
<dc:identifier>doi:10.1101/2024.02.25.581985</dc:identifier>
<dc:title><![CDATA[Comparative study of enriched dopaminergic neurons from siblings with Gaucher disease discordant for parkinsonism]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.02.26.582055v1?rss=1">
<title>
<![CDATA[
Alpha-synuclein aggregates trigger anti-viral immune pathways and RNA editing in human astrocytes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.02.26.582055v1?rss=1"
</link>
<description><![CDATA[
RNA editing is a post transcriptional mechanism that targets changes in RNA transcripts to modulate innate immune responses. We report the role of astrocyte specific, ADAR1 mediated RNA editing in neuroinflammation in Parkinsons disease. We generated hiPSC-derived astrocytes, neurons and co-cultures and exposed them to small soluble alpha-synuclein aggregates. Oligomeric alpha-synuclein triggered an inflammatory glial state associated with TLR activation, viral responses, and cytokine secretion. This reactive state resulted in loss of neurosupportive functions, and the induction of neuronal toxicity. Notably, interferon response pathways were activated leading to upregulation, and isoform switching of the RNA deaminase enzyme, ADAR1. ADAR1 mediates A-to-I RNA editing, and increases in RNA editing were observed in inflammatory pathways in cells, as well as in post-mortem human PD brain. Aberrant, or dysregulated, ADAR1 responses and RNA editing may lead to sustained inflammatory reactive states in astrocytes triggered by alpha-synuclein aggregation, and this may drive the neuroinflammatory cascade in Parkinsons.
]]></description>
<dc:creator>D'Sa, K.</dc:creator>
<dc:creator>Choi, M. L.</dc:creator>
<dc:creator>Wagen, A. Z.</dc:creator>
<dc:creator>Seto-Salvia, N.</dc:creator>
<dc:creator>Kopach, O.</dc:creator>
<dc:creator>Evans, J. R.</dc:creator>
<dc:creator>Rodrigues, M.</dc:creator>
<dc:creator>Lopez-Garcia, P.</dc:creator>
<dc:creator>Ghareeb, A.</dc:creator>
<dc:creator>Bayne, J.</dc:creator>
<dc:creator>Grant-Peters, M.</dc:creator>
<dc:creator>Garcia-Ruiz, S.</dc:creator>
<dc:creator>Chen, Z.</dc:creator>
<dc:creator>Rodriques, S.</dc:creator>
<dc:creator>Athauda, D.</dc:creator>
<dc:creator>Gustavsson, E.</dc:creator>
<dc:creator>Gagliano Taliun, S. A.</dc:creator>
<dc:creator>Reynolds, R. H.</dc:creator>
<dc:creator>Young, G.</dc:creator>
<dc:creator>Strohbuecker, S.</dc:creator>
<dc:creator>Warner, T.</dc:creator>
<dc:creator>Rusakov, D. A.</dc:creator>
<dc:creator>Patani, R.</dc:creator>
<dc:creator>Bryant, C.</dc:creator>
<dc:creator>Klenerman, D. A.</dc:creator>
<dc:creator>Gandhi, S.</dc:creator>
<dc:creator>Ryten, M.</dc:creator>
<dc:date>2024-02-28</dc:date>
<dc:identifier>doi:10.1101/2024.02.26.582055</dc:identifier>
<dc:title><![CDATA[Alpha-synuclein aggregates trigger anti-viral immune pathways and RNA editing in human astrocytes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.02.26.582098v1?rss=1">
<title>
<![CDATA[
Adoptive transfer of mitochondrial antigen-specific CD8+ T-cells in mice causes parkinsonism and compromises the dopamine system 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.02.26.582098v1?rss=1"
</link>
<description><![CDATA[
The progressive degeneration of dopamine (DA) neurons drives motor symptoms in Parkinsons disease (PD). Whether this neuronal degeneration is due to cell-autonomous dysfunctions in DA neurons or to death signals generated by other cell types is a key problem to address. Recent evidence suggests that loss of function of the protein PINK1, linked to early-onset forms of PD, enhances the presentation of self-derived mitochondrial antigens, which induces the response of autoreactive CD8+ T cells. Whether mitochondrial antigen-specific CD8+ T cells alone are sufficient to induce nigrostriatal dysfunction has not been directly tested. Here we performed adoptive transfer of mitochondrial antigen-specific CD8+ T cells into wild-type or PINK1-deficient mice. We provide evidence for the entry and persistence of such cells in the brain and show that this leads to levodopa-reversible motor dysfunctions and partial degeneration of the nigrostriatal DA system in both genotypes. These findings establish that brain entry of autoreactive CD8+ T cells is sufficient to drive nigrostriatal degeneration and parkinsonian motor deficits, providing the most direct support to date for the hypothesis that an adaptive immune attack plays a key role in PD-like neurodegeneration.
]]></description>
<dc:creator>Elemeery, M. N.</dc:creator>
<dc:creator>Tchung, A.</dc:creator>
<dc:creator>Boulet, S.</dc:creator>
<dc:creator>Mukherjee, S.</dc:creator>
<dc:creator>Giguere, N.</dc:creator>
<dc:creator>Daudelin, J.-F.</dc:creator>
<dc:creator>Hetu-Arbour, R.</dc:creator>
<dc:creator>Matheoud, D.</dc:creator>
<dc:creator>STRATTON, J. A.</dc:creator>
<dc:creator>Labrecque, N.</dc:creator>
<dc:creator>Trudeau, L.-E.</dc:creator>
<dc:date>2024-02-28</dc:date>
<dc:identifier>doi:10.1101/2024.02.26.582098</dc:identifier>
<dc:title><![CDATA[Adoptive transfer of mitochondrial antigen-specific CD8+ T-cells in mice causes parkinsonism and compromises the dopamine system]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.02.28.582356v1?rss=1">
<title>
<![CDATA[
Proportion and distribution of neurotransmitter-defined cell types in the ventral tegmental area and substantia nigra pars compacta 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.02.28.582356v1?rss=1"
</link>
<description><![CDATA[
Most studies on the ventral tegmental area (VTA) and substantia nigra pars compacta (SNc) have focused on dopamine neurons and their role in processes such as motivation, learning, movement, and associated disorders such as addiction and Parkinsons disease. However there has been increasing attention on other VTA and SNc cell types that release GABA, glutamate, or a combination of neurotransmitters. Yet the relative distributions and proportions of neurotransmitter-defined cell types across VTA and SNc has remained unclear. Here, we used fluorescent in situ hybridization in male and female mice to label VTA and SNc neurons that expressed mRNA encoding the canonical vesicular transporters for dopamine, GABA, or glutamate: vesicular monoamine transporter (VMAT2), vesicular GABA transporter (VGAT), and vesicular glutamate transporter (VGLUT2). Within VTA, we found that no one type was particularly more abundant, instead we observed similar numbers of VMAT2+ (44%), VGAT+ (37%) and VGLUT2+ (41%) neurons. In SNc we found that a slight majority of neurons expressed VMAT2 (54%), fewer were VGAT+ (42%), and VGLUT2+ neurons were least abundant (16%). Moreover, 20% of VTA neurons and 10% of SNc neurons expressed more than one vesicular transporter, including 45% of VGLUT2+ neurons. We also assessed within VTA and SNc subregions and found remarkable heterogeneity in cell-type composition. And by quantifying density across both anterior-posterior and medial-lateral axes we generated heatmaps to visualize the distribution of each cell type. Our data complement recent single-cell RNAseq studies and support a more diverse landscape of neurotransmitter-defined cell types in VTA and SNc than is typically appreciated.
]]></description>
<dc:creator>Conrad, W. S.</dc:creator>
<dc:creator>Oriol, L.</dc:creator>
<dc:creator>Faget, L.</dc:creator>
<dc:creator>Hnasko, T. S.</dc:creator>
<dc:date>2024-02-28</dc:date>
<dc:identifier>doi:10.1101/2024.02.28.582356</dc:identifier>
<dc:title><![CDATA[Proportion and distribution of neurotransmitter-defined cell types in the ventral tegmental area and substantia nigra pars compacta]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.02.28.582420v1?rss=1">
<title>
<![CDATA[
Comparison of Alternative pre-mRNA Splicing and Gene Expression Patterns in Midbrain Lineage Cells Carrying Familial Parkinson's Disease Mutations 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.02.28.582420v1?rss=1"
</link>
<description><![CDATA[
Parkinsons disease (PD) arises from genetic and environmental factors. Human genetics has identified mutations in [~]20 inherited familial genes linked to monogenic forms of PD. To investigate the effects of individual familial PD mutations, human pluripotent embryonic stem cells (hPSCs) carrying 12 distinct familial PD mutations were differentiated into midbrain lineage cells, including dopaminergic (mDA) neurons. Global gene expression and pre-mRNA splicing patterns were analyzed in midbrain cultures carrying pathogenic PD mutations in the PRKN, SNCA, LRRK2, PINK1, DNAJC6, FBXO7, SYNJ1, DJ1, VPS13C, ATP13A2 and GBA1 genes. We have grouped the analysis of these familial PD mutations to genes expressed in mDA neurons and whose pre-mRNA splicing changes are linked to known PD defects in transport, cytoskeleton, lysosomes and mitochondria. Importantly, we have also shown that subsets of these splicing changes overlap with changes found in PD patient postmortem brains. Mutation-specific pre-mRNA isoforms may function as both diagnostic biomarkers for familial PD-associated genotypes and promising therapeutic targets.
]]></description>
<dc:creator>Lee, Y. J.</dc:creator>
<dc:creator>Syed, K.</dc:creator>
<dc:creator>Busquets, O.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Dunnack, J.</dc:creator>
<dc:creator>Sahagun, A.</dc:creator>
<dc:creator>Bateup, H.</dc:creator>
<dc:creator>Soldner, F.</dc:creator>
<dc:creator>Hockemeyer, D.</dc:creator>
<dc:creator>Rio, D.</dc:creator>
<dc:date>2024-03-03</dc:date>
<dc:identifier>doi:10.1101/2024.02.28.582420</dc:identifier>
<dc:title><![CDATA[Comparison of Alternative pre-mRNA Splicing and Gene Expression Patterns in Midbrain Lineage Cells Carrying Familial Parkinson's Disease Mutations]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.02.28.582460v1?rss=1">
<title>
<![CDATA[
GLP-1 receptor agonism ameliorates Parkinsons disease through modulation of neuronal insulin signalling and glial suppression 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.02.28.582460v1?rss=1"
</link>
<description><![CDATA[
Neuronal insulin resistance is linked to the pathogenesis of Parkinsons disease through unclear, but potentially targetable, mechanisms. We delineated neuronal and glial mechanisms of insulin resistance and glucagon-like 1 peptide (GLP-1) receptor agonism in human iPSC models of synucleinopathy, and corroborated our findings in patient samples from a Phase 2 trial of a GLP-1R agonist in Parkinsons (NCT01971242). Human iPSC models of synucleinopathy exhibit neuronal insulin resistance and dysfunctional insulin signalling, which is associated with inhibition of the neuroprotective Akt pathways, and increased expression of the MAPK-associated p38 and JNK stress pathways. Ultimately, this imbalance is associated with cellular stress, impaired proteostasis, accumulation of -synuclein, and neuronal loss. The GLP-1R agonist exenatide led to restoration of insulin signalling, associated with restoration of Akt signalling and suppression of the MAPK pathways in neurons. GLP-1R agonism reverses the neuronal toxicity associated with the synucleinopathy, through reduction of oxidative stress, improved mitochondrial and lysosomal function, reduced aggregation of -synuclein, and enhanced neuronal viability. GLP-1R agonism further suppresses synuclein induced inflammatory states in glia, leading to neuroprotection through non cell autonomous effects. In the exenatide-PD2 clinical trial, exenatide treatment was associated with clinical improvement in individuals with higher baseline MAPK expression (and thus insulin resistance). Exenatide treatment led to a reduction of -synuclein aggregates, and a reduction in inflammatory cytokine IL-6. Taken together, our patient platform defines the mechanisms of GLP-1R action in neurons and astrocytes, identifies the population likely to benefit from GLP-1R agonism, and highlights the utility of GLP-1R agonism as a disease modifying strategy in synucleinopathies.
]]></description>
<dc:creator>Athauda, D.</dc:creator>
<dc:creator>Evans, J. R.</dc:creator>
<dc:creator>Sanchez, L.</dc:creator>
<dc:creator>Virdi, G.</dc:creator>
<dc:creator>Lopez Garcia, P.</dc:creator>
<dc:creator>Wernick, A.</dc:creator>
<dc:creator>Wagen, A.</dc:creator>
<dc:creator>D'Sa, K.</dc:creator>
<dc:creator>Lachica, J.</dc:creator>
<dc:creator>Strohbuecker, S.</dc:creator>
<dc:creator>Vecchi, G.</dc:creator>
<dc:creator>Leighton, C.</dc:creator>
<dc:creator>Saleeb, R. S.</dc:creator>
<dc:creator>OShaughnessy, J.</dc:creator>
<dc:creator>Toomey, C.</dc:creator>
<dc:creator>Vijiaratnam, N.</dc:creator>
<dc:creator>Girges, C.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Mustapic, M.</dc:creator>
<dc:creator>Ismail, K.</dc:creator>
<dc:creator>Davies, M.</dc:creator>
<dc:creator>Kapogiannis, D.</dc:creator>
<dc:creator>Choi, M. L.</dc:creator>
<dc:creator>Ryten, M.</dc:creator>
<dc:creator>Horrocks, M. H.</dc:creator>
<dc:creator>Greig, N. H.</dc:creator>
<dc:creator>Foltynie, T.</dc:creator>
<dc:creator>Gandhi, S.</dc:creator>
<dc:date>2024-02-28</dc:date>
<dc:identifier>doi:10.1101/2024.02.28.582460</dc:identifier>
<dc:title><![CDATA[GLP-1 receptor agonism ameliorates Parkinsons disease through modulation of neuronal insulin signalling and glial suppression]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.02.28.582561v1?rss=1">
<title>
<![CDATA[
LRRK2 G2019S mutation suppresses differentiation of Th9 and Treg cells via JAK/STAT3 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.02.28.582561v1?rss=1"
</link>
<description><![CDATA[
The Leucine-rich repeat kinase-2 (LRRK2) G2019S mutation, resulting in aberrantly enhanced kinase activity, is one of the well-recognized genetic risk factors in Parkinsons Disease (PD). Increased LRRK2 activity was also observed in immune cells from PD patients. Emerging results have also unveiled an upsurge in -synuclein (-syn)-specific CD4+ T cell responses in PD patients. Given that LRRK2 mutations in PD are germline mutations, there are unmet meets to explore whether LRRK2 G2019S mutation contributes to the pathogenesis of PD via altering CD4+ T-cell functions. To fill this knowledge gap, we generated a new T cell receptor (TCR) transgenic mouse strain bearing LRRK2 G2019S knock-in mutation, OT-II/LRRK2 (Refer to Mut). As CD4+ T cells from OT-II mice specifically recognize ovalbumin, this new strain enables us to explore the impact of LRRK2 G2019S mutation on T-cell functions in an antigen-specific manner. We found that the abundance and proliferation of major immune subsets in spleen tissue from Mut mice are comparable to wild-type (OT-II, Refer to WT) control. However, when we characterized T cell differentiation in these two strains, T cells derived from Mut mice displayed increased Th2 differentiation (IL-4) and decreased Th9 (IL-9) and Treg (Foxp3+ %) differentiation. LRRK2 G2019S mutation significantly altered the expression levels of master transcription factors (TFs) for T cell differentiation. Specifically, Mut T cells displayed an increase in mRNA expression of Gata3 (TF for Th2), a decrease in expression of Irf4 and Foxp3 (TFs for Th9 and Treg, respectively). Mechanistically, LRRK2 mutation decreased IL-9 production and Treg cell population through the JAK/STAT3 signaling. In conclusion, LRRK2 plays a critical role in regulating T cell differentiation, warranting further studies to evaluate the impacts of altered T cell differentiation led by LRRK2 mutation in dopaminergic neuron damages.
]]></description>
<dc:creator>Zheng, N.</dc:creator>
<dc:creator>Jaffery, R.</dc:creator>
<dc:creator>Guerrero, A.</dc:creator>
<dc:creator>Hou, J.</dc:creator>
<dc:creator>Pan, Y.</dc:creator>
<dc:creator>Zhou, F.</dc:creator>
<dc:creator>Chen, S.</dc:creator>
<dc:creator>Xu, C.</dc:creator>
<dc:creator>Egan, N. A.</dc:creator>
<dc:creator>Bohat, R.</dc:creator>
<dc:creator>Chen, K.</dc:creator>
<dc:creator>Schwarzschild, M. A.</dc:creator>
<dc:creator>Chen, X.</dc:creator>
<dc:creator>Peng, W.</dc:creator>
<dc:date>2024-03-03</dc:date>
<dc:identifier>doi:10.1101/2024.02.28.582561</dc:identifier>
<dc:title><![CDATA[LRRK2 G2019S mutation suppresses differentiation of Th9 and Treg cells via JAK/STAT3]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.03.03.583145v1?rss=1">
<title>
<![CDATA[
Deep sequencing of proteotoxicity modifier genes uncovers a Presenilin-2/beta-amyloid-actin genetic risk module shared among alpha-synucleinopathies 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.03.03.583145v1?rss=1"
</link>
<description><![CDATA[
Whether neurodegenerative diseases linked to misfolding of the same protein share genetic risk drivers or whether different protein-aggregation pathologies in neurodegeneration are mechanistically related remains uncertain. Conventional genetic analyses are underpowered to address these questions. Through careful selection of patients based on protein aggregation phenotype (rather than clinical diagnosis) we can increase statistical power to detect associated variants in a targeted set of genes that modify proteotoxicities. Genetic modifiers of alpha-synuclein ([a]S) and beta-amyloid (A{beta}) cytotoxicity in yeast are enriched in risk factors for Parkinsons disease (PD) and Alzheimers disease (AD), respectively. Here, along with known AD/PD risk genes, we deeply sequenced exomes of 430 [a]S/A{beta} modifier genes in patients across alpha-synucleinopathies (PD, Lewy body dementia and multiple system atrophy). Beyond known PD genes GBA1 and LRRK2, rare variants AD genes (CD33, CR1 and PSEN2) and A{beta} toxicity modifiers involved in RhoA/actin cytoskeleton regulation (ARGHEF1, ARHGEF28, MICAL3, PASK, PKN2, PSEN2) were shared risk factors across synucleinopathies. Actin pathology occurred in iPSC synucleinopathy models and RhoA downregulation exacerbated [a]S pathology. Even in sporadic PD, the expression of these genes was altered across CNS cell types. Genome-wide CRISPR screens revealed the essentiality of PSEN2 in both human cortical and dopaminergic neurons, and PSEN2 mutation carriers exhibited diffuse brainstem and cortical synucleinopathy independent of AD pathology. PSEN2 contributes to a common-risk signal in PD GWAS and regulates [a]S expression in neurons. Our results identify convergent mechanisms across synucleinopathies, some shared with AD.
]]></description>
<dc:creator>Nazeen, S.</dc:creator>
<dc:creator>Wang, X.</dc:creator>
<dc:creator>Zielinski, D.</dc:creator>
<dc:creator>Lam, I.</dc:creator>
<dc:creator>Xu, P.</dc:creator>
<dc:creator>Ethier, E.</dc:creator>
<dc:creator>Strom, R.</dc:creator>
<dc:creator>Zanella, C. A.</dc:creator>
<dc:creator>Nithianandam, V.</dc:creator>
<dc:creator>Ritter, D.</dc:creator>
<dc:creator>Henderson, A.</dc:creator>
<dc:creator>Saurat, N.</dc:creator>
<dc:creator>Nutter-Upham, A.</dc:creator>
<dc:creator>Benyamini, H.</dc:creator>
<dc:creator>Copty, J.</dc:creator>
<dc:creator>Ravishankar, S.</dc:creator>
<dc:creator>Morrow, A.</dc:creator>
<dc:creator>Mitchel, J.</dc:creator>
<dc:creator>Neavin, D.</dc:creator>
<dc:creator>Gupta, R.</dc:creator>
<dc:creator>Farbehi, N.</dc:creator>
<dc:creator>Grundman, J.</dc:creator>
<dc:creator>Myers, R. H.</dc:creator>
<dc:creator>Scherzer, C. R.</dc:creator>
<dc:creator>Trojanowski, J. Q.</dc:creator>
<dc:creator>Van Deerlin, V. M.</dc:creator>
<dc:creator>Cooper, A. A.</dc:creator>
<dc:creator>Lee, E. B.</dc:creator>
<dc:creator>Erlich, Y.</dc:creator>
<dc:creator>Lindquist, S.</dc:creator>
<dc:creator>Peng, J.</dc:creator>
<dc:creator>Geschwind, D. H.</dc:creator>
<dc:creator>Powell, J.</dc:creator>
<dc:creator>Studer, L.</dc:creator>
<dc:creator>Feany, M. B.</dc:creator>
<dc:creator>Sunyaev, S. R.</dc:creator>
<dc:creator>Khurana, V.</dc:creator>
<dc:date>2024-03-05</dc:date>
<dc:identifier>doi:10.1101/2024.03.03.583145</dc:identifier>
<dc:title><![CDATA[Deep sequencing of proteotoxicity modifier genes uncovers a Presenilin-2/beta-amyloid-actin genetic risk module shared among alpha-synucleinopathies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.10.12.562142v1?rss=1">
<title>
<![CDATA[
Parkinsonism Sac domain mutation in Synaptojanin-1 affects ciliary properties in iPSC-derived dopaminergic neurons 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.10.12.562142v1?rss=1"
</link>
<description><![CDATA[
Synaptojanin-1 (SJ1) is a major neuronal-enriched PI(4,5)P2 4- and 5-phosphatase implicated in the shedding of endocytic factors during endocytosis. A mutation (R258Q) that impairs selectively its 4-phosphatase activity causes Parkinsonism in humans and neurological defects in mice (SJ1RQKI mice). Studies of these mice showed, besides an abnormal assembly state of endocytic factors at synapses, the presence of dystrophic nerve terminals selectively in a subset of nigro-striatal dopamine (DA)-ergic axons, suggesting a special lability of DA neurons to the impairment of SJ1 function. Here we have further investigated the impact of SJ1 on DA neurons using iPSC-derived SJ1 KO and SJ1RQKI DA neurons and their isogenic controls. In addition to the expected enhanced clustering of endocytic factors in nerve terminals, we observed in both SJ1 mutant neuronal lines increased cilia length. Further analysis of cilia of SJ1RQDA neurons revealed abnormal accumulation of the Ca2+ channel Cav1.3 and of ubiquitin chains, suggesting an impaired clearing of proteins from cilia which may result from an endocytic defect at the ciliary base, where a focal concentration of SJ1 was observed. We suggest that SJ1 may contribute to the control of ciliary protein dynamics in DA neurons, with implications on cilia-mediated signaling.
]]></description>
<dc:creator>Mohd Rafiq, N.</dc:creator>
<dc:creator>Fujise, K.</dc:creator>
<dc:creator>Rosenfeld, M.</dc:creator>
<dc:creator>Xu, P.</dc:creator>
<dc:creator>Wu, Y.</dc:creator>
<dc:creator>De Camilli, P.</dc:creator>
<dc:date>2023-10-13</dc:date>
<dc:identifier>doi:10.1101/2023.10.12.562142</dc:identifier>
<dc:title><![CDATA[Parkinsonism Sac domain mutation in Synaptojanin-1 affects ciliary properties in iPSC-derived dopaminergic neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-10-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.12.13.571527v1?rss=1">
<title>
<![CDATA[
Isolation and Lipidomic Profiling of Neuronal Lipid Droplets: Unveiling the Lipid Landscape for insights into Neurodegenerative Disorders 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.12.13.571527v1?rss=1"
</link>
<description><![CDATA[
Recent advances have expanded the role of lipid droplets (LDs) beyond passive lipid storage, implicating their involvement in various metabolic processes across mammalian tissues. Neuronal LDs, long debated in existence, have been identified in several neural structures, raising questions about their contribution to neurodegenerative disorders. Elucidating the specific chemical makeup of these organelles within neurons is critical for understanding their implication in neural pathologies. This study outlines an improved methodology to stimulate and isolate mature LDs from cultured primary neurons, offering insights into their unique lipid-protein composition. Integrating this method with high-throughput techniques may unveil disease-specific alterations in lipid metabolism, providing avenues for potential therapeutic interventions.
]]></description>
<dc:creator>Kumar, M.</dc:creator>
<dc:creator>Knapp, J.</dc:creator>
<dc:creator>Gupta, K.</dc:creator>
<dc:creator>Ryan, T. A.</dc:creator>
<dc:date>2023-12-14</dc:date>
<dc:identifier>doi:10.1101/2023.12.13.571527</dc:identifier>
<dc:title><![CDATA[Isolation and Lipidomic Profiling of Neuronal Lipid Droplets: Unveiling the Lipid Landscape for insights into Neurodegenerative Disorders]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.02.19.581062v1?rss=1">
<title>
<![CDATA[
Inhibition of Indirect Pathway Activity Causes Abnormal Decision-Making In a Mouse Model of Impulse Control Disorder in Parkinson's Disease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.02.19.581062v1?rss=1"
</link>
<description><![CDATA[
Healthy action selection relies on the coordinated activity of striatal direct and indirect pathway neurons. In Parkinsons disease (PD), in which loss of midbrain dopamine neurons is associated with progressive motor and cognitive deficits, this coordination is disrupted. Dopamine replacement therapy can remediate motor symptoms, but can also cause impulse control disorder (ICD), which is characterized by pathological gambling, hypersexuality, and/or compulsive shopping. The cellular and circuit mechanisms of ICD remain unknown. Here we developed a mouse model of PD/ICD, in which ICD-like behavior was assayed with a delay discounting task. We found that in parkinsonian mice, the dopamine agonist pramipexole drove more pronounced delay discounting, as well as disrupted firing in both direct and indirect pathway neurons. We found that chemogenetic inhibition of indirect pathway neurons in parkinsonian mice drove similar phenotypes. Together, these findings provide a new mouse model and insights into ICD pathophysiology.
]]></description>
<dc:creator>Zhuang, X.</dc:creator>
<dc:creator>Lemak, J.</dc:creator>
<dc:creator>Sridhar, S.</dc:creator>
<dc:creator>Nelson, A. B.</dc:creator>
<dc:date>2024-02-22</dc:date>
<dc:identifier>doi:10.1101/2024.02.19.581062</dc:identifier>
<dc:title><![CDATA[Inhibition of Indirect Pathway Activity Causes Abnormal Decision-Making In a Mouse Model of Impulse Control Disorder in Parkinson's Disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-02-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.03.26.586828v1?rss=1">
<title>
<![CDATA[
Lysosomal storage disease proteo/lipidomic profiling using nMOST links ferritinophagy with mitochondrial iron deficiencies in cells lacking NPC2 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.03.26.586828v1?rss=1"
</link>
<description><![CDATA[
Lysosomal storage diseases (LSDs) comprise [~]50 monogenic disorders marked by the buildup of cellular material in lysosomes, yet systematic global molecular phenotyping of proteins and lipids is lacking. We present a nanoflow-based multi-omic single-shot technology (nMOST) workflow that quantifies HeLa cell proteomes and lipidomes from over two dozen LSD mutants. Global cross-correlation analysis between lipids and proteins identified autophagy defects, notably the accumulation of ferritinophagy substrates and receptors, especially in NPC1-/- and NPC2-/- mutants, where lysosomes accumulate cholesterol. Autophagic and endocytic cargo delivery failures correlated with elevated lyso-phosphatidylcholine species and multi-lamellar structures visualized by cryo-electron tomography. Loss of mitochondrial cristae, MICOS- complex components, and OXPHOS components rich in iron-sulfur cluster proteins in NPC2-/- cells was largely alleviated when iron was provided through the transferrin system. This study reveals how lysosomal dysfunction affects mitochondrial homeostasis and underscores nMOST as a valuable discovery tool for identifying molecular phenotypes across LSDs.
]]></description>
<dc:creator>Harper, J. W.</dc:creator>
<dc:creator>Kraus, F.</dc:creator>
<dc:creator>He, Y.</dc:creator>
<dc:creator>Swarup, S.</dc:creator>
<dc:creator>Overmyer, K. A.</dc:creator>
<dc:creator>Brenner, J.</dc:creator>
<dc:creator>Capitanio, C.</dc:creator>
<dc:creator>Bieber, A.</dc:creator>
<dc:creator>Jen, A.</dc:creator>
<dc:creator>Nightingale, N. M.</dc:creator>
<dc:creator>Anderson, B. J.</dc:creator>
<dc:creator>Lee, C.</dc:creator>
<dc:creator>Paulo, J. A.</dc:creator>
<dc:creator>Smith, I. R.</dc:creator>
<dc:creator>Plitzko, J. M.</dc:creator>
<dc:creator>Schulman, B. A.</dc:creator>
<dc:creator>Wilfling, F.</dc:creator>
<dc:creator>Coon, J. J.</dc:creator>
<dc:date>2024-03-27</dc:date>
<dc:identifier>doi:10.1101/2024.03.26.586828</dc:identifier>
<dc:title><![CDATA[Lysosomal storage disease proteo/lipidomic profiling using nMOST links ferritinophagy with mitochondrial iron deficiencies in cells lacking NPC2]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.03.27.586699v1?rss=1">
<title>
<![CDATA[
Tonic dendritic GABA release by substantia nigra dopaminergic neurons 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.03.27.586699v1?rss=1"
</link>
<description><![CDATA[
Recent studies have demonstrated the importance of extrastriatal dopamine release in the emergence of the network dysfunction underlying motor deficits in Parkinsons disease (PD). To better characterize the actions of dopamine on substantia nigra pars reticulata (SNr) GABAergic neurons, optogenetic and electrophysiological tools were used in ex vivo mouse brain slices to monitor synaptic transmission arising from globus pallidus externa (GPe) neurons. As predicted by previous work, activation of D2 dopamine receptors (D2Rs) suppressed GABA release evoked by stimulation of GPe axons. However, D2R activation also suppressed a tonic, GABAA receptor-mediated inhibition of SNr spiking. D2R-mediated inhibition of tonic GABA release led to a roughly 30% increase in SNr spiking rate. Chemogenetic inhibition of GPe terminals or excitation of astrocytes did not affect tonic GABA release in the SNr. In contrast, chemogenetic inhibition of dopaminergic neurons or knocking down the expression of aldehyde dehydrogenase 1A1 (ALDH1A1) blunted tonic GABAergic signaling. Antagonizing D1 dopamine receptors on the terminals of striatonigral neurons also modestly increased SNr spiking. Lastly, in a progressive mouse model of PD targeting dopaminergic neurons, the tonic inhibition of SNr neurons by GABA release also was lost. Taken together, these observations suggest that dopamine and GABA are co-released by the dendrites of ALDH1A1-expressing dopaminergic neurons that course through the SNr. The co-release of these transmitters could serve to promote movement by making SNr neurons less responsive to phasic activity arising from the indirect pathway circuitry and by lowering basal spiking rates.
]]></description>
<dc:creator>Simmons, D. V.</dc:creator>
<dc:creator>Ding, J.</dc:creator>
<dc:creator>Khakh, B.</dc:creator>
<dc:creator>Awatramani, R. V.</dc:creator>
<dc:creator>Surmeier, D. J.</dc:creator>
<dc:date>2024-03-30</dc:date>
<dc:identifier>doi:10.1101/2024.03.27.586699</dc:identifier>
<dc:title><![CDATA[Tonic dendritic GABA release by substantia nigra dopaminergic neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-03-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.04.05.588321v1?rss=1">
<title>
<![CDATA[
Chronic hyperactivation of midbrain dopamine neurons causes preferential dopamine neuron degeneration 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.04.05.588321v1?rss=1"
</link>
<description><![CDATA[
Parkinsons disease (PD) is characterized by the death of substantia nigra (SNc) dopamine (DA) neurons, but the pathophysiological mechanisms that precede and drive their death remain unknown. The activity of DA neurons is likely altered in PD, but we understand little about if or how chronic changes in activity may contribute to degeneration. To address this question, we developed a chemogenetic (DREADD) mouse model to chronically increase DA neuron activity, and confirmed this increase using ex vivo electrophysiology. Chronic hyperactivation of DA neurons resulted in prolonged increases in locomotor activity during the light cycle and decreases during the dark cycle, consistent with chronic changes in DA release and circadian disturbances. We also observed early, preferential degeneration of SNc projections, recapitulating the PD hallmarks of selective vulnerability of SNc axons and the comparative resilience of ventral tegmental area axons. This was followed by eventual loss of midbrain DA neurons. Continuous DREADD activation resulted in a sustained increase in baseline calcium levels, supporting a role for increased calcium in the neurodegeneration process. Finally, spatial transcriptomics from DREADD mice examining midbrain DA neurons and striatal targets, and cross-validation with human patient samples, provided insights into potential mechanisms of hyperactivity-induced toxicity and PD. Our results thus reveal the preferential vulnerability of SNc DA neurons to increased neural activity, and support a potential role for increased neural activity in driving degeneration in PD.
]]></description>
<dc:creator>Rademacher, K. L.</dc:creator>
<dc:creator>Doric, Z.</dc:creator>
<dc:creator>Haddad, D.</dc:creator>
<dc:creator>Mamaligas, A.</dc:creator>
<dc:creator>Liao, S.-C.</dc:creator>
<dc:creator>Creed, R.</dc:creator>
<dc:creator>Kano, K.</dc:creator>
<dc:creator>Chatterton, Z.</dc:creator>
<dc:creator>Fu, Y.</dc:creator>
<dc:creator>Garcia, J. H.</dc:creator>
<dc:creator>Vance, V.</dc:creator>
<dc:creator>Sei, Y.</dc:creator>
<dc:creator>Kreitzer, A.</dc:creator>
<dc:creator>Halliday, G.</dc:creator>
<dc:creator>Nelson, A.</dc:creator>
<dc:creator>Margolis, E.</dc:creator>
<dc:creator>Nakamura, K. L.</dc:creator>
<dc:date>2024-04-10</dc:date>
<dc:identifier>doi:10.1101/2024.04.05.588321</dc:identifier>
<dc:title><![CDATA[Chronic hyperactivation of midbrain dopamine neurons causes preferential dopamine neuron degeneration]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2021.04.20.440527v1?rss=1">
<title>
<![CDATA[
Transient dopamine neuron activity precedes and encodes the vigor of contralateral movements 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2021.04.20.440527v1?rss=1"
</link>
<description><![CDATA[
Dopamine neurons (DANs) in the substantia nigra pars compacta (SNc) have been related to movement vigor, and loss of these neurons leads to bradykinesia in Parkinsons disease. However, it remains unclear whether DANs encode a general motivation signal or modulate movement kinematics. We imaged activity of SNc DANs in mice trained in a novel operant task which relies on individual forelimb sequences. We uncovered that a similar proportion of SNc DANs increased their activity before ipsi- vs. contralateral sequences. However, the magnitude of this activity was higher for contralateral actions, and was related to contralateral but not ipsilateral sequence length. In contrast, the activity of reward-related DANs, largely distinct from those modulated by movement, was not lateralized. Finally, unilateral dopamine depletion impaired contralateral, but not ipsilateral, sequence length. These results indicate that movement-initiation DANs encode more than a general motivation signal, and invigorate kinematic aspects of contralateral movements.

TeaserTransient activity in substantia nigra compacta dopamine neurons encodes contralateral, but not ipsilateral action vigor.
]]></description>
<dc:creator>Mendonca, M. D.</dc:creator>
<dc:creator>Alves da Silva, J.</dc:creator>
<dc:creator>Hernandez, L. F.</dc:creator>
<dc:creator>Castela, I.</dc:creator>
<dc:creator>Obeso, J.</dc:creator>
<dc:creator>Costa, R. M.</dc:creator>
<dc:date>2021-04-20</dc:date>
<dc:identifier>doi:10.1101/2021.04.20.440527</dc:identifier>
<dc:title><![CDATA[Transient dopamine neuron activity precedes and encodes the vigor of contralateral movements]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2021-04-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.03.18.533302v1?rss=1">
<title>
<![CDATA[
escheR: Unified multi-dimensional visualizations with Gestalt principles 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.03.18.533302v1?rss=1"
</link>
<description><![CDATA[
The creation of effective visualizations is a fundamental component of data analysis. In biomedical research, new challenges are emerging to visualize multi-dimensional data in a 2D space, but current data visualization tools have limited capabilities. To address this problem, we leverage Gestalt principles to improve the design and interpretability of multi-dimensional data in 2D data visualizations, layering aesthetics to display multiple variables. The proposed visualization can be applied to spatially-resolved transcriptomics data, but also broadly to data visualized in 2D space, such as embedding visualizations. We provide an open source R package escheR, which is built off of the state-of-the-art ggplot2 visualization framework and can be seamlessly integrated into genomics toolboxes and workflows.

Availability and implementationThe open source R package escheR is freely available on Bioconductor (bioconductor.org/packages/escheR).
]]></description>
<dc:creator>Guo, B.</dc:creator>
<dc:creator>Hicks, S. C.</dc:creator>
<dc:date>2023-03-23</dc:date>
<dc:identifier>doi:10.1101/2023.03.18.533302</dc:identifier>
<dc:title><![CDATA[escheR: Unified multi-dimensional visualizations with Gestalt principles]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-03-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.06.26.546578v1?rss=1">
<title>
<![CDATA[
SynBot: An open-source image analysis software for automated quantification of synapses 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.06.26.546578v1?rss=1"
</link>
<description><![CDATA[
The formation of precise numbers of neuronal connections, known as synapses, is crucial for brain function. Therefore, synaptogenesis mechanisms have been one of the main focuses of neuroscience. Immunohistochemistry is a common tool for visualizing synapses. Thus, quantifying the numbers of synapses from light microscopy images enables screening the impacts of experimental manipulations on synapse development. Despite its utility, this approach is paired with low throughput analysis methods that are challenging to learn and results are variable between experimenters, especially when analyzing noisy images of brain tissue. We developed an open-source ImageJ-based software, SynBot, to address these technical bottlenecks by automating the analysis. SynBot incorporates the advanced algorithms ilastik and SynQuant for accurate thresholding for synaptic puncta identification, and the code can easily be modified by users. The use of this software will allow for rapid and reproducible screening of synaptic phenotypes in healthy and diseased nervous systems.

MotivationLight microscopy imaging of pre- and post-synaptic proteins from neurons in tissue or in vitro allows for the effective identification of synaptic structures. Previous methods for quantitative analysis of these images were time-consuming, required extensive user training, and the source code could not be easily modified. Here, we describe SynBot, a new open-source tool that automates the synapse quantification process, decreases the requirement for user training, and allows for easy modifications to the code.
]]></description>
<dc:creator>Savage, J. T.</dc:creator>
<dc:creator>Ramirez, J.</dc:creator>
<dc:creator>Risher, W. C.</dc:creator>
<dc:creator>Irala, D.</dc:creator>
<dc:creator>Eroglu, C.</dc:creator>
<dc:date>2023-06-28</dc:date>
<dc:identifier>doi:10.1101/2023.06.26.546578</dc:identifier>
<dc:title><![CDATA[SynBot: An open-source image analysis software for automated quantification of synapses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-06-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.09.14.557673v1?rss=1">
<title>
<![CDATA[
Linking Gba1 E326K mutation to microglia activation and mild age-dependent dopaminergic Neurodegeneration 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.09.14.557673v1?rss=1"
</link>
<description><![CDATA[
Mutations in the GBA1 gene have been identified as a prevalent genetic risk factor for Parkinsons disease (PD). GBA1 mutations impair enzymatic activity, leading to lysosomal dysfunction and elevated levels of -synuclein (-syn). While most research has primarily focused on GBA1s role in promoting synucleinopathy, emerging evidence suggests that neuroinflammation may be a key pathogenic alteration caused by GBA1 deficiency. To examine the molecular mechanism underlying GBA1 deficiency-mediated neuroinflammation, we generated Gba1 E326K knock-in (KI) mice using the CRISPR/Cas9 technology, which is linked to an increased risk of PD and dementia with Lewy bodies (DLB). In the ventral midbrain and hippocampus of 24-month-old Gba1 E326K KI mice, we found a moderate decline in GBA1 enzymatic activity, a buildup of glucosylceramide, and an increase in microglia density. Furthermore, we observed increased levels of pro-inflammatory cytokines and formation of reactive astrocytes in primary microglia and astrocytes, respectively, cultured from Gba1 E326K KI mice following treatment with pathologic -syn preformed fibrils (PFF). Additionally, the gut inoculation of -syn PFF in Gba1 E326K KI mice significantly enhanced the accumulation of Lewy bodies in the dentate gyrus of the hippocampus, accompanied by aggravated neuroinflammation and exacerbated non-motor symptoms. This research significantly enhances our understanding of the Gba1 E326K mutations involvement in neuroinflammation and the cell-to-cell transmission of pathogenic -syn in the brain, thereby opening new therapeutic avenues.
]]></description>
<dc:creator>Kweon, S. H.</dc:creator>
<dc:creator>Ryu, H. G.</dc:creator>
<dc:creator>Park, H.</dc:creator>
<dc:creator>Lee, S.</dc:creator>
<dc:creator>Kim, N.</dc:creator>
<dc:creator>Kwon, S.-H.</dc:creator>
<dc:creator>Ma, S.</dc:creator>
<dc:creator>Kim, S.</dc:creator>
<dc:creator>Ko, H. S.</dc:creator>
<dc:date>2023-09-14</dc:date>
<dc:identifier>doi:10.1101/2023.09.14.557673</dc:identifier>
<dc:title><![CDATA[Linking Gba1 E326K mutation to microglia activation and mild age-dependent dopaminergic Neurodegeneration]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-09-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.11.02.565364v1?rss=1">
<title>
<![CDATA[
LRRK2 kinase activity is necessary for development and regeneration in Nematostella vectensis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.11.02.565364v1?rss=1"
</link>
<description><![CDATA[
BackgroundThe starlet sea anemone, Nematostella vectensis, is an emerging model organism with a high regenerative capacity, which was recently found to possess an orthologue to the human LRRK2 gene (nvLRRK2). The leucine rich repeat kinase 2 (LRRK2) gene, when mutated, is the most common cause of inherited Parkinsons Disease (PD). Its protein product (LRRK2) has implications in a variety of cellular processes, however, the full function of LRRK2 is not well established. Current research is focusing on understanding the function of LRRK2, including both its physiological role as well as its pathobiological underpinnings.

MethodsWe used bioinformatics to determine the cross-species conservation of LRRK2, then applied drugs targeting the kinase activity of LRRK2 to examine its function in development, homeostasis and regeneration in Nematostella vectensis.

ResultsAn in-silico characterization and phylogenetic analysis of nvLRRK2 comparing it to human LRRK2 highlighted key conserved motifs and residues. In vivo analyses inhibiting the kinase function of this enzyme demonstrated a role of nvLRRK2 in development and regeneration of N. vectensis. These findings implicate a developmental role of LRRK2 in Nematostella, adding to the expanding knowledge of its physiological function.

ConclusionsOur work introduces a new model organism with which to study LRRK biology. We show a necessity for LRRK2 in development and regeneration. Given the short generation time, genetic trackability and in vivo imaging capabilities, this work introduces Nematostella vectensis as a new model in which to study genes linked to neurodegenerative diseases such as Parkinsons.
]]></description>
<dc:creator>Holmes, G.</dc:creator>
<dc:creator>Ferguson, S. R.</dc:creator>
<dc:creator>Lewis, P. A.</dc:creator>
<dc:creator>Echeverri, K.</dc:creator>
<dc:date>2023-11-04</dc:date>
<dc:identifier>doi:10.1101/2023.11.02.565364</dc:identifier>
<dc:title><![CDATA[LRRK2 kinase activity is necessary for development and regeneration in Nematostella vectensis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.11.14.567123v1?rss=1">
<title>
<![CDATA[
Development of LRRK2 designed ankyrin-repeat proteins 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.11.14.567123v1?rss=1"
</link>
<description><![CDATA[
Leucine rich repeat kinase 2 (LRRK2) is a large multidomain protein containing two catalytic domains, a kinase and a GTPase, as well protein interactions domains, including a WD40 domain. The association of increased LRRK2 kinase activity with both the familial and sporadic forms of Parkinsons disease (PD) has led to intense interest in determining its cellular function. However, small molecule probes that can bind to LRRK2 and report on or affect its activity are needed. Here, we identified a series of high-affinity LRRK2-binding designed ankyrin-repeat proteins (DARPins). One of these DARPins (E11) bound to the LRRK2 WD40 domain with high affinity. LRRK2 bound to DARPin E11 showed improved behavior on cryo-EM grids, resulting in higher resolution LRRK2 structures. DARPin E11 did not affect the catalytic activity of a truncated form of LRRK2 in vitro but decreased the phosphorylation of Rab8A, a LRRK2 substrate, in cells. We also found that DARPin E11 disrupts the formation of microtubule-associated LRRK2 filaments in cells, which are known to require WD40-based dimerization. Thus, DARPin E11 is a new tool to explore the function and dysfunction of LRRK2 and guide the development of LRRK2 kinase inhibitors that target the WD40 domain instead of the kinase.
]]></description>
<dc:creator>Dederer, V.</dc:creator>
<dc:creator>Sanz Murillo, M.</dc:creator>
<dc:creator>Karasmanis, E. P.</dc:creator>
<dc:creator>Hatch, K. S.</dc:creator>
<dc:creator>Chatterjee, D.</dc:creator>
<dc:creator>Preuss, F.</dc:creator>
<dc:creator>Abdul Azeez, K. R.</dc:creator>
<dc:creator>Nguyen, L. V.</dc:creator>
<dc:creator>Galicia, C.</dc:creator>
<dc:creator>Dreier, B.</dc:creator>
<dc:creator>Pluckthun, A.</dc:creator>
<dc:creator>Versees, W.</dc:creator>
<dc:creator>Mathea, S.</dc:creator>
<dc:creator>Leschziner, A. E.</dc:creator>
<dc:creator>Reck-Peterson, S. L.</dc:creator>
<dc:creator>Knapp, S.</dc:creator>
<dc:date>2023-11-15</dc:date>
<dc:identifier>doi:10.1101/2023.11.14.567123</dc:identifier>
<dc:title><![CDATA[Development of LRRK2 designed ankyrin-repeat proteins]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.01.25.577280v1?rss=1">
<title>
<![CDATA[
Adult-Onset Deletion of ATP13A2 in Mice Induces Progressive Nigrostriatal Pathway Dopaminergic Degeneration and Lysosomal Abnormalities 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.01.25.577280v1?rss=1"
</link>
<description><![CDATA[
Although most cases of Parkinsons disease (PD) are sporadic, mutations in over 20 genes are known to cause heritable forms of PD. A surprising number of familial PD-linked genes and PD risk genes are involved in intracellular trafficking and protein degradation. Recessive loss-of-function mutations in ATP13A2, a lysosomal transmembrane P5B-type ATPase and polyamine exporter, can cause early-onset familial PD. Familial ATP13A2 mutations are also linked to related neurodegenerative diseases, including Kufor-Rakeb syndrome (KRS), hereditary spastic paraplegias (HSPs), neuronal ceroid lipofuscinosis, and amyotrophic lateral sclerosis (ALS). Given the severe effects of ATP13A2 mutations in humans, it is surprising that ATP13A2 knockout (KO) mice fail to exhibit neurodegeneration even at advanced ages. This discrepancy between human subjects and rodents makes it challenging to study the neuropathological effects of ATP13A2 loss in vivo. Germline deletion of ATP13A2 in rodents may trigger the upregulation of compensatory pathways during embryonic development that mask the full neurotoxic effects of ATP13A2 loss in the brain. To explore this idea, we selectively deleted ATP13A2 in the adult mouse brain by the unilateral delivery of an AAV-Cre vector into the substantia nigra of young adult mice carrying conditional loxP-flanked ATP13A2 KO alleles. We observe a progressive loss of striatal dopaminergic nerve terminals at 3 and 10 months after AAV-Cre delivery. Cre-injected mice also exhibit robust dopaminergic neuronal degeneration in the substantia nigra at 10 months. Adult-onset ATP13A2 KO also recreates many of the phenotypes observed in aged germline ATP13A2 KO mice, including lysosomal abnormalities, p62-positive inclusions, and neuroinflammation. Our study demonstrates that the adult-onset homozygous deletion of ATP13A2 in the nigrostriatal pathway produces robust and progressive dopaminergic neurodegeneration that serves as a useful in vivo model of ATP13A2-related neurodegenerative diseases.
]]></description>
<dc:creator>Erb, M. L.</dc:creator>
<dc:creator>Sipple, K.</dc:creator>
<dc:creator>Levine, N.</dc:creator>
<dc:creator>Chen, X.</dc:creator>
<dc:creator>Moore, D. J.</dc:creator>
<dc:date>2024-01-26</dc:date>
<dc:identifier>doi:10.1101/2024.01.25.577280</dc:identifier>
<dc:title><![CDATA[Adult-Onset Deletion of ATP13A2 in Mice Induces Progressive Nigrostriatal Pathway Dopaminergic Degeneration and Lysosomal Abnormalities]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-01-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.04.02.587827v1?rss=1">
<title>
<![CDATA[
SARM1 activation induces reversible mitochondrial dysfunction and can be prevented in human neurons by antisense oligonucleotides 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.04.02.587827v1?rss=1"
</link>
<description><![CDATA[
SARM1 is a key regulator of a conserved program of axon degeneration increasingly linked to human neurodegenerative diseases. Pathological SARM1 activation causes rapid NAD consumption, disrupting cellular homeostasis and leading to axon degeneration. In this study, we develop antisense oligonucleotides targeting human SARM1, demonstrating robust neuroprotection against morphological, metabolic, and mitochondrial impairment in human iPSC-derived dopamine neurons induced by the lethal neurotoxin vacor, a potent SARM1 activator. Furthermore, our findings reveal that axon fragmentation can be prevented, and mitochondrial dysfunction reversed using the NAD precursor nicotinamide, a form of vitamin B3, even after SARM1 activation has occurred, when neurons are already unhealthy. This research identifies ASOs as a promising therapeutic strategy to block SARM1, and provides an extensive characterisation and further mechanistic insights that demonstrate the reversibility of SARM1 toxicity in human neurons. It also identifies the SARM1 activator vacor as a specific and reversible neuroablative agent in human neurons.
]]></description>
<dc:creator>Loreto, A.</dc:creator>
<dc:creator>Cramb, K. M. L.</dc:creator>
<dc:creator>McDermott, L. A.</dc:creator>
<dc:creator>Antoniou, C.</dc:creator>
<dc:creator>Cirilli, I.</dc:creator>
<dc:creator>Caiazza, M. C.</dc:creator>
<dc:creator>Merlini, E.</dc:creator>
<dc:creator>Arthur-Farra, P.</dc:creator>
<dc:creator>Mock, E. D.</dc:creator>
<dc:creator>Zhao, H. T.</dc:creator>
<dc:creator>Bennett, D. L.</dc:creator>
<dc:creator>Orsomando, G.</dc:creator>
<dc:creator>Coleman, M. P.</dc:creator>
<dc:creator>Wade-Martins, R.</dc:creator>
<dc:date>2024-04-03</dc:date>
<dc:identifier>doi:10.1101/2024.04.02.587827</dc:identifier>
<dc:title><![CDATA[SARM1 activation induces reversible mitochondrial dysfunction and can be prevented in human neurons by antisense oligonucleotides]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.04.22.590542v1?rss=1">
<title>
<![CDATA[
Gut-Initiated Alpha Synuclein Fibrils Drive Parkinson's Disease Phenotypes: Temporal Mapping of non-Motor Symptoms and REM Sleep Behavior Disorder 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.04.22.590542v1?rss=1"
</link>
<description><![CDATA[
Parkinsons disease (PD) is characterized by progressive motor as well as less recognized non-motor symptoms that arise often years before motor manifestation, including sleep and gastrointestinal disturbances. Despite the heavy burden on the patients quality of life, these non-motor manifestations are poorly understood. To elucidate the temporal dynamics of the disease, we employed a mouse model involving injection of alpha-synuclein (Syn) pre-formed fibrils (PFF) in the duodenum and antrum as a gut-brain model of Parkinsonism. Using anatomical mapping of Syn-PFF propagation and behavioral and physiological characterizations, we unveil a correlation between post-injection time the temporal dynamics of Syn propagation and non-motor/motor manifestations of the disease. We highlight the concurrent presence of Syn aggregates in key brain regions, expressing acetylcholine or dopamine, involved in sleep duration, wakefulness, and particularly REM-associated atonia corresponding to REM behavioral disorder-like symptoms. This study presents a novel and in-depth exploration into the multifaceted nature of PD, unraveling the complex connections between -synucleinopathies, gut-brain connectivity, and the emergence of non-motor phenotypes.

One Sentence SummaryPropagation of Syn from the gut-to-the-brain induces RBD and several non-motor and motor phenotypes of Parkinsons disease.
]]></description>
<dc:creator>Dautan, D.</dc:creator>
<dc:creator>Paslawski, W.</dc:creator>
<dc:creator>Montejo, S.</dc:creator>
<dc:creator>Doyon, D.</dc:creator>
<dc:creator>Marangiu, R.</dc:creator>
<dc:creator>Kaplitt, M. G.</dc:creator>
<dc:creator>Chen, R.</dc:creator>
<dc:creator>Dawson, V. L.</dc:creator>
<dc:creator>Zhang, X.</dc:creator>
<dc:creator>Dawson, T. M.</dc:creator>
<dc:creator>Svenningsson, P.</dc:creator>
<dc:date>2024-04-26</dc:date>
<dc:identifier>doi:10.1101/2024.04.22.590542</dc:identifier>
<dc:title><![CDATA[Gut-Initiated Alpha Synuclein Fibrils Drive Parkinson's Disease Phenotypes: Temporal Mapping of non-Motor Symptoms and REM Sleep Behavior Disorder]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.05.09.593320v1?rss=1">
<title>
<![CDATA[
Reward perseveration is shaped by GABAA-mediated dopamine pauses 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.05.09.593320v1?rss=1"
</link>
<description><![CDATA[
When animals unexpectedly fail, their dopamine neurons undergo phasic inhibition that canonically drives extinction learning--a cognitive-flexibility mechanism for discarding outdated strategies. However, the existing evidence equates natural and artificial phasic inhibition, despite their spatiotemporal differences. Addressing this gap, we targeted a GABAA-receptor antagonist precisely to dopamine neurons, yielding three unexpected findings. First, this intervention blocked natural phasic inhibition selectively, leaving tonic activity unaffected. Second, blocking natural phasic inhibition accelerated extinction learning--opposite to canonical mechanisms. Third, our approach selectively benefitted perseverative mice, restoring rapid extinction without affecting new reward learning. Our findings reveal that extinction learning is rapid by default and slowed by natural phasic inhibition--challenging foundational learning theories, while delineating a synaptic mechanism and therapeutic target for cognitive rigidity.
]]></description>
<dc:creator>Burwell, S. C. V.</dc:creator>
<dc:creator>Yan, H.</dc:creator>
<dc:creator>Lim, S. S. X.</dc:creator>
<dc:creator>Shields, B. C.</dc:creator>
<dc:creator>TADROSS, M. R.</dc:creator>
<dc:date>2024-05-09</dc:date>
<dc:identifier>doi:10.1101/2024.05.09.593320</dc:identifier>
<dc:title><![CDATA[Reward perseveration is shaped by GABAA-mediated dopamine pauses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.04.26.591350v1?rss=1">
<title>
<![CDATA[
A comparative analysis of Parkinson's disease and inflammatory bowel disease gut microbiomes highlights shared depletions in key butyrate-producing bacteria 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.04.26.591350v1?rss=1"
</link>
<description><![CDATA[
Epidemiological studies reveal that a diagnosis of inflammatory bowel disease (IBD) is associated with an increased risk of developing Parkinsons disease (PD). The presence of gut dysbiosis has been documented in both PD and IBD patients, however it is currently unknown how alterations in the gut microbiome may contribute to the epidemiological link between both diseases. To identify shared and distinct features of the PD and IBD microbiome, we performed the first joint analysis of 54 PD, 26 IBD, and 16 healthy control gut metagenomes recruited from clinics at the University of Florida, and directly compared the gut microbiomes from PD and IBD persons. Larger, publicly available PD and IBD metagenomic datasets were also analyzed to validate and extend our findings. Depletions in short-chain fatty acid (SCFA) producing bacteria, including Roseburia intestinalis, Faecalibacterium prausnitzii, Anaerostipes hadrus, and Eubacterium rectale, as well as depletions in SCFA synthesis pathways, were demonstrated across PD and IBD datasets. We posit that direct comparison of PD and IBD gut microbiomes will be important in identifying features within the IBD gut which may be associated with PD. The data revealed a consistent depletion in SCFA-producing bacteria across both PD and IBD, suggesting that loss of these microbes may influence the pathophysiology of both disease states.
]]></description>
<dc:creator>Krueger, M. E.</dc:creator>
<dc:creator>Boles, J. S.</dc:creator>
<dc:creator>Simon, Z. D.</dc:creator>
<dc:creator>Alvarez, S. D.</dc:creator>
<dc:creator>McFarland, N. R.</dc:creator>
<dc:creator>Okun, M. S.</dc:creator>
<dc:creator>Zimmermann, E. M.</dc:creator>
<dc:creator>Forsmark, C. E.</dc:creator>
<dc:creator>Tansey, M. G.</dc:creator>
<dc:date>2024-04-29</dc:date>
<dc:identifier>doi:10.1101/2024.04.26.591350</dc:identifier>
<dc:title><![CDATA[A comparative analysis of Parkinson's disease and inflammatory bowel disease gut microbiomes highlights shared depletions in key butyrate-producing bacteria]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-04-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.05.09.593334v1?rss=1">
<title>
<![CDATA[
Mouse α-synuclein fibrils are structurally and functionally distinct from human fibrils associated with Lewy body diseases 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.05.09.593334v1?rss=1"
</link>
<description><![CDATA[
Short AbstractThe intricate process of -synuclein aggregation and fibrillization hold pivotal roles in Parkinsons disease (PD) and multiple system atrophy (MSA). While mouse -synuclein can fibrillize in vitro, whether these fibrils commonly used in research to induce this process or form can reproduce structures in the human brain remains unknown. Here we report the first atomic structure of mouse -synuclein fibrils, which was solved in parallel by two independent teams. The structure shows striking similarity to MSA-amplified and PD-associated E46K fibrils. However, mouse -synuclein fibrils display altered packing arrangements, reduced hydrophobicity, heightened fragmentation sensitivity, and evoke only weak immunological responses. Furthermore, mouse -synuclein fibrils exhibit exacerbated pathological spread in neurons and humanized -synuclein mice. These findings provide new insights into the structural underpinnings of -synuclein pathogenicity and emphasize a need to reassess the role of mouse -synuclein fibrils in the development of related diagnostic probes and therapeutic interventions.
]]></description>
<dc:creator>Sokratian, A.</dc:creator>
<dc:creator>Zhou, Y.</dc:creator>
<dc:creator>Tatli, M.</dc:creator>
<dc:creator>Burbidge, K. J.</dc:creator>
<dc:creator>Xu, E.</dc:creator>
<dc:creator>Viverette, E.</dc:creator>
<dc:creator>Duda, A. M.</dc:creator>
<dc:creator>Yuan, Y.</dc:creator>
<dc:creator>Li, H.</dc:creator>
<dc:creator>Strader, S.</dc:creator>
<dc:creator>Patel, N.</dc:creator>
<dc:creator>Shiell, L.</dc:creator>
<dc:creator>Malankhanova, T.</dc:creator>
<dc:creator>Chen, O.</dc:creator>
<dc:creator>Mazzulli, J. R.</dc:creator>
<dc:creator>Perera, L.</dc:creator>
<dc:creator>Stahlberg, H.</dc:creator>
<dc:creator>Borgnia, M. J.</dc:creator>
<dc:creator>Bartesaghi, A.</dc:creator>
<dc:creator>Lashuel, H.</dc:creator>
<dc:creator>West, A.</dc:creator>
<dc:date>2024-05-09</dc:date>
<dc:identifier>doi:10.1101/2024.05.09.593334</dc:identifier>
<dc:title><![CDATA[Mouse α-synuclein fibrils are structurally and functionally distinct from human fibrils associated with Lewy body diseases]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.05.15.594309v1?rss=1">
<title>
<![CDATA[
Genetic screening and metabolomics identify glial adenosine metabolism as a therapeutic target in Parkinson's disease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.05.15.594309v1?rss=1"
</link>
<description><![CDATA[
Parkinsons disease (PD) is the second most common neurodegenerative disorder and lacks disease-modifying therapies. We developed a Drosophila model for identifying novel glial-based therapeutic targets for PD. In this model, human -synuclein is expressed in neurons and individual genes are independently knocked down in glia. We performed a forward genetic screen, knocking down the entire Drosophila kinome in glia in -synuclein expressing flies. Among the top hits were five genes (Ak1, Ak6, Adk1, Adk2, and awd) involved in adenosine metabolism. Knockdown of each gene improved locomotor dysfunction, rescued neurodegeneration, and increased brain adenosine levels. We determined that the mechanism of neuroprotection involves adenosine itself, as opposed to a downstream metabolite. We dove deeper into the mechanism for one gene, Ak1, finding rescue of dopaminergic neuron loss, - synuclein aggregation, and bioenergetic dysfunction after glial Ak1 knockdown. We performed metabolomics in Drosophila and in human PD patients, allowing us to comprehensively characterize changes in purine metabolism and identify potential biomarkers of dysfunctional adenosine metabolism in people. These experiments support glial adenosine as a novel therapeutic target in PD.
]]></description>
<dc:creator>Sodders, M. J.</dc:creator>
<dc:creator>Avila-Pacheco, J.</dc:creator>
<dc:creator>Okorie, E. C.</dc:creator>
<dc:creator>Shen, M.</dc:creator>
<dc:creator>Kumari, N.</dc:creator>
<dc:creator>Marathi, A.</dc:creator>
<dc:creator>Lakhani, M.</dc:creator>
<dc:creator>Bullock, K.</dc:creator>
<dc:creator>Pierce, K.</dc:creator>
<dc:creator>Dennis, C.</dc:creator>
<dc:creator>Jeanfavre, S.</dc:creator>
<dc:creator>Sarkar, S.</dc:creator>
<dc:creator>Scherzer, C. R.</dc:creator>
<dc:creator>Clish, C.</dc:creator>
<dc:creator>Olsen, A. L.</dc:creator>
<dc:date>2024-05-15</dc:date>
<dc:identifier>doi:10.1101/2024.05.15.594309</dc:identifier>
<dc:title><![CDATA[Genetic screening and metabolomics identify glial adenosine metabolism as a therapeutic target in Parkinson's disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.05.17.594681v1?rss=1">
<title>
<![CDATA[
Tagless LysoIP method for molecular profiling of lysosomal content in clinical samples 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.05.17.594681v1?rss=1"
</link>
<description><![CDATA[
Lysosomes are implicated in a wide spectrum of human diseases including monogenic lysosomal storage disorders (LSDs), age-associated neurodegeneration and cancer. Profiling lysosomal content using tag-based lysosomal immunoprecipitation (LysoTagIP) in cell and animal models allowed major discoveries in the field, however studying lysosomal dysfunction in human patients remains challenging. Here, we report the development of the "tagless LysoIP method" to enable rapid enrichment of lysosomes, via immunoprecipitation, using the endogenous integral lysosomal membrane protein TMEM192, directly from clinical samples and human cell lines (e.g. induced Pluripotent Stem Cell (iPSCs) derived neurons). Isolated lysosomes are intact and suitable for subsequent multimodal omics analyses. To validate our approach, we employed the tagless LysoIP to enrich lysosomes from peripheral blood mononuclear cells (PBMCs) derived from fresh blood from patients with CLN3 disease, a neurodegenerative LSD. Metabolic profiling of isolated lysosomes showed massive accumulation of glycerophosphodiesters (GPDs) in patients lysosomes. Interestingly, a patient with a milder phenotype and genotype displayed lower accumulation of lysosomal GPDs, consistent with their potential role as disease biomarkers. Altogether, the tagless LysoIP provides a framework to study native lysosomes from patient samples, identify novel biomarkers and discover human-relevant disease mechanisms.
]]></description>
<dc:creator>Saarela, D.</dc:creator>
<dc:creator>Lis, P.</dc:creator>
<dc:creator>Gomes, S.</dc:creator>
<dc:creator>Nirujogi, R. S.</dc:creator>
<dc:creator>Dong, W.</dc:creator>
<dc:creator>Rawat, E.</dc:creator>
<dc:creator>Glendinning, S.</dc:creator>
<dc:creator>Zeneviciute, K.</dc:creator>
<dc:creator>Bagnoli, E.</dc:creator>
<dc:creator>Fasimoye, R.</dc:creator>
<dc:creator>Lin, C.</dc:creator>
<dc:creator>Nyame, K.</dc:creator>
<dc:creator>Boros, F. A.</dc:creator>
<dc:creator>Zunke, F.</dc:creator>
<dc:creator>Lamoliatte, F.</dc:creator>
<dc:creator>Elshani, S.</dc:creator>
<dc:creator>Jaconnelli, M.</dc:creator>
<dc:creator>Jans, J. J. M.</dc:creator>
<dc:creator>Huisman, M. A.</dc:creator>
<dc:creator>Posern, C.</dc:creator>
<dc:creator>Westermann, L. M.</dc:creator>
<dc:creator>Schulz, A.</dc:creator>
<dc:creator>van Hasselt, P. M.</dc:creator>
<dc:creator>Alessi, D. R.</dc:creator>
<dc:creator>Abu-Remaileh, M.</dc:creator>
<dc:creator>Sammler, E.</dc:creator>
<dc:date>2024-05-19</dc:date>
<dc:identifier>doi:10.1101/2024.05.17.594681</dc:identifier>
<dc:title><![CDATA[Tagless LysoIP method for molecular profiling of lysosomal content in clinical samples]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.07.26.501466v1?rss=1">
<title>
<![CDATA[
Cell type directed design of synthetic enhancers 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.07.26.501466v1?rss=1"
</link>
<description><![CDATA[
Transcriptional enhancers act as docking stations for combinations of transcription factors and thereby regulate spatiotemporal activation of their target genes. A single enhancer, of a few hundred base pairs in length, can autonomously and independently of its location and orientation drive cell-type specific expression of a gene or transgene. It has been a long-standing goal in the field to decode the regulatory logic of an enhancer and to understand the details of how spatiotemporal gene expression is encoded in an enhancer sequence. Recently, deep learning models have yielded unprecedented insight into the enhancer code, and well-trained models are reaching a level of understanding that may be close to complete. As a consequence, we hypothesized that deep learning models can be used to guide the directed design of synthetic, cell type specific enhancers, and that this process would allow for a detailed tracing of all enhancer features at nucleotide-level resolution. Here we implemented and compared three different design strategies, each built on a deep learning model: (1) directed sequence evolution; (2) directed iterative motif implanting; and (3) generative design. We evaluated the function of fully synthetic enhancers to specifically target Kenyon cells in the fruit fly brain using transgenic animals. We then exploited this concept further by creating "dual-code" enhancers that target two cell types, and minimal enhancers smaller than 50 base pairs that are fully functional. By examining the trajectories followed during state space searches towards functional enhancers, we could accurately define the enhancer code as the optimal strength, combination, and relative distance of TF activator motifs, and the absence of TF repressor motifs. Finally, we applied the same three strategies to successfully design human enhancers. In conclusion, enhancer design guided by deep learning leads to better understanding of how enhancers work and shows that their code can be exploited to manipulate cell states.
]]></description>
<dc:creator>Taskiran, I. I.</dc:creator>
<dc:creator>Spanier, K. I.</dc:creator>
<dc:creator>Christiaens, V.</dc:creator>
<dc:creator>Mauduit, D.</dc:creator>
<dc:creator>Aerts, S.</dc:creator>
<dc:date>2022-07-27</dc:date>
<dc:identifier>doi:10.1101/2022.07.26.501466</dc:identifier>
<dc:title><![CDATA[Cell type directed design of synthetic enhancers]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-07-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.08.24.554646v1?rss=1">
<title>
<![CDATA[
Neither alpha-synuclein-preformed fibrils derived from patients with GBA1 mutations nor the host murine genotype significantly influence seeding efficacy in the mouse olfactory bulb. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.08.24.554646v1?rss=1"
</link>
<description><![CDATA[
Parkinsons disease (PD) is a neurodegenerative disease characterized by progressive motor symptoms and alpha-synuclein (syn) aggregation in the nervous system. For unclear reasons, PD patients with certain GBA mutations (GBA-PD) have a more aggressive clinical progression. Two testable hypotheses that can potentially account for this phenomenon are that GBA1 mutations promote syn spread or drive the generation of highly pathogenic syn polymorphs (i.e., strains). We tested these hypotheses by treating homozygous GBA1 D409V knockin (KI) mice with human -syn-preformed fibrils (PFFs) and treating wild-type mice (WT) with several syn-PFF polymorphs amplified from brain autopsy samples collected from patients with idiopathic PD and GBA-PD patients with either homozygous or heterozygous GBA1 mutations. Robust phosphorylated-syn (PSER129) positive pathology was observed at the injection site (i.e., the olfactory bulb granular layer) and throughout the brain six months following PFF injection. The PFF seeding efficiency and degree of spread were similar regardless of the mouse genotype or PFF polymorphs. We found that PFFs amplified from the human brain, regardless of patient genotype, were generally more effective seeders than wholly synthetic PFFs (i.e., non-amplified); however, PFF concentration differed between these two studies, and this might also account for the observed differences. To investigate whether the molecular composition of pathology differed between different seeding conditions, we permed Biotinylation by Antibody Recognition on PSER129 (BAR-PSER129). We found that for BAR-PSER129, the endogenous PSER129 pool dominated identified interactions, and thus, very few potential interactions were explicitly identified for seeded pathology. However, we found Dctn2 interaction was shared across all PFF conditions, and Nckap1 and Ap3b2 were unique to PFFs amplified from GBA-PD brains of heterozygous mutation carriers. In conclusion, both the genotype and syn strain had little effect on overall seeding efficacy and global PSER129-interactions.
]]></description>
<dc:creator>Killinger, B.</dc:creator>
<dc:creator>Kordower, J. H.</dc:creator>
<dc:creator>Walton, S.</dc:creator>
<dc:creator>Fenyi, A.</dc:creator>
<dc:creator>Tittle, T.</dc:creator>
<dc:creator>Choi, S.</dc:creator>
<dc:creator>Sidransky, E.</dc:creator>
<dc:creator>Paul, G.</dc:creator>
<dc:creator>Melki, R.</dc:creator>
<dc:date>2023-08-25</dc:date>
<dc:identifier>doi:10.1101/2023.08.24.554646</dc:identifier>
<dc:title><![CDATA[Neither alpha-synuclein-preformed fibrils derived from patients with GBA1 mutations nor the host murine genotype significantly influence seeding efficacy in the mouse olfactory bulb.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-08-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.11.20.567854v1?rss=1">
<title>
<![CDATA[
Aggregation inhibits alpha-synuclein dephosphorylation resulting in the observation of pathological enrichment 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.11.20.567854v1?rss=1"
</link>
<description><![CDATA[
Alpha-synuclein (syn) is an intrinsically disordered protein that aggregates in the brain in several neurodegenerative diseases collectively called synucleinopathies. Phosphorylation of syn at serine 129 (PSER129) was considered rare in the healthy human brain but is enriched in pathological syn aggregates and is used as a specific marker for disease inclusions. However, recent observations challenge this assumption by demonstrating that PSER129 results from neuronal activity and can be readily detected in the non-diseased mammalian brain. Here, we investigated experimental conditions under which two distinct PSER129 pools, namely endogenous-PSER129 and aggregated-PSER129, could be detected and differentiated in the mammalian brain. Results showed that in the wild-type (WT) mouse brain, perfusion fixation conditions greatly influenced the detection of endogenous-PSER129, with endogenous-PSER129 being nearly undetectable after delayed perfusion fixation (30-minute and 1-hour postmortem interval). Exposure to anesthetics (e.g., Ketamine or xylazine) before perfusion did not significantly influence endogenous-PSER129 detection or levels. In situ, non-specific phosphatase calf alkaline phosphatase (CIAP) selectively dephosphorylated endogenous-PSER129 while syn preformed fibril (PFF)-seeded aggregates and genuine disease aggregates (Lewy pathology and Papp-Lantos bodies in Parkinsons disease and multiple systems atrophy brain, respectively) were resistant to CIAP-mediated dephosphorylation. The phosphatase resistance of aggregates was abolished by sample denaturation, and CIAP-resistant PSER129 was closely associated with proteinase K (PK)-resistant syn (i.e., a marker of aggregation). CIAP pretreatment allowed for highly specific detection of seeded syn aggregates in a mouse model that accumulates non-aggregated-PSER129. We conclude that syn aggregates are impervious to phosphatases, and CIAP pretreatment increases detection specificity for aggregated-PSER129, particularly in well-preserved biological samples (e.g., perfusion fixed or flash-frozen mammalian tissues) where there is a high probability of interference from endogenous-PSER129. Our findings have important implications for the mechanism of PSER129-accumulation in the synucleinopathy brain and provide a simple experimental method to differentiate endogenous-from aggregated PSER129.

Significance StatementPhosphorylated alpha-synuclein (PSER129) was widely regarded as a sensitive, specific marker for pathological aggregates in synucleinopathies until recent data demonstrated that PSER129 is abundant in the healthy mammalian nervous system and results from normal neuronal activity. Differentiating pathological (i.e., aggregated PSER129) and biological (non-aggregated PSER129) has thus become of critical importance to the field. Here, we describe our discovery that aggregated-PSER129 is impervious to enzymatic dephosphorylation. We leverage this discovery to develop a technique (CIAP-PSER129) to detect normal or pathological PSER129 selectively. Our technique allowed us to unambiguously differentiate pathological inclusions in brain regions and mouse models where excessive non-aggregated PSER129 severely limits the sensitivity of aggregate detection. CIAP-PSER129 is nondestructive and compatible with most downstream assays, including mass spectrometry-based peptide identification. These findings have important implications and utility for the synucleinopathy field and may have applicability to other neuropathological proteins (e.g., tau).
]]></description>
<dc:creator>Choi, S. G.</dc:creator>
<dc:creator>Prada, D. A. G.</dc:creator>
<dc:creator>Tittle, T.</dc:creator>
<dc:creator>Kordower, J. H.</dc:creator>
<dc:creator>Melki, R.</dc:creator>
<dc:creator>Killinger, B. A.</dc:creator>
<dc:date>2023-11-21</dc:date>
<dc:identifier>doi:10.1101/2023.11.20.567854</dc:identifier>
<dc:title><![CDATA[Aggregation inhibits alpha-synuclein dephosphorylation resulting in the observation of pathological enrichment]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-11-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.05.20.594995v1?rss=1">
<title>
<![CDATA[
Impaired Cortico-Amygdala Functional Connectivity Can Drive Social Behavior Deficits in Synucleinopathies 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.05.20.594995v1?rss=1"
</link>
<description><![CDATA[
The small molecule protein -synuclein forms insoluble aggregates in a group of neurological disorders, including Parkinsons disease and dementia with Lewy bodies (DLB), which are collectively called synucleinopathies. In PD and DLB, the amygdala has been identified as a particularly susceptible region in the brain for the deposition of Lewy-like -synuclein aggregates. Though -synuclein aggregation is closely associated with neurodegeneration, there is a poor correlation between neurodegeneration in the amygdala and the clinical features of PD/DLB. We hypothesize that, prior to neurodegeneration, -synuclein aggregation disrupts functional cortical modulation of the amygdala circuits, leading to emotion dysregulation in synucleinopathies. In the present study, we combined electrophysiology, optogenetics, mouse model of synucleinopathies, and behavioral analysis to test this hypothesis. Using an -synuclein preformed fibrils (PFFs)-based mouse model of synucleinopathies, we reported dynamic changes in the levels of -synuclein pathology in the basolateral amygdala (BLA). Such dynamic changes of pathology associated with a decreased cortico-BLA connection strength prior to a significant loss of cortical axon terminals. In parallel to the reduced cortico-BLA connection, PFFs-injected mice manifested impaired social preference behavior. The impaired sociability of PFFs-injected mice could be rescued by chemogenetic stimulation of cortico-BLA inputs. Altogether, we presented a series of evidence to delineate key circuit events associated with -synuclein pathology development in the amygdala circuits. The present work highlights the necessity of a thorough investigation of functional consequences of -synuclein aggregation to advance our understand of pathophysiology of synucleinopathies and development of effective therapies.
]]></description>
<dc:creator>Zhou, W.</dc:creator>
<dc:creator>Daniels, S.</dc:creator>
<dc:creator>Singh, V.</dc:creator>
<dc:creator>Menard, M.</dc:creator>
<dc:creator>Galvis, M. E.</dc:creator>
<dc:creator>Chu, H.-Y.</dc:creator>
<dc:date>2024-05-20</dc:date>
<dc:identifier>doi:10.1101/2024.05.20.594995</dc:identifier>
<dc:title><![CDATA[Impaired Cortico-Amygdala Functional Connectivity Can Drive Social Behavior Deficits in Synucleinopathies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.05.23.595566v1?rss=1">
<title>
<![CDATA[
Decreased cellular excitability of pyramidal tract neurons in primary motor cortex leads to paradoxically increased network activity in simulated parkinsonian motor cortex 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.05.23.595566v1?rss=1"
</link>
<description><![CDATA[
Primary motor cortex (M1) layer 5B pyramidal tract (PT5B) neurons develop intrinsic pathology in rodent and primate Parkinsons disease (PD) models. We used computer simulation to predict how decreased PT5B neuron excitability, identified with current injection in vitro, would change activity patterns of the M1 network. Using NEURON/NetPyNE, we implemented computer simulations of PT5B neurons based on control and 6-OHDA-treated mouse slice data. Parkinsonian PT5B neurons, in an otherwise unmodified simulated M1 network, produced major changes in LFP oscillatory power: an order of magnitude increase in beta band power around 15 Hz in the rest state. This demonstrated that relatively small changes in PT5B neuron excitability would alter oscillatory patterns of activity throughout the M1 circuit, increasing beta band power, a signature of PD pathophysiology. Dysfunction in PT5B neurons, the final-common-pathway to brainstem and spinal cord, provides a new target to treat PD motor symptoms.
]]></description>
<dc:creator>Doherty, D. W.</dc:creator>
<dc:creator>Chen, L.</dc:creator>
<dc:creator>Smith, Y.</dc:creator>
<dc:creator>Wichmann, T.</dc:creator>
<dc:creator>Chu, H.-y.</dc:creator>
<dc:creator>Lytton, W. W.</dc:creator>
<dc:date>2024-05-23</dc:date>
<dc:identifier>doi:10.1101/2024.05.23.595566</dc:identifier>
<dc:title><![CDATA[Decreased cellular excitability of pyramidal tract neurons in primary motor cortex leads to paradoxically increased network activity in simulated parkinsonian motor cortex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.05.28.596142v1?rss=1">
<title>
<![CDATA[
Functional efficacy of the MAO-B inhibitor safinamide in murine substantia nigra pars compacta dopaminergic neurons in vitro: a comparative study with tranylcypromine 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.05.28.596142v1?rss=1"
</link>
<description><![CDATA[
Safinamide (SAF) is currently used to treat Parkinsons disease (PD) symptoms based on its theoretical ability to potentiate the dopamine (DA) signal, blocking monoamine oxidase (MAO) B. The present work aims to highlight the functional relevance of SAF as an enhancer of the DA signal, by evaluating its ability to prolong recovery from DA-mediated firing inhibition of DAergic neurons of the substantia nigra pars compacta (SNpc), compared to another MAO antagonist, tranylcypromine (TCP). Using multielectrode array (MEA) and single electrode extracellular recordings of spontaneous spikes from presumed SNpc DAergic cells in vitro, we show that SAF (30 uM) mildly prolongs the DA-mediated firing inhibition, as opposed to the profound effect of TCP (10 uM). In patch-clamp recordings, we found that SAF (30 uM) significantly reduced the number of spikes evoked by depolarizing currents in SNpc DAergic neurons, in a sulpiride (1 uM) independent manner. According to our results, SAF marginally potentiates the DA signal in SNpc DAergic neurons, while exerting an inhibitory effect on the postsynaptic excitability acting on membrane conductances. Thus, we propose that the therapeutic effects of SAF in PD patients partially depends on MAO inhibition, while other MAO-independent sites of action could be more relevant.
]]></description>
<dc:creator>Zarrilli, B.</dc:creator>
<dc:creator>Giacomet, C.</dc:creator>
<dc:creator>Cossa, F.</dc:creator>
<dc:creator>Federici, M.</dc:creator>
<dc:creator>Berretta, N.</dc:creator>
<dc:creator>Mercuri, N. B.</dc:creator>
<dc:date>2024-06-01</dc:date>
<dc:identifier>doi:10.1101/2024.05.28.596142</dc:identifier>
<dc:title><![CDATA[Functional efficacy of the MAO-B inhibitor safinamide in murine substantia nigra pars compacta dopaminergic neurons in vitro: a comparative study with tranylcypromine]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.05.30.596437v1?rss=1">
<title>
<![CDATA[
The diversity of SNCA transcripts in neurons, and its impact on antisense oligonucleotide therapeutics 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.05.30.596437v1?rss=1"
</link>
<description><![CDATA[
The role of the SNCA gene locus in driving Parkinsons disease (PD) through rare and common genetic variation is well-recognized, but the transcriptional diversity of SNCA in vulnerable cell types remains unclear. We performed SNCA long-read RNA sequencing in human dopaminergic neurons and show that annotated SNCA transcripts account for only 5% of expression. Rather, the majority of expression (75%) at the SNCA locus originates from transcripts with alternative 5 and 3 untranslated regions. Importantly, 10% originates from transcripts encoding open reading frames not previously annotated, which are translated and detectable in human postmortem brain. Defining the 3 untranslated regions enabled the rational design of antisense oligonucleotides targeting the majority of SNCA transcripts, leading to the effective reversal of PD pathology, including protein aggregation, mitochondrial dysfunction, and toxicity. Resolving the complexity of the SNCA transcriptional landscape impacts RNA therapies and highlights differences in protein isoforms and their contribution to disease.
]]></description>
<dc:creator>Evans, J. R.</dc:creator>
<dc:creator>Gustavsson, E. K.</dc:creator>
<dc:creator>Doykov, I.</dc:creator>
<dc:creator>Murphy, D.</dc:creator>
<dc:creator>Virdi, G.</dc:creator>
<dc:creator>Lachica, J.</dc:creator>
<dc:creator>Rontgen, A.</dc:creator>
<dc:creator>Murtada, M. H.</dc:creator>
<dc:creator>Pang, C. W.</dc:creator>
<dc:creator>Macpherson, H.</dc:creator>
<dc:creator>Wernick, A. I.</dc:creator>
<dc:creator>Toomey, C. E.</dc:creator>
<dc:creator>Athauda, D.</dc:creator>
<dc:creator>Choi, M.</dc:creator>
<dc:creator>Hardy, J.</dc:creator>
<dc:creator>Wood, N. W.</dc:creator>
<dc:creator>Vendruscolo, M.</dc:creator>
<dc:creator>Mills, K.</dc:creator>
<dc:creator>Heywood, W.</dc:creator>
<dc:creator>Ryten, M.</dc:creator>
<dc:creator>Gandhi, S.</dc:creator>
<dc:date>2024-05-31</dc:date>
<dc:identifier>doi:10.1101/2024.05.30.596437</dc:identifier>
<dc:title><![CDATA[The diversity of SNCA transcripts in neurons, and its impact on antisense oligonucleotide therapeutics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.06.06.596654v1?rss=1">
<title>
<![CDATA[
Refinement of efficient encodings of movement in the dorsolateral striatum throughout learning 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.06.06.596654v1?rss=1"
</link>
<description><![CDATA[
The striatum is required for normal action selection, movement, and sensorimotor learning. Although action-specific striatal ensembles have been well documented, it is not well understood how these ensembles are formed and how their dynamics may evolve throughout motor learning. Here we used longitudinal 2-photon Ca2+ imaging of dorsal striatal neurons in head-fixed mice as they learned to self-generate locomotion. We observed a significant activation of both direct- and indirect-pathway spiny projection neurons (dSPNs and iSPNs, respectively) during early locomotion bouts and sessions that gradually decreased over time. For dSPNs, onset- and offset-ensembles were gradually refined from active motion-nonspecific cells. iSPN ensembles emerged from neurons initially active during opponent actions before becoming onset- or offset-specific. Our results show that as striatal ensembles are progressively refined, the number of active nonspecific striatal neurons decrease and the overall efficiency of the striatum information encoding for learned actions increases.
]]></description>
<dc:creator>Jaidar, O.</dc:creator>
<dc:creator>Albarran, E.</dc:creator>
<dc:creator>Albarran, E.</dc:creator>
<dc:creator>Wu, Y.-W.</dc:creator>
<dc:creator>Ding, J. B.</dc:creator>
<dc:date>2024-06-06</dc:date>
<dc:identifier>doi:10.1101/2024.06.06.596654</dc:identifier>
<dc:title><![CDATA[Refinement of efficient encodings of movement in the dorsolateral striatum throughout learning]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.06.07.597975v1?rss=1">
<title>
<![CDATA[
α-Synuclein Overexpression and the Microbiome Shape the Gut and Brain Metabolome in Mice 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.06.07.597975v1?rss=1"
</link>
<description><![CDATA[
Pathological forms of the protein -synuclein contribute to a family of disorders termed synucleinopathies, which includes Parkinsons disease (PD). Most cases of PD are believed to arise from gene-environment interactions. Microbiome composition is altered in PD, and gut bacteria are causal to symptoms and pathology in animal models. To explore how the microbiome may impact PD-associated genetic risks, we quantitatively profiled nearly 630 metabolites from 26 biochemical classes in the gut, plasma, and brain of -synuclein-overexpressing (ASO) mice with or without microbiota. We observe tissue-specific changes driven by genotype, microbiome, and their interaction. Many differentially expressed metabolites in ASO mice are also dysregulated in human PD patients, including amine oxides, bile acids and indoles. Notably, levels of the microbial metabolite trimethylamine N-oxide (TMAO) strongly correlate from the gut to the plasma to the brain, identifying a product of gene-environment interactions that may influence PD-like outcomes in mice. TMAO is elevated in the blood and cerebral spinal fluid of PD patients. These findings uncover broad metabolomic changes that are influenced by the intersection of host genetics and the microbiome in a mouse model of PD.
]]></description>
<dc:creator>Morais, L. H.</dc:creator>
<dc:creator>Boktor, J. C.</dc:creator>
<dc:creator>MahmoudianDehkordi, S.</dc:creator>
<dc:creator>Kaddurah-Daouk, R.</dc:creator>
<dc:creator>Mazmanian, S. K.</dc:creator>
<dc:date>2024-06-10</dc:date>
<dc:identifier>doi:10.1101/2024.06.07.597975</dc:identifier>
<dc:title><![CDATA[α-Synuclein Overexpression and the Microbiome Shape the Gut and Brain Metabolome in Mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.06.08.598070v1?rss=1">
<title>
<![CDATA[
Lysosome damage triggers acute formation of ER to lysosomes membrane tethers mediated by the bridge-like lipid transport protein VPS13C 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.06.08.598070v1?rss=1"
</link>
<description><![CDATA[
Based on genetic studies, lysosome dysfunction is thought to play a pathogenetic role in Parkinsons disease (PD). Here we show that VPS13C, a bridge-like lipid transport protein and a PD gene, is a sensor of lysosome stress/damage. Upon lysosome membrane perturbation, VPS13C rapidly relocates from the cytosol to the surface of lysosomes where it tethers their membranes to the ER. This recruitment depends on Rab7 and requires a signal at the damaged lysosome surface that releases an inhibited state of VPS13C which hinders access of its VAB domain to lysosome-bound Rab7. While another PD protein, LRRK2, is also recruited to stressed/damaged lysosomes, its recruitment occurs at much later stages and by different mechanisms. Given the role of VPS13 proteins in bulk lipid transport, these findings suggest that lipid delivery to lysosomes by VPS13C is part of an early protective response to lysosome damage.
]]></description>
<dc:creator>Wang, X.</dc:creator>
<dc:creator>Xu, P.</dc:creator>
<dc:creator>Bentley-DeSousa, A.</dc:creator>
<dc:creator>Hancock-Cerutti, W.</dc:creator>
<dc:creator>Cai, S.</dc:creator>
<dc:creator>Johnson, B. T.</dc:creator>
<dc:creator>Tonelli, F.</dc:creator>
<dc:creator>Talaia, G.</dc:creator>
<dc:creator>Alessi, D. R.</dc:creator>
<dc:creator>Ferguson, S. M.</dc:creator>
<dc:creator>De Camilli, P.</dc:creator>
<dc:date>2024-06-08</dc:date>
<dc:identifier>doi:10.1101/2024.06.08.598070</dc:identifier>
<dc:title><![CDATA[Lysosome damage triggers acute formation of ER to lysosomes membrane tethers mediated by the bridge-like lipid transport protein VPS13C]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.06.10.598366v1?rss=1">
<title>
<![CDATA[
Activity-Dependent Remodeling of Corticostriatal Axonal Boutons During Motor Learning 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.06.10.598366v1?rss=1"
</link>
<description><![CDATA[
Motor skill learning induces long-lasting synaptic plasticity at not only the inputs, such as dendritic spines1-4, but also at the outputs to the striatum of motor cortical neurons5,6. However, very little is known about the activity and structural plasticity of corticostriatal axons during learning in the adult brain. Here, we used longitudinal in vivo two-photon imaging to monitor the activity and structure of thousands of corticostriatal axonal boutons in the dorsolateral striatum in awake mice. We found that learning a new motor skill induces dynamic regulation of axonal boutons. The activities of motor corticostriatal axonal boutons exhibited selectivity for rewarded movements (RM) and un-rewarded movements (UM). Strikingly, boutons on the same axonal branches showed diverse responses during behavior. Motor learning significantly increased the fraction of RM boutons and reduced the heterogeneity of bouton activities. Moreover, motor learning-induced profound structural dynamism in boutons. By combining structural and functional imaging, we identified that newly formed axonal boutons are more likely to exhibit selectivity for RM and are stabilized during motor learning, while UM boutons are selectively eliminated. Our results highlight a novel form of plasticity at corticostriatal axons induced by motor learning, indicating that motor corticostriatal axonal boutons undergo dynamic reorganization that facilitates the acquisition and execution of motor skills.
]]></description>
<dc:creator>Sheng, M.</dc:creator>
<dc:creator>Lu, D.</dc:creator>
<dc:creator>Sheng, K.</dc:creator>
<dc:creator>Ding, J.</dc:creator>
<dc:date>2024-06-10</dc:date>
<dc:identifier>doi:10.1101/2024.06.10.598366</dc:identifier>
<dc:title><![CDATA[Activity-Dependent Remodeling of Corticostriatal Axonal Boutons During Motor Learning]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.06.11.598416v1?rss=1">
<title>
<![CDATA[
Endogenous LRRK2 and PINK1 function in a convergent neuroprotective ciliogenesis pathway in the brain 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.06.11.598416v1?rss=1"
</link>
<description><![CDATA[
Mutations in LRRK2 and PINK1 are associated with familial Parkinsons disease (PD). LRRK2 phosphorylates Rab GTPases within the Switch II domain whilst PINK1 directly phosphorylates Parkin and ubiquitin and indirectly induces phosphorylation of a subset of Rab GTPases. Herein we have crossed LRRK2 [R1441C] mutant knock-in mice with PINK1 knock-out (KO) mice and report that loss of PINK1 does not impact endogenous LRRK2-mediated Rab phosphorylation nor do we see significant effect of mutant LRRK2 on PINK1-mediated Rab and ubiquitin phosphorylation. In addition, we observe that a pool of the Rab-specific, PPM1H phosphatase, is transcriptionally up-regulated and recruited to damaged mitochondria, independent of PINK1 or LRRK2 activity. Parallel signalling of LRRK2 and PINK1 pathways is supported by assessment of motor behavioural studies that show no evidence of genetic interaction in crossed mouse lines. Previously we showed loss of cilia in LRRK2 R1441C mice and herein we show that PINK1 KO mice exhibit a ciliogenesis defect in striatal cholinergic interneurons and astrocytes that interferes with Hedgehog induction of glial derived-neurotrophic factor (GDNF) transcription. This is not exacerbated in double mutant LRRK2 and PINK1 mice. Overall, our analysis indicates that LRRK2 activation and/or loss of PINK1 function along parallel pathways to impair ciliogenesis, suggesting a convergent mechanism towards PD. Our data suggests that reversal of defects downstream of ciliogenesis offers a common therapeutic strategy for LRRK2 or PINK1 PD patients whereas LRRK2 inhibitors that are currently in clinical trials are unlikely to benefit PINK1 PD patients.
]]></description>
<dc:creator>Bagnoli, E.</dc:creator>
<dc:creator>Lin, Y.-E.</dc:creator>
<dc:creator>Burel, S.</dc:creator>
<dc:creator>Jaimon, E.</dc:creator>
<dc:creator>Antico, O.</dc:creator>
<dc:creator>Themistokleous, C.</dc:creator>
<dc:creator>Nikoloff, J.</dc:creator>
<dc:creator>Morella, I.</dc:creator>
<dc:creator>Watzlawik, J.</dc:creator>
<dc:creator>Fiesel, F.</dc:creator>
<dc:creator>Springer, W.</dc:creator>
<dc:creator>Tonelli, F.</dc:creator>
<dc:creator>Brooks, S.</dc:creator>
<dc:creator>Dunnett, S.</dc:creator>
<dc:creator>Brambilla, R.</dc:creator>
<dc:creator>Alessi, D. R.</dc:creator>
<dc:creator>Pfeffer, S. R.</dc:creator>
<dc:creator>Muqit, M.</dc:creator>
<dc:date>2024-06-11</dc:date>
<dc:identifier>doi:10.1101/2024.06.11.598416</dc:identifier>
<dc:title><![CDATA[Endogenous LRRK2 and PINK1 function in a convergent neuroprotective ciliogenesis pathway in the brain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.06.12.598718v1?rss=1">
<title>
<![CDATA[
Mitochondrial damage triggers concerted degradation of negative regulators of neuronal autophagy 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.06.12.598718v1?rss=1"
</link>
<description><![CDATA[
Mutations in genes that regulate mitophagy, a key mitochondrial quality control pathway, are causative for neurological disorders including Parkinsons. Here, we identify a novel stress response pathway activated by mitochondrial damage that regulates mitophagy in neurons. We find that increasing levels of mitochondrial stress triggers a graded, concerted response that induces proteasomal degradation of negative regulators of autophagy. These include Myotubularin-related phosphatase 5 (MTMR5), MTMR2 and Rubicon. This  Mitophagic Stress Response (MitoSR) pathway is neuron-specific and acts in parallel to the classical Pink1/Parkin-mediated mitophagy pathway. While MTMR5/MTMR2 inhibits autophagosome biogenesis, we find that Rubicon inhibits lysosomal function and thus blocks autophagosome maturation. Targeted depletion of these negative regulators is sufficient to enhance mitophagy, promoting autophagosome biogenesis and facilitating the fusion of mitophagosomes with lysosomes. Our work suggests that therapeutic activation of the MitoSR pathway to induce degradation of negative regulators of autophagy may enhance mitochondrial quality control in stressed neurons.
]]></description>
<dc:creator>Basak, B.</dc:creator>
<dc:creator>Holzbaur, E. L. F.</dc:creator>
<dc:date>2024-06-13</dc:date>
<dc:identifier>doi:10.1101/2024.06.12.598718</dc:identifier>
<dc:title><![CDATA[Mitochondrial damage triggers concerted degradation of negative regulators of neuronal autophagy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.06.13.598820v1?rss=1">
<title>
<![CDATA[
Mitochondrial oxidant stress promotes alpha-synuclein aggregation and spreading in mice with mutated glucocerebrosidase 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.06.13.598820v1?rss=1"
</link>
<description><![CDATA[
Mutations of the glucocerebrosidase-encoding gene, GBA1, are common risk factors for Parkinsons disease. Although only a minority of mutation-carrying individuals develops the disease, the mechanisms of neuronal vulnerability predisposing to pathology conversion remain largely unclear. In this study, heterozygous expression of a common glucocerebrosidase variant, namely the L444P mutation, was found to exacerbate -synuclein aggregation and spreading in a mouse model of Parkinson-like pathology targeting neurons of the medullary vagal system. These neurons are primary sites of -synuclein lesions in Parkinsons disease and were shown here to become more vulnerable to oxidative stress after L444P expression. Nitrative burden paralleled the enhanced formation of reactive oxygen species within vagal neurons expressing mutated glucocerebrosidase, as indicated by pronounced accumulation of nitrated -synuclein. A causal relationship linked mutation-induced oxidative stress to enhanced -synuclein pathology that could indeed be rescued by neuronal overexpression of the mitochondrial antioxidant enzyme superoxide dismutase 2. Further evidence supported a key involvement of mitochondria as sources of reactive oxygen species as well as targets of oxidative and nitrative damage within L444P-expressing neurons. Scavenging of oxygen species by superoxide dismutase 2 effectively counteracted deleterious nitrative reactions and prevented nitrated -synuclein burden. Taken together, these findings support the conclusion that enhanced vulnerability to mitochondrial oxidative stress conferred by glucocerebrosidase mutations should be considered an important mechanism predisposing to Parkinsons disease pathology, particularly in brain regions targeted by -synuclein aggregation and involved in -synuclein spreading.
]]></description>
<dc:creator>La Vitola, P.</dc:creator>
<dc:creator>Szego, E. M.</dc:creator>
<dc:creator>Pinto-Costa, R.</dc:creator>
<dc:creator>Rollar, A.</dc:creator>
<dc:creator>Harbachova, E.</dc:creator>
<dc:creator>Schapira, A. H. V.</dc:creator>
<dc:creator>Ulusoy, A.</dc:creator>
<dc:creator>Di Monte, D. A.</dc:creator>
<dc:date>2024-06-13</dc:date>
<dc:identifier>doi:10.1101/2024.06.13.598820</dc:identifier>
<dc:title><![CDATA[Mitochondrial oxidant stress promotes alpha-synuclein aggregation and spreading in mice with mutated glucocerebrosidase]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.06.15.599140v1?rss=1">
<title>
<![CDATA[
Network nature of ligand-receptor interactions underlies disease comorbidity in the brain 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.06.15.599140v1?rss=1"
</link>
<description><![CDATA[
Neurodegenerative disorders have overlapping symptoms and have high comorbidity rates, but this is not reflected in overlaps of risk genes. We have investigated whether ligand-receptor interactions (LRIs) are a mechanism by which distinct genes associated with disease risk can impact overlapping outcomes. We found that LRIs are likely disrupted in neurological disease and that the ligand-receptor networks associated with neurological diseases have substantial overlaps. Specifically, 96.8% of LRIs associated with disease risk are interconnected in a single LR network. These ligands and receptors are enriched for roles in inflammatory pathways and highlight the role of glia in cross-disease risk. Disruption to this LR network due to disease-associated processes (e.g. differential transcript use, protein misfolding) is likely to contribute to disease progression and risk of comorbidity. Our findings have implications for drug development, as they highlight the potential benefits and risks of pursuing cross-disease drug targets.
]]></description>
<dc:creator>Grant-Peters, M.</dc:creator>
<dc:creator>Fairbrother-Browne, A.</dc:creator>
<dc:creator>Hicks, A.</dc:creator>
<dc:creator>Guo, B.</dc:creator>
<dc:creator>Reynolds, R. H.</dc:creator>
<dc:creator>Huuki-Myers, L. A.</dc:creator>
<dc:creator>Eagles, N.</dc:creator>
<dc:creator>Brenton, J.</dc:creator>
<dc:creator>Garcia-Ruiz, S.</dc:creator>
<dc:creator>Wood, N.</dc:creator>
<dc:creator>Gandhi, S.</dc:creator>
<dc:creator>Martinowich, K.</dc:creator>
<dc:creator>Maynard, K.</dc:creator>
<dc:creator>Collado-Torres, L.</dc:creator>
<dc:creator>Ryten, M.</dc:creator>
<dc:date>2024-06-16</dc:date>
<dc:identifier>doi:10.1101/2024.06.15.599140</dc:identifier>
<dc:title><![CDATA[Network nature of ligand-receptor interactions underlies disease comorbidity in the brain]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.06.17.599289v1?rss=1">
<title>
<![CDATA[
Pathogenic LRRK2 mutations cause loss of primary cilia and neuroprotective Neurturin in striatal Parvalbumin interneurons 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.06.17.599289v1?rss=1"
</link>
<description><![CDATA[
Parkinsons disease-associated, activating mutations in LRRK2 kinase block primary cilia formation in cell culture and in specific cell types in the brain. In the striatum that is important for movement control, about half of astrocytes and cholinergic interneurons, but not the predominant medium spiny neurons, lose their primary cilia. Here we show that Parvalbumin interneurons that are inhibitory regulators of movement also lose primary cilia. Without cilia, these neurons are not able to respond to Sonic hedgehog signals that normally induce the expression of Patched protein, and their numbers decrease. In addition, glial cell line-derived neurotrophic factor-related Neurturin expression is significantly decreased. These experiments highlight the importance of Parvalbumin neurons in cilia-dependent, neuroprotective signaling pathways and show that LRRK2 activation decreases Neurturin production, resulting in less neuroprotection for dopamine neurons.

SummaryParvalbumin interneurons in the dorsal striatum lose primary cilia in mice harboring Parkinsons-associated, activating mutations in LRRK2 kinase, resulting in loss of Hedgehog signaling and decreased production of neuroprotective, Glial cell line-derived neurotrophic factor-related Neurturin to support dopamine neurons.
]]></description>
<dc:creator>Lin, Y.-E.</dc:creator>
<dc:creator>Jaimon, E.</dc:creator>
<dc:creator>Tonelli, F.</dc:creator>
<dc:creator>Alessi, D. R.</dc:creator>
<dc:creator>Pfeffer, S. R.</dc:creator>
<dc:date>2024-06-17</dc:date>
<dc:identifier>doi:10.1101/2024.06.17.599289</dc:identifier>
<dc:title><![CDATA[Pathogenic LRRK2 mutations cause loss of primary cilia and neuroprotective Neurturin in striatal Parvalbumin interneurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.06.18.599639v1?rss=1">
<title>
<![CDATA[
Transcriptional pathobiology and multi-omics predictors for Parkinson's disease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.06.18.599639v1?rss=1"
</link>
<description><![CDATA[
Early diagnosis and biomarker discovery to bolster the therapeutic pipeline for Parkinsons disease (PD) are urgently needed. In this study, we leverage the large-scale whole-blood total RNA-seq dataset from the Accelerating Medicine Partnership in Parkinsons Disease (AMP PD) program to identify PD-associated RNAs, including both known genes and novel circular RNAs (circRNA) and enhancer RNAs (eRNAs). There were 1,111 significant marker RNAs, including 491 genes, 599 eRNAs, and 21 circRNAs, that were first discovered in the PPMI cohort (FDR < 0.05) and confirmed in the PDBP/BioFIND cohorts (nominal p < 0.05). Functional enrichment analysis showed that the PD-associated genes are involved in neutrophil activation and degranulation, as well as the TNF-alpha signaling pathway. We further compare the PD-associated genes in blood with those in post-mortem brain dopamine neurons in our BRAINcode cohort. 44 genes show significant changes with the same direction in both PD brain neurons and PD blood, including neuroinflammation-associated genes IKBIP, CXCR2, and NFKBIB. Finally, we built a novel multi-omics machine learning model to predict PD diagnosis with high performance (AUC = 0.89), which was superior to previous studies and might aid the decision-making for PD diagnosis in clinical practice. In summary, this study delineates a wide spectrum of the known and novel RNAs linked to PD and are detectable in circulating blood cells in a harmonized, large-scale dataset. It provides a generally useful computational framework for further biomarker development and early disease prediction.

Significance statementEarly and accurate diagnosis of Parkinsons disease (PD) is urgently needed. However, biomarkers for early detection of PD are still lacking. Also, the limit of sample size remains one of the main pitfalls of current PD biomarker studies. We employed an analysis of large-scale whole-blood RNA-seq data. By identifying 1,111 significant marker RNAs, we establish a robust foundation for early PD detection, which implicated in neutrophil activation, degranulation, and TNF-alpha signaling, offer unprecedented insights into PD pathogenesis. Our multi-omics machine learning model, boasting an AUC of 0.89, outperforms previous studies, promising a transformative tool for precise PD diagnosis in clinical settings. This study marks a pivotal step toward enhanced biomarker development and early disease prediction.
]]></description>
<dc:creator>Hu, R.</dc:creator>
<dc:creator>Wang, R.</dc:creator>
<dc:creator>Yuan, J.</dc:creator>
<dc:creator>Lin, Z.</dc:creator>
<dc:creator>Hutchins, E.</dc:creator>
<dc:creator>Landin, B.</dc:creator>
<dc:creator>Liao, Z.</dc:creator>
<dc:creator>Liu, G.</dc:creator>
<dc:creator>Scherzer, C. R.</dc:creator>
<dc:creator>Dong, X.</dc:creator>
<dc:date>2024-06-21</dc:date>
<dc:identifier>doi:10.1101/2024.06.18.599639</dc:identifier>
<dc:title><![CDATA[Transcriptional pathobiology and multi-omics predictors for Parkinson's disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.06.20.599918v1?rss=1">
<title>
<![CDATA[
MICROENDOSCOPIC CALCIUM IMAGING IN SUPPLEMENTARY MOTOR AREA AND PRIMARY MOTOR CORTEX OF RHESUS MACAQUES AT REST AND DURING ARM MOVEMENT 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.06.20.599918v1?rss=1"
</link>
<description><![CDATA[
The study of motor cortices in non-human primates is relevant to our understanding of human motor control, both in healthy conditions and in movement disorders. Calcium imaging and miniature microscopes allow the study of multiple genetically identified neurons with excellent spatial resolution. We used this method to examine activity patterns of projection neurons in deep layers of the supplementary motor (SMA) and primary motor areas (M1) in four rhesus macaques. We implanted gradient index lenses and expressed GCaMP6f to image calcium transients while the animals were at rest or engaged in an arm reaching task. We tracked the activity of SMA and M1 neurons across conditions, examined cell pairs for synchronous activity, and assessed whether SMA and M1 neuronal activation followed specific sequential activation patterns. We demonstrate the value of in vivo calcium imaging for studying patterns of activity in groups of corticofugal neurons in SMA and M1.

HIGHLIGHTSO_LIUse of one-photon miniature microscopes and microendoscopic calcium imaging to study the activity of cortical projection neurons in the supplementary motor area (SMA) and primary motor cortex (M1) of rhesus macaques at rest or performing simple arm reaches.
C_LIO_LICalcium transients were related to arm reaches and showed directional sensitivity in a proportion of cells in SMA and M1.
C_LIO_LISubsets of cell pairs showed coactivation in SMA and M1 during rest and reaching tasks. The strength of coactivity was not related to the distance between cells.
C_LIO_LISMA and M1 neurons displayed sequential activation patterns.
C_LIO_LIWe demonstrated that microendoscopic calcium imaging can be used to assess dynamic activity within genetically identified cell populations in deep layers of SMA and M1.
C_LI
]]></description>
<dc:creator>Martel, A.-C.</dc:creator>
<dc:creator>Pittard, D.</dc:creator>
<dc:creator>Devergnas, A.</dc:creator>
<dc:creator>Risk, B.</dc:creator>
<dc:creator>Nassi, J.</dc:creator>
<dc:creator>Yu, W.</dc:creator>
<dc:creator>Downer, J.</dc:creator>
<dc:creator>Wichmann, T.</dc:creator>
<dc:creator>Galvan, A.</dc:creator>
<dc:date>2024-06-25</dc:date>
<dc:identifier>doi:10.1101/2024.06.20.599918</dc:identifier>
<dc:title><![CDATA[MICROENDOSCOPIC CALCIUM IMAGING IN SUPPLEMENTARY MOTOR AREA AND PRIMARY MOTOR CORTEX OF RHESUS MACAQUES AT REST AND DURING ARM MOVEMENT]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.07.09.602700v1?rss=1">
<title>
<![CDATA[
Loss of the lysosomal lipid flippase ATP10B leads to progressive dopaminergic neurodegeneration and Parkinsonian motor deficits 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.07.09.602700v1?rss=1"
</link>
<description><![CDATA[
BackgroundATP10B, a transmembrane lipid flippase located in late endosomes and lysosomes, facilitates the export of glucosylceramide and phosphatidylcholine by coupling this process to ATP hydrolysis. Recently, loss-of-function mutations in the ATP10B gene have been identified in Parkinsons disease patients, pointing to ATP10B as a candidate genetic risk factor. Previous studies have shown compromised lysosomal functionality upon ATP10B knockdown in human cell lines and primary cortical neurons. However, its role in vivo and specifically in the nigrostriatal dopaminergic system remains poorly understood.

MethodsTo investigate the role ATP10B in PD neuropathology, we induced ATP10B knockdown specifically in substantia nigra pars compacta neurons of rats using viral vector technology. Two different microRNA-based shRNA constructs targeting distinct regions of the ATP10B mRNA were used to cross-validate the findings. Behavioral evaluation, dopamine transporter 18F-FE-PE2I positron emission tomography imaging and neuropathological examination of the nigrostriatal pathway at one year post-injection were conducted. Additionally, midbrain neuronal cultures derived from ATP10B knock-out human induced pluripotent stem cells clones were used to study the impact of ATP10B loss in dopaminergic neurons in a more translational model.

ResultsATP10B knockdown in rat brain induced Parkinsonian motor deficits, and longitudinal striatal dopamine transporter 18F-FE-PE2I PET imaging revealed a progressive decrease in binding potential. Immunohistochemical analysis conducted one year post-injection confirmed the loss of dopaminergic terminals in the striatum, alongside a loss of dopaminergic neurons in the substantia nigra pars compacta. The expression of LAMP1, LAMP2a, cathepsin B and glucocerebrosidase was studied by immunofluorescence in the surviving dopaminergic neurons. A decrease in lysosomal numbers and an increase in lysosomal volume were observed more consistently in one of the knockdown constructs. The vulnerability of dopaminergic neurons to ATP10B loss-of-function was also observed in midbrain neuronal cultures derived from ATP10B knock-out human induced pluripotent stem cells clones, which showed a significant reduction in TH-positive neurons.

ConclusionTaken together, our findings demonstrate that ATP10B depletion detrimentally impacts the viability of dopaminergic neurons both in vivo and in vitro. Moreover, a broader impact on the functionality of the nigrostriatal pathway was evidenced as rats with ATP10B knockdown exhibited motor impairments similar to those observed in PD patients.
]]></description>
<dc:creator>Sanchiz-Calvo, M.</dc:creator>
<dc:creator>Coccia, E.</dc:creator>
<dc:creator>Cawthorne, C.</dc:creator>
<dc:creator>Parfitt, G.</dc:creator>
<dc:creator>Van Laere, K.</dc:creator>
<dc:creator>Torre-Muruzabal, T.</dc:creator>
<dc:creator>Cabezudo, D.</dc:creator>
<dc:creator>Tsafaras, G.</dc:creator>
<dc:creator>Cascalho, A.</dc:creator>
<dc:creator>Van Den Haute, C.</dc:creator>
<dc:creator>Vangheluwe, P.</dc:creator>
<dc:creator>Blanchard, J.</dc:creator>
<dc:creator>Bentea, E.</dc:creator>
<dc:creator>Baekelandt, V.</dc:creator>
<dc:date>2024-07-13</dc:date>
<dc:identifier>doi:10.1101/2024.07.09.602700</dc:identifier>
<dc:title><![CDATA[Loss of the lysosomal lipid flippase ATP10B leads to progressive dopaminergic neurodegeneration and Parkinsonian motor deficits]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.07.01.601474v1?rss=1">
<title>
<![CDATA[
The Neurolipid Atlas: a lipidomics resource for neurodegenerative diseases uncovers cholesterol as a regulator of astrocyte reactivity impaired by ApoE4 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.07.01.601474v1?rss=1"
</link>
<description><![CDATA[
Lipid changes in the brain have been implicated in many neurodegenerative diseases including Alzheimers Disease (AD), Parkinsons disease and Amyotrophic Lateral Sclerosis. To facilitate comparative lipidomic research across brain-diseases we established a data commons named the Neurolipid Atlas, that we have pre-populated with novel human, mouse and isogenic induced pluripotent stem cell (iPSC)-derived lipidomics data for different brain diseases. We show that iPSC-derived neurons, microglia and astrocytes display distinct lipid profiles that recapitulate in vivo lipotypes. Leveraging multiple datasets, we show that the AD risk gene ApoE4 drives cholesterol ester (CE) accumulation in human astrocytes recapitulating CE accumulation measured in the human AD brain. Multi-omic interrogation of iPSC-derived astrocytes revealed that cholesterol plays a major role in astrocyte interferon-dependent pathways such as the immunoproteasome and major histocompatibility complex (MHC) class I antigen presentation. We show that through enhanced cholesterol esterification ApoE4 suppresses immune activation of astrocytes. Our novel data commons, available at neurolipidatlas.com, provides a user-friendly tool and knowledge base for a better understanding of lipid dyshomeostasis in neurodegenerative diseases.
]]></description>
<dc:creator>Feringa, F. M.</dc:creator>
<dc:creator>Koppes-den Hertog, S. J.</dc:creator>
<dc:creator>Wang, L.</dc:creator>
<dc:creator>Derks, R. J. E.</dc:creator>
<dc:creator>Kruijff, I.</dc:creator>
<dc:creator>Erlebach, L.</dc:creator>
<dc:creator>Heijneman, J.</dc:creator>
<dc:creator>Miramontes, R.</dc:creator>
<dc:creator>Pömpner, N.</dc:creator>
<dc:creator>Blomberg, N.</dc:creator>
<dc:creator>Olivier-Jimenez, D.</dc:creator>
<dc:creator>Johanson, L. E.</dc:creator>
<dc:creator>Cammack, A. J.</dc:creator>
<dc:creator>Giblin, A.</dc:creator>
<dc:creator>Toomey, C. E.</dc:creator>
<dc:creator>Rose, I. V. L.</dc:creator>
<dc:creator>Yuan, H.</dc:creator>
<dc:creator>Ward, M. E.</dc:creator>
<dc:creator>Isaacs, A.</dc:creator>
<dc:creator>Kampmann, M.</dc:creator>
<dc:creator>Kronenberg-Versteeg, D.</dc:creator>
<dc:creator>Lashley, T.</dc:creator>
<dc:creator>Thompson, L. M.</dc:creator>
<dc:creator>Ori, A.</dc:creator>
<dc:creator>Mohammed, Y.</dc:creator>
<dc:creator>Giera, M.</dc:creator>
<dc:creator>van der Kant, R. H. N.</dc:creator>
<dc:date>2024-07-03</dc:date>
<dc:identifier>doi:10.1101/2024.07.01.601474</dc:identifier>
<dc:title><![CDATA[The Neurolipid Atlas: a lipidomics resource for neurodegenerative diseases uncovers cholesterol as a regulator of astrocyte reactivity impaired by ApoE4]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.06.25.600616v1?rss=1">
<title>
<![CDATA[
Presynaptic GABAA receptors control integration of nicotinic input onto dopaminergic axons in the striatum 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.06.25.600616v1?rss=1"
</link>
<description><![CDATA[
Axons of dopaminergic neurons express gamma-aminobutyric acid type-A receptors (GABAARs) and nicotinic acetylcholine receptors (nAChRs) which are both independently positioned to shape striatal dopamine release. Using electrophysiology and calcium imaging, we investigated how interactions between GABAARs and nAChRs influence dopaminergic axon excitability. Direct axonal recordings showed that benzodiazepine application suppresses subthreshold axonal input from cholinergic interneurons (CINs). In imaging experiments, we used the first temporal derivative of presynaptic calcium signals to distinguish between direct- and nAChR-evoked activity in dopaminergic axons. We found that GABAAR antagonism with gabazine selectively enhanced nAChR-evoked axonal signals. Acetylcholine release was unchanged in gabazine suggesting that GABAARs located on dopaminergic axons, but not CINs, mediated this enhancement. Unexpectedly, we found that a widely used GABAAR antagonist, picrotoxin, inhibits axonal nAChRs and should be used cautiously for striatal circuit analysis. Overall, we demonstrate that GABAARs on dopaminergic axons regulate integration of nicotinic input to shape presynaptic excitability.
]]></description>
<dc:creator>Brill-Weil, S. G.</dc:creator>
<dc:creator>Kramer, P. F.</dc:creator>
<dc:creator>Yanez, A.</dc:creator>
<dc:creator>Clever, F. H.</dc:creator>
<dc:creator>Zhang, R.</dc:creator>
<dc:creator>Khaliq, Z. M.</dc:creator>
<dc:date>2024-06-25</dc:date>
<dc:identifier>doi:10.1101/2024.06.25.600616</dc:identifier>
<dc:title><![CDATA[Presynaptic GABAA receptors control integration of nicotinic input onto dopaminergic axons in the striatum]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.06.18.598931v1?rss=1">
<title>
<![CDATA[
A rewiring of the earliest immune events leading to T-cell mediated disease following intestinal infection in a PINK1KO mouse model of Parkinsons disease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.06.18.598931v1?rss=1"
</link>
<description><![CDATA[
Parkinsons disease (PD) is characterized by a protracted period of non-motor symptoms, including gastrointestinal (GI) dysfunction, which can precede the development of the cardinal motor deficits by decades. This long prodrome of disease is highly suggestive of immune cell involvement in the initiation of disease, but currently the field lacks robust model systems to study such mechanisms. It has been hypothesized that pathology may be first initiated in the periphery due to environmental triggers, such as pathogens that enter the GI tract. We further speculate that the impact of such pathogens on the immune system could be exacerbated in genetically predisposed individuals. Our group has developed a GI-targeted pathogen-induced PD mouse model system in PINK1 KO mice with Gram-negative bacterial infections and found that T cells are a major player in driving PD-like motor symptoms at late stages following infection. Herein, we now map the initiating immune events at the site of infection at the earliest stages with the goal of shedding light on the earliest mechanisms triggering immune-mediated pathological processes relevant to PD. Using unbiased single cell sequencing, we demonstrate that myeloid cells are the earliest dysregulated immune cell type in PINK1 KO infected mice at 1-week post-infection, followed by a dysregulated T cell response shortly after, at 2 weeks post-infection. We find that these myeloid cells have an enhanced proinflammatory profile, are more mature, and develop enhanced capacity for antigen presentation. Using unbiased prediction analysis, our data suggest that cytotoxic T cells and myeloid cells are particularly poised for interacting with each other, and we identify possible direct cell-cell interaction pathways that might be implicated. Taken together, deciphering the earliest immune mechanisms in the periphery underpinning PD autoimmunity will be instrumental in the development of effective therapeutic targeting strategies before irrevocable neuronal damage ensues.
]]></description>
<dc:creator>Recinto, S.</dc:creator>
<dc:creator>Kazanova, A.</dc:creator>
<dc:creator>MacDonald, A.</dc:creator>
<dc:creator>Afanasiev, E.</dc:creator>
<dc:creator>Premachandran, S.</dc:creator>
<dc:creator>Allot, A.</dc:creator>
<dc:creator>Bessaiah, H.</dc:creator>
<dc:creator>Cordeiro, B.</dc:creator>
<dc:creator>Mukherjee, S.</dc:creator>
<dc:creator>Pei, J.</dc:creator>
<dc:creator>McBride, H. M.</dc:creator>
<dc:creator>Trudeau, L.-E.</dc:creator>
<dc:creator>Gruenheid, S.</dc:creator>
<dc:creator>Stratton, J. A.</dc:creator>
<dc:date>2024-06-22</dc:date>
<dc:identifier>doi:10.1101/2024.06.18.598931</dc:identifier>
<dc:title><![CDATA[A rewiring of the earliest immune events leading to T-cell mediated disease following intestinal infection in a PINK1KO mouse model of Parkinsons disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2022.10.31.514594v1?rss=1">
<title>
<![CDATA[
Excessive Firing of Dyskinesia-Associated Striatal Direct Pathway Neurons is Gated By Dopamine and Excitatory Synaptic Input 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2022.10.31.514594v1?rss=1"
</link>
<description><![CDATA[
The striatum integrates dopaminergic and glutamatergic inputs to select preferred versus alternative actions, but the precise mechanisms remain unclear. One way to study action selection is when it breaks down. Here, we explored the cellular and synaptic mechanisms of levodopa-induced dyskinesia (LID), a complication of Parkinsons disease therapy characterized by involuntary movements. We used an activity-dependent tool (FosTRAP) in conjunction with a mouse model of LID to investigate functionally distinct subsets of striatal direct pathway medium spiny neurons (dMSNs). In vivo, levodopa differentially activates dyskinesia-associated (TRAPed) dMSNs compared to other dMSNs. This activation is likely to be driven by two cellular mechanisms we identified through ex vivo electrophysiology: higher sensitivity to dopamine and stronger excitatory input from the motor cortex and thalamus. Together, these findings suggest how intrinsic and synaptic properties of heterogeneous dMSN subpopulations integrate to support action selection.
]]></description>
<dc:creator>Ryan, M. B.</dc:creator>
<dc:creator>Girasole, A. E.</dc:creator>
<dc:creator>McGregor, M. M.</dc:creator>
<dc:creator>Brakaj, R.</dc:creator>
<dc:creator>Paletzki, R. F.</dc:creator>
<dc:creator>Gerfen, C. R.</dc:creator>
<dc:creator>Nelson, A. B.</dc:creator>
<dc:date>2022-11-01</dc:date>
<dc:identifier>doi:10.1101/2022.10.31.514594</dc:identifier>
<dc:title><![CDATA[Excessive Firing of Dyskinesia-Associated Striatal Direct Pathway Neurons is Gated By Dopamine and Excitatory Synaptic Input]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2022-11-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.01.17.524433v1?rss=1">
<title>
<![CDATA[
GEARBOCS: An Adeno Associated Virus Tool for In Vivo Gene Editing in Astrocytes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.01.17.524433v1?rss=1"
</link>
<description><![CDATA[
CRISPR/Cas9-based genome engineering enables rapid and precise gene manipulations in the CNS. Here, we developed a non-invasive astrocyte-specific method utilizing a single AAV vector, which we named GEARBOCS (Gene Editing in AstRocytes Based On CRISPR/Cas9 System). We verified GEARBOCS specificity to mouse cortical astrocytes and demonstrated its utility for three types of gene manipulations: knockout (KO); tagging (TagIn); and reporter knock-in (GeneTrap) strategies. Next, we deployed GEARBOCS in two test cases. First, we determined that astrocytes are a necessary source of the synaptogenic factor Sparcl1 for thalamocortical synapse maintenance in the mouse primary visual cortex. Second, we determined that cortical astrocytes express the synaptic vesicle associated Vamp2 protein and found that it is required for maintaining excitatory and inhibitory synapse numbers in the visual cortex. These results show that the GEARBOCS strategy provides a fast and efficient means to study astrocyte biology in vivo.

MotivationAstrocytes are indispensable for brain development, function, and health. However, molecular tools to study astrocyte biology and function in vivo have been largely limited to genetically modified mice. Here, we developed a CRISPR/Cas9-based gene editing strategy within a single AAV vector that enables efficient genome manipulations in astrocytes. We designed and optimized this easy-to-use viral tool to understand gene expression, protein localization and function in astrocytes in vivo.
]]></description>
<dc:creator>Sivadasan Bindu, D.</dc:creator>
<dc:creator>Tan, C. X.</dc:creator>
<dc:creator>Savage, J. T.</dc:creator>
<dc:creator>Eroglu, C.</dc:creator>
<dc:date>2023-01-19</dc:date>
<dc:identifier>doi:10.1101/2023.01.17.524433</dc:identifier>
<dc:title><![CDATA[GEARBOCS: An Adeno Associated Virus Tool for In Vivo Gene Editing in Astrocytes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-01-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.05.15.594341v1?rss=1">
<title>
<![CDATA[
Rapid modulation of striatal cholinergic interneurons and dopamine release by satellite astrocytes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.05.15.594341v1?rss=1"
</link>
<description><![CDATA[
Astrocytes are increasingly thought to have underestimated and important roles in modulating neuronal circuits. Astrocytes in striatum can regulate dopamine transmission by governing the extracellular tone of axonal neuromodulators, including GABA and adenosine. However, here we reveal that striatal astrocytes occupy a cell type-specific anatomical and functional relationship with cholinergic interneurons (ChIs), through which they rapidly excite ChIs and govern dopamine release via nicotinic acetylcholine receptors on subsecond timescales. We identify that ChI somata are in unexpectedly close proximity to astrocyte somata, in mouse and human, forming a "soma-to-soma" satellite-like configuration not typically observed for other striatal neurons. Transient depolarization of astrocytes in mouse striatum reversibly regulated ChI excitability by decreasing extracellular calcium. These findings reveal a privileged satellite astrocyte-interneuron interaction for striatal ChIs operating on subsecond timescales via regulation of extracellular calcium dynamics to shape downstream striatal circuit activity and dopamine signaling.
]]></description>
<dc:creator>Stedehouder, J.</dc:creator>
<dc:creator>Roberts, B. M.</dc:creator>
<dc:creator>Raina, S.</dc:creator>
<dc:creator>Bossi, S.</dc:creator>
<dc:creator>Liu, A. K. L.</dc:creator>
<dc:creator>Doig, N. M.</dc:creator>
<dc:creator>McGerty, K.</dc:creator>
<dc:creator>Magill, P. J.</dc:creator>
<dc:creator>Parkkinen, L.</dc:creator>
<dc:creator>Cragg, S. J.</dc:creator>
<dc:date>2024-05-15</dc:date>
<dc:identifier>doi:10.1101/2024.05.15.594341</dc:identifier>
<dc:title><![CDATA[Rapid modulation of striatal cholinergic interneurons and dopamine release by satellite astrocytes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.05.15.594419v1?rss=1">
<title>
<![CDATA[
Cortico-amygdala synaptic structural abnormalities produced by templated aggregation of α-synuclein 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.05.15.594419v1?rss=1"
</link>
<description><![CDATA[
Parkinsons disease (PD) and Dementia with Lewy bodies (DLB) are characterized by neuronal -synuclein (-syn) inclusions termed Lewy Pathology, which are abundant in the amygdala. The basolateral amygdala (BLA), in particular, receives projections from the thalamus and cortex. These projections play a role in cognition and emotional processing, behaviors which are impaired in -synucleinopathies. To understand if and how pathologic -syn impacts the BLA requires animal models of -syn aggregation. Injection of -synuclein pre-formed fibrils (PFFs) into the striatum induces robust -synuclein aggregation in excitatory neurons in the BLA that corresponds with reduced contextual fear conditioning. At early time points after aggregate formation, cortico-amygdala excitatory transmission is abolished. The goal of this project was to determine if -syn inclusions in the BLA induce synaptic degeneration and/or morphological changes. In this study, we used C57BL/6J mice injected bilaterally with PFFs in the dorsal striatum to induce -syn aggregate formation in the BLA. A method was developed using immunofluorescence and three-dimensional reconstruction to analyze excitatory cortico-amygdala and thalamo-amygdala presynaptic terminals closely juxtaposed to postsynaptic densities. The abundance and morphology of synapses were analyzed at 6- or 12-weeks post-injection of PFFs. -Syn aggregate formation in the BLA did not cause a significant loss of synapses, but cortico-amygdala and thalamo-amygdala presynaptic terminals and postsynaptic densities with aggregates of -synuclein show increased volumes, similar to previous findings in human DLB cortex, and in non-human primate models of PD. Transmission electron microscopy showed that PFF-injected mice showed reduced intervesicular distances similar to a recent study showing phospho-serine-129 -synuclein increases synaptic vesicle clustering. Thus, pathologic -synuclein causes major alterations to synaptic architecture in the BLA, potentially contributing to behavioral impairment and amygdala dysfunction observed in synucleinopathies.
]]></description>
<dc:creator>Gcwensa, N. Z.</dc:creator>
<dc:creator>Russell, D. L.</dc:creator>
<dc:creator>Long, K. Y.</dc:creator>
<dc:creator>Brzozowski, C. F.</dc:creator>
<dc:creator>Liu, X.</dc:creator>
<dc:creator>Gamble, K. L.</dc:creator>
<dc:creator>Cowell, R. M.</dc:creator>
<dc:creator>Volpicelli-Daley, L.</dc:creator>
<dc:date>2024-05-15</dc:date>
<dc:identifier>doi:10.1101/2024.05.15.594419</dc:identifier>
<dc:title><![CDATA[Cortico-amygdala synaptic structural abnormalities produced by templated aggregation of α-synuclein]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.05.21.594886v1?rss=1">
<title>
<![CDATA[
Genetically Encoded and Modular SubCellular Organelle Probes (GEM-SCOPe) reveal widespread lysosomal and mitochondrial dysfunction in human PRKN knockout astrocytes and neurons 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.05.21.594886v1?rss=1"
</link>
<description><![CDATA[
Cellular processes including lysosomal and mitochondrial dysfunction are implicated in the development of many diseases. Quantitative visualization of mitochondria and lysosomes is crucial to understand how these organelles are dysregulated during disease. To address a gap in live-imaging tools, we developed GEM-SCOPe (Genetically Encoded and Modular SubCellular Organelle Probes), a modular toolbox of fluorescent markers designed to inform on localization, distribution, turnover, and oxidative stress of specific organelles. We expressed GEM-SCOPe in differentiated astrocytes and neurons from a human pluripotent stem cell PRKN-knockout model of Parkinsons disease and identified disease-associated changes in proliferation, lysosomal distribution, mitochondrial transport and turnover, and reactive oxygen species. We demonstrate GEM-SCOPe is a powerful panel that provide critical insight into the subcellular mechanisms underlying Parkinsons disease in human cells. GEM-SCOPe can be expanded upon and applied to a diversity of cellular models to glean an understanding of the mechanisms that promote disease onset and progression.
]]></description>
<dc:creator>Goldman, C.</dc:creator>
<dc:creator>Kareva, T.</dc:creator>
<dc:creator>Sarrafha, L.</dc:creator>
<dc:creator>Saharsrabudhe, A.</dc:creator>
<dc:creator>Schuldt, B. R.</dc:creator>
<dc:creator>Ahfeldt, T.</dc:creator>
<dc:creator>Blanchard, J. W.</dc:creator>
<dc:date>2024-05-21</dc:date>
<dc:identifier>doi:10.1101/2024.05.21.594886</dc:identifier>
<dc:title><![CDATA[Genetically Encoded and Modular SubCellular Organelle Probes (GEM-SCOPe) reveal widespread lysosomal and mitochondrial dysfunction in human PRKN knockout astrocytes and neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-05-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.06.05.597615v1?rss=1">
<title>
<![CDATA[
Internalized α-synuclein fibrils become truncated and resist degradation in neurons while glial cells rapidly degrade α-synuclein fibrils. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.06.05.597615v1?rss=1"
</link>
<description><![CDATA[
Parkinsons disease (PD) and other -synucleinopathies are characterized by the intracellular aggregates of -synuclein (S) believed to spread via the cell-to-cell transmission. To understand the contributions of various brain cells to the spreading of S pathology, we examined the metabolism of S aggregates in neuronal and glial cells. In neurons, while the full-length S rapidly disappeared following S PFF uptake, truncated S accumulated with a half-life of days rather than hours. Epitope mapping and fractionation studies indicate that S fibrils internalized by neurons was truncated at the C-terminal region and remained insoluble. In contrast, microglia and astrocytes rapidly metabolized S fibrils as the half-lives of S fibrils in these glial cells were <6 hours. Differential uptake and processing of S fibrils by neurons and glia was recapitulated in vivo where injection of fluorescently labeled S fibrils initially accumulated in glial cells followed by rapid clearance while neurons stably accumulated S fibrils at slower rate. Immunolocalization and subcellular fractionation studies show that internalized S PFF is initially localized to endosomes followed by lysosomes. The lysosome is largely responsible for the degradation of internalized S PFF as the inhibition of lysosomal function leads to the stabilization of S in all cell types. Significantly, S PFF causes lysosomal dysfunction in neurons. In summary, we show that neurons are inefficient in metabolizing internalized S aggregates, partially because S aggregates cause lysosomal dysfunction, potentially generating aggregation-prone truncated S. In contrast, glial cells may protect neurons from S aggregates by rapidly clearing S aggregates.
]]></description>
<dc:creator>Karim, M. R.</dc:creator>
<dc:creator>GAsparini, E.</dc:creator>
<dc:creator>Tiegs, E.</dc:creator>
<dc:creator>Schlichte, R.</dc:creator>
<dc:creator>Vermilyea, S. C.</dc:creator>
<dc:creator>Lee, M. K.</dc:creator>
<dc:date>2024-06-08</dc:date>
<dc:identifier>doi:10.1101/2024.06.05.597615</dc:identifier>
<dc:title><![CDATA[Internalized α-synuclein fibrils become truncated and resist degradation in neurons while glial cells rapidly degrade α-synuclein fibrils.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.06.06.597594v1?rss=1">
<title>
<![CDATA[
LRRK2 mediates haloperidol-induced changes in indirect pathway striatal projection neurons 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.06.06.597594v1?rss=1"
</link>
<description><![CDATA[
Haloperidol is used to manage psychotic symptoms in several neurological disorders through mechanisms that involve antagonism of dopamine D2 receptors that are highly expressed in the striatum. Significant side effects of haloperidol, known as extrapyramidal symptoms, lead to motor deficits similar to those seen in Parkinsons disease and present a major challenge in clinical settings. The underlying molecular mechanisms responsible for these side effects remain poorly understood. Parkinsons disease-associated LRRK2 kinase has an important role in striatal physiology and a known link to dopamine D2 receptor signaling. Here, we systematically explore convergent signaling of haloperidol and LRRK2 through pharmacological or genetic inhibition of LRRK2 kinase, as well as knock-in mouse models expressing pathogenic mutant LRRK2 with increased kinase activity. Behavioral assays show that LRRK2 kinase inhibition ameliorates haloperidol-induced motor changes in mice. A combination of electrophysiological and anatomical approaches reveals that LRRK2 kinase inhibition interferes with haloperidol-induced changes, specifically in striatal neurons of the indirect pathway. Proteomic studies and targeted intracellular pathway analyses demonstrate that haloperidol induces a similar pattern of intracellular signaling as increased LRRK2 kinase activity. Our study suggests that LRRK2 kinase plays a key role in striatal dopamine D2 receptor signaling underlying the undesirable motor side effects of haloperidol. This work opens up new therapeutic avenues for dopamine-related disorders, such as psychosis, also furthering our understanding of Parkinsons disease pathophysiology.

SummaryChen et al. demonstrate that haloperidol mediated changes in the striatal indirect pathway neurons and circuits are linked to Parkinsons disease associated LRRK2. Inhibiting LRRK2 kinase activity ameliorates the motoric side effects of haloperidol, suggesting a potential approach to alleviating the unwanted side effects of antipsychotics.
]]></description>
<dc:creator>Chen, C.</dc:creator>
<dc:creator>Masotti, M.</dc:creator>
<dc:creator>Shepard, N.</dc:creator>
<dc:creator>Promes, V.</dc:creator>
<dc:creator>Tombesi, G.</dc:creator>
<dc:creator>Arango, D.</dc:creator>
<dc:creator>Manzoni, C.</dc:creator>
<dc:creator>Greggio, E.</dc:creator>
<dc:creator>Hilfiker, S.</dc:creator>
<dc:creator>Kozorovitskiy, Y.</dc:creator>
<dc:creator>Parisiadou, L.</dc:creator>
<dc:date>2024-06-08</dc:date>
<dc:identifier>doi:10.1101/2024.06.06.597594</dc:identifier>
<dc:title><![CDATA[LRRK2 mediates haloperidol-induced changes in indirect pathway striatal projection neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.06.06.597807v1?rss=1">
<title>
<![CDATA[
Molecular and spatial transcriptomic classification of midbrain dopamine neurons and their alterations in a LRRK2G2019S model of Parkinson's disease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.06.06.597807v1?rss=1"
</link>
<description><![CDATA[
Several studies have revealed that midbrain dopamine (DA) neurons, even within a single neuroanatomical area, display heterogeneous properties. In parallel, studies using single cell profiling techniques have begun to cluster DA neurons into subtypes based on their molecular signatures. Recent work has shown that molecularly defined DA subtypes within the substantia nigra (SNc) display distinctive anatomic and functional properties, and differential vulnerability in Parkinsons disease (PD). Based on these provocative results, a granular understanding of these putative subtypes and their alterations in PD models, is imperative. We developed an optimized pipeline for single-nuclear RNA sequencing (snRNA-seq) and generated a high-resolution hierarchically organized map revealing 20 molecularly distinct DA neuron subtypes belonging to three main families. We integrated this data with spatial MERFISH technology to map, with high definition, the location of these subtypes in the mouse midbrain, revealing heterogeneity even within neuroanatomical sub-structures. Finally, we demonstrate that in the preclinical LRRK2G2019S knock-in mouse model of PD, subtype organization and proportions are preserved. Transcriptional alterations occur in many subtypes including those localized to the ventral tier SNc, where differential expression is observed in synaptic pathways, which might account for previously described DA release deficits in this model. Our work provides an advancement of current taxonomic schemes of the mouse midbrain DA neuron subtypes, a high-resolution view of their spatial locations, and their alterations in a prodromal mouse model of PD.

Teaser: Using snRNASeq and MERFISH we identified midbrain DA subtypes, mapped their spatial location, and identified alterations in a LRRK2 model
]]></description>
<dc:creator>Gaertner, Z.</dc:creator>
<dc:creator>Oram, C.</dc:creator>
<dc:creator>Schneeweis, A.</dc:creator>
<dc:creator>Schonfeld, E.</dc:creator>
<dc:creator>Bolduc, C.</dc:creator>
<dc:creator>Chen, C.</dc:creator>
<dc:creator>Dombeck, D. A.</dc:creator>
<dc:creator>Parisiadou, L.</dc:creator>
<dc:creator>Poulin, J. F.</dc:creator>
<dc:creator>Awatramani, R.</dc:creator>
<dc:date>2024-06-08</dc:date>
<dc:identifier>doi:10.1101/2024.06.06.597807</dc:identifier>
<dc:title><![CDATA[Molecular and spatial transcriptomic classification of midbrain dopamine neurons and their alterations in a LRRK2G2019S model of Parkinson's disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.06.18.599606v1?rss=1">
<title>
<![CDATA[
Cryo-electron tomography reveals the microtubule-bound form of inactive LRRK2 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.06.18.599606v1?rss=1"
</link>
<description><![CDATA[
Parkinsons Disease (PD) is the second most common neurodegenerative disorder. Mutations in leucine-rich repeat kinase 2 (LRRK2), a multi-domain protein containing both a kinase and a GTPase, are a leading cause of the familial form of PD. Pathogenic LRRK2 mutations increase LRRK2 kinase activity. While the bulk of LRRK2 is found in the cytosol, the protein associates with membranes where its Rab GTPase substrates are found, and under certain conditions, with microtubules. Integrative structural studies using single-particle cryo-electron microscopy (cryo-EM) and in situ cryo-electron tomography (cryo-ET) have revealed the architecture of microtubule-associated LRRK2 filaments, and that formation of these filaments requires LRRK2s kinase to be in the active-like conformation. However, whether LRRK2 can interact with and form filaments on microtubules in its autoinhibited state, where the kinase domain is in the inactive conformation and the N-terminal LRR domain covers the kinase active site, was not known. Using cryo-ET, we show that full-length LRRK2 can oligomerize on microtubules in its autoinhibited state. Both WT-LRRK2 and PD-linked LRRK2 mutants formed filaments on microtubules. While these filaments are stabilized by the same interfaces seen in the active-LRRK2 filaments, we observed a new interface involving the N-terminal repeats that were disordered in the active-LRRK2 filaments. The helical parameters of the autoinhibited-LRRK2 filaments are different from those reported for the active-LRRK2 filaments. Finally, the autoinhibited-LRRK2 filaments are shorter and less regular, suggesting they are less stable.
]]></description>
<dc:creator>Chen, S.</dc:creator>
<dc:creator>Basiashvili, T.</dc:creator>
<dc:creator>Hutchings, J.</dc:creator>
<dc:creator>Murillo, M. S.</dc:creator>
<dc:creator>Suarez, A. V.</dc:creator>
<dc:creator>Louro, J. A.</dc:creator>
<dc:creator>Leschziner, A. E.</dc:creator>
<dc:creator>Villa, E.</dc:creator>
<dc:date>2024-06-20</dc:date>
<dc:identifier>doi:10.1101/2024.06.18.599606</dc:identifier>
<dc:title><![CDATA[Cryo-electron tomography reveals the microtubule-bound form of inactive LRRK2]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.06.18.599622v1?rss=1">
<title>
<![CDATA[
Dissociation of novel open loop from ventral putamen to motor areas from classic closed loop in humans II: task-based function 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.06.18.599622v1?rss=1"
</link>
<description><![CDATA[
Humans ubiquitously increase the speed of their movements when motivated by incentives (i.e., capturing reward or avoiding loss). The complex interplay between incentivization and motor output is pertinent for unpacking the functional profiles of different circuits that link the basal ganglia with motor cortical areas. Here, we analyzed the functional profile of nodes forming two circuits involving putamen and motor cortical areas: the traditional "closed-loop circuit" (CLC) from sensorimotor dorsal putamen (PUTd) and a putative "open-loop circuit" (OLC) from ventral putamen (PUTv). Establishing differential function between CLC and OLC is particularly relevant for therapeutic approaches to Parkinsons disease, where OLC function is hypothesized to be relatively spared by the disease process. In a large sample fMRI study, 68 healthy controls executed speeded reaches with a joystick under different levels of incentivization to accurately hit precision targets. We dissociated effects of "incentive per se" (i.e., changes in brain activity when an upcoming movement obtains a reward or avoids a loss) from "RT effects" (i.e., brain activity that directly scales with adjustments to movement initiation time). Incentive per se was observed across sites in both CLC and OLC. However, RT effects were primarily in nodes of the OLC and motor sites, consistent with the hypothesized anatomy and function of OLC. Our findings additionally suggest valence might mediate when incentives recruit OLC to more prominent control of motor behavior.
]]></description>
<dc:creator>Dundon, N. M.</dc:creator>
<dc:creator>Rizor, E.</dc:creator>
<dc:creator>Stasiak, J.</dc:creator>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Sabugo, K.</dc:creator>
<dc:creator>Villaneuva, C.</dc:creator>
<dc:creator>Barandon, P.</dc:creator>
<dc:creator>Bostan, A. C.</dc:creator>
<dc:creator>Lapate, R. C.</dc:creator>
<dc:creator>Grafton, S. T.</dc:creator>
<dc:date>2024-06-22</dc:date>
<dc:identifier>doi:10.1101/2024.06.18.599622</dc:identifier>
<dc:title><![CDATA[Dissociation of novel open loop from ventral putamen to motor areas from classic closed loop in humans II: task-based function]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.07.10.602947v1?rss=1">
<title>
<![CDATA[
Spatially organized striatum-wide acetylcholine dynamics for the learning and extinction of Pavlovian cues and actions 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.07.10.602947v1?rss=1"
</link>
<description><![CDATA[
Striatal acetylcholine (ACh) signaling has been proposed to counteract reinforcement signals to promote extinction and behavioral flexibility. ACh dips to cues and rewards may open a temporal window for associative plasticity to occur, while elevations may promote extinction. Changes in multi-phasic striatal ACh signals have been widely reported during learning, but how and where signals are distributed to enable region-specific plasticity for the learning and degradation of cue-reward associations is poorly understood. We used array fiber photometry in mice to investigate how ACh release across the striatum evolves during learning and extinction of Pavlovian associations. We report a topographic organization of opposing changes in ACh release to cues, rewards, and consummatory actions across distinct striatum regions. We localized reward prediction error encoding in particular phases of the ACh dynamics to a specific region of the anterior dorsal striatum (aDS). Positive prediction errors in the aDS were expressed in ACh dips, and negative prediction errors in long latency ACh elevations. Silencing aDS ACh release impaired behavioral extinction, suggesting a role for ACh elevations in down-regulating cue-reward associations. Dopamine release in aDS dipped for cues during extinction, but glutamate input onto cholinergic interneurons did not change, suggesting an intrastriatal mechanism for the emergence of ACh elevations. Our large scale measurements indicate how and where ACh dynamics can shape region-specific plasticity to gate learning and promote extinction of Pavlovian associations.
]]></description>
<dc:creator>Bouabid, S.</dc:creator>
<dc:creator>Zhang, L.</dc:creator>
<dc:creator>Vu, M.-A. T.</dc:creator>
<dc:creator>Tang, K.</dc:creator>
<dc:creator>Graham, B. M.</dc:creator>
<dc:creator>Noggle, C. A.</dc:creator>
<dc:creator>Howe, M. W.</dc:creator>
<dc:date>2024-07-16</dc:date>
<dc:identifier>doi:10.1101/2024.07.10.602947</dc:identifier>
<dc:title><![CDATA[Spatially organized striatum-wide acetylcholine dynamics for the learning and extinction of Pavlovian cues and actions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.07.24.604995v1?rss=1">
<title>
<![CDATA[
Motor Cortical Neuronal Hyperexcitability Associated with α-Synuclein Aggregation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.07.24.604995v1?rss=1"
</link>
<description><![CDATA[
Dysfunction of the cerebral cortex is thought to underlie motor and cognitive impairments in Parkinson disease (PD). While cortical function is known to be suppressed by abnormal basal ganglia output following dopaminergic degeneration, it remains to be determined how the deposition of Lewy pathology disrupts cortical circuit integrity and function. Moreover, it is also unknown whether cortical Lewy pathology and midbrain dopaminergic degeneration interact to disrupt cortical function in late-stage. To begin to address these questions, we injected -synuclein (Syn) preformed fibrils (PFFs) into the dorsolateral striatum of mice to seed Syn pathology in the cortical cortex and induce degeneration of midbrain dopaminergic neurons. Using this model system, we reported that Syn aggregates accumulate in the motor cortex in a layer- and cell-subtype-specific pattern. Particularly, intratelencephalic neurons (ITNs) showed earlier accumulation and greater extent of Syn aggregates relative to corticospinal neurons (CSNs). Moreover, we demonstrated that the intrinsic excitability and inputs resistance of Syn aggregates-bearing ITNs in the secondary motor cortex (M2) are increased, along with a noticeable shrinkage of cell bodies and loss of dendritic spines. Last, neither the intrinsic excitability of CSNs nor their thalamocortical input was altered by a partial striatal dopamine depletion associated with Syn pathology. Our results documented motor cortical neuronal hyperexcitability associated with Syn aggregation and provided a novel mechanistic understanding of cortical circuit dysfunction in PD.
]]></description>
<dc:creator>Chen, L.</dc:creator>
<dc:creator>Chehade, H. D.</dc:creator>
<dc:creator>Chu, H.-Y.</dc:creator>
<dc:date>2024-07-24</dc:date>
<dc:identifier>doi:10.1101/2024.07.24.604995</dc:identifier>
<dc:title><![CDATA[Motor Cortical Neuronal Hyperexcitability Associated with α-Synuclein Aggregation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-07-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.07.31.606089v1?rss=1">
<title>
<![CDATA[
Restoration of striatal neuroprotective pathways by kinase inhibitor treatment of Parkinsons linked-LRRK2 mutant mice 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.07.31.606089v1?rss=1"
</link>
<description><![CDATA[
Parkinsons disease-associated, activating mutations in Leucine Rich Repeat Kinase 2 (LRRK2) block primary cilia formation in cholinergic and parvalbumin interneurons and astrocytes in the striatum, decreasing the production of GDNF and NRTN neuroprotective factors that normally support dopaminergic neuron viability. We show here that 3 month-dietary administration of the MLi-2 LRRK2 kinase inhibitor restores primary cilia and the Hedgehog-responsive production of neuroprotective GDNF and NRTN by these neurons; cilia are also restored on cholinergic neurons of the pedunculopontine nucleus. Importantly, we detect recovery of striatal dopaminergic processes and decreased stress-triggered Hedgehog signaling by nigral dopaminergic neurons. Thus, pathogenic LRRK2-driven cilia loss is reversible in post-mitotic neurons and astrocytes, which suggests that early administration of specific LRRK2 inhibitors may have significant therapeutic benefit for patients in the future.

One Sentence SummaryKinase inhibitor restores cilia, Hedgehog signaling, neuroprotective factors and dopamine processes in Parkinsons linked-LRRK2 mouse striatum
]]></description>
<dc:creator>Jaimon, E.</dc:creator>
<dc:creator>Lin, Y.-E.</dc:creator>
<dc:creator>Tonelli, F.</dc:creator>
<dc:creator>Antico, O.</dc:creator>
<dc:creator>Alessi, D. R.</dc:creator>
<dc:creator>Pfeffer, S. R.</dc:creator>
<dc:date>2024-08-03</dc:date>
<dc:identifier>doi:10.1101/2024.07.31.606089</dc:identifier>
<dc:title><![CDATA[Restoration of striatal neuroprotective pathways by kinase inhibitor treatment of Parkinsons linked-LRRK2 mutant mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.08.13.607784v1?rss=1">
<title>
<![CDATA[
Experimental and Computational Methods for Allelic Imbalance Analysis from Single-Nucleus RNA-seq Data 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.08.13.607784v1?rss=1"
</link>
<description><![CDATA[
Single-cell RNA-seq (scRNA-seq) is emerging as a powerful tool for understanding gene function across diverse cells. Recently, this has included the use of allele-specific expression (ASE) analysis to better understand how variation in the human genome affects RNA expression at the single-cell level. We reasoned that because intronic reads are more prevalent in single-nucleus RNA-Seq (snRNA-Seq), and introns are under lower purifying selection and thus enriched for genetic variants, that snRNA-seq should facilitate single-cell analysis of ASE. Here we demonstrate how experimental and computational choices can improve the results of allelic imbalance analysis. We explore how experimental choices, such as RNA source, read length, sequencing depth, genotyping, etc., impact the power of ASE-based methods. We developed a new suite of computational tools to process and analyze scRNA-seq and snRNA-seq for ASE. As hypothesized, we extracted more ASE information from reads in intronic regions than those in exonic regions and show how read length can be set to increase power. Additionally, hybrid selection improved our power to detect allelic imbalance in genes of interest. We also explored methods to recover allele-specific isoform expression levels from both long- and short-read snRNA-seq. To further investigate ASE in the context of human disease, we applied our methods to a Parkinsons disease cohort of 94 individuals and show that ASE analysis had more power than eQTL analysis to identify significant SNP/gene pairs in our direct comparison of the two methods. Overall, we provide an end-to-end experimental and computational approach for future studies.
]]></description>
<dc:creator>Simmons, S. K.</dc:creator>
<dc:creator>Adiconis, X.</dc:creator>
<dc:creator>Haywood, N.</dc:creator>
<dc:creator>Parker, J.</dc:creator>
<dc:creator>Lin, Z.</dc:creator>
<dc:creator>Liao, Z.</dc:creator>
<dc:creator>Tuncali, I.</dc:creator>
<dc:creator>Al'Khafaji, A.</dc:creator>
<dc:creator>Shin, A.</dc:creator>
<dc:creator>Jagadeesh, K.</dc:creator>
<dc:creator>Gosik, K.</dc:creator>
<dc:creator>Gatzen, M.</dc:creator>
<dc:creator>Smith, J. T.</dc:creator>
<dc:creator>El Kodsi, D. N.</dc:creator>
<dc:creator>Kuras, Y.</dc:creator>
<dc:creator>Baecher-Allan, C.</dc:creator>
<dc:creator>Serrano, G. E.</dc:creator>
<dc:creator>Beach, T. G.</dc:creator>
<dc:creator>Garimella, K.</dc:creator>
<dc:creator>Rozenblatt-Rosen, O.</dc:creator>
<dc:creator>Regev, A.</dc:creator>
<dc:creator>Dong, X.</dc:creator>
<dc:creator>Scherzer, C.</dc:creator>
<dc:creator>Levin, J. Z.</dc:creator>
<dc:date>2024-08-16</dc:date>
<dc:identifier>doi:10.1101/2024.08.13.607784</dc:identifier>
<dc:title><![CDATA[Experimental and Computational Methods for Allelic Imbalance Analysis from Single-Nucleus RNA-seq Data]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.08.19.608634v1?rss=1">
<title>
<![CDATA[
Peripheral Blood Immune Cells from Individuals with Parkinson's Disease or Inflammatory Bowel Disease Share Deficits in Iron Storage and Transport that are Modulated by Non-Steroidal Anti-Inflammatory Drugs 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.08.19.608634v1?rss=1"
</link>
<description><![CDATA[
Parkinsons Disease (PD) is a multisystem disorder in which dysregulated neuroimmune crosstalk and inflammatory relay via the gut-blood-brain axis have been implicated in PD pathogenesis. Although alterations in circulating inflammatory cytokines and reactive oxygen species (ROS) have been associated with PD, no biomarkers have been identified that predict clinical progression or disease outcome. Gastrointestinal (GI) dysfunction, which involves perturbation of the underlying immune system, is an early and often-overlooked symptom that affects up to 80% of individuals living with PD. Interestingly, 50-70% of individuals with inflammatory bowel disease (IBD), a GI condition that has been epidemiologically linked to PD, display chronic illness-induced anemia -- which drives toxic accumulation of iron in the gut. Ferroptotic (or iron loaded) cells have small and dysmorphic mitochondria--suggesting that mitochondrial dysfunction is a consequence of iron accumulation. In pro-inflammatory environments, iron accumulates in immune cells, suggesting a possible connection and/or synergy between iron dysregulation and immune cell dysfunction. Peripheral blood mononuclear cells (PBMCs) recapitulate certain PD-associated neuropathological and inflammatory signatures and can act as communicating messengers in the gut-brain axis. Additionally, this communication can be modulated by several environmental factors; specifically, our data further support existing literature demonstrating a role for non-steroidal anti-inflammatory drugs (NSAIDs) in modulating immune transcriptional states in inflamed individuals. A mechanism linking chronic gut inflammation to iron dysregulation and mitochondrial function within peripheral immune cells has yet to be identified in conferring risk for PD. To that end, we isolated PBMCs and simultaneously evaluated their directed transcriptome and bioenergetic status, to investigate if iron dysregulation and mitochondrial sensitization are linked in individuals living with PD or IBD because of chronic underlying remittent immune activation. We have identified shared features of peripheral inflammation and immunometabolism in individuals living with IBD or PD that may contribute to the epidemiological association reported between IBD and risk for PD.
]]></description>
<dc:creator>Bolen, M. L.</dc:creator>
<dc:creator>Gomes, B. N.</dc:creator>
<dc:creator>Gill, B.</dc:creator>
<dc:creator>Menees, K. B.</dc:creator>
<dc:creator>Staley, H.</dc:creator>
<dc:creator>Jernigan, J.</dc:creator>
<dc:creator>Tansey, M. G.</dc:creator>
<dc:date>2024-08-19</dc:date>
<dc:identifier>doi:10.1101/2024.08.19.608634</dc:identifier>
<dc:title><![CDATA[Peripheral Blood Immune Cells from Individuals with Parkinson's Disease or Inflammatory Bowel Disease Share Deficits in Iron Storage and Transport that are Modulated by Non-Steroidal Anti-Inflammatory Drugs]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.08.28.609967v1?rss=1">
<title>
<![CDATA[
Reconstitution of BNIP3/NIX-mediated autophagy reveals two pathways and hierarchical flexibility of the initiation machinery 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.08.28.609967v1?rss=1"
</link>
<description><![CDATA[
Selective autophagy is a lysosomal degradation pathway that is critical for maintaining cellular homeostasis by disposing of harmful cellular material. While the mechanisms by which soluble cargo receptors recruit the autophagy machinery are becoming increasingly clear, the principles governing how organelle-localized transmembrane cargo receptors initiate selective autophagy remain poorly understood. Here, we demonstrate that transmembrane cargo receptors can initiate autophagosome biogenesis not only by recruiting the upstream FIP200/ULK1 complex but also via a WIPI-ATG13 complex. This latter pathway is employed by the BNIP3/NIX receptors to trigger mitophagy. Additionally, other transmembrane mitophagy receptors, including FUNDC1 and BCL2L13, exclusively use the FIP200/ULK1 complex, while FKBP8 and the ER-phagy receptor TEX264 are capable of utilizing both pathways to initiate autophagy. Our study defines the molecular rules for initiation by transmembrane cargo receptors, revealing remarkable flexibility in the assembly and activation of the autophagy machinery, with significant implications for therapeutic interventions.
]]></description>
<dc:creator>Adriaenssens, E.</dc:creator>
<dc:creator>Schaar, S.</dc:creator>
<dc:creator>Cooke, A. S. I.</dc:creator>
<dc:creator>Stuke, J. F. M.</dc:creator>
<dc:creator>Sawa-Makarska, J.</dc:creator>
<dc:creator>Nguyen, T. N.</dc:creator>
<dc:creator>Ren, X.</dc:creator>
<dc:creator>Schuschnig, M.</dc:creator>
<dc:creator>Romanov, J.</dc:creator>
<dc:creator>Khuu, G.</dc:creator>
<dc:creator>Lazarou, M.</dc:creator>
<dc:creator>Hummer, G.</dc:creator>
<dc:creator>Hurley, J. H.</dc:creator>
<dc:creator>Martens, S.</dc:creator>
<dc:date>2024-08-28</dc:date>
<dc:identifier>doi:10.1101/2024.08.28.609967</dc:identifier>
<dc:title><![CDATA[Reconstitution of BNIP3/NIX-mediated autophagy reveals two pathways and hierarchical flexibility of the initiation machinery]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.08.28.610129v1?rss=1">
<title>
<![CDATA[
Dissociation of putative open loop circuit from ventral putamen to motor cortical areas in humans I: high-resolution connectomics 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.08.28.610129v1?rss=1"
</link>
<description><![CDATA[
Human movement is partly organized and executed by cortico-basal ganglia-thalamic closed-loop circuits (CLCs), wherein motor cortical areas both send inputs to and receive feedback from the basal ganglia, particularly the dorsal putamen (PUTd). These networks are compromised in Parkinsons disease (PD) due to neurodegeneration of dopaminergic inputs primarily to PUTd. Yet, fluid movement in PD can sporadically occur, especially when induced by emotionally arousing events. Rabies virus tracing in non-human primates has identified a potential alternative motor pathway, wherein the ventral putamen (PUTv) receives inputs from subcortical limbic areas (such as amygdala nuclei) and sends outputs to motor cortical areas putatively via the nucleus basalis of Meynert (NBM). We hypothesize that this separable open loop circuit (OLC) may exist in humans and explain the preservation of movement after CLC degradation. Here, we provide evidence for the normal human OLC with ultra-high field (7T), multi-echo functional magnetic resonance imaging. We acquired resting-state functional connectivity (FC) scans from 21 healthy adults (avg. age = 29, 12M/9F, all right-handed) and mapped left-hemisphere seed-to-voxel connectivity to assess PUTv FC with putative subcortical nodes and motor cortical areas. We found that putative OLC node (basolateral amygdala, NBM) FC was greater with PUTv (p < 0.05), while CLC subcortical seed (ventrolateral nucleus of thalamus) FC was greater with PUTd (p<0.01). Striatal FC patterns varied across cortical motor areas, with cingulate (p < 0.0001) and supplementary (p < 0.0001) motor areas showing greater FC with PUTv vs. nucleus accumbens. SMA had greater FC with PUTd vs. PUTv (p < 0.0001), while cingulate and primary motor areas showed no significant differences in FC between PUTd and PUTv (p > 0.1). Collectively, these results suggest that PUTv is functionally connected to motor cortical areas and may be integrated into a separable motor OLC with subcortical limbic inputs.
]]></description>
<dc:creator>Rizor, E. J.</dc:creator>
<dc:creator>Dundon, N. M.</dc:creator>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Stasiak, J.</dc:creator>
<dc:creator>Li, T.</dc:creator>
<dc:creator>Bostan, A.</dc:creator>
<dc:creator>Lapate, R. C.</dc:creator>
<dc:creator>Grafton, S. T.</dc:creator>
<dc:date>2024-08-29</dc:date>
<dc:identifier>doi:10.1101/2024.08.28.610129</dc:identifier>
<dc:title><![CDATA[Dissociation of putative open loop circuit from ventral putamen to motor cortical areas in humans I: high-resolution connectomics]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.08.29.610310v1?rss=1">
<title>
<![CDATA[
Movement-related activity in the internal globus pallidus of the parkinsonian macaque 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.08.29.610310v1?rss=1"
</link>
<description><![CDATA[
Although the basal ganglia (BG) plays a central role in the motor symptoms of Parkinsons disease, few studies have investigated the influence of parkinsonism on movement-related activity in the BG. Here, we studied the perimovement activity of neurons in globus pallidus internus (GPi) of non-human primates during performance of a choice reaction time reaching task before and after the induction of parkinsonism by administration of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). Neuronal responses, including increases or decreases in firing rate, were equally common in the parkinsonian brain as seen prior to MPTP and the distribution of different response types was largely unchanged. The slowing of behavioral reaction times and movement durations following the induction of parkinsonism was accompanied by a prolongation of the time interval between neuronal response onset and movement initiation. Neuronal responses were also reduced in magnitude and prolonged in duration after the induction of parkinsonism. Importantly, those two effects were more pronounced among decrease-type responses, and they persisted after controlling for MPTP-induced changes in the between-trial variability in response timing. Following MPTP the trial-to-trial timing of neuronal responses also became uncoupled from the time of movement onset and more variable in general. Overall, the effects of MPTP on temporal features of GPi responses were related to the severity of parkinsonian motor impairments whereas changes in response magnitude and duration did not reflect symptom severity consistently. These findings point to a previously underappreciated potential role for abnormalities in the timing of GPi task-related activity in the generation of parkinsonian motor signs.

New & NoteworthyAlthough the globus pallidus internus (GPi) plays a central role in the cardinal symptoms of Parkinsons disease (PD), how parkinsonism alters the movement-related activity of GPi neurons remains understudied. Using a monkey model of PD, we found that: 1) the timing of GPi responses became uncoupled from movement onset. And 2) responses, especially decrease-type responses, became attenuated and prolonged. These abnormalities in GPi perimovement activity may contribute to the generation of parkinsonian motor signs.
]]></description>
<dc:creator>Kase, D.</dc:creator>
<dc:creator>Zimnik, A. J.</dc:creator>
<dc:creator>Han, Y.</dc:creator>
<dc:creator>Harsch, D. R.</dc:creator>
<dc:creator>Bacha, S.</dc:creator>
<dc:creator>Cox, K. M.</dc:creator>
<dc:creator>Bostan, A. C.</dc:creator>
<dc:creator>Richardson, R. M.</dc:creator>
<dc:creator>Turner, R. S.</dc:creator>
<dc:date>2024-08-30</dc:date>
<dc:identifier>doi:10.1101/2024.08.29.610310</dc:identifier>
<dc:title><![CDATA[Movement-related activity in the internal globus pallidus of the parkinsonian macaque]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-30</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.08.30.610524v1?rss=1">
<title>
<![CDATA[
Temporal control of acute protein aggregate turnover by UBE3C and NRF1-dependent proteasomal pathways 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.08.30.610524v1?rss=1"
</link>
<description><![CDATA[
A hallmark of neurodegenerative diseases is the progressive loss of proteostasis, leading to the accumulation of misfolded proteins or protein aggregates, with subsequent cytotoxicity. To combat this toxicity, cells have evolved degradation pathways (ubiquitin-proteasome system and autophagy) that detect and degrade misfolded proteins. However, studying the underlying cellular pathways and mechanisms has remained a challenge, as formation of many types of protein aggregates is asynchronous, with individual cells displaying distinct kinetics, thereby hindering rigorous time-course studies. Here, we merge a kinetically tractable and synchronous agDD-GFP system for aggregate formation with targeted gene knockdowns, to uncover degradation mechanisms used in response to acute aggregate formation. We find that agDD-GFP forms amorphous aggregates by cryo-electron tomography at both early and late stages of aggregate formation. Aggregate turnover occurs in a proteasome-dependent mechanism in a manner that is dictated by cellular aggregate burden, with no evidence of the involvement of autophagy. Lower levels of misfolded agDD-GFP, enriched in oligomers, utilizes UBE3C-dependent proteasomal degradation in a pathway that is independent of RPN13 ubiquitylation by UBE3C. Higher aggregate burden activates the NRF1 transcription factor to increase proteasome subunit transcription, and subsequent degradation capacity of cells. Loss or gain of NRF1 function alters the turnover of agDD-GFP under conditions of high aggregate burden. Together, these results define the role of UBE3C in degradation of this class of misfolded aggregation-prone proteins and reveals a role for NRF1 in proteostasis control in response to widespread protein aggregation.
]]></description>
<dc:creator>Hickey, K. L.</dc:creator>
<dc:creator>Panov, A.</dc:creator>
<dc:creator>Whelan, E. M.</dc:creator>
<dc:creator>Schaefer, T.</dc:creator>
<dc:creator>Mizrak, A.</dc:creator>
<dc:creator>Kopito, R. R.</dc:creator>
<dc:creator>Baumeister, W.</dc:creator>
<dc:creator>Fernandez-Busnadiego, R.</dc:creator>
<dc:creator>Harper, W.</dc:creator>
<dc:date>2024-09-01</dc:date>
<dc:identifier>doi:10.1101/2024.08.30.610524</dc:identifier>
<dc:title><![CDATA[Temporal control of acute protein aggregate turnover by UBE3C and NRF1-dependent proteasomal pathways]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.08.30.610586v1?rss=1">
<title>
<![CDATA[
CHCHD2 mutant mice display mitochondrial protein accumulation and disrupted energy metabolism 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.08.30.610586v1?rss=1"
</link>
<description><![CDATA[
Mutations in the mitochondrial cristae protein CHCHD2 lead to a late-onset autosomal dominant form of Parkinsons disease (PD) which closely resembles idiopathic PD, providing the opportunity to gain new insights into the mechanisms of mitochondrial dysfunction contributing to PD. To begin to address this, we used CRISPR genome-editing to generate CHCHD2 T61I point mutant mice. CHCHD2 T61I mice had normal viability, and had only subtle motor deficits with no signs of premature dopaminergic (DA) neuron degeneration. Nonetheless, CHCHD2 T61I mice exhibited robust molecular changes in the brain including increased CHCHD2 insolubility, accumulation of CHCHD2 protein preferentially in the substantia nigra (SN), and elevated levels of -synuclein. Metabolic analyses revealed an increase in glucose metabolism through glycolysis relative to the TCA cycle with increased respiratory exchange ratio, and immune-electron microscopy revelated disrupted mitochondria in DA neurons. Moreover, spatial genomics revealed decreased expression of mitochondrial complex I and III respiratory chain proteins, while proteomics revealed increased respiratory chain and other mitochondrial protein-protein interactions. As such, the CHCHD2 T61I point-mutation mice exhibit robust mitochondrial disruption and a consequent metabolic shift towards glycolysis. These findings thus establish CHCHD2 T61I mice as a new model for mitochondrial-based PD, and implicate disrupted respiratory chain function as a likely causative driver.
]]></description>
<dc:creator>Liao, S.-C.</dc:creator>
<dc:creator>Kano, K.</dc:creator>
<dc:creator>Phanse, S.</dc:creator>
<dc:creator>Nguyen, M.</dc:creator>
<dc:creator>Margolis, E. B.</dc:creator>
<dc:creator>Fu, Y.</dc:creator>
<dc:creator>Meng, J.</dc:creator>
<dc:creator>Moutaoufik, M. T.</dc:creator>
<dc:creator>Chatterton, Z.</dc:creator>
<dc:creator>Aoki, H.</dc:creator>
<dc:creator>Simms, J.</dc:creator>
<dc:creator>Hsieh, I.</dc:creator>
<dc:creator>Suteja, F.</dc:creator>
<dc:creator>Sei, Y.</dc:creator>
<dc:creator>Huang, E. J.</dc:creator>
<dc:creator>McAvoy, K.</dc:creator>
<dc:creator>Manfredi, G.</dc:creator>
<dc:creator>Halliday, G.</dc:creator>
<dc:creator>Babu, M.</dc:creator>
<dc:creator>Nakamura, K.</dc:creator>
<dc:date>2024-09-01</dc:date>
<dc:identifier>doi:10.1101/2024.08.30.610586</dc:identifier>
<dc:title><![CDATA[CHCHD2 mutant mice display mitochondrial protein accumulation and disrupted energy metabolism]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.06.20.599774v1?rss=1">
<title>
<![CDATA[
Cortical synaptic vulnerabilities revealed in a α-synuclein aggregation model of Parkinson's disease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.06.20.599774v1?rss=1"
</link>
<description><![CDATA[
-Synuclein aggregates are a hallmark of multiple neurodegenerative disorders, including Parkinsons disease and dementia with Lewy bodies. Cortical pathology in these diseases correlates with cognitive decline. This pathology may progressively impact synaptic connectivity, and we tested this in the frontal cortex in a -synucleinopathy model. High-resolution imaging showed that intracortical VGLUT1-positive excitatory synapses became increasingly vulnerable, exhibiting early loss near aggregates. The remaining intracortical excitatory synapses were structurally disrupted as measured by high-resolution imaging and electron microscopy. These effects were accompanied by synaptic -syn accumulation. Ontology analysis of gene expression changes in pathology-bearing neurons showed altered synapse-related processes, with upregulated genes indicating candidate resilience mechanisms. Analyses informed by neuroanatomical connectivity provided additional insights. Mesoscans determined that synapse loss is correlated between cortical layer V and layer II/III, which includes neurons projecting to layer V. Analysis across cortical hemispheres supported a vulnerability of intratelencephalic (IT) projection neurons in layer V. In contrast to intracortical synapses, long-range VGLUT2+ excitatory synapses were spared. Inhibitory VGAT+ synapses were modestly impacted, with a late-stage reduction in postsynaptic specializations. Electrophysiological recordings validated impaired excitatory transmission. This study reveals progressive, synapse-type specific vulnerabilities and resilience and characterizes intracortical synapses as cellular locus of disrupted connectivity in -synucleinopathy.
]]></description>
<dc:creator>Sah, S.</dc:creator>
<dc:creator>Sauerbeck, A. D.</dc:creator>
<dc:creator>Gupta, J.</dc:creator>
<dc:creator>Perez-Acuna, D.</dc:creator>
<dc:creator>Reiber, J. E.</dc:creator>
<dc:creator>Russell, D.</dc:creator>
<dc:creator>Goralski, T.</dc:creator>
<dc:creator>Henderson, M.</dc:creator>
<dc:creator>Volpicelli-Daley, L. A.</dc:creator>
<dc:creator>Higley, M. J.</dc:creator>
<dc:creator>Kummer, T. T.</dc:creator>
<dc:creator>Biederer, T.</dc:creator>
<dc:date>2024-06-21</dc:date>
<dc:identifier>doi:10.1101/2024.06.20.599774</dc:identifier>
<dc:title><![CDATA[Cortical synaptic vulnerabilities revealed in a α-synuclein aggregation model of Parkinson's disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-06-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.08.27.609924v1?rss=1">
<title>
<![CDATA[
Behavioral screening defines three molecular Parkinsonism subgroups in Drosophila 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.08.27.609924v1?rss=1"
</link>
<description><![CDATA[
Parkinsonism is defined by motor dysfunction, but the specific upstream molecular causes of these clinical symptoms can vary widely. We hypothesize that these causes converge onto a limited number of core cellular pathways. To investigate this, we created a new collection of 24 genetically very well-controlled animal models of familial forms of parkinsonism. Using unbiased behavioral screening and machine learning we identified three clusters of mutants that converge on (1) mitochondrial function; (2) retromer/vesicle trafficking; and (3) proteostasis/autophagy. Genes within each cluster have a similar genetic interaction profile and compounds that target specific molecular pathways ameliorate dopaminergic neuron dysfunction in a cluster-specific manner. This suggests that familial parkinsonism can be stratified into three broad functional groups and our findings pave the way for targeted biomarker discovery and drug development.
]]></description>
<dc:creator>Kaempf, N.</dc:creator>
<dc:creator>Valadas, J. D. S.</dc:creator>
<dc:creator>Robberechts, P.</dc:creator>
<dc:creator>Schoovaerts, N.</dc:creator>
<dc:creator>Praschberger, R.</dc:creator>
<dc:creator>Ortega, A.</dc:creator>
<dc:creator>Kilic, A.</dc:creator>
<dc:creator>Chabot, D.</dc:creator>
<dc:creator>Pech, U.</dc:creator>
<dc:creator>Kuenen, S.</dc:creator>
<dc:creator>Vilain, S.</dc:creator>
<dc:creator>Baz, E.-S.</dc:creator>
<dc:creator>Singh, J.</dc:creator>
<dc:creator>Davis, J.</dc:creator>
<dc:creator>Liu, S.</dc:creator>
<dc:creator>Verstreken, P.</dc:creator>
<dc:date>2024-08-27</dc:date>
<dc:identifier>doi:10.1101/2024.08.27.609924</dc:identifier>
<dc:title><![CDATA[Behavioral screening defines three molecular Parkinsonism subgroups in Drosophila]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.09.03.610925v1?rss=1">
<title>
<![CDATA[
The mitochondrial unfolded protein response promotes senescence in human microglia by increasing S-adenosylmethionine availability for polyamine synthesis. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.09.03.610925v1?rss=1"
</link>
<description><![CDATA[
Mitochondria have developed a specialized mitochondrial unfolded protein response (UPRmt) to maintain proteostasis and promote recovery under stress conditions. Research in simple organisms has demonstrated that UPRmt activation in glial cells promotes proteostasis through beneficial non-cell-autonomous communication with neurons. However, the role of mitochondrial stress responses in the human brain remains unclear. To address this knowledge gap, we profiled the cell type-specific roles of the UPRmt using human induced pluripotent stem cell-derived neuronal and glial cultures and brain organoids. We found that UPRmt activation induces metabolic rewiring in human microglia, resulting in a senescence phenotype mediated by S-adenosylmethionine availability for polyamine synthesis. Additionally, UPRmt disrupted microglial intercellular communication, leading to microglia-mediated dysfunction of phagocytic pathways and increased inflammatory signaling. Using microglia-brain-assembloids, we observed distinct contributions by microglia to brain senescence and neurodegenerative disease processes driven by mitochondrial stress responses. These findings underscore the profound impact of defects in mitochondrial proteostasis on intercellular networks during brain aging and disease.
]]></description>
<dc:creator>Perez Jimenez, M. J.</dc:creator>
<dc:creator>Bertoli, F.</dc:creator>
<dc:creator>Raji, H.</dc:creator>
<dc:creator>Lam, A.</dc:creator>
<dc:creator>Bosch, M.</dc:creator>
<dc:creator>Weissleder, C.</dc:creator>
<dc:creator>Nemazanyy, I.</dc:creator>
<dc:creator>Kalb, S.</dc:creator>
<dc:creator>Hirschberg, I.</dc:creator>
<dc:creator>Brunetti, D.</dc:creator>
<dc:creator>Heckenbach, I.</dc:creator>
<dc:creator>Scheibye-Knudsen, M.</dc:creator>
<dc:creator>Deleidi, M.</dc:creator>
<dc:date>2024-09-03</dc:date>
<dc:identifier>doi:10.1101/2024.09.03.610925</dc:identifier>
<dc:title><![CDATA[The mitochondrial unfolded protein response promotes senescence in human microglia by increasing S-adenosylmethionine availability for polyamine synthesis.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.09.09.611943v1?rss=1">
<title>
<![CDATA[
Presynapses are mitophagy pit stops that prevent axon degeneration 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.09.09.611943v1?rss=1"
</link>
<description><![CDATA[
Defects in neuronal mitophagy have been linked to neurodegenerative diseases including Parkinsons disease. However, despite the importance of mitophagy in neuronal homeostasis, the mechanistic basis for neurodegeneration when mitophagy is defective is unclear. Here, using human neurons, we discover that presynapses are mitophagy pit stops for damaged axonal mitochondria. We show that while mitochondrial damage and PINK1/Parkin activation events are distributed throughout axons, mitophagy initiation and autophagosome formation are restricted to presynapses, which we show contain the machineries required for mitophagy. Being the primary sites of axonal mitophagy, presynapses were vulnerable when PINK1/Parkin mitophagy was defective. We observed local cytochrome c release within presynapses from an accumulation of damaged mitochondria. This resulted in downstream degradative caspase activation, defining a mechanism for neurodegeneration. Pharmacological rescue of axon degeneration was achieved through synthetic upregulation of receptor mediated mitophagy with the clinically approved compound Roxadustat, revealing a potential therapeutic avenue for disease.
]]></description>
<dc:creator>Lam, W. K.</dc:creator>
<dc:creator>Lindblom, R. S. J.</dc:creator>
<dc:creator>Milky, B.</dc:creator>
<dc:creator>Mazzachi, P.</dc:creator>
<dc:creator>Hadian-Jazi, M.</dc:creator>
<dc:creator>Kung, C.</dc:creator>
<dc:creator>Khuu, G.</dc:creator>
<dc:creator>Uoselis, L.</dc:creator>
<dc:creator>Nguyen, T. N.</dc:creator>
<dc:creator>Skulsuppaisarn, M.</dc:creator>
<dc:creator>Saunders, T. L.</dc:creator>
<dc:creator>Schmidt, M. F.</dc:creator>
<dc:creator>Dewson, G.</dc:creator>
<dc:creator>Fogel, A. I.</dc:creator>
<dc:creator>Bardy, C.</dc:creator>
<dc:creator>Lazarou, M.</dc:creator>
<dc:date>2024-09-09</dc:date>
<dc:identifier>doi:10.1101/2024.09.09.611943</dc:identifier>
<dc:title><![CDATA[Presynapses are mitophagy pit stops that prevent axon degeneration]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.09.17.613365v1?rss=1">
<title>
<![CDATA[
Type-II kinase inhibitors that target Parkinson's Disease-associated LRRK2 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.09.17.613365v1?rss=1"
</link>
<description><![CDATA[
Aberrant increases in kinase activity of leucine-rich repeat kinase 2 (LRRK2) are associated with Parkinsons disease (PD). Numerous LRRK2-selective type-I kinase inhibitors have been developed and some have entered clinical trials. In this study, we present the first LRRK2-selective type-II kinase inhibitors. Targeting the inactive conformation of LRRK2 is functionally distinct from targeting the active-like conformation using type-I inhibitors. We designed these inhibitors using a combinatorial chemistry approach fusing selective LRRK2 type-I and promiscuous type-II inhibitors by iterative cycles of synthesis supported by structural biology and activity testing. Our current lead structures are selective and potent LRRK2 inhibitors. Through cellular assays, cryo-electron microscopy structural analysis, and in vitro motility assays, we show that our inhibitors stabilize the open, inactive kinase conformation. These new conformation-specific compounds will be invaluable as tools to study LRRK2s function and regulation, and expand the potential therapeutic options for PD.
]]></description>
<dc:creator>Raig, N. D.</dc:creator>
<dc:creator>Surridge, K. J.</dc:creator>
<dc:creator>Sanz-Murillo, M.</dc:creator>
<dc:creator>Dederer, V.</dc:creator>
<dc:creator>Kramer, A.</dc:creator>
<dc:creator>Schwalm, M. P.</dc:creator>
<dc:creator>Elson, L.</dc:creator>
<dc:creator>Chatterjee, D.</dc:creator>
<dc:creator>Mathea, S.</dc:creator>
<dc:creator>Hanke, T.</dc:creator>
<dc:creator>Leschziner, A. E.</dc:creator>
<dc:creator>Reck-Peterson, S. L.</dc:creator>
<dc:creator>Knapp, S.</dc:creator>
<dc:date>2024-09-18</dc:date>
<dc:identifier>doi:10.1101/2024.09.17.613365</dc:identifier>
<dc:title><![CDATA[Type-II kinase inhibitors that target Parkinson's Disease-associated LRRK2]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.09.26.615247v1?rss=1">
<title>
<![CDATA[
Comparative Analysis of AAV Serotypes for Transduction of Olfactory Sensory Neurons 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.09.26.615247v1?rss=1"
</link>
<description><![CDATA[
Olfactory sensory neurons within the nasal epithelium detect volatile odorants and relay odor information to the central nervous system. Unlike other sensory inputs, olfactory sensory neurons interface with the external environment and project their axons directly into the central nervous system. The use of adeno-associated viruses to target these neurons has garnered interest for applications in gene therapy, probing olfactory sensory neuron biology, and modeling disease. To date, there is no consensus on the optimal AAV serotype for efficient and selective transduction of olfactory sensory neurons in vivo. Here we utilized serial confocal imaging and single-nucleus RNA sequencing to evaluate the efficacy of 11 different AAV serotypes in transducing murine olfactory sensory neurons via non-invasive nasal inoculation. Our results reveal that AAV1, while highly effective, exhibited broad tropism, whereas AAV-DJ/8 showed the greatest specificity for olfactory sensory neurons.
]]></description>
<dc:creator>Belfort, B. D.</dc:creator>
<dc:creator>Jia, J. D.</dc:creator>
<dc:creator>Garza, A. R.</dc:creator>
<dc:creator>Insalaco, A. M.</dc:creator>
<dc:creator>McGinnis, J. P.</dc:creator>
<dc:creator>Pekarek, B. T.</dc:creator>
<dc:creator>Ortiz, J.</dc:creator>
<dc:creator>Tepe, B.</dc:creator>
<dc:creator>Chen, H.</dc:creator>
<dc:creator>Liu, Z.</dc:creator>
<dc:creator>Arenkiel, B. R.</dc:creator>
<dc:date>2024-09-28</dc:date>
<dc:identifier>doi:10.1101/2024.09.26.615247</dc:identifier>
<dc:title><![CDATA[Comparative Analysis of AAV Serotypes for Transduction of Olfactory Sensory Neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.09.28.615015v1?rss=1">
<title>
<![CDATA[
Bridge-like lipid transfer protein 3A (BLTP3A) is associated with membranes of the late endocytic pathway and is an effector of CASM 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.09.28.615015v1?rss=1"
</link>
<description><![CDATA[
Recent studies have identified a family of rod-shaped proteins thought to mediate lipid transfer at intracellular membrane contacts by a bridge-like mechanism. We show one such protein, bridge-like lipid transfer protein 3A (BLTP3A)/UHRF1BP1 binds VAMP7 vesicles via its C-terminal region and anchors them to lysosomes via its chorein domain containing N-terminal region to Rab7. Upon lysosome damage, BLTP3A-positive vesicles rapidly (within minutes) dissociate from lysosomes. Lysosome damage is known to activate the CASM (Conjugation of ATG8 to Single Membranes) pathway leading to lipidation and recruitment to lysosomes of mammalian ATG8 (mATG8) proteins. We find that this process drives the reassociation of BLTP3A with damaged lysosomes via an interaction of its LIR motif with mATG8 which coincides with a dissociation from the vesicles. Our findings reveal that BLTP3A is an effector of CASM, potentially as part of a mechanism to help repair or minimize lysosome damage.

TeaserBLTP3A is localized on membranes of the late endocytic system and is CASM effector, pointing to a role of this proteins in the response to lysosome damage
]]></description>
<dc:creator>Hanna, M. G.</dc:creator>
<dc:creator>Rodriguez Cruz, H. O.</dc:creator>
<dc:creator>Fujise, K.</dc:creator>
<dc:creator>Zhuoning, L.</dc:creator>
<dc:creator>Monetti, M.</dc:creator>
<dc:creator>De Camilli, P.</dc:creator>
<dc:date>2024-09-28</dc:date>
<dc:identifier>doi:10.1101/2024.09.28.615015</dc:identifier>
<dc:title><![CDATA[Bridge-like lipid transfer protein 3A (BLTP3A) is associated with membranes of the late endocytic pathway and is an effector of CASM]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.10.14.618312v1?rss=1">
<title>
<![CDATA[
DIO-SPOTlight Transgenic Mouse to Functionally Monitor Protein Synthesis Regulated by the Integrated Stress Response 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.10.14.618312v1?rss=1"
</link>
<description><![CDATA[
The integrated stress response (ISR) is a core pathway for maintaining cellular proteostasis and a key regulator of translation in processes beyond the cellular response to stress. For example, the ISR regulates developmental axonogenesis, learning and memory, and synaptic plasticity in the brain. One barrier to uncovering ISR roles in health and disease is the challenge of monitoring its activity. The transient nature of regulatory phosphorylation events and lack of transgenic ISR reporter mouse lines make visually capturing the molecular hallmarks of ISR activation in specific cell types especially difficult. We recently developed the SPOTlight (Selective Phospho-eIF2 Open reading frame Tracking light) reporter, which uniquely provides a readout of the functional state of protein synthesis initiation dynamics that are regulated by the ISR. Here, we report the generation of a transgenic mouse line with Cre-dependent expression of SPOTlight. This resource enables selective visualization of ISR-regulated functional activity across genetically defined cell populations body-wide. Using a pan-neuronal Cre line (Nestin-Cre), we demonstrate the reporters performance and applications for cell-specific discovery, live tissue assessments and quantitative comparisons across broad physical space. We also specifically investigated the extent to which the property of steady-state basal ISR activation, recently described in dorsal striatal cholinergic interneurons, extends to other classes of cholinergic neurons and provide a CNS-wide atlas of SPOTlight activity in these cells. The DIO-SPOTlight mouse enables a wide range of studies in all organ systems and functional monitoring opportunities not previously accessible.
]]></description>
<dc:creator>Oliver, M. L.</dc:creator>
<dc:creator>Caffall, Z. F.</dc:creator>
<dc:creator>Eatman, C. B.</dc:creator>
<dc:creator>Faw, T. D.</dc:creator>
<dc:creator>Calakos, N.</dc:creator>
<dc:date>2024-10-15</dc:date>
<dc:identifier>doi:10.1101/2024.10.14.618312</dc:identifier>
<dc:title><![CDATA[DIO-SPOTlight Transgenic Mouse to Functionally Monitor Protein Synthesis Regulated by the Integrated Stress Response]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.10.14.618295v1?rss=1">
<title>
<![CDATA[
Cell-Type Resolved Protein Atlas of Brain Lysosomes Identifies SLC45A1-Associated Disease as a Lysosomal Disorder 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.10.14.618295v1?rss=1"
</link>
<description><![CDATA[
Mutations in lysosomal genes cause neurodegeneration and neurological lysosomal storage disorders (LSDs). Despite their essential role in brain homeostasis, the cell-type-specific composition and function of lysosomes remain poorly understood. Here, we report a quantitative protein atlas of the lysosome from mouse neurons, astrocytes, oligodendrocytes, and microglia. We identify dozens of novel lysosomal proteins and reveal the diversity of the lysosomal composition across brain cell types. Notably, we discovered SLC45A1, mutations in which cause a monogenic neurological disease, as a neuron-specific lysosomal protein. Loss of SLC45A1 causes lysosomal dysfunction in vitro and in vivo. Mechanistically, SLC45A1 plays a dual role in lysosomal sugar transport and stabilization of V1 subunits of the V-ATPase. SLC45A1 deficiency depletes the V1 subunits, elevates lysosomal pH, and disrupts iron homeostasis causing mitochondrial dysfunction. Altogether, our work redefines SLC45A1-associated disease as a LSD and establishes a comprehensive map to study lysosome biology at cell-type resolution in the brain and its implications for neurodegeneration.
]]></description>
<dc:creator>Ghoochani, A.</dc:creator>
<dc:creator>Heiby, J. C.</dc:creator>
<dc:creator>Rawat, E. S.</dc:creator>
<dc:creator>Medoh, U. N.</dc:creator>
<dc:creator>Di Fraia, D.</dc:creator>
<dc:creator>Dong, W.</dc:creator>
<dc:creator>Gastou, M.</dc:creator>
<dc:creator>Nyame, K.</dc:creator>
<dc:creator>Laqtom, N. N.</dc:creator>
<dc:creator>Gomez-Ospina, N.</dc:creator>
<dc:creator>Ori, A.</dc:creator>
<dc:creator>Abu-Remaileh, M.</dc:creator>
<dc:date>2024-10-15</dc:date>
<dc:identifier>doi:10.1101/2024.10.14.618295</dc:identifier>
<dc:title><![CDATA[Cell-Type Resolved Protein Atlas of Brain Lysosomes Identifies SLC45A1-Associated Disease as a Lysosomal Disorder]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.10.11.617939v1?rss=1">
<title>
<![CDATA[
Striatal lateral inhibition regulates action selection in a mouse model of levodopa-induced dyskinesia 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.10.11.617939v1?rss=1"
</link>
<description><![CDATA[
Striatal medium spiny neurons (MSNs) integrate multiple external inputs to shape motor output. In addition, MSNs form local inhibitory synaptic connections with one another. The function of striatal lateral inhibition is unknown, but one possibility is in selecting an intended action while suppressing alternatives. Action selection is disrupted in several movement disorders, including levodopa-induced dyskinesia (LID), a complication of Parkinsons disease (PD) therapy characterized by involuntary movements. Here, we identify chronic changes in the strength of striatal lateral inhibitory synapses in a mouse model of PD/LID. These synapses are also modulated by acute dopamine signaling. Chemogenetic suppression of lateral inhibition originating from dopamine D2 receptor-expressing MSNs lowers the threshold to develop involuntary movements in vivo, supporting a role in motor control. By examining the role of lateral inhibition in basal ganglia function and dysfunction, we expand the framework surrounding the role of striatal microcircuitry in action selection.
]]></description>
<dc:creator>Twedell, E. L.</dc:creator>
<dc:creator>Bair-Marshall, C. J.</dc:creator>
<dc:creator>Girasole, A. E.</dc:creator>
<dc:creator>Scaria, L. K.</dc:creator>
<dc:creator>Sridhar, S.</dc:creator>
<dc:creator>Nelson, A. B.</dc:creator>
<dc:date>2024-10-12</dc:date>
<dc:identifier>doi:10.1101/2024.10.11.617939</dc:identifier>
<dc:title><![CDATA[Striatal lateral inhibition regulates action selection in a mouse model of levodopa-induced dyskinesia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.10.07.617117v1?rss=1">
<title>
<![CDATA[
Dopamine and cortical iPSC-derived neurons with different Parkinsonian mutations show variation in lysosomal and mitochondrial dysfunction: implications for protein deposition versus selective cell loss 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.10.07.617117v1?rss=1"
</link>
<description><![CDATA[
BackgroundMutations causing Parkinsons disease (PD) give diverse pathological phenotypes whose cellular correlates remain to be determined. For example, those with PRKN loss of function mutations have significantly earlier selective vulnerability of dopamine neurons, those with SNCA mutations have increased alpha-synuclein deposition, while those with LRRK2 mutations have additional deposition of tau. Yet all three mutation types are implicated in mitochondrial and/or lysosomal dysfunction. Direct comparison of cell models with these mutations would clarify the relative cellular dysfunctions associated with these different pathological phenotypes.

MethodsAn unbiased high-content imaging platform using orthogonal probes to assess both lysosomal and mitochondrial dysfunction, along with alpha-synuclein and tau protein deposition was established using induced pluripotent stem cell (iPSC) derived cortical and ventral midbrain neurons. Three mutation types, SNCA A53T, LRRK2 R1441G and PRKN loss of function (lof), were selected as exemplars of divergent PD pathological phenotypes and compared to each other, and to control iPSC from subjects without PD.

ResultsDifferent PD mutations caused cell type specific dysfunctions, likely to impact on both selective neuronal vulnerability and the pathologies observed in PD. Comparison of dopamine neurons identified that both lysosomal and mitochondrial dysfunction were predominant with PRKN lof mutations, whereas immunofluorescent staining revealed that SNCA A53T and LRRK2 R1441G mutations had increased tau deposition. In contrast, cortical neurons with SNCA and LRRK2 mutations both had mitochondrial and autophagy impairments without protein deposition, with LRRK2 cells additionally showing decreased glucocerebrosidase activity and increased alpha-synuclein phosphorylation.

ConclusionsLysosomal and mitochondrial dysfunction are predominant in dopamine neurons with PRKN lof mutations, and may drive the early selective loss of dopamine neurons in PRKN mutation carriers. More subtle cellular abnormalities in the SNCA A53T cell lines are likely to predispose to alpha-synuclein aggregation and tau protein deposition over time. The LRRK2 R1441G may also predispose to tau deposition, but despite substantial lysosomal dysfunction with increased alpha-synuclein phosphorylation, pathological alpha-synuclein accumulations were not observed. Understanding the mechanistic differences in how lysosomal and mitochondrial dysfunction impact on PD pathogenesis in different disease subtypes may be important for therapeutic development.
]]></description>
<dc:creator>Chedid, J.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Labrador-Garrido, A.</dc:creator>
<dc:creator>Abu-Bonsrah, D.</dc:creator>
<dc:creator>Pavan, C.</dc:creator>
<dc:creator>Fraser, T.</dc:creator>
<dc:creator>Zhong, M.</dc:creator>
<dc:creator>Johnston, J. A.</dc:creator>
<dc:creator>Thompson, L. H.</dc:creator>
<dc:creator>Kirik, D.</dc:creator>
<dc:creator>Parish, C.</dc:creator>
<dc:creator>Halliday, G.</dc:creator>
<dc:creator>Sue, C.</dc:creator>
<dc:creator>Wali, G.</dc:creator>
<dc:creator>Dzamko, N.</dc:creator>
<dc:date>2024-10-11</dc:date>
<dc:identifier>doi:10.1101/2024.10.07.617117</dc:identifier>
<dc:title><![CDATA[Dopamine and cortical iPSC-derived neurons with different Parkinsonian mutations show variation in lysosomal and mitochondrial dysfunction: implications for protein deposition versus selective cell loss]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.10.07.616997v1?rss=1">
<title>
<![CDATA[
LRRK2-mutant microglia trigger dopaminergic neurodegeneration when activated by neuromelanin 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.10.07.616997v1?rss=1"
</link>
<description><![CDATA[
Parkinsons disease (PD) is a progressive, incurable neurodegenerative disorder characterized by the loss of neuromelanin (NM)-containing dopamine neurons (DAn) in the substantia nigra of the midbrain. Non-neuronal cells are increasingly recognized as contributors to PD. We generated human microglia-like cells (hMG) from induced pluripotent stem cells (iPSC) derived from patients with LRRK2 PD-causing mutations, gene-corrected isogenic controls, and healthy donors. While neither genotype induced neurodegeneration in healthy DAn, LRRK2 hMG become hyperreactive to LPS stimulation, exhibiting increased cytokine expression, reactive oxygen species, and phagocytosis. When exposed to NM-containing particles, but not -synuclein fibrils, LRRK2 hMG trigger DAn degeneration, in a process that is prevented by pre-treatment with the immunomodulatory drug ivermectin. Finally, post-mortem analysis of midbrain tissue of LRRK2-PD patients show increased microglia activation around NM-containing neurons, confirming our in vitro findings. Overall, our work highlights NM-activated microglias role in PD progression, and provides a model for testing therapeutic targets.
]]></description>
<dc:creator>Blasco-Agell, L.</dc:creator>
<dc:creator>Espinal, M. P.</dc:creator>
<dc:creator>Testa, V.</dc:creator>
<dc:creator>Roch, G.</dc:creator>
<dc:creator>Montero-Mu, J.</dc:creator>
<dc:creator>Fern, I.</dc:creator>
<dc:creator>Baruffi, V.</dc:creator>
<dc:creator>Gonzalez-Sepulveda, M.</dc:creator>
<dc:creator>Richaud-Patin, Y.</dc:creator>
<dc:creator>Jim, S.</dc:creator>
<dc:creator>Cuadros, T.</dc:creator>
<dc:creator>Cladera-Sastre, J. M.</dc:creator>
<dc:creator>Compte, J.</dc:creator>
<dc:creator>Otero, M. J.</dc:creator>
<dc:creator>Tolosa, E.</dc:creator>
<dc:creator>Raya, A.</dc:creator>
<dc:creator>Vila, M.</dc:creator>
<dc:creator>Consiglio, A.</dc:creator>
<dc:date>2024-10-07</dc:date>
<dc:identifier>doi:10.1101/2024.10.07.616997</dc:identifier>
<dc:title><![CDATA[LRRK2-mutant microglia trigger dopaminergic neurodegeneration when activated by neuromelanin]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.10.22.619706v1?rss=1">
<title>
<![CDATA[
Mitochondrial fission controls astrocyte morphogenesis and organization in the cortex 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.10.22.619706v1?rss=1"
</link>
<description><![CDATA[
Dysfunctional mitochondrial dynamics are a hallmark of devastating neurodevelopmental disorders such as childhood refractory epilepsy. However, the role of glial mitochondria in proper brain development is not well understood. We show that astrocyte mitochondria undergo extensive fission while populating astrocyte distal branches during postnatal cortical development. Loss of mitochondrial fission regulator, Dynamin-related protein 1 (Drp1), decreases mitochondrial localization to distal astrocyte processes, and this mitochondrial mislocalization reduces astrocyte morphological complexity. Functionally, astrocyte-specific conditional deletion of Drp1 induces astrocyte reactivity and disrupts astrocyte organization in the cortex. These morphological and organizational deficits are accompanied by loss of perisynaptic astrocyte process (PAP) proteins such as gap junction protein Connexin 43. These findings uncover a crucial role for mitochondrial fission in coordinating astrocytic morphogenesis and organization, revealing the regulation of astrocytic mitochondria dynamics as a critical step in neurodevelopment.

SummaryDuring cortical astrocyte morphogenesis, mitochondria fragment and decrease in size to populate distal astrocyte processes. Drp1-mediated mitochondrial fission is necessary for peripheral astrocyte process formation. Astrocyte-specific Drp1 loss induces astrocyte reactivity, disrupts cortical astrocyte organization, and dysregulates PAP proteins including gap-junction protein Connexin 43 abundance.
]]></description>
<dc:creator>Salazar, M. P. R.</dc:creator>
<dc:creator>Kolanukuduru, S.</dc:creator>
<dc:creator>Ramirez, V.</dc:creator>
<dc:creator>Lyu, B.</dc:creator>
<dc:creator>Sejourne, G.</dc:creator>
<dc:creator>Sesaki, H.</dc:creator>
<dc:creator>Yu, G.</dc:creator>
<dc:creator>Eroglu, C.</dc:creator>
<dc:date>2024-10-23</dc:date>
<dc:identifier>doi:10.1101/2024.10.22.619706</dc:identifier>
<dc:title><![CDATA[Mitochondrial fission controls astrocyte morphogenesis and organization in the cortex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.10.13.617729v1?rss=1">
<title>
<![CDATA[
Assembloid model to study loop circuits of the human nervous system 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.10.13.617729v1?rss=1"
</link>
<description><![CDATA[
Neural circuits connecting the cerebral cortex, the basal ganglia and the thalamus are fundamental networks for sensorimotor processing and their dysfunction has been consistently implicated in neuropsychiatric disorders1-9. These recursive, loop circuits have been investigated in animal models and by clinical neuroimaging, however, direct functional access to developing human neurons forming these networks has been limited. Here, we use human pluripotent stem cells to reconstruct an in vitro cortico-striatal-thalamic-cortical circuit by creating a four-part loop assembloid. More specifically, we generate regionalized neural organoids that resemble the key elements of the cortico-striatal-thalamic-cortical circuit, and functionally integrate them into loop assembloids using custom 3D-printed biocompatible wells. Volumetric and mesoscale calcium imaging, as well as extracellular recordings from individual parts of these assembloids reveal the emergence of synchronized patterns of neuronal activity. In addition, a multi-step rabies retrograde tracing approach demonstrate the formation of neuronal connectivity across the network in loop assembloids. Lastly, we apply this system to study heterozygous loss of ASH1L gene associated with autism spectrum disorder and Tourette syndrome and discover aberrant synchronized activity in disease model assembloids. Taken together, this human multi-cellular platform will facilitate functional investigations of the cortico-striatal-thalamic-cortical circuit in the context of early human development and in disease conditions.
]]></description>
<dc:creator>Miura, Y.</dc:creator>
<dc:creator>Kim, J.-i.</dc:creator>
<dc:creator>Jurjut, O.</dc:creator>
<dc:creator>Kelley, K. W.</dc:creator>
<dc:creator>Yang, X.</dc:creator>
<dc:creator>Chen, X.</dc:creator>
<dc:creator>Thete, M. V.</dc:creator>
<dc:creator>Revah, O.</dc:creator>
<dc:creator>Cui, B.</dc:creator>
<dc:creator>Pachitariu, M.</dc:creator>
<dc:creator>Pasca, S. P.</dc:creator>
<dc:date>2024-10-14</dc:date>
<dc:identifier>doi:10.1101/2024.10.13.617729</dc:identifier>
<dc:title><![CDATA[Assembloid model to study loop circuits of the human nervous system]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.09.24.614704v1?rss=1">
<title>
<![CDATA[
Endo-IP and Lyso-IP Toolkit for Endolysosomal Profiling of Human Induced Neurons 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.09.24.614704v1?rss=1"
</link>
<description><![CDATA[
Plasma membrane protein degradation and recycling is regulated by the endolysosomal system, wherein endosomes bud from the plasma membrane into the cytosol and mature into degradative lysosomes. As such, the endolysosomal system plays a critical role in determining the abundance of proteins on the cell surface, influencing cellular identity and function. Highly polarized cells, like neurons, rely on the endolysosomal system for axonal and dendritic specialization and synaptic compartmentalization. The importance of this system to neuronal function is reflected by the prevalence of risk variants in components of the system in several neurodegenerative diseases, ranging from Parkinsons to Alzheimers disease. Nevertheless, our understanding of endocytic cargo and core endolysosomal machinery in neurons is limited, in part due to technical limitations. Here, we developed a toolkit for capturing EEA1-postive endosomes (Endo-IP) and TMEM192-positive lysosomes (Lyso-IP) in stem cell-derived induced neurons (iNeurons). We demonstrated its utility by revealing the endolysosomal protein landscapes for cortical-like iNeurons and stem cells. This allowed us to globally profile endocytic cargo, identifying hundreds of transmembrane proteins, including neurogenesis and synaptic proteins, as well as endocytic cargo with predicted SNX17 or SNX27 recognition motifs. By contrast, parallel lysosome profiling reveals a simpler protein repertoire, reflecting in part temporally controlled recycling or degradation for many endocytic targets. This system will facilitate mechanistic interrogation of endolysosomal components found as risk factors in neurodegenerative disease.
]]></description>
<dc:creator>Hundley, F. V.</dc:creator>
<dc:creator>Gonzalez-Lozano, M.</dc:creator>
<dc:creator>Gottschalk, L. M.</dc:creator>
<dc:creator>Cook, A. N. K.</dc:creator>
<dc:creator>Zhang, J.</dc:creator>
<dc:creator>Paulo, J. A.</dc:creator>
<dc:creator>Harper, J. W.</dc:creator>
<dc:date>2024-09-26</dc:date>
<dc:identifier>doi:10.1101/2024.09.24.614704</dc:identifier>
<dc:title><![CDATA[Endo-IP and Lyso-IP Toolkit for Endolysosomal Profiling of Human Induced Neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.09.06.611664v1?rss=1">
<title>
<![CDATA[
Abnormal hyperactivity of specific striatal ensembles encodes distinct dyskinetic behaviors revealed by high-resolution clustering 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.09.06.611664v1?rss=1"
</link>
<description><![CDATA[
L-DOPA-induced dyskinesia (LID) is a debilitating complication of dopamine replacement therapy in Parkinso[n]s disease and the most common hyperkinetic disorder of basal ganglia origin. Abnormal activity of striatal D1 and D2 spiny projection neurons (SPNs) is critical for LID, yet the link between SPN activity patterns and specific dyskinetic movements remains unknown. To explore this, we developed a novel method for clustering movements based on high-resolution motion sensors and video recordings. In a mouse model of LID, this method identified two main dyskinesia types and pathological rotations, all absent during normal behavior. Using single-cell resolution imaging, we found that specific sets of both D1 and D2-SPNs were abnormally active during these pathological movements. Under baseline conditions, the same SPN sets were active during behaviors sharing physical features with LID movements. These findings indicate that ensembles of behavior-encoding D1- and D2-SPNs form new combinations of hyperactive neurons mediating specific dyskinetic movements.
]]></description>
<dc:creator>Alcacer, C.</dc:creator>
<dc:creator>Klaus, A.</dc:creator>
<dc:creator>Mendonca, M.</dc:creator>
<dc:creator>Abalde, S.</dc:creator>
<dc:creator>Cenci, M. A.</dc:creator>
<dc:creator>Costa, R. M.</dc:creator>
<dc:date>2024-09-08</dc:date>
<dc:identifier>doi:10.1101/2024.09.06.611664</dc:identifier>
<dc:title><![CDATA[Abnormal hyperactivity of specific striatal ensembles encodes distinct dyskinetic behaviors revealed by high-resolution clustering]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-09-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.08.28.610136v1?rss=1">
<title>
<![CDATA[
Segregated basal ganglia output pathways correspond to genetically divergent neuronal subclasses 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.08.28.610136v1?rss=1"
</link>
<description><![CDATA[
The basal ganglia control multiple sensorimotor behaviors though anatomically segregated and topographically organized subcircuits with outputs to specific downstream circuits. However, it is unclear how the anatomical organization of basal ganglia output circuits relates to the molecular diversity of cell types. Here, we demonstrate that the major output nucleus of the basal ganglia, the substantia nigra pars reticulata (SNr) is comprised of transcriptomically distinct subclasses that reflect its distinct progenitor lineages. We show that these subclasses are topographically organized within SNr, project to distinct targets in the midbrain and hindbrain, and receive inputs from different striatal subregions. Finally, we show that these mouse subclasses are also identifiable in human SNr neurons, suggesting that the genetic organization of SNr is evolutionarily conserved. These findings provide a unifying logic for how the developmental specification of diverse SNr neurons relates to the anatomical organization of basal ganglia circuits controlling specialized downstream brain regions.
]]></description>
<dc:creator>Mendelsohn, A.</dc:creator>
<dc:creator>Nikoobakht, L.</dc:creator>
<dc:creator>Bikoff, J.</dc:creator>
<dc:creator>Costa, R.</dc:creator>
<dc:date>2024-08-29</dc:date>
<dc:identifier>doi:10.1101/2024.08.28.610136</dc:identifier>
<dc:title><![CDATA[Segregated basal ganglia output pathways correspond to genetically divergent neuronal subclasses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-08-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.10.29.620938v1?rss=1">
<title>
<![CDATA[
A Protocol for Neuralized Murine Olfactory Organoids 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.10.29.620938v1?rss=1"
</link>
<description><![CDATA[
Chronic olfactory dysfunction can be associated with parkinsonism, dementia, demyelinating disorders and schizophrenia. The olfactory epithelium (OE) represents an interface between the environment and the central nervous system. Mounting evidence implicates environmental factors in neurodegenerative disease processes, necessitating investigations into their interactions with the hosts genome. In Parkinson disease, hyposmia often precedes motor symptoms, raising the possibility that the OE could be involved in disease initiation. We previously demonstrated abundant -synuclein expression in mammalian OE as well as aggregate formation in the olfactory nerve. Current in vitro models of OE are limited, relying primarily on post-mitotic cultures established from biopsies. To address this gap, we present a method for generating olfactory organoids of OE from adult mice. These organoids comprise neuronal and non-neuronal cell types, including sustentacular cells, thus encompassing structural elements of OE in situ. Expression of the olfactory sensory neuron marker OMP and Parkinsons-linked -synuclein was also detected in olfactory organoids, highlighting their potential usefulness to mechanistic research. We established OE organoids that were kept in culture for up to 3 weeks. In addition, we inoculated organoids with the neurotropic vesicular stomatitis virus to model infections. We conclude that this olfactory organoid model system offers a new platform for studying airborne environmental factors in their interactions with a genetically defined host; this, to study OE biology and enable the exploration of disease processes within olfactory tissue.
]]></description>
<dc:creator>Ozgun, A.</dc:creator>
<dc:creator>Suman, P.</dc:creator>
<dc:creator>Coulombe, J.</dc:creator>
<dc:creator>aSCENT-PD Investigators,</dc:creator>
<dc:creator>Brown, E. G.</dc:creator>
<dc:creator>Tomlinson, J. J.</dc:creator>
<dc:creator>Woulfe, J. M.</dc:creator>
<dc:creator>Schlossmacher, M. G.</dc:creator>
<dc:date>2024-10-31</dc:date>
<dc:identifier>doi:10.1101/2024.10.29.620938</dc:identifier>
<dc:title><![CDATA[A Protocol for Neuralized Murine Olfactory Organoids]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.10.30.620894v1?rss=1">
<title>
<![CDATA[
Hydrophobic tails enable diverse functions of the extracellular chaperone clusterin 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.10.30.620894v1?rss=1"
</link>
<description><![CDATA[
Clusterin, a conserved secretory glycoprotein abundant in blood plasma and cerebrospinal fluid, functions as a molecular chaperone and apolipoprotein (Wyatt et al. 2013, Raulin et al. 2022). Dysregulation of clusterin is linked to late-onset Alzheimers disease, cardiovascular pathology and cancer (Rohne et al. 2016, Satapathy and Wilson 2021, Wilson et al. 2023). Despite its prominent role in extracellular proteostasis, the chaperone mechanism of clusterin has remained unclear. Here we present crystal structures of human clusterin, revealing a discontinuous three-domain architecture. Structure-based mutational analysis demonstrated that two intrinsically disordered, hydrophobic peptide tails enable diverse clusterin activities. Resembling the N-terminal substrate binding regions of so-called small heat shock proteins, these sequences mediate clusterins chaperone function in suppressing amyloid-{beta}, tau and -synuclein aggregation. In conjunction with highly conserved surface areas, the tail segments also participate in clusterin binding to very low density lipoprotein receptor (VLDLR) and cellular uptake. Moreover, the disordered tails cooperate with an adjacent amphipathic helix in lipoprotein formation, but remain accessible for chaperone function in the lipoprotein complex. The remarkable versatility of these sequences allows clusterin to function alone or bound to lipid in maintaining solubility of aberrant extracellular proteins and facilitating their clearance by endocytosis and lysosomal degradation.
]]></description>
<dc:creator>Yuste-Checa, P.</dc:creator>
<dc:creator>Carvajal, A. I.</dc:creator>
<dc:creator>Chenchen, M.</dc:creator>
<dc:creator>Paatz, S.</dc:creator>
<dc:creator>Hartl, F.-U.</dc:creator>
<dc:creator>Bracher, A.</dc:creator>
<dc:date>2024-10-31</dc:date>
<dc:identifier>doi:10.1101/2024.10.30.620894</dc:identifier>
<dc:title><![CDATA[Hydrophobic tails enable diverse functions of the extracellular chaperone clusterin]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.10.22.619411v1?rss=1">
<title>
<![CDATA[
Network analysis of α-synuclein pathology progression reveals p21-activated kinases as regulators of vulnerability 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.10.22.619411v1?rss=1"
</link>
<description><![CDATA[
-Synuclein misfolding and progressive accumulation drives a pathogenic process in Parkinsons disease. To understand cellular and network vulnerability to -synuclein pathology, we developed a framework to quantify network-level vulnerability and identify new therapeutic targets at the cellular level. Full brain -synuclein pathology was mapped in mice over 9 months. Empirical pathology data was compared to theoretical pathology estimates from a diffusion model of pathology progression along anatomical connections. Unexplained variance in the model enabled us to derive regional vulnerability that we compared to regional gene expression. We identified gene expression patterns that relate to regional vulnerability, including 12 kinases that were enriched in vulnerable regions. Among these, an inhibitor of group II PAKs demonstrated protection from neuron death and -synuclein pathology, even after delayed compound treatment. This study provides a framework for the derivation of cellular vulnerability from network-based studies and identifies a promising therapeutic pathway for Parkinsons disease.



O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=198 SRC="FIGDIR/small/619411v1_ufig1.gif" ALT="Figure 1">
View larger version (60K):
org.highwire.dtl.DTLVardef@10ac986org.highwire.dtl.DTLVardef@134b55eorg.highwire.dtl.DTLVardef@16f4581org.highwire.dtl.DTLVardef@3c491a_HPS_FORMAT_FIGEXP  M_FIG C_FIG HIGHLIGHTSO_LILongitudinal -synuclein pathology assessment in 1046 brain regions over 9 months
C_LIO_LILinear diffusion modeling derivation of network vulnerability to -synuclein pathology
C_LIO_LIPANGEA: assessment of over 19,000 genes in 302 brain regions
C_LIO_LIGroup II PAK inhibitor prevents -synuclein pathology and neuron death
C_LI
]]></description>
<dc:creator>Vatsa, N.</dc:creator>
<dc:creator>Brynildsen, J. K.</dc:creator>
<dc:creator>Goralski, T. M.</dc:creator>
<dc:creator>Kurgat, K.</dc:creator>
<dc:creator>Meyerdirk, L.</dc:creator>
<dc:creator>Breton, L.</dc:creator>
<dc:creator>DeWeerd, D.</dc:creator>
<dc:creator>Brasseur, L.</dc:creator>
<dc:creator>Turner, L.</dc:creator>
<dc:creator>Becker, K.</dc:creator>
<dc:creator>Gallik, K. L.</dc:creator>
<dc:creator>Bassett, D. S.</dc:creator>
<dc:creator>Henderson, M. X.</dc:creator>
<dc:date>2024-10-22</dc:date>
<dc:identifier>doi:10.1101/2024.10.22.619411</dc:identifier>
<dc:title><![CDATA[Network analysis of α-synuclein pathology progression reveals p21-activated kinases as regulators of vulnerability]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.10.24.620078v1?rss=1">
<title>
<![CDATA[
RGS10 Attenuates Systemic Immune Dysregulation Induced by Chronic Inflammatory Stress 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.10.24.620078v1?rss=1"
</link>
<description><![CDATA[
Regulator of G-protein signaling 10 (RGS10), a key homeostatic regulator of immune cells, has been implicated in multiple diseases associated with aging and chronic inflammation including Parkinsons Disease (PD). Interestingly, subjects with idiopathic PD display reduced levels of RGS10 in subsets of peripheral immune cells. Additionally, individuals with PD have been shown to have increased activated peripheral immune cells in cerebral spinal fluid (CSF) compared to age-matched healthy controls. However, it is unknown whether CSF-resident peripheral immune cells in individuals with PD also exhibit decreased levels of RGS10. Therefore, we performed an analysis of RGS10 levels in the proteomic database of the CSF from the Michael J. Fox Foundation Parkinsons Progression Markers Initiative (PPMI) study. We found that RGS10 levels are decreased in the CSF of individuals with PD compared to healthy controls and prodromal individuals. Moreover, we find that RGS10 levels decrease with age but not PD progression and that males have less RGS10 than females in PD. Importantly, studies have established an association between chronic systemic inflammation (CSI) and neurodegenerative diseases, such as PD, and known sources of CSI have been identified as risk factors for developing PD; however, the role of peripheral immune cell dysregulation in this process has been underexplored. As RGS10 levels are decreased in the CSF and circulating peripheral immune cells of individuals with PD, we hypothesized that RGS10 regulates peripheral immune cell responses to CSI prior to the onset of neurodegeneration. To test this, we induced CSI for 6 weeks in C57BL6/J mice and RGS10 KO mice to assess circulating and CNS-associated peripheral immune cell responses. We found that RGS10 deficiency synergizes with CSI to induce a bias for inflammatory and cytotoxic cell populations, a reduction in antigen presentation in peripheral blood immune cells, as well as in and around the brain that is most notable in males. These results highlight RGS10 as an important regulator of the systemic immune response to CSI and implicate RGS10 as a potential contributor to the development of immune dysregulation in PD.
]]></description>
<dc:creator>Jernigan, J. E.</dc:creator>
<dc:creator>Staley, H. A.</dc:creator>
<dc:creator>Baty, Z.</dc:creator>
<dc:creator>Bolen, M. L.</dc:creator>
<dc:creator>Gomes, B. N.</dc:creator>
<dc:creator>Holt, J.</dc:creator>
<dc:creator>Cole, C. L.</dc:creator>
<dc:creator>Neighbarger, N. K.</dc:creator>
<dc:creator>Dheeravath, K.</dc:creator>
<dc:creator>Merchak, A. R.</dc:creator>
<dc:creator>Menees, K. B.</dc:creator>
<dc:creator>Coombes, S. A.</dc:creator>
<dc:creator>Tansey, M. G.</dc:creator>
<dc:date>2024-10-29</dc:date>
<dc:identifier>doi:10.1101/2024.10.24.620078</dc:identifier>
<dc:title><![CDATA[RGS10 Attenuates Systemic Immune Dysregulation Induced by Chronic Inflammatory Stress]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.11.06.622357v1?rss=1">
<title>
<![CDATA[
RNApysoforms: Fast rendering interactive visualization of RNA isoform structure and expression in Python 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.11.06.622357v1?rss=1"
</link>
<description><![CDATA[
MotivationAlternative splicing generates multiple RNA isoforms from a single gene, enriching genetic diversity and impacting gene function. Effective visualization of these isoforms and their expression patterns is crucial but challenging due to limitations in existing tools. Traditional genome browsers lack programmability, while other tools offer limited customization, produce static plots, or cannot simultaneously display structures and expression levels. RNApysoforms was developed to address these gaps by providing a Python-based package that enables concurrent visualization of RNA isoform structures and expression data. Leveraging plotly and polars libraries, it offers an interactive, customizable, and faster-rendering framework suitable for web applications, enhancing the analysis and dissemination of RNA isoform research.

Availability and implementationRNApysoforms is a Python package available at (https://github.com/UK-SBCoA-EbbertLab/RNApysoforms) via an open-source MIT license. It can be easily installed using the piip package installer for Python. Thorough documentation and usage vignettes are available at: https://rna-pysoforms.readthedocs.io/en/latest/.
]]></description>
<dc:creator>Heberle, B. A.</dc:creator>
<dc:creator>Page, M. L.</dc:creator>
<dc:creator>Gustavsson, E. K.</dc:creator>
<dc:creator>Ryten, M.</dc:creator>
<dc:creator>Ebbert, M. T. W.</dc:creator>
<dc:date>2024-11-08</dc:date>
<dc:identifier>doi:10.1101/2024.11.06.622357</dc:identifier>
<dc:title><![CDATA[RNApysoforms: Fast rendering interactive visualization of RNA isoform structure and expression in Python]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.11.07.621551v1?rss=1">
<title>
<![CDATA[
Autophagic stress activates distinct compensatory secretory pathways in neurons 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.11.07.621551v1?rss=1"
</link>
<description><![CDATA[
Autophagic dysfunction is a hallmark of neurodegenerative disease, leaving neurons vulnerable to the accumulation of damaged organelles and proteins. However, the late onset of diseases suggests that compensatory quality control mechanisms may be engaged to delay the deleterious effects induced by compromised autophagy. Neurons expressing common familial Parkinsons disease (PD)-associated mutations in LRRK2 kinase exhibit defective autophagy. Here, we demonstrate that both primary murine neurons and human iPSC-derived neurons harboring pathogenic LRRK2 upregulate the secretion of extracellular vesicles. We used unbiased proteomics to characterize the secretome of LRRK2G2019S neurons and found that autophagic cargos including mitochondrial proteins were enriched. Based on these observations, we hypothesized that autophagosomes are rerouted toward secretion when cell-autonomous degradation is compromised, likely to mediate clearance of undegraded cellular waste. Immunoblotting confirmed the release of autophagic cargos and immunocytochemistry demonstrated that secretory autophagy was upregulated in LRRK2G2019S neurons. We also found that LRRK2G2019S neurons upregulate the release of exosomes containing miRNAs. Live-cell imaging confirmed that this upregulation of exosomal release was dependent on hyperactive LRRK2 activity, while pharmacological experiments indicate that this release staves off apoptosis. Finally, we show that markers of both vesicle populations are upregulated in plasma from mice expressing pathogenic LRRK2. In sum, we find that neurons expressing pathogenic LRRK2 upregulate the compensatory release of secreted autophagosomes and exosomes, to mediate waste disposal and transcellular communication, respectively. We propose that this increased secretion contributes to the maintenance of cellular homeostasis, delaying neurodegenerative disease progression over the short term while potentially contributing to increased neuroinflammation over the longer term.

SIGNIFICANCEA hallmark feature of many neurodegenerative diseases is autophagy dysfunction, resulting in the accumulation of damaged proteins and organelles that is detrimental to neuronal health. The late onset of neurodegenerative diseases, however, suggests alternative quality control mechanisms may delay neuronal degeneration. Here, we demonstrate that neurons expressing a Parkinsons Disease-causing mutation upregulate the release of two extracellular vesicle populations. First, we observe the increased expulsion of secreted autophagosomes to mediate cellular waste disposal. Second, we observe the increased release of exosomes, likely to facilitate transcellular communication. Thus, we propose that increases in secretory autophagy and exosome release are a homeostatic response in neurons undergoing chronic stress.
]]></description>
<dc:creator>Palumbos, S. D.</dc:creator>
<dc:creator>Popolow, J.</dc:creator>
<dc:creator>Goldsmith, J.</dc:creator>
<dc:creator>Holzbaur, E.</dc:creator>
<dc:date>2024-11-07</dc:date>
<dc:identifier>doi:10.1101/2024.11.07.621551</dc:identifier>
<dc:title><![CDATA[Autophagic stress activates distinct compensatory secretory pathways in neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.10.13.618101v1?rss=1">
<title>
<![CDATA[
Amyloid-beta, alpha-synuclein and tau aggregated co-pathologies enhance neuropathology and neuroinflammation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.10.13.618101v1?rss=1"
</link>
<description><![CDATA[
Alzheimers (AD) and Parkinson disease (PD) pathology often co-occur. Amyloid-{beta} and phosphorylated tau are found in 30-50% of idiopathic PD cases, while -synuclein inclusions are present in 50% of AD cases. These co-pathologies are linked to increased mortality and earlier onset of cognitive decline. Immune activation is a hallmark of these neurodegenerative diseases, but current models primarily examine each pathology in isolation. How these co-pathologies drive inflammation and neuronal loss remains poorly understood. We therefore developed a mouse model combining tau, amyloid-{beta}, and -synuclein. We found that co-pathologies synergistically trigger an amplified neuroimmune response, with expanded populations of CD4+ and CD8+ tissue-resident memory T cells and CD68+ microglia, compared to single pathologies. These changes were abundant in the hippocampus and cortex, regions with elevated protein pathology load and enhanced neuronal loss. Our findings demonstrate that co-pathologies enhance proteinopathy and synergistically enhance immune activation and neurodegeneration, suggesting that combinatorial therapeutic strategies that target both co-pathologies and inflammation, may be disease modifying.

SummaryWebster et al. demonstrate that co-occurring Alzheimers and Parkinson disease protein pathologies, common in cognitively impaired patient populations, amplify proteinopathy and synergistically enhance CNS neuroinflammatory responses and neurodegeneration. This work supports the need for combinatorial therapeutic strategies and positions neuroinflammation as an important link for co-pathology enhanced neurodegeneration.
]]></description>
<dc:creator>Webster, J. M.</dc:creator>
<dc:creator>Stone, W. J.</dc:creator>
<dc:creator>Yang, Y.-T.</dc:creator>
<dc:creator>Miller, A. T.</dc:creator>
<dc:creator>Childers, G. M.</dc:creator>
<dc:creator>Corbin-Stein, N. J.</dc:creator>
<dc:creator>Mudium, N.</dc:creator>
<dc:creator>Molina, J.</dc:creator>
<dc:creator>Won, W.-J.</dc:creator>
<dc:creator>Sandoval, I. M.</dc:creator>
<dc:creator>Tyrrell, D. J.</dc:creator>
<dc:creator>Manfredsson, F. P.</dc:creator>
<dc:creator>Kordower, J. H.</dc:creator>
<dc:creator>Harms, A. S.</dc:creator>
<dc:date>2024-10-13</dc:date>
<dc:identifier>doi:10.1101/2024.10.13.618101</dc:identifier>
<dc:title><![CDATA[Amyloid-beta, alpha-synuclein and tau aggregated co-pathologies enhance neuropathology and neuroinflammation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-10-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.10.31.621388v1?rss=1">
<title>
<![CDATA[
Thalamic integration of basal ganglia and cerebellar circuits during motor learning 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.10.31.621388v1?rss=1"
</link>
<description><![CDATA[
The ability to control movement and learn new motor skills is one of the fundamental functions of the brain. The basal ganglia (BG) and the cerebellum (CB) are two key brain regions involved in controlling movement, and neuronal plasticity within these two regions is crucial for acquiring new motor skills. However, how these regions interact to produce a cohesive unified motor output remains elusive. Here, we discovered that a subset of neurons in the motor thalamus receive converging synaptic inputs from both BG and CB. By performing multi-site fiber photometry in mice learning motor tasks, we found that motor thalamus neurons integrate BG and CB signals and show distinct movement-related activity. Lastly, we found a critical role of these thalamic neurons and their BG and CB inputs in motor learning and control. These results identify the thalamic convergence of BG and CB and its crucial role in integrating movement signals.

HighlightsO_LIIndividual neurons in motor thalamus receive converging synaptic input from SNr and DCN projections.
C_LIO_LIThalamic neurons with SNr and DCN input are concentrated at the border between VM and VAL thalamic nuclei.
C_LIO_LIThalamic neurons functionally integrate SNr and DCN activity and adapt with motor learning.
C_LIO_LIThalamic neurons and their inputs from SNr and DCN are critical for learning and executing motor tasks.
C_LI
]]></description>
<dc:creator>Roth, R. H.</dc:creator>
<dc:creator>Muniak, M. A.</dc:creator>
<dc:creator>Huang, C. J.</dc:creator>
<dc:creator>Hwang, F.-J.</dc:creator>
<dc:creator>Sun, Y.</dc:creator>
<dc:creator>Min, C.</dc:creator>
<dc:creator>Mao, T.</dc:creator>
<dc:creator>Ding, J. B.</dc:creator>
<dc:date>2024-11-01</dc:date>
<dc:identifier>doi:10.1101/2024.10.31.621388</dc:identifier>
<dc:title><![CDATA[Thalamic integration of basal ganglia and cerebellar circuits during motor learning]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.11.04.621665v1?rss=1">
<title>
<![CDATA[
Corticonigral projections recruit substantia nigra pars lateralis dopaminergic neurons for auditory threat memories 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.11.04.621665v1?rss=1"
</link>
<description><![CDATA[
Dopaminergic neurons (DANs) in the lateral substantia nigra project to the tail of striatum (TS), which is involved in threat conditioning. Auditory cortex also contributes to threatening behaviors, but whether it directly interacts with midbrain DANs and how these interactions might influence threat conditioning remain unclear. Here, functional mapping revealed robust excitatory input from auditory and temporal association cortexes to substantia nigra pars lateralis (SNL) DANs, but not to pars compacta (SNc) DANs. SNL DANs exhibited unique firing patterns, with irregular pacemaking and higher maximal firing, reflecting different channel complements than SNc DANs. Behaviorally, inhibiting cortex to SNL projections impaired memory retrieval during auditory threat conditioning. Thus, we demonstrate robust corticonigral projections to SNL DANs, contrasting with previous observations of sparse cortical input to substantia nigra DANs. These findings distinguish SNL DANs from other nigral populations, highlighting their role in threatening behaviors and expanding knowledge of cortex to midbrain interactions.
]]></description>
<dc:creator>Sansalone, L.</dc:creator>
<dc:creator>Evans, R. C.</dc:creator>
<dc:creator>Twedell, E.</dc:creator>
<dc:creator>Zhang, R.</dc:creator>
<dc:creator>Khaliq, Z. M.</dc:creator>
<dc:date>2024-11-05</dc:date>
<dc:identifier>doi:10.1101/2024.11.04.621665</dc:identifier>
<dc:title><![CDATA[Corticonigral projections recruit substantia nigra pars lateralis dopaminergic neurons for auditory threat memories]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.11.20.624489v1?rss=1">
<title>
<![CDATA[
Ancestry-specific gene expression in peripheral monocytes mediates risk of neurodegenerative disease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.11.20.624489v1?rss=1"
</link>
<description><![CDATA[
It is hypothesised that peripheral immune states responding to regional environmental triggers contribute to central neurodegeneration. Region-specific genetic selection pressures require this hypothesis to be assessed in an ancestry specific manner. Here we utilise genome-wide association studies and expression quantitative trait loci from African, East Asian and European ancestries to show that genes causing neurodegeneration are preferentially expressed in innate rather than adaptive immune cells, and that expression of these genes mediates the risk of neurodegenerative disease in monocytes in an ancestry-specific manner.
]]></description>
<dc:creator>Wagen, A. Z.</dc:creator>
<dc:creator>Reynolds, R. H.</dc:creator>
<dc:creator>Nee Foo, J.</dc:creator>
<dc:creator>Fairbrother-Browne, A.</dc:creator>
<dc:creator>Gustavsson, E. K.</dc:creator>
<dc:creator>Gagliano-Turin, S. A.</dc:creator>
<dc:creator>Wood, N. W.</dc:creator>
<dc:creator>Blauwendraat, C. K.</dc:creator>
<dc:creator>Gandhi, S.</dc:creator>
<dc:creator>Ryten, M.</dc:creator>
<dc:date>2024-11-22</dc:date>
<dc:identifier>doi:10.1101/2024.11.20.624489</dc:identifier>
<dc:title><![CDATA[Ancestry-specific gene expression in peripheral monocytes mediates risk of neurodegenerative disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.11.22.624040v1?rss=1">
<title>
<![CDATA[
CNV-Finder: Streamlining Copy Number Variation Discovery 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.11.22.624040v1?rss=1"
</link>
<description><![CDATA[
Copy Number Variations (CNVs) play pivotal roles in the etiology of complex diseases and are variable across diverse populations. Understanding the association between CNVs and disease susceptibility is significant in disease genetics research and often requires analysis of large sample sizes. One of the most cost-effective and scalable methods for detecting CNVs is based on normalized signal intensity values, such as Log R Ratio (LRR) and B Allele Frequency (BAF), from Illumina genotyping arrays. In this study, we present CNV-Finder, a novel pipeline integrating deep learning techniques on array data, specifically a Long Short-Term Memory (LSTM) network, to expedite the large-scale identification of CNVs within predefined genomic regions. This facilitates efficient prioritization of samples for time-consuming or costly subsequent analyses such as Multiplex Ligation-dependent Probe Amplification (MLPA), short-read, and long-read whole genome sequencing. We incorporate four genes to establish our methods--Parkin (PRKN), Leucine Rich Repeat And Ig Domain Containing 2 (LINGO2), Microtubule Associated Protein Tau (MAPT), and alpha-Synuclein (SNCA)--which may be relevant to neurological diseases such as Alzheimers disease (AD), Parkinsons disease (PD), Progressive Supranuclear Palsy (PSP), or related disorders such as essential tremor (ET). By training our models on expert-annotated samples and validating them across diverse cohorts, including those from the Global Parkinsons Genetics Program (GP2) and additional dementia-specific databases, we demonstrate the efficacy of CNV-Finder in accurately detecting deletions and duplications. Our pipeline outputs app-compatible files for visualization within CNV-Finders interactive web application. This interface enables researchers to review predictions and filter displayed samples by model prediction values, LRR range, and variant count in order to explore or confirm results. Our pipeline integrates this human feedback to enhance model performance and reduce false positive rates. Through a series of comprehensive analyses and validations using visual inspection, MLPA, short-read, and long-read sequencing data, we demonstrate the robustness and adaptability of CNV-Finder in identifying CNVs with regions of varied size, probe density, and noise. Our findings highlight the significance of contextual understanding and human expertise in enhancing the precision of CNV identification, particularly in complex genomic regions like 17q21.31. The CNV-Finder pipeline is a scalable, publicly available resource for the scientific community, available on GitHub (https://github.com/GP2code/CNV-Finder; DOI 10.5281/zenodo.14182563). CNV-Finder not only expedites accurate candidate identification but also significantly reduces the manual workload for researchers, enabling future targeted validation and downstream analyses in regions or phenotypes of interest.
]]></description>
<dc:creator>Kuznetsov, N.</dc:creator>
<dc:creator>Daida, K.</dc:creator>
<dc:creator>Makarious, M. B.</dc:creator>
<dc:creator>Al-Mubarak, B.</dc:creator>
<dc:creator>Atterling Brolin, K.</dc:creator>
<dc:creator>Malik, L.</dc:creator>
<dc:creator>Kouam, C.</dc:creator>
<dc:creator>Baker, B.</dc:creator>
<dc:creator>Ostrozovicova, M.</dc:creator>
<dc:creator>Andersh, K. M.</dc:creator>
<dc:creator>Kung, P.-J.</dc:creator>
<dc:creator>Mecheri, Y.</dc:creator>
<dc:creator>Tay, Y.-W.</dc:creator>
<dc:creator>Soundous Malek, B.</dc:creator>
<dc:creator>Al Tassan, N.</dc:creator>
<dc:creator>Teresa Perinan, M.</dc:creator>
<dc:creator>Hong, S.</dc:creator>
<dc:creator>Koretsky, M.</dc:creator>
<dc:creator>Sargeant, L.</dc:creator>
<dc:creator>Levine, K.</dc:creator>
<dc:creator>Blauwendraat, C.</dc:creator>
<dc:creator>Billingsley, K. J.</dc:creator>
<dc:creator>Bandres-Ciga, S.</dc:creator>
<dc:creator>Leonard, H. L.</dc:creator>
<dc:creator>Morris, H. R.</dc:creator>
<dc:creator>Singleton, A. B.</dc:creator>
<dc:creator>Nalls, M. A.</dc:creator>
<dc:creator>Vitale, D.</dc:creator>
<dc:creator>The Global Parkinson's Genetics Program,</dc:creator>
<dc:date>2024-11-23</dc:date>
<dc:identifier>doi:10.1101/2024.11.22.624040</dc:identifier>
<dc:title><![CDATA[CNV-Finder: Streamlining Copy Number Variation Discovery]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.11.26.625468v1?rss=1">
<title>
<![CDATA[
LRRK2 G2019S mutation incites increased cell-intrinsic neutrophil effector functions and intestinal inflammation in a model of infectious colitis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.11.26.625468v1?rss=1"
</link>
<description><![CDATA[
Parkinsons Disease (PD) is a progressive, neurodegenerative disorder characterised by motor and non-motor symptoms. Emerging evidence suggests a link between PD and gastrointestinal dysfunction. Constipation is frequently observed years prior to development of motor dysfunction in PD, and people with inflammatory bowel disease (IBD) are more likely to develop PD. Mutations in the leucine-rich repeat kinase 2 gene (LRRK2) account for approximately 1% of all PD cases and are associated with increased risk for IBD. Among them, LRRK2 Gly2019Ser (G2019S), located within the kinase domain, is the most common PD-associated mutation and increases kinase activity. It is unknown how LRRK2 mutation affects susceptibility to intestinal inflammation or pathogenesis of PD. Using single cell RNA sequencing (scRNAseq), we demonstrate that LRRK2 G2019S mutation promotes a dysregulated gene profile, especially within neutrophil, monocyte and {gamma}{delta} T cell populations, following Citrobacter rodentium infection in mice. Transcriptionally, LRRK2 G2019S neutrophils have a greater pro- inflammatory type I and II IFN response compared to those of WT mice. This is accompanied by an increase in neutrophil numbers in the lamina propria in LRRK2 G2019S mice. We also uncover cell-intrinsic functional defects in LRRK2 G2019S neutrophils, including increased chemotaxis, degranulation and neutrophil extracellular traps (NETosis) formation. Increased neutrophil infiltration is associated with an upregulation in Th17 immune responses, which may together contribute to the observed increase in colon pathology during infection. These findings increase our understanding of the role of PD-associated genes in immune cells and their contribution to immune dysregulation. Understanding the early perturbations driven by the LRRK2 G2019S mutation in gastrointestinal pathology may facilitate the development of biomarkers for early diagnosis and intervention in PD.
]]></description>
<dc:creator>Pei, J.</dc:creator>
<dc:creator>Oliveira, N. L.</dc:creator>
<dc:creator>Recinto, S. J.</dc:creator>
<dc:creator>Kazanova, A.</dc:creator>
<dc:creator>Queiroz-Junior, C. M.</dc:creator>
<dc:creator>Li, Z.</dc:creator>
<dc:creator>Couto, K.</dc:creator>
<dc:creator>Westfall, S.</dc:creator>
<dc:creator>King, I. L.</dc:creator>
<dc:creator>Ribeiro, C. T.</dc:creator>
<dc:creator>Milnerwood, A.</dc:creator>
<dc:creator>Desjardins, M.</dc:creator>
<dc:creator>Thanabalasuriar, A.</dc:creator>
<dc:creator>STRATTON, J. A.</dc:creator>
<dc:creator>Gruenheid, S.</dc:creator>
<dc:date>2024-12-02</dc:date>
<dc:identifier>doi:10.1101/2024.11.26.625468</dc:identifier>
<dc:title><![CDATA[LRRK2 G2019S mutation incites increased cell-intrinsic neutrophil effector functions and intestinal inflammation in a model of infectious colitis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.12.14.628386v1?rss=1">
<title>
<![CDATA[
The pyrethroid insecticide deltamethrin disrupts neuropeptide and monoamine signaling pathways in the gastrointestinal tract 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.12.14.628386v1?rss=1"
</link>
<description><![CDATA[
Enteroendocrine cells (EECs) are a rare cell type of the intestinal epithelium. Various subtypes of EECs produce distinct repertoires of monoamines and neuropeptides which modulate intestinal motility and other physiologies. EECs also possess neuron-like properties, suggesting a potential vulnerability to ingested environmental neurotoxicants. One such group of toxicants are pyrethroids, a class of prevalent insecticides used residentially and agriculturally. Pyrethroids agonize voltage-gated sodium channels (VGSCs), inducing neuronal excitotoxicity, and affect the function of monoamine-producing neurons. Given their anatomical location at the interface with the environment and their expression of VGSCs, EECs likely represent a vulnerable cell-type to oral pyrethroid exposure. In this study, we used the EEC cell line, STC-1 cells, to evaluate the effects of the common pyrethroid deltamethrin on the functional status of EECs. We find that deltamethrin impacts both expression of serotonergic pathways and inhibits the adrenergic-evoked release of an EEC hormone, GLP-1, in vitro. In a mouse model of oral exposure, we found that deltamethrin induced an acute, yet transient, loss of intestinal motility, in both fed and fasted conditions. This constipation phenotype was accompanied by a significant decrease in peripheral serotonin production and an inhibition of nutrient-evoked intestinal hormone release. Together, these data demonstrate that deltamethrin alters monoaminergic signaling pathways in EECs and regulates intestinal motility. This work demonstrates a mechanistic link between pyrethroid exposure and intestinal impacts relevant to pyrethroid-associated diseases, including inflammatory bowel disease, neurodegenerative disease, and metabolic disorders.
]]></description>
<dc:creator>White, A. C.</dc:creator>
<dc:creator>Krout, I. N.</dc:creator>
<dc:creator>Mouhi, S.</dc:creator>
<dc:creator>Chang, J.</dc:creator>
<dc:creator>Kelly, S. D.</dc:creator>
<dc:creator>Caudle, W. M.</dc:creator>
<dc:creator>Sampson, T.</dc:creator>
<dc:date>2024-12-17</dc:date>
<dc:identifier>doi:10.1101/2024.12.14.628386</dc:identifier>
<dc:title><![CDATA[The pyrethroid insecticide deltamethrin disrupts neuropeptide and monoamine signaling pathways in the gastrointestinal tract]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.12.15.627490v1?rss=1">
<title>
<![CDATA[
Exploring the relationship between GBA1 host genotype and gut microbiome in the GBA1L444P/WT mouse model: Implications for Parkinson disease pathogenesis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.12.15.627490v1?rss=1"
</link>
<description><![CDATA[
BackgroundHeterozygous variants in GBA1 are the commonest genetic risk factor for Parkinson disease (PD) but penetrance is incomplete. GBA1 dysfunction can cause gastrointestinal disturbances and microbiome changes in preclinical models. Mounting evidence suggests that the microbiota-gut-brain axis is potentially implicated in PD pathogenesis. Whether the gut microbiome composition is influenced by host GBA1 genetics in heterozygosis has never been explored.

ObjectivesTo evaluate whether heterozygosity for the GBA1 pathogenic L444P variant can cause perturbations in gut microbiome composition.

MethodsFaecal samples collected from GBA1L444P/WT and GBA1WT/WT mice at 3 and 6 months of age were analysed through shotgun metagenomic sequencing.

ResultsNo differences in - and {beta}-diversity were detected between genotyped groups, at either time points. Overall, we found a little variation of the gut microbiome composition and functional potential between GBA1L444P/WT and GBA1WT/WT mice over time.

ConclusionHost GBA1 genotype does not impact gut microbiome structure and composition in the presented GBA1L444P/WT mouse model. Studies investigating the effect of a second hit on gut physiology and microbiome composition could explain the partial penetrance of GBA1 variants in PD.
]]></description>
<dc:creator>Menozzi, E.</dc:creator>
<dc:creator>Geiger, M.</dc:creator>
<dc:creator>Meslier, V.</dc:creator>
<dc:creator>Fierli, F.</dc:creator>
<dc:creator>Gilles, M.</dc:creator>
<dc:creator>Chau, K.-Y.</dc:creator>
<dc:creator>David, A.</dc:creator>
<dc:creator>Shahar Golan, R.</dc:creator>
<dc:creator>Famechon, A.</dc:creator>
<dc:creator>Koletsi, S.</dc:creator>
<dc:creator>Morabito, C.</dc:creator>
<dc:creator>Quinquis, B.</dc:creator>
<dc:creator>Pons, N.</dc:creator>
<dc:creator>Ehrlich, S. D.</dc:creator>
<dc:creator>Macnaughtan, J.</dc:creator>
<dc:creator>Almeida, M.</dc:creator>
<dc:creator>Schapira, A. H.</dc:creator>
<dc:date>2024-12-15</dc:date>
<dc:identifier>doi:10.1101/2024.12.15.627490</dc:identifier>
<dc:title><![CDATA[Exploring the relationship between GBA1 host genotype and gut microbiome in the GBA1L444P/WT mouse model: Implications for Parkinson disease pathogenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.12.18.629251v1?rss=1">
<title>
<![CDATA[
The gut microbiome promotes mitochondrial respiration in the brain of a Parkinson's disease mouse model 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.12.18.629251v1?rss=1"
</link>
<description><![CDATA[
The pathophysiology of Parkinsons disease (PD) involves gene-environment interactions that impair various cellular processes such as autophagy, lysosomal function, or mitochondrial dysfunction. Specifically, mitochondria-associated gene mutations increase PD risk, mitochondrial respiration is altered in the PD brain, and mitochondrial-damaging toxins cause PD-like motor and gastrointestinal symptoms in animal models. The gut microbiome is altered in PD patients and represents an environmental risk, however a relationship between mitochondrial function and the microbiome in PD has not been previously established. Herein, we report that striatal mitochondria are functionally overactive in -synuclein-overexpressing (ASO) mice, a model of PD, and that microbiome depletion restores respiration and mitochondria-associated gene expression patterns to wild-type levels. ASO mice harboring a complex microbiome produce increased reactive oxygen species in the striatum whereas germ-free counterparts express elevated levels of antioxidant proteins that may buffer against oxidative damage. Indeed, antioxidant treatment improves motor performance in ASO mice and, remarkably, blocking oxidant scavenging in germ-free mice induces -synuclein-dependent motor deficits. Thus, the gut microbiome increases mitochondrial respiration and oxidative stress in the brain, which enhances motor symptoms in a mouse model of PD.
]]></description>
<dc:creator>Morais, L. H.</dc:creator>
<dc:creator>Stiles, L.</dc:creator>
<dc:creator>Freeman, M.</dc:creator>
<dc:creator>Oguienko, A. D.</dc:creator>
<dc:creator>Hoang, J. D.</dc:creator>
<dc:creator>Jones, J.</dc:creator>
<dc:creator>Quan, B.</dc:creator>
<dc:creator>Devine, J.</dc:creator>
<dc:creator>Bois, J. S.</dc:creator>
<dc:creator>Chou, T.-F.</dc:creator>
<dc:creator>Trinh, J.</dc:creator>
<dc:creator>Picard, M.</dc:creator>
<dc:creator>Gradinaru, V.</dc:creator>
<dc:creator>Mazmanian, S. K.</dc:creator>
<dc:date>2024-12-21</dc:date>
<dc:identifier>doi:10.1101/2024.12.18.629251</dc:identifier>
<dc:title><![CDATA[The gut microbiome promotes mitochondrial respiration in the brain of a Parkinson's disease mouse model]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.12.20.629776v1?rss=1">
<title>
<![CDATA[
A Vulnerable Subtype of Dopaminergic Neurons Drives Early Motor Deficits in Parkinson's Disease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.12.20.629776v1?rss=1"
</link>
<description><![CDATA[
In Parkinsons disease (PD), dopaminergic neurons (DANs) in the midbrain gradually degenerate, with ventral substantia nigra pars compacta (SNc) DANs exhibiting greater vulnerability. However, it remains unclear whether specific molecular subtypes of ventral SNc DANs are more susceptible to degeneration in PD, and if they contribute to the early motor symptoms associated with the disease. We identified a subtype of Sox6+ DANs, Anxa1+, which are selectively lost earlier than other DANs, and with a time course that aligns with the development of motor symptoms in MitoPark mice. We generated a knock-in Cre mouse line for Anxa1+ DANs and showed differential anatomical inputs and outputs of this population. Further, we found that the inhibition of transmitter release in Anxa1+ neurons led to bradykinesia and tremor. Therefore, Anxa1+ is not only a biomarker of a selectively vulnerable subtype of DANs, but is also sufficient to drive early motor symptoms in Parkinsons disease.
]]></description>
<dc:creator>Fushiki, A.</dc:creator>
<dc:creator>Ng, D.</dc:creator>
<dc:creator>Lewis, Z. R.</dc:creator>
<dc:creator>Yadav, A.</dc:creator>
<dc:creator>Saraiva, T.</dc:creator>
<dc:creator>Hammand, L. A.</dc:creator>
<dc:creator>Wirblich, C.</dc:creator>
<dc:creator>Tasic, B.</dc:creator>
<dc:creator>Menon, V.</dc:creator>
<dc:creator>Silva, J. A. d.</dc:creator>
<dc:creator>Costa, R. M.</dc:creator>
<dc:date>2024-12-21</dc:date>
<dc:identifier>doi:10.1101/2024.12.20.629776</dc:identifier>
<dc:title><![CDATA[A Vulnerable Subtype of Dopaminergic Neurons Drives Early Motor Deficits in Parkinson's Disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.12.22.629963v1?rss=1">
<title>
<![CDATA[
Anxa1+ dopamine neuron vulnerability defines prodromal Parkinson's disease bradykinesia and procedural motor learning impairment 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.12.22.629963v1?rss=1"
</link>
<description><![CDATA[
Progressive degeneration of dopamine neurons (DANs) defines Parkinsons disease (PD). However, the identity and function of the most vulnerable DAN populations in prodromal PD remain undefined. Here, we identify substantia nigra DANs with Annexin A1 (Anxa1) expression as selectively vulnerable across multiple prodromal PD models and significantly reduced in patient-derived DANs. We found that Anxa1+ DANs have a unique functional profile, as they do not signal reward or reinforce actions, and they are not necessary for motivated behavior. Instead, activity of Anxa1+ DAN axons correlates with vigorous movements during self-paced exploration, yet their silencing only disrupts a subset of action sequences that mirror a PD bradykinesia profile. Importantly, Anxa1+ DANs are essential for procedural learning in a maze task and for motor learning of dexterous actions. These findings establish the early vulnerability of Anxa1+ DANs in PD, whose function can explain prodromal bradykinesia and impairments in procedural motor learning.
]]></description>
<dc:creator>Mantas, I.</dc:creator>
<dc:creator>Contestabile, A.</dc:creator>
<dc:creator>Skara, V.</dc:creator>
<dc:creator>Loiseau, C.</dc:creator>
<dc:creator>Santos, I.</dc:creator>
<dc:creator>Cramb, K.</dc:creator>
<dc:creator>Filograna, R.</dc:creator>
<dc:creator>Wade-Martins, R.</dc:creator>
<dc:creator>Magill, P.</dc:creator>
<dc:creator>Meletis, K.</dc:creator>
<dc:date>2024-12-22</dc:date>
<dc:identifier>doi:10.1101/2024.12.22.629963</dc:identifier>
<dc:title><![CDATA[Anxa1+ dopamine neuron vulnerability defines prodromal Parkinson's disease bradykinesia and procedural motor learning impairment]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.12.26.630411v1?rss=1">
<title>
<![CDATA[
Sub-second characterization of locomotor activities of mouse models of Parkinsonism 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.12.26.630411v1?rss=1"
</link>
<description><![CDATA[
The degeneration of midbrain dopamine (DA) neurons disrupts the neural control of natural behavior, such as walking, posture, and gait in Parkinsons disease. While some aspects of motor symptoms can be managed by dopamine replacement therapies, others respond poorly. Recent advancements in machine learning-based technologies offer opportunities to better understand the organizing principles of behavior modules at fine time scales and its dependence on dopaminergic modulation. In the present study, we applied the motion sequencing (MoSeq) platform to study the spontaneous locomotor activities of neurotoxin and genetic mouse models of Parkinsonism as the midbrain DA neurons progressively degenerate. We also evaluated the treatment efficacy of levodopa (L-DOPA) on behavioral modules at fine time scales. We revealed robust changes in the kinematics and usage of the behavioral modules that encode spontaneous locomotor activity. Further analysis demonstrates that fast behavioral modules with higher velocities were more vulnerable to loss of DA and preferentially affected at early stages of Parkinsonism. Last, L-DOPA effectively improved the velocity, but not the usage and transition probability, of behavioral modules in Parkinsonian animals. In conclusion, the hypokinetic phenotypes in Parkinsonism involve the decreased velocities of behavioral modules and their disrupted temporal organization during movement. Moreover, we showed that the therapeutic effect of L-DOPA is mainly mediated by its effect on the velocities of behavior modules at fine time scales. This work documents robust changes in the velocity, usage, and temporal organization of behavioral modules and their responsiveness to dopaminergic treatment under the Parkinsonian state.

Significance StatementParkinsons disease is the second largest neurodegenerative disease without a cure. Detection of subtle Parkinsonian signs is critical for disease-modification by applying early interventions. The present work explores the possibility of using machine learning-based approaches for early detection of subtle behavioral changes in Parkinsonian animals and evaluating the therapeutic efficacy of dopaminergic medications.
]]></description>
<dc:creator>Berezhnoi, D.</dc:creator>
<dc:creator>Chehade, H. D.</dc:creator>
<dc:creator>Simms, G.</dc:creator>
<dc:creator>Chen, L.</dc:creator>
<dc:creator>Chu, H.-Y.</dc:creator>
<dc:date>2024-12-26</dc:date>
<dc:identifier>doi:10.1101/2024.12.26.630411</dc:identifier>
<dc:title><![CDATA[Sub-second characterization of locomotor activities of mouse models of Parkinsonism]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-26</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.01.01.630988v1?rss=1">
<title>
<![CDATA[
GPNMB is a biomarker for lysosomal dysfunction and is secreted via LRRK2-modulated lysosomal exocytosis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.01.01.630988v1?rss=1"
</link>
<description><![CDATA[
Genome-wide association studies have identified Glycoprotein Nmb (GPNMB) as a risk factor for Parkinsons Disease. The risk allele increases GPNMB transcription and GPNMB protein levels in the CSF highlighting GPMNB as a potential biomarker for Parkinsons Disease. However, a lack of knowledge of GPNMBs function and mechanism of secretion have hindered an interpretation of secreted GPNMB levels. In this study, we assessed the mechanism of GPNMB secretion by macrophages, the primary cell type expressing GPNMB in the brain. We show that GPNMB is secreted in response to lysosomal stress via lysosomal exocytosis and highlight the Parkinsons Disease risk factor LRRK2 as a strong modulator of GPNMB secretion.
]]></description>
<dc:creator>Bogacki, E. C.</dc:creator>
<dc:creator>Longmore, G.</dc:creator>
<dc:creator>Lewis, P. A.</dc:creator>
<dc:creator>Herbst, S.</dc:creator>
<dc:date>2025-01-01</dc:date>
<dc:identifier>doi:10.1101/2025.01.01.630988</dc:identifier>
<dc:title><![CDATA[GPNMB is a biomarker for lysosomal dysfunction and is secreted via LRRK2-modulated lysosomal exocytosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.01.02.631090v1?rss=1">
<title>
<![CDATA[
State-dependent modulation of spiny projection neurons controls levodopa-induced dyskinesia in a mouse model of Parkinson's disease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.01.02.631090v1?rss=1"
</link>
<description><![CDATA[
In the later stages of Parkinsons disease (PD), patients often manifest levodopa-induced dyskinesia (LID), compromising their quality of life. The pathophysiology underlying LID is poorly understood, and treatment options are limited. To move toward filling this treatment gap, the intrinsic and synaptic changes in striatal spiny projection neurons (SPNs) triggered by the sustained elevation of dopamine (DA) during dyskinesia were characterized using electrophysiological, pharmacological, molecular and behavioral approaches. Our studies revealed that the intrinsic excitability and functional corticostriatal connectivity of SPNs in dyskinetic mice oscillate between the on- and off-states of LID in a cell- and state-specific manner. Although triggered by levodopa, these rapid oscillations in SPN properties depended on both dopaminergic and cholinergic signaling. In a mouse PD model, disrupting M1 muscarinic receptor signaling specifically in iSPNs or deleting its downstream signaling partner CalDAG-GEFI blunted the levodopa-induced oscillation in functional connectivity, enhanced the beneficial effects of levodopa and attenuated LID severity.
]]></description>
<dc:creator>Zhai, S.</dc:creator>
<dc:creator>Cui, Q.</dc:creator>
<dc:creator>Wokosin, D.</dc:creator>
<dc:creator>Sun, L.</dc:creator>
<dc:creator>Tkatch, T.</dc:creator>
<dc:creator>Crittenden, J. R.</dc:creator>
<dc:creator>Graybiel, A. M.</dc:creator>
<dc:creator>Surmeier, D. J.</dc:creator>
<dc:date>2025-01-02</dc:date>
<dc:identifier>doi:10.1101/2025.01.02.631090</dc:identifier>
<dc:title><![CDATA[State-dependent modulation of spiny projection neurons controls levodopa-induced dyskinesia in a mouse model of Parkinson's disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.01.09.632283v1?rss=1">
<title>
<![CDATA[
Transgenic A53T mice have astrocytic α-synuclein aggregates in dopamine and striatal regions 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.01.09.632283v1?rss=1"
</link>
<description><![CDATA[
AimsParkinsons disease is considered biologically a neuronal -synuclein disease, largely ignoring the more widespread -synuclein deposition that occurs in astrocytes, with the aim of this study to identify whether astrocytes accumulate small -synuclein aggregates before or after neurons.

MethodsFixed serial midbrain and striatal sections from M83 A53T transgenic mouse model of Parkinsons disease and wild-type controls were histologically processed for multiplex labelling of -synuclein and astrocytic markers and astrocyte quantitation performed on digital images using QuPath software.

ResultsThe density of astrocytes within the substantia nigra pars compacta was approximately 30% greater compared with other sampled regions (P<0.005). Small aggregates of -synuclein were observed in astrocytic processes, including in wild-type mice where a quarter of all astrocytes had an obvious -synuclein aggregate. Compared to wild-type, A53T transgenic astrocytes had significantly enlarged somas (P<0.001) with more processes (P<0.001) consistent with a reactive phenotype. The A53T transgenic mice had more than double the numbers of astrocytes (P<0.001) and 2.5 times more astrocytes with -synuclein aggregates compared to wild-type mice (P<0.001).

ConclusionsThese data suggest that small -synuclein aggregates are normally cleared by astrocytes and that the substantia nigra pars compacta requires more astrocytic support for this function than other midbrain dopaminergic regions or the striatum. This adds another vulnerability factor to those already known for the substantia nigra with early deficits in clearance of small -synuclein aggregates by astrocytes associated with an increased astrocytic reactivity in the A53T transgenic mouse model.

Key PointsO_LIThe substantia nigra pars compacta contains a higher density of astrocytes than the ventral tegmental area or striatum, indicating a greater reliance on astrocytic function and a greater vulnerability to astrocyte dysfunction
C_LIO_LISmall aggregates of -synuclein were observed in wild-type midbrain and striatal astrocytes, indicating normal clearance of -synuclein by these astrocytes
C_LIO_LIMidbrain and striatal astrocytes from A53T transgenic astrocytes have more than double the number of astrocytes and more astrocytes containing -synuclein aggregates which have a reactive morphological phenotype.
C_LI
]]></description>
<dc:creator>Peat, C.</dc:creator>
<dc:creator>Prasad, A.</dc:creator>
<dc:creator>Finkelstein, D. I.</dc:creator>
<dc:creator>Sue, C.</dc:creator>
<dc:creator>Johnston, J.</dc:creator>
<dc:creator>Parish, C.</dc:creator>
<dc:creator>Thompson, L.</dc:creator>
<dc:creator>Kirik, D.</dc:creator>
<dc:creator>Halliday, G.</dc:creator>
<dc:date>2025-01-12</dc:date>
<dc:identifier>doi:10.1101/2025.01.09.632283</dc:identifier>
<dc:title><![CDATA[Transgenic A53T mice have astrocytic α-synuclein aggregates in dopamine and striatal regions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.01.13.632710v1?rss=1">
<title>
<![CDATA[
Alpha-Synuclein aggregates inhibit ESCRT-III through sequestration and collateral degradation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.01.13.632710v1?rss=1"
</link>
<description><![CDATA[
-Synuclein aggregation is a hallmark of Parkinsons disease and related synucleinopathies. Extracellular -synuclein fibrils enter naive cells via endocytosis, followed by transit into the cytoplasm to seed endogenous -synuclein aggregation. Intracellular aggregates sequester numerous proteins, including subunits of the ESCRT-III system for endolysosome membrane repair, but the toxic effects of these events remain poorly understood. Using cellular models and in vitro reconstitution, we found that -synuclein fibrils interact with an -helix common to ESCRT-III proteins. This interaction results in sequestration of ESCRT-III subunits and triggers their proteasomal destruction in a process of "collateral degradation." These twin mechanisms deplete the available ESCRT-III pool, initiating a toxic feedback loop. The ensuing loss of ESCRT function compromises endolysosome membranes, thereby facilitating escape of aggregate seeds into the cytoplasm, which in turn increases aggregation and ESCRT-III sequestration. We suggest that collateral degradation and triggering of self-perpetuating systems could be general mechanisms of sequestration-induced proteotoxicity.

HIGHLIGHTSO_LI-Synuclein fibrils bind and sequester ESCRT-III endolysosome repair proteins
C_LIO_LIAn -helical segment common to ESCRT-III mediates fibril-selective interaction
C_LIO_LIFibril-bound ESCRT-III subunits undergo "collateral degradation" via the proteasome
C_LIO_LIESCRT-III depletion damages endolysosomes and worsens -synuclein aggregation
C_LI
]]></description>
<dc:creator>Sitron, C. S.</dc:creator>
<dc:creator>Trinkaus, V. A.</dc:creator>
<dc:creator>Galesic, A. G.</dc:creator>
<dc:creator>Garhammer, M.</dc:creator>
<dc:creator>Yuste Checa, P.</dc:creator>
<dc:creator>Dransfeld, U.</dc:creator>
<dc:creator>Feigenbutz, D.</dc:creator>
<dc:creator>Zhang, J.</dc:creator>
<dc:creator>Dudanova, I.</dc:creator>
<dc:creator>Harper, J. W.</dc:creator>
<dc:creator>Hartl, F. U.</dc:creator>
<dc:date>2025-01-13</dc:date>
<dc:identifier>doi:10.1101/2025.01.13.632710</dc:identifier>
<dc:title><![CDATA[Alpha-Synuclein aggregates inhibit ESCRT-III through sequestration and collateral degradation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.01.17.633315v1?rss=1">
<title>
<![CDATA[
LINE-1 retrotransposons regulate the exit of human pluripotency and early brain development 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.01.17.633315v1?rss=1"
</link>
<description><![CDATA[
Long interspersed nuclear element 1 (L1) retrotransposons represent a vast source of divergent genetic information. However, mechanistic analysis of whether and how L1s contribute to human developmental programs is lacking, in part due to the challenges associated with specific profiling and manipulation of human L1 expression. Here we show that thousands of hominoid-specific L1 integrants are expressed in human induced pluripotent stem cells and cerebral organoids. The activity of individual L1 promoters is surprisingly divergent and correlates with an active epigenetic state. Efficient on-target CRISPRi silencing of L1s revealed nearly a hundred co-opted L1-derived chimeric transcripts and L1 silencing resulted in changes in neural differentiation programs and reduced cerebral organoid size. Together, these data implicate L1s and L1-derived transcripts in hominoid-specific CNS developmental processes.
]]></description>
<dc:creator>Adami, A.</dc:creator>
<dc:creator>Garza, R.</dc:creator>
<dc:creator>Gerdes, P.</dc:creator>
<dc:creator>Johansson, P. A.</dc:creator>
<dc:creator>Dorazehi, F.</dc:creator>
<dc:creator>Koutounidou, S.</dc:creator>
<dc:creator>Castilla-Vallmanya, L.</dc:creator>
<dc:creator>Atacho, D. A. M.</dc:creator>
<dc:creator>Sharma, Y.</dc:creator>
<dc:creator>Johansson, J. G.</dc:creator>
<dc:creator>Tam, O.</dc:creator>
<dc:creator>Kirkeby, A.</dc:creator>
<dc:creator>Barker, R. A.</dc:creator>
<dc:creator>Gale Hammell, M.</dc:creator>
<dc:creator>Douse, C. H.</dc:creator>
<dc:creator>Jakobsson, J.</dc:creator>
<dc:date>2025-01-17</dc:date>
<dc:identifier>doi:10.1101/2025.01.17.633315</dc:identifier>
<dc:title><![CDATA[LINE-1 retrotransposons regulate the exit of human pluripotency and early brain development]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.01.20.633968v1?rss=1">
<title>
<![CDATA[
Lysosomal Glucocerebrosidase is needed for ciliary Hedgehog signaling: A convergent pathway to Parkinsons disease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.01.20.633968v1?rss=1"
</link>
<description><![CDATA[
Mutations in LRRK2 and GBA1 are the most common genetic causes of familial Parkinsons disease. Previously, we showed that pathogenic LRRK2 mutations inhibit primary cilia formation in rare interneurons and astrocytes of the mouse and human dorsal striatum. This blocks Hedgehog signaling and reduces synthesis of neuroprotective GDNF and NRTN, which support neurons vulnerable in PD. Here we show that GBA1 mutations also impair Hedgehog signaling through a distinct mechanism. Loss of GBA1 activity decreases accessible cholesterol in primary cilia of cultured cells, thereby disrupting Hedgehog signaling. In the mouse striatum, Gba1 mutations result in reduced Hedgehog-induced Gdnf RNA expression in cholinergic interneurons, despite having no detectable impact on cilia formation. Also, both Lrrk2 and Gba1 mutations suppress Hedgehog-induced Bdnf expression in striatal astrocytes. These findings underscore the role of Hedgehog signaling in the nigrostriatal circuit and reveal a convergent mechanism by which distinct mutations may contribute to PD pathogenesis.
]]></description>
<dc:creator>Nair, S. V.</dc:creator>
<dc:creator>Jaimon, E.</dc:creator>
<dc:creator>Adhikari, A.</dc:creator>
<dc:creator>Nikoloff, J.</dc:creator>
<dc:creator>Pfeffer, S. R.</dc:creator>
<dc:date>2025-01-21</dc:date>
<dc:identifier>doi:10.1101/2025.01.20.633968</dc:identifier>
<dc:title><![CDATA[Lysosomal Glucocerebrosidase is needed for ciliary Hedgehog signaling: A convergent pathway to Parkinsons disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.01.20.633975v1?rss=1">
<title>
<![CDATA[
Standardised TruAI automated quantification of intracellular neuromelanin granules in human brain tissue sections 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.01.20.633975v1?rss=1"
</link>
<description><![CDATA[
AimsTo standardise and automate the quantitation of human-unique neuromelanin granules in catecholamine neurons in post-mortem tissue sections from healthy individuals at different ages to understand any changes in these granules with age.

Methods5-6 {micro}m-thick fixed and paraffin-embedded transverse midbrain tissue sections were supplied from 47 cases from three brain banks following ethical approvals. Sections were prepared and automated digital images acquired. Standardisation and automation of the quantification of neuromelanin granules was performed using the TruAI feature of the Olympus VS200 desktop platform. Comparisons between stained and unstained sections as well as correlations with age were performed.

ResultsThe automated platform reliably identified both stained and unstained intracellular neuromelanin granules and extracellular pigments, showing high reproducibility in measurements across laboratories using different tissue processing methods. Extraneuronal pigments were significantly smaller than intracellular neuromelanin granules. Sections processed for haematoxylin and eosin staining impacted the size and colour of both neuromelanin and the neurons containing neuromelanin. Haematoxylin made neuromelanin bluer, and the increased tissue processing made the intracellular area occupied by neuromelanin smaller in younger people. There was an increase in neuromelanin optical density and colour change (browner) with age.

ConclusionsThe TruAI automated platform reliably quantifies individual neuromelanin granules in catecholamine neurons. Extraneuronal pigments are considerably smaller in size than intracellular neuromelanin, and intracellular neuromelanin changes its properties with age. The darkening and colour change of intracellular neuromelanin suggests an increase in eumelanin over time in healthy individuals. These changes can be reliably identified using the automated platform.

Key pointsO_LIStandardised, reliable TruAI automated quantitation of intracellular neuromelanin granules in human brain sections gives novel insights into their structure and function
C_LIO_LIExtraneuronal pigments, including neuromelanin granules released from dying pigmented neurons, are significantly smaller than intracellular neuromelanin granules
C_LIO_LIIntracellular neuromelanin granules change their properties with healthy ageing becoming darker, consistent with more antioxidant eumelanin
C_LI
]]></description>
<dc:creator>Filimontseva, A.</dc:creator>
<dc:creator>Cuadros, T.</dc:creator>
<dc:creator>Chatterton, Z.</dc:creator>
<dc:creator>Compte, J.</dc:creator>
<dc:creator>Fu, Y.</dc:creator>
<dc:creator>Vila, M.</dc:creator>
<dc:creator>Halliday, G.</dc:creator>
<dc:date>2025-01-23</dc:date>
<dc:identifier>doi:10.1101/2025.01.20.633975</dc:identifier>
<dc:title><![CDATA[Standardised TruAI automated quantification of intracellular neuromelanin granules in human brain tissue sections]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-01-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.01.30.635139v1?rss=1">
<title>
<![CDATA[
Micro-invasive probes for monitoring electrical and chemical neural activity in nonhuman primates 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.01.30.635139v1?rss=1"
</link>
<description><![CDATA[
We leveraged carbon fiber materials for creating sensors that provide dual neurochemical and electrical neural activity recording at micro-invasive (10 {micro}m) spatial footprints proximal to recording sites, and enabling these measurements from deep brain targets of primates with conventional cranial chambers. These shaft-assisted micro-invasive probes (s-{micro}IPs) are approximately 10 {micro}m in diameter along the distal length (1 - 15 mm) immediately surrounding the targeted recording site. This micro-invasive portion ensures that the recording site is isolated from tissue damage induced by the wider shaft portion of the device. The shaft (150 - 165 {micro}m in diameter) within the device stiffens the remaining length of the probe (> 100 mm), and provides compatibility with standard intracranial insertion protocols (e.g., guide tubes and chamber setups) that require a sufficiently rigid and long shaft for deep brain insertion in monkeys. The s-{micro}IP was further expanded to provide dual-channel chemical and electrical neural activity recording with micrometer spatial resolution. Measurements of reward- and movement-related dopamine, spikes, and local field potentials were made from single and dual-channel s-{micro}IPs implanted in task-performing monkeys. Recordings from chronically implanted s-{micro}IPs display the capability of functional multi-modal (chemical and electrical) neural activity measurements over 1-year post-implantation from micro-invasive devices.
]]></description>
<dc:creator>Amjad, U.</dc:creator>
<dc:creator>Mahajan, S.</dc:creator>
<dc:creator>Choi, J.</dc:creator>
<dc:creator>Shrivastav, R.</dc:creator>
<dc:creator>Murray, R.</dc:creator>
<dc:creator>Somich, A.</dc:creator>
<dc:creator>Coyne, O.</dc:creator>
<dc:creator>Schwerdt, H. N.</dc:creator>
<dc:date>2025-02-05</dc:date>
<dc:identifier>doi:10.1101/2025.01.30.635139</dc:identifier>
<dc:title><![CDATA[Micro-invasive probes for monitoring electrical and chemical neural activity in nonhuman primates]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-05</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.02.07.636106v1?rss=1">
<title>
<![CDATA[
EndoMAPV1, a Structural Protein Complex Landscape of Human Endosomes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.02.07.636106v1?rss=1"
</link>
<description><![CDATA[
Early/sorting endosomes are dynamic organelles that play key roles in proteome control by triaging plasma membrane proteins for either recycling or degradation in the lysosome1,2,3. These events are coordinated by numerous transiently-associated regulatory complexes and integral membrane components that contribute to organelle identity during endosome maturation4. While a subset of the several hundred protein components and cargoes known to associate with endosomes have been studied at the biochemical and/or structural level, interaction partners and higher order molecular assemblies for many endosomal components remain unknown. Here, we combine cross-linking and native gel mass spectrometry5-8 of purified early endosomes with AlphaFold9,10 and computational analysis to create a systematic human endosomal structural interactome. We present dozens of structural models for endosomal protein pairs and higher order assemblies supported by experimental cross-links from their native subcellular context, suggesting structural mechanisms for previously reported regulatory processes. Using induced neurons, we validate two candidate complexes whose interactions are supported by crosslinks and structural predictions: TMEM230 as a subunit of ATP8/11 lipid flippases11 and TMEM9/9B as subunits of CLCN3/4/5 chloride-proton antiporters12. This resource and its accompanying structural network viewer provide an experimental framework for understanding organellar structural interactomes and large-scale validation of structural predictions.
]]></description>
<dc:creator>Gonzalez-Lozano, M.</dc:creator>
<dc:creator>Schmid, E. W.</dc:creator>
<dc:creator>Whelan, E. M.</dc:creator>
<dc:creator>Jiang, Y.</dc:creator>
<dc:creator>Paulo, J. A.</dc:creator>
<dc:creator>Walter, J.</dc:creator>
<dc:creator>Harper, J. W.</dc:creator>
<dc:date>2025-02-09</dc:date>
<dc:identifier>doi:10.1101/2025.02.07.636106</dc:identifier>
<dc:title><![CDATA[EndoMAPV1, a Structural Protein Complex Landscape of Human Endosomes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.02.09.637107v1?rss=1">
<title>
<![CDATA[
Cholesterol-mediated Lysosomal Dysfunction in APOE4 Astrocytes Promotes α-Synuclein Pathology in Human Brain Tissue 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.02.09.637107v1?rss=1"
</link>
<description><![CDATA[
The pathological hallmark of neurodegenerative disease is the aberrant post-translational modification and aggregation of proteins leading to the formation of insoluble protein inclusions. Genetic factors like APOE4 are known to increase the prevalence and severity of tau, amyloid, and -Synuclein inclusions. However, the human brain is largely inaccessible during this process, limiting our mechanistic understanding. Here, we developed an iPSC-based 3D model that integrates neurons, glia, myelin, and cerebrovascular cells into a functional human brain tissue (miBrain). Like the human brain, we found pathogenic phosphorylation and aggregation of -Synuclein is increased in the APOE4 miBrain. Combinatorial experiments revealed that lipid-droplet formation in APOE4 astrocytes impairs the degradation of -synuclein and leads to a pathogenic transformation that seeds neuronal inclusions of -Synuclein. Collectively, this study establishes a robust model for investigating protein inclusions in human brain tissue and highlights the role of astrocytes and cholesterol in APOE4-mediated pathologies, opening therapeutic opportunities.
]]></description>
<dc:creator>Mesentier-Louro, L. A.</dc:creator>
<dc:creator>Goldman, C.</dc:creator>
<dc:creator>Ndayisaba, A.</dc:creator>
<dc:creator>Buonfiglioli, A.</dc:creator>
<dc:creator>Rooklin, R. B.</dc:creator>
<dc:creator>Schuldt, B. R.</dc:creator>
<dc:creator>Uchitelev, A.</dc:creator>
<dc:creator>Khurana, V.</dc:creator>
<dc:creator>Blanchard, J. W.</dc:creator>
<dc:date>2025-02-09</dc:date>
<dc:identifier>doi:10.1101/2025.02.09.637107</dc:identifier>
<dc:title><![CDATA[Cholesterol-mediated Lysosomal Dysfunction in APOE4 Astrocytes Promotes α-Synuclein Pathology in Human Brain Tissue]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.02.10.636943v1?rss=1">
<title>
<![CDATA[
Aseptic, semi-sealed cranial chamber implants for chronic multi-channel neurochemical and electrophysiological neural recording in nonhuman primates 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.02.10.636943v1?rss=1"
</link>
<description><![CDATA[
We developed an implantable neural interface for monitoring both chemical and electrical forms of brain activity in monkeys that maintains aseptic properties for year-long periods while leveraging the modular functions (e.g., sensor moveability) provided by a chamber system. Invasive electrophysiological recordings, especially in subcortical structures from nonhuman primates usually involves implanting electrodes into the brain through a skull-mounted chamber. These electrodes may be attached temporarily for several hours of recording, or permanently. Permanent attachments are favorable to allow for sealing the chamber completely from externally originating pathogenic species that can infiltrate and compromise the health of the animal. A sealed chamber also reduces the need for frequent chamber cleaning required to minimize the accumulation of pathogenic organisms. However, neurochemical measurements require specialized electrodes with extremely fragile carbon fiber tips and are not compatible with recently developed sealed chamber systems. Here, we leveraged osseointegrating materials and hermetic sealing strategies to enable both neurochemical and electrical neural activity measurements from a sealed chamber with an aspirating port for culturing chamber fluid to ensure an aseptic environment. The system was shown to provide successful recordings of neural activity in two monkeys while maintaining negative bacteria culture results for over a year post-implant.
]]></description>
<dc:creator>Choi, J.</dc:creator>
<dc:creator>Amjad, U.</dc:creator>
<dc:creator>Murray, R.</dc:creator>
<dc:creator>Shrivastav, R.</dc:creator>
<dc:creator>Teichert, T.</dc:creator>
<dc:creator>Goodell, B.</dc:creator>
<dc:creator>Schaeffer, D. J.</dc:creator>
<dc:creator>Oluoch, J. K.</dc:creator>
<dc:creator>Schwerdt, H. N.</dc:creator>
<dc:date>2025-02-11</dc:date>
<dc:identifier>doi:10.1101/2025.02.10.636943</dc:identifier>
<dc:title><![CDATA[Aseptic, semi-sealed cranial chamber implants for chronic multi-channel neurochemical and electrophysiological neural recording in nonhuman primates]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.02.10.637505v1?rss=1">
<title>
<![CDATA[
Molecularly-guided spatial proteomics captures single-cell identity and heterogeneity of the nervous system 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.02.10.637505v1?rss=1"
</link>
<description><![CDATA[
Single-cell proteomics is an emerging field with significant potential to characterize heterogeneity within biological tissues. It offers complementary insights to single-cell transcriptomics by revealing unbiased proteomic changes downstream of the transcriptome. Recent advancements have focused on enhancing proteome coverage and depth, mostly in cultured cell lines, and a few recent studies have explored the potential of analyzing tissue micro-samples but were limited to homogenous peripheral tissues. In this current work, we utilize the power of spatial single cell-proteomics through immunostaining-guided laser capture microdissection (LCM) coupled with LC-MS to investigate the heterogenous central nervous system. We used this method to compare neuronal populations from cortex and substantia nigra, two brain regions associated with motor and cognitive function and various neurological disorders. Moreover, we used the technique to understand the neuroimmune changes associated with stab wound injury. Finally, we focus our application on the peripheral nervous system, where we compare the proteome of the myenteric plexus cell ganglion to the nerve bundle. This study demonstrates the utility of spatial single-cell proteomics in neuroscience research toward understanding fundamental biology and the molecular drivers of neurological conditions.
]]></description>
<dc:creator>Dutta, S.</dc:creator>
<dc:creator>Pang, M.</dc:creator>
<dc:creator>Coughlin, G. M.</dc:creator>
<dc:creator>Gudavalli, S.</dc:creator>
<dc:creator>Roukes, M. L.</dc:creator>
<dc:creator>Chou, T.-F.</dc:creator>
<dc:creator>Gradinaru, V.</dc:creator>
<dc:date>2025-02-11</dc:date>
<dc:identifier>doi:10.1101/2025.02.10.637505</dc:identifier>
<dc:title><![CDATA[Molecularly-guided spatial proteomics captures single-cell identity and heterogeneity of the nervous system]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.02.10.637521v1?rss=1">
<title>
<![CDATA[
Specific cell states underlie complex tissue regeneration in spiny mice 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.02.10.637521v1?rss=1"
</link>
<description><![CDATA[
SUMMARY STATEMENTComparing regenerative vs. fibrotic healing, we identify injury-induced cell states associated with persistent cell cycle progression and complex tissue regeneration in mammals.

Cell proliferation is an elemental feature of epimorphic regeneration in vertebrate taxa. We previously reported that in contrast to fibrotic repair observed in laboratory mouse (Mus) strains, highly regenerative spiny mice (Acomys spp.) exhibit cell cycle progression and cell proliferation to faithfully replace missing tissue. However, little is known about proliferation dynamics, and specific cell types and states that may contribute to complex tissue regeneration in mammals. Using temporal pulse-chase experiments, we show that stromal cells in Acomys dimidiatus rapidly re-enter the cell cycle in response to injury and maintain tight spatiotemporal control of cell cycle progression to restrict the proliferative population to a distal area relative to the injury. Conversely, Mus stromal cells incorporate thymidine analogs without cell division supporting an S-phase arrest after D10. Deploying immunostaining and scRNA-seq, we identify several key cell types (CRABP1+, SMA+) differentially associated with regenerating versus scar tissue. Importantly, our single cell data revealed distinct gene expression profiles for cross-species stromal cell types, identifying cell states specific for regenerative or fibrotic healing. While CRABP1+ fibroblasts are enriched in Acomys ears before and after injury, similar fibroblasts enriched in young, postnatal Mus ears remain unable to promote regeneration. Our data underscore the finely regulated dynamics of proliferating cells during regeneration and emphasize that regeneration depends on multiple factors including the presence of specific cell types and the ability of cells to acquire specific states.

Key Conclusions-Differentiated cells in Acomys, Mus and Danio re-enter the cell cycle in response to injury, while homeostatic cycling cells contribute to blastema formation in Ambystoma
-Pulse-chase thymidine analog labeling shows tight spatiotemporal control of proliferating stromal cells during regeneration in Acomys.
-Following injury, CRABP1 and SMA are expressed in distinct stromal cell populations in Acomys but are co-expressed in Mus stromal cell populations.
-Species-specific cell states underlie regenerative and fibrotic repair
-CRABP1+ cells are lost during embryonic development in Mus ear pinna but are retained in Acomys to adulthood.
-Young neonatal Mus with abundant CRABP1+ cells still fail to execute regenerative healing
]]></description>
<dc:creator>Rivadeneira, E. O.</dc:creator>
<dc:creator>Allen, R. S.</dc:creator>
<dc:creator>Adam, M.</dc:creator>
<dc:creator>Seifert, A. W.</dc:creator>
<dc:date>2025-02-11</dc:date>
<dc:identifier>doi:10.1101/2025.02.10.637521</dc:identifier>
<dc:title><![CDATA[Specific cell states underlie complex tissue regeneration in spiny mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.03.03.641233v1?rss=1">
<title>
<![CDATA[
Postsynaptic adaptations in direct pathway muscarinic M4-receptor signaling follow the temporal and regional pattern of dopaminergic degeneration 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.03.03.641233v1?rss=1"
</link>
<description><![CDATA[
In Parkinson’s disease (PD), imbalances in dorsal striatum (DSt) output pathways leading to motor dysfunction are thought to be driven by the loss of dopamine (DA) itself and the disruption of its coordinated modulation with acetylcholine (ACh). While the gradual decline of DA across striatal regions over time is a defining characteristic of PD, less is known about the adaptive and/or pathological alterations in cholinergic signaling that develop throughout disease progression in response to DA loss. Here, we examined changes in cholinergic modulation of striatal direct pathway medium spiny neurons (dMSNs) in mice that were partially or completely depleted of DA, in order to model early and advanced stages of PD. We found a reduction in muscarinic M4 receptor signaling that began in the dorsolateral striatum (DLS) following a partial loss of DA, yet was not evident in the dorsomedial region (DMS) until the dopaminergic lesion was nearly complete. Combining electrophysiological, pharmacological and 2-photon imaging approaches, we determined that this decrease was the result of reduced postsynaptic M4 receptor function, and was not accounted for by changes in ACh release or clearance. Replacing the partial loss of endogenous DA with levodopa could not rescue the dysfunctional M4 receptors. Together, these findings reveal how changes in cholinergic modulation closely follow the temporal and regional pattern of dopaminergic degeneration, which is critical for understanding their shared role in PD progression, and for developing alternative therapeutic interventions.
]]></description>
<dc:creator>Nielsen, B.</dc:creator>
<dc:creator>Ford, C.</dc:creator>
<dc:date>2025-3-10</dc:date>
<dc:identifier>doi:10.1101/2025.03.03.641233</dc:identifier>
<dc:title><![CDATA[Postsynaptic adaptations in direct pathway muscarinic M4-receptor signaling follow the temporal and regional pattern of dopaminergic degeneration]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-3-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.03.19.644182v1?rss=1">
<title>
<![CDATA[
PPM1M, a LRRK2-counteracting, phosphoRab12-preferring phosphatase with potential link to Parkinsons disease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.03.19.644182v1?rss=1"
</link>
<description><![CDATA[
Leucine-rich repeat kinase 2 (LRRK2) phosphorylates a subset of Rab GTPases that regulate receptor trafficking; activating mutations in LRRK2 are linked to Parkinsons disease. Rab phosphorylation is a transient event that can be reversed by phosphatases, including PPM1H, that acts on phosphoRab8A and phosphoRab10. Here we report a phosphatome-wide siRNA screen that identified PPM1M as a phosphoRab12-preferring phosphatase that also acts on phosphoRab8A and phosphoRab10. Upon knockout from cells or mice, PPM1M displays selectivity for phosphoRab12. As shown previously for mice harboring LRRK2 pathway mutations, knockout of Ppm1m leads to primary cilia loss in striatal cholinergic interneurons. We have also identified a rare PPM1M mutation in patients with Parkinsons disease that is catalytically inactive when tested in vitro and in cells. These findings identify PPM1M as a key player in the LRRK2 signaling pathway and provide a new therapeutic target for the possible benefit of patients with Parkinsons disease.

TeaserParkinsons linked Rab phosphorylation is reversed by PPM1M; the inactive D440N variant is implicated in rare patient cases.
]]></description>
<dc:creator>Chiang, C. Y.</dc:creator>
<dc:creator>Pratuseviciute, N.</dc:creator>
<dc:creator>Lin, Y.-E.</dc:creator>
<dc:creator>Adhikari, A.</dc:creator>
<dc:creator>Yeshaw, W. M.</dc:creator>
<dc:creator>Flitton, C.</dc:creator>
<dc:creator>Sherpa, P. L.</dc:creator>
<dc:creator>Tonelli, F.</dc:creator>
<dc:creator>Rektorova, I.</dc:creator>
<dc:creator>Lynch, T.</dc:creator>
<dc:creator>Siuda, J.</dc:creator>
<dc:creator>Rudzinska-Bar, M.</dc:creator>
<dc:creator>Pulyk, O.</dc:creator>
<dc:creator>Bauer, P.</dc:creator>
<dc:creator>Beetz, C.</dc:creator>
<dc:creator>Dickson, D. W.</dc:creator>
<dc:creator>Ross, O. A.</dc:creator>
<dc:creator>Wszolek, Z.</dc:creator>
<dc:creator>Klein, C.</dc:creator>
<dc:creator>Zimprich, A.</dc:creator>
<dc:creator>Alessi, D. R.</dc:creator>
<dc:creator>Sammler, E. M.</dc:creator>
<dc:creator>Pfeffer, S. R.</dc:creator>
<dc:date>2025-03-19</dc:date>
<dc:identifier>doi:10.1101/2025.03.19.644182</dc:identifier>
<dc:title><![CDATA[PPM1M, a LRRK2-counteracting, phosphoRab12-preferring phosphatase with potential link to Parkinsons disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.03.24.645001v1?rss=1">
<title>
<![CDATA[
In situ cryo-ET visualization of mitochondrial depolarization and mitophagic engulfment 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.03.24.645001v1?rss=1"
</link>
<description><![CDATA[
Defective mitochondrial quality control in response to loss of mitochondrial membrane polarization is implicated in Parkinsons disease by mutations in PINK1 and PRKN. Application of in situ cryo-electron tomography (cryo-ET) made it possible to visualize the consequences of mitochondrial depolarization at higher resolution than heretofore attainable. Parkin-expressing U2OS cells were treated with the depolarizing agents oligomycin and antimycin A (OA), subjected to cryo-FIB milling, and mitochondrial structure was characterized by in situ cryo-ET. Phagophores were visualized in association with mitochondrial fragments. Bridge-like lipid transporter (BLTP) densities potentially corresponding to ATG2A were seen connected to mitophagic phagophores. Mitochondria in OA-treated cells were fragmented and devoid of matrix calcium phosphate crystals. The intermembrane gap of cristae was narrowed and the intermembrane volume reduced, and some fragments were devoid of cristae. A subpopulation of ATP synthases re-localized from cristae to the inner boundary membrane (IBM) apposed to the outer membrane (OMM). The structure of the dome-shaped prohibitin complex, a dodecamer of PHB1-PHB2 dimers, was determined in situ by sub-tomogram averaging in untreated and treated cells and found to exist in open and closed conformations, with the closed conformation is enriched by OA treatment. These findings provide a set of native snapshots of the manifold nano-structural consequences of mitochondrial depolarization and provide a baseline for future in situ dissection of Parkin-dependent mitophagy.
]]></description>
<dc:creator>Rose, K.</dc:creator>
<dc:creator>Herrmann, E.</dc:creator>
<dc:creator>Kakudji, E. V.</dc:creator>
<dc:creator>Lizarrondo, J.</dc:creator>
<dc:creator>Celebi, A. Y.</dc:creator>
<dc:creator>Wilfling, F.</dc:creator>
<dc:creator>Lewis, S. C.</dc:creator>
<dc:creator>Hurley, J. H.</dc:creator>
<dc:date>2025-03-25</dc:date>
<dc:identifier>doi:10.1101/2025.03.24.645001</dc:identifier>
<dc:title><![CDATA[In situ cryo-ET visualization of mitochondrial depolarization and mitophagic engulfment]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.03.24.645038v1?rss=1">
<title>
<![CDATA[
ATG2A engages Rab1a and ARFGAP1 positive membranes during autophagosome biogenesis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.03.24.645038v1?rss=1"
</link>
<description><![CDATA[
Autophagosomes form from seed membranes that expand through bulk-lipid transport via the bridge-like lipid transporter ATG2. The origins of the seed membranes and their relationship to the lipid transport machinery are poorly understood. Using proximity labeling and a variety of fluorescence microscopy techniques, we show that ATG2A localizes to extra-Golgi ARFGAP1 puncta during autophagosome biogenesis. ARFGAP1 itself is dispensable during macroautophagy, but among other proteins associating to these membranes, we find that RAB1 is essential. ATG2A co-immunoprecipitates strongly, albeit indirectly, with RAB1A, and siRNA-mediated depletion of RAB1A/B blocks autophagy downstream of LC3B lipidation, similar to ATG2A depletion. Further, when either autophagosome formation or the early secretory pathway is perturbed, ARFGAP1 and RAB1A accumulate at ectopic locations with autophagic machinery. Our results indicate that ATG2A engages a RAB1A complex on select early secretory membranes in support of autophagosome biogenesis.

Significance StatementThis study elucidates the role of early secretory membranes in autophagosome biogenesis. The authors demonstrate that RAB1/ARFGAP1 positive membranes are essential to autophagy and are recruited to the phagophore assembly site at an early step of autophagosome biogenesis. These membranes interact with the bridge-like lipid transport protein ATG2A and are positive for LC3B and WIPI2, suggesting that RAB1 membranes are a direct source for autophagosome formation.
]]></description>
<dc:creator>Fuller, D. M.</dc:creator>
<dc:creator>Wu, Y.</dc:creator>
<dc:creator>Schueder, F.</dc:creator>
<dc:creator>Rasool, B.</dc:creator>
<dc:creator>Nag, S.</dc:creator>
<dc:creator>Korfhage, J. L.</dc:creator>
<dc:creator>Garcia-Milian, R.</dc:creator>
<dc:creator>Melnyk, K. D.</dc:creator>
<dc:creator>Bewersdorf, J.</dc:creator>
<dc:creator>De Camilli, P.</dc:creator>
<dc:creator>Melia, T.</dc:creator>
<dc:date>2025-03-25</dc:date>
<dc:identifier>doi:10.1101/2025.03.24.645038</dc:identifier>
<dc:title><![CDATA[ATG2A engages Rab1a and ARFGAP1 positive membranes during autophagosome biogenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.11.20.624606v1?rss=1">
<title>
<![CDATA[
AI-directed voxel extraction and volume EM identify intrusions as sites of mitochondrial contact 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.11.20.624606v1?rss=1"
</link>
<description><![CDATA[
Membrane contact sites (MCS) establish organelle interactomes in cells to enable communication and exchange of materials. Volume electron microscopy (vEM) is ideally suited for MCS analyses, but semantic segmentation of large vEM datasets remains challenging. Recent adoption of artificial intelligence (AI) for segmentation has greatly enhanced our analysis capabilities. However, we show that organelle boundaries, which are important for defining MCS, are the least confident predictions made by AI. We outline a segmentation strategy termed AI-directed Voxel Extraction (AIVE), that combines AI predictions with image electron signals to confidently segment membrane boundaries irrespective of the AI model used. We demonstrate the precision conferred by AIVE by applying it to the quantitative analysis of organelle interactomes from multiple FIB-SEM datasets. Through AIVE, we discover a previously unknown category of mitochondrial contact that we term the mitochondrial intrusion. We hypothesise that intrusions serve as anchors that stabilize MCS and promote organelle communication.
]]></description>
<dc:creator>Padman, B. S.</dc:creator>
<dc:creator>Lindblom, R. S. J.</dc:creator>
<dc:creator>Lazarou, M.</dc:creator>
<dc:date>2024-11-21</dc:date>
<dc:identifier>doi:10.1101/2024.11.20.624606</dc:identifier>
<dc:title><![CDATA[AI-directed voxel extraction and volume EM identify intrusions as sites of mitochondrial contact]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-11-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2024.12.24.630165v1?rss=1">
<title>
<![CDATA[
A single Citrobacter rodentium infection in Pink1 knockout and wild type mice leads to regional blood-brain-barrier perturbation and glial activation without dopamine neuron axon terminal loss 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2024.12.24.630165v1?rss=1"
</link>
<description><![CDATA[
A growing body of research supports the hypothesis of links between immune system activation and the development of Parkinsons disease (PD). A recent study revealed that repeated gastrointestinal infection with Citrobacter rodentium can lead to PD-like motor dysfunction in Pink1 knockout (KO) mice and immune cell entry in the brain. With the objective of better understanding the mechanisms leading to immune attack of the brain in this model, we evaluated the hypothesis that such mild infections are sufficient to increase blood brain barrier (BBB) permeability and cause brain inflammation. Pink1 wild-type (WT) and KO mice were infected with Citrobacter rodentium and at day 13 and 26 post infection, we conducted gadolinium-enhanced magnetic resonance imaging (MRI) to identify signs of BBB permeability changes. Quantification of MRI results provided evidence of increased blood-brain barrier permeability in both WT and Pink1 KO mice at 26 days after the infection in the striatum, dentate gyrus, somatosensory cortex, and thalamus. This was not accompanied by any change in global expression of tight-junction proteins or in markers of the integrity of the dopamine (DA) system in the striatum at both time points. However, chronic microglial activation was detected at day 26 post infection, accompanied by an elevation of the inflammatory mediators eotaxin, IFN{gamma}, CXCL9, IL-17 and MIP-2 in the striatum, accompanied by an elevation of IL-17 and CXCL1 in the serum of Pink1 KO mice. Neutrophil infiltration in the brain of infected mice was also noted at day 26 post infection, as revealed by immune cell profiling by flow cytometry. Finally, a bulk RNA-seq transcriptome analysis revealed that gene sets related to synaptic function were particularly influenced by the infection and that inflammation-related genes were upregulated by the infection in the Pink1 KO mice. Our results support the hypothesis that even after mild gastro-intestinal infection, increased BBB permeability could contribute to perturbations of brain homeostasis including altered expression of synaptic genes, increased microglial activation and the establishment of a chronic state of brain inflammation. Such perturbations could potentially act as a first hit for subsequent induction of PD pathology in the context of aging in genetically susceptible individuals.

Author summaryWe hypothesize that immune system activation is linked to the development of Parkinsons disease (PD). Previously work revealed that repeated gastrointestinal infections with Citrobacter rodentium causes PD-like symptoms and immune cell invasion in the brain of Pink1 knockout (KO) mice. In the current study, we tested whether a single mild gut infection alters blood-brain barrier (BBB) permeability and causes brain inflammation. We infected Pink1 WT and KO mice with Citrobacter rodentium and used gadolinium-enhanced MRI to detect BBB permeability changes at 13- and 26-days post-infection. Results showed increased BBB permeability in specific brain regions at 26 days. While tight-junction and dopamine (DA)-related proteins remained unchanged, we observed altered expression of synaptic genes, chronic microglial activation, elevated inflammatory markers, and neutrophil infiltration in the brain. Our findings suggest that even mild gastrointestinal infections can increase BBB permeability, which could be permissive for immune cell entry into the brain and potentially interact with other mechanisms triggering PD pathophysiology, particularly in genetically susceptible individuals.
]]></description>
<dc:creator>Mukherjee, S.</dc:creator>
<dc:creator>Grouza, V.</dc:creator>
<dc:creator>Tchung, A.</dc:creator>
<dc:creator>Even, A.</dc:creator>
<dc:creator>Yaqubi, M.</dc:creator>
<dc:creator>Tuznik, M.</dc:creator>
<dc:creator>Cannon, T.</dc:creator>
<dc:creator>Recinto, S. J.</dc:creator>
<dc:creator>Gavino, C.</dc:creator>
<dc:creator>Bourque, M.-J.</dc:creator>
<dc:creator>McBride, H. M.</dc:creator>
<dc:creator>Desjardins, M.</dc:creator>
<dc:creator>Gruenheid, S.</dc:creator>
<dc:creator>Stratton, J. A.</dc:creator>
<dc:creator>Rudko, D. A.</dc:creator>
<dc:creator>Trudeau, L.-E.</dc:creator>
<dc:date>2024-12-24</dc:date>
<dc:identifier>doi:10.1101/2024.12.24.630165</dc:identifier>
<dc:title><![CDATA[A single Citrobacter rodentium infection in Pink1 knockout and wild type mice leads to regional blood-brain-barrier perturbation and glial activation without dopamine neuron axon terminal loss]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2024-12-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.03.20.644460v1?rss=1">
<title>
<![CDATA[
Soluble Immune Factor Profiles in Blood and CSF Associated with LRRK2 Mutations and Parkinson's Disease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.03.20.644460v1?rss=1"
</link>
<description><![CDATA[
Background and ObjectivesMutations in the Leucine-rich repeat kinase 2 (LRRK2) gene are one of the most common genetic causes of Parkinsons disease (PD) and are linked to immune dysregulation in both the central nervous system and periphery. However, peripheral and central profiles of soluble immune factors associated with LRRK2 mutations and PD have not been comprehensively characterized. Using serum and CSF samples from the LRRK2 Cohort Consortium (LCC), this study aimed to probe a broad range of soluble immune biomarkers associated with LRRK2 mutations and PD.

MethodsWe investigated the levels of soluble immune regulators in the serum (n=651) and cerebrospinal fluid (CSF, n=129) of LRRK2 mutation carriers and non-carriers, both with and without PD. A total of 65 cytokines, chemokines, growth factors, and soluble receptors were assessed by Luminex immunoassay. A multivariable robust linear model was used to determine levels associated with LRRK2 mutations and PD status, adjusting for age, sex, and sample cohort. Correlations were assessed using the Spearman correlation coefficient. LRRK2 G2019S knock-in mice were used to validate the associations identified in the LCC.

ResultsIn this extensive discovery cohort, we identified several elevated serum immune regulatory factors associated with LRRK2 mutations. In particular, serum stromal cell-derived factor-1 alpha (SDF-1 alpha) levels, as supported by findings in LRRK2 G2019S knock-in mice, and tumor necrosis factor receptor II (TNF-RII) were significantly increased after multiple comparison adjustment. In contrast, LRRK2 mutations were associated with reduced soluble immune markers, including BAFF, CD40-Ligand, I-TAC, MIP-3 alpha, NGF beta, and IL-27 in CSF. Those with clinically diagnosed PD, with or without LRRK2 mutations, did not show strong signals in serum but reduced inflammatory analytes in CSF, including MIF, MMP-1, CD30, Tweak, and SDF-1 alpha. In addition, we found that the serum levels of these soluble immune factors display varied correlations with their corresponding CSF levels.

DiscussionThis study highlights distinct immune profiles associated with LRRK2 mutations and PD in the periphery and CNS. Serum levels of SDF-1alpha and TNF-RII were elevated in LRRK2 mutation carriers, while CSF immune markers were reduced. In PD, irrespective of LRRK2 status, reduced CSF inflammatory analytes and weak serum signals were observed. These results provide insight into immune dysregulation linked to LRRK2 mutations. If replicable in independent datasets, they offer potential avenues for biomarker and therapeutic exploration.
]]></description>
<dc:creator>Jaffery, R.</dc:creator>
<dc:creator>Zhao, Y.</dc:creator>
<dc:creator>Ahmed, S.</dc:creator>
<dc:creator>Schumacher, J. G.</dc:creator>
<dc:creator>Ahn, J.</dc:creator>
<dc:creator>Shi, L.</dc:creator>
<dc:creator>Wang, Y.</dc:creator>
<dc:creator>Tan, Y.</dc:creator>
<dc:creator>Chen, K.</dc:creator>
<dc:creator>Tawbi, H.</dc:creator>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Schwarzschild, M. A.</dc:creator>
<dc:creator>Peng, W.</dc:creator>
<dc:creator>Chen, X.</dc:creator>
<dc:date>2025-03-24</dc:date>
<dc:identifier>doi:10.1101/2025.03.20.644460</dc:identifier>
<dc:title><![CDATA[Soluble Immune Factor Profiles in Blood and CSF Associated with LRRK2 Mutations and Parkinson's Disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.04.01.646648v1?rss=1">
<title>
<![CDATA[
iNeurons are sweet, maybe too sweet? Exploring the impact of media composition on PINK1-dependent mitophagy 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.04.01.646648v1?rss=1"
</link>
<description><![CDATA[
Parkinsons disease associated proteins PINK1 and Parkin collaboratively regulate stress-induced mitophagy. While in vitro human neuronal cultures are valuable for studying the roles of PINK1 and Parkin in a disease-relevant context, the impact of culture conditions on these processes remains largely underexplored. Here, it is shown that human induced neurons (iNeurons) cultured in N2B27 and BrainPhys medium exhibit distinct PINK1-Parkin dependent mitophagy phenotypes. Specifically, BrainPhys-cultured iNeurons show greater resistance to PINK1-dependent mitophagy initiation, linked to a reduction in glucose availability and reduced PINK1 protein availabilities, leading to decreases in stress-induced and basal mitophagy fluxes. These findings highlight the critical impact of culture conditions on mitophagy dynamics and emphasise the need to account for media-specific differences when using in vitro models to investigate mitophagy mechanisms in human neurons.
]]></description>
<dc:creator>O'Callaghan, B.</dc:creator>
<dc:creator>Melandri, D.</dc:creator>
<dc:creator>Soltic, D.</dc:creator>
<dc:creator>Cosker, K.</dc:creator>
<dc:creator>Soutar, M. P. M.</dc:creator>
<dc:creator>Plun-Favreau, H.</dc:creator>
<dc:date>2025-04-03</dc:date>
<dc:identifier>doi:10.1101/2025.04.01.646648</dc:identifier>
<dc:title><![CDATA[iNeurons are sweet, maybe too sweet? Exploring the impact of media composition on PINK1-dependent mitophagy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.04.02.646792v1?rss=1">
<title>
<![CDATA[
HyDrop v2: Scalable atlas construction for training sequence-to-function models 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.04.02.646792v1?rss=1"
</link>
<description><![CDATA[
Deciphering cis-regulatory logic underlying cell type identity is a fundamental question in biology. Single-cell chromatin accessibility (scATAC-seq) data has enabled training of sequence-to-function deep learning models allowing decoding of enhancer logic and design of synthetic enhancers. Training such models requires large amounts of high-quality training data across species, organs, development, aging, and disease. To facilitate the cost-effective generation of large scATAC-seq atlases for model training, we developed a new version of the open-source microfluidic system HyDrop with increased sensitivity and scale: HyDrop v2. We generated HyDrop v2 atlases for the mouse cortex and Drosophila embryo development and compared them to atlases generated on commercial platforms. HyDrop v2 data integrates seamlessly with commercially available chromatin accessibility methods (10x Genomics). Differentially accessible regions and motif enrichment across cell types are equivalent between HyDrop-v2 and 10x atlases. Sequence-to-function models trained on either atlas are comparable as well in terms of enhancer predictions, sequence explainability, and transcription factor footprinting. By offering accessible data generation, enhancer models trained on HyDrop-v2 and mixed atlases can contribute to unraveling cell-type specific regulatory elements in health and disease.
]]></description>
<dc:creator>Dickmanken, H.</dc:creator>
<dc:creator>Wojno, M.</dc:creator>
<dc:creator>Theunis, K.</dc:creator>
<dc:creator>Eksi, E. C.</dc:creator>
<dc:creator>Mahieu, L.</dc:creator>
<dc:creator>Christiaens, V.</dc:creator>
<dc:creator>Kempynck, N.</dc:creator>
<dc:creator>De Rop, F. V.</dc:creator>
<dc:creator>Roels, N.</dc:creator>
<dc:creator>Spanier, K. I.</dc:creator>
<dc:creator>Vandepoel, R.</dc:creator>
<dc:creator>Hulselmans, G. J.</dc:creator>
<dc:creator>Poovathingal, S.</dc:creator>
<dc:creator>Aerts, S.</dc:creator>
<dc:date>2025-04-04</dc:date>
<dc:identifier>doi:10.1101/2025.04.02.646792</dc:identifier>
<dc:title><![CDATA[HyDrop v2: Scalable atlas construction for training sequence-to-function models]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.04.02.646812v1?rss=1">
<title>
<![CDATA[
CREsted: modeling genomic and synthetic cell type-specific enhancers across tissues and species 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.04.02.646812v1?rss=1"
</link>
<description><![CDATA[
Sequence-based deep learning models have become the state of the art for the analysis of the genomic regulatory code. Particularly for transcriptional enhancers, deep learning models excel at deciphering sequence features and grammar that underlie their spatiotemporal activity. To enable end-to-end enhancer modeling and design, we developed a software and modeling package, called CREsted. It combines preprocessing starting from single-cell ATAC-seq data; modeling with a choice of several architectures for training classification and regression models on either topics or pseudobulk peak heights; sequence design using multiple strategies; and downstream analysis through a collection of tools to locate transcription factor (TF) binding sites, infer the effect of a TF (activating or repressing) on enhancer accessibility, decipher enhancer grammar, and score gene loci. We demonstrate CREsted using a mouse cortex model that we validate using the BICCN collection of in vivo validated mouse brain enhancers. Classical enhancers in immune cells, including the IFNB1 enhanceosome are revisited using a PBMC model, and we assess the accuracy of TF binding site predictions with ChIP-seq. Additionally, we use CREsted to compare mesenchymal-like cancer cell states between tumor types; and we investigate different fine-tuning strategies of Borzoi within CREsted, comparing their performance and explainability with CREsted models trained from scratch. Finally, we train a CREsted model on a scATAC-seq atlas of zebrafish development and use this to design and in vivo validate cell type-specific synthetic enhancers in three tissues. For varying datasets, we demonstrate that CREsted facilitates efficient training and analyses, enabling scrutinization of the enhancer logic and design of synthetic enhancers across tissues and species. CREsted is available at https://crested.readthedocs.io.
]]></description>
<dc:creator>Kempynck, N.</dc:creator>
<dc:creator>De Winter, S.</dc:creator>
<dc:creator>Blaauw, C. H.</dc:creator>
<dc:creator>Konstantakos, V.</dc:creator>
<dc:creator>Dieltiens, S.</dc:creator>
<dc:creator>Eksi, E. C.</dc:creator>
<dc:creator>Bercier, V.</dc:creator>
<dc:creator>Taskiran, I. I.</dc:creator>
<dc:creator>Hulselmans, G.</dc:creator>
<dc:creator>Spanier, K.</dc:creator>
<dc:creator>Christiaens, V.</dc:creator>
<dc:creator>Van Den Bosch, L.</dc:creator>
<dc:creator>Mahieu, L.</dc:creator>
<dc:creator>Aerts, S.</dc:creator>
<dc:date>2025-04-03</dc:date>
<dc:identifier>doi:10.1101/2025.04.02.646812</dc:identifier>
<dc:title><![CDATA[CREsted: modeling genomic and synthetic cell type-specific enhancers across tissues and species]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.04.18.649338v1?rss=1">
<title>
<![CDATA[
Sex-dimorphic effects of neuromelanin buildup in rodent nigral dopamine neurons: implications for sex-biased vulnerability in Parkinson's disease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.04.18.649338v1?rss=1"
</link>
<description><![CDATA[
Neuromelanin (NM) is a dark pigment accumulating with age in human substantia nigra pars compacta (SNpc) dopamine (DA) neurons, conferring the dark look that inspired nigral areas name. Despite NM has long been associated with Parkinsons disease (PD), as melanized neurons favorably degenerate during disease development, NM functions within SNpc DA neurons are still mostly elusive. Here, by exploiting an NM-producing rat model generated by viral vector-induced expression of human Tyrosinase (hTyr), we inspected NM impact on nigral DA neurons survival and activity, on mitochondrial functionality of SNpc, and behaviors resembling non-motor and motor PD symptoms. Our data reveal sex dimorphism in NM effects on nigrostriatal dopamine circuit, with sex-biased alterations in neuronal firing activity and underlying intrinsic currents, nigral mitochondrial functions, and non-motor PD symptoms (anxiety). In conclusion, this study discloses unrealized NM effects within nigral DA neurons, advancing our comprehension of sex-specific features shaping sex-biased vulnerability to PD.
]]></description>
<dc:creator>D'Addario, S. L.</dc:creator>
<dc:creator>Massaro Cenere, M.</dc:creator>
<dc:creator>Cossa, F.</dc:creator>
<dc:creator>Scaricamazza, S.</dc:creator>
<dc:creator>Nesci, V.</dc:creator>
<dc:creator>D'Ottavio, G.</dc:creator>
<dc:creator>Caprioli, D.</dc:creator>
<dc:creator>Ferri, A.</dc:creator>
<dc:creator>Vila, M.</dc:creator>
<dc:creator>Ledonne, A.</dc:creator>
<dc:creator>Mercuri, N. B.</dc:creator>
<dc:date>2025-04-23</dc:date>
<dc:identifier>doi:10.1101/2025.04.18.649338</dc:identifier>
<dc:title><![CDATA[Sex-dimorphic effects of neuromelanin buildup in rodent nigral dopamine neurons: implications for sex-biased vulnerability in Parkinson's disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.04.21.649867v1?rss=1">
<title>
<![CDATA[
Differential memory enrichment of cytotoxic CD4 T cells in Parkinson's disease patients reactive to α-synuclein 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.04.21.649867v1?rss=1"
</link>
<description><![CDATA[
Parkinsons disease (PD) is a complex neurodegenerative disease with a largely unknown etiology. Although the loss of dopaminergic neurons in the substantia nigra pars compacta is the pathological hallmark of PD, neuroinflammation also plays a fundamental role in PD pathology. We have previously reported that PD patients have increased frequencies of T cell reactive to peptides from -synuclein (-syn). However, not all PD participants respond to -syn. Furthermore, we have previously found that CD4 T cells from PD participants responding to -syn (PD_R) are transcriptionally distinct from PD participants not responding to -syn (PD_NR). To gain further insight into the pathology of PD_R participants, we investigated surface protein expression of 11 proteins whose genes had previously been found to be differentially expressed when comparing PD_R and healthy control participants not responding to -syn (HC_NR). We found that Cadherin EGF LAG seven-pass G-type receptor 2 (CELSR2) was expressed on a significantly higher proportion of CD4 effector memory T cells (TEM) in PD_R compared to HC_NR. Single-cell RNA sequencing analysis of cells expressing or not expressing CELSR2 revealed that PD_R participants have elevated frequencies of activated TEM subsets and an almost complete loss of cytotoxic TEM cells. Flow cytometry analyses confirmed that Granulysin+ CD4 cytotoxic TEM cells are reduced in PD_R. Taken together, these results provide further insight into the perturbation of T cell subsets in PD_R, and highlights the need for further investigation into the role of Granulysin+ CD4 cytotoxic TEM in PD pathology.
]]></description>
<dc:creator>Freuchet, A.</dc:creator>
<dc:creator>Johansson, E.</dc:creator>
<dc:creator>Frazier, A.</dc:creator>
<dc:creator>Litvan, I.</dc:creator>
<dc:creator>Goldman, J. G.</dc:creator>
<dc:creator>Alcalay, R. N.</dc:creator>
<dc:creator>Sulzer, D.</dc:creator>
<dc:creator>Lindestam Arlehamn, C. S.</dc:creator>
<dc:creator>Sette, A.</dc:creator>
<dc:date>2025-04-24</dc:date>
<dc:identifier>doi:10.1101/2025.04.21.649867</dc:identifier>
<dc:title><![CDATA[Differential memory enrichment of cytotoxic CD4 T cells in Parkinson's disease patients reactive to α-synuclein]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.04.21.649871v1?rss=1">
<title>
<![CDATA[
T cell responses towards PINK1 and α-synuclein are elevated in prodromal Parkinson's disease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.04.21.649871v1?rss=1"
</link>
<description><![CDATA[
A role of the immune system in Parkinsons disease (PD) progression has long been suspected due to the increased frequency of activated glial cells and infiltrating T cells into the substantia nigra. It was previously reported that PD donors have increased T cell responses towards PINK1 and -synuclein (-syn), two Lewy body-associated proteins. Further, T cell reactivity towards -syn was highest closer to disease onset, highlighting that autoreactive T cells might play a role in PD pathogenesis. However, whether T cell autoreactivity is present during prodromal PD is unknown. Here, we investigated T cell responses towards PINK1 and -syn in donors at high risk of developing PD (i.e. prodromal PD: genetic risk, hyposmia, and or REM sleep behavior disorder), in comparison to PD and healthy control donors. T cell reactivity to these two autoantigens was detected in prodromal PD at levels comparable to those detected in individuals with clinically diagnosed PD. Aligned with the increased incidence of PD in males, we found that males with PD, but not females, had elevated T cell reactivity compared to healthy controls. However, among prodromal PD donors, males and females had elevated T cell responses. These differing trends in reactivity highlights the need for further studies of the impact of biological sex on neuroinflammation and PD progression.
]]></description>
<dc:creator>Johansson, E.</dc:creator>
<dc:creator>Freuchet, A.</dc:creator>
<dc:creator>Williams, G. P.</dc:creator>
<dc:creator>Michealis, T.</dc:creator>
<dc:creator>Frazier, A.</dc:creator>
<dc:creator>Litvan, I.</dc:creator>
<dc:creator>Goldman, J. G.</dc:creator>
<dc:creator>Alcalay, R. N.</dc:creator>
<dc:creator>Standaert, D. G.</dc:creator>
<dc:creator>Amara, A. W.</dc:creator>
<dc:creator>Stover, N.</dc:creator>
<dc:creator>Fon, E. A.</dc:creator>
<dc:creator>Postuma, R. B.</dc:creator>
<dc:creator>Sidney, J.</dc:creator>
<dc:creator>Sulzer, D.</dc:creator>
<dc:creator>Lindestam Arlehamn, C. S.</dc:creator>
<dc:creator>Sette, A.</dc:creator>
<dc:date>2025-04-24</dc:date>
<dc:identifier>doi:10.1101/2025.04.21.649871</dc:identifier>
<dc:title><![CDATA[T cell responses towards PINK1 and α-synuclein are elevated in prodromal Parkinson's disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-04-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2023.12.23.573214v1?rss=1">
<title>
<![CDATA[
Spatial genomics of AAVs reveals mechanism of transcriptional crosstalk that enables targeted delivery of large genetic cargo 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2023.12.23.573214v1?rss=1"
</link>
<description><![CDATA[
Integrating cell type-specific regulatory elements (e.g. enhancers) with recombinant adeno-associated viruses (AAVs) can provide broad and efficient genetic access to specific cell types. However, the packaging capacity of AAVs restricts the size of both the enhancers and the cargo that can be delivered. Transcriptional crosstalk offers a novel paradigm for cell type-specific expression of large cargo, by separating distally-acting regulatory elements into a second AAV genome. Here, we identify and profile transcriptional crosstalk in AAV genomes carrying 11 different enhancers active in mouse brain. To understand transcriptional crosstalk, we develop spatial genomics methods to identify and localize AAV genomes and their concatemeric forms in cultured cells and in tissue. Using these methods, we construct detailed views of the dynamics of AAV transduction and demonstrate that transcriptional crosstalk is dependent upon concatemer formation. Finally, we leverage transcriptional crosstalk to drive expression of a large Cas9 cargo in a cell type-specific manner with systemically-administered engineered AAVs and demonstrate AAV-delivered, minimally-invasive, cell type-specific gene editing in wildtype animals that recapitulates known disease phenotypes.

HighlightsO_LITranscriptional crosstalk between enhancers and promoters delivered in trans by AAVs is a generalized phenomenon.
C_LIO_LISpatial genomics techniques, AAV-Zombie and SpECTr, reveal that AAV genome concatemerization facilitates transcriptional crosstalk.
C_LIO_LITranscriptional crosstalk can be leveraged for minimally-invasive, targeted AAV delivery of large cargo, including machinery for CRISPR-based gene editing and manipulation.
C_LIO_LITranscriptional crosstalk enables cell-type specific gene disruption in wildtype animals, recapitulating behavioural phenotypes of genetic knockouts.
C_LI
]]></description>
<dc:creator>Coughlin, G. M.</dc:creator>
<dc:creator>Borsos, M.</dc:creator>
<dc:creator>Appling, N.</dc:creator>
<dc:creator>Barcelona, B. H.</dc:creator>
<dc:creator>Mayfield, A. M. H.</dc:creator>
<dc:creator>Mackey, E. D.</dc:creator>
<dc:creator>Eser, R. A.</dc:creator>
<dc:creator>Chen, X.</dc:creator>
<dc:creator>Kumar, S. R.</dc:creator>
<dc:creator>Gradinaru, V.</dc:creator>
<dc:date>2023-12-24</dc:date>
<dc:identifier>doi:10.1101/2023.12.23.573214</dc:identifier>
<dc:title><![CDATA[Spatial genomics of AAVs reveals mechanism of transcriptional crosstalk that enables targeted delivery of large genetic cargo]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2023-12-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.05.09.653147v1?rss=1">
<title>
<![CDATA[
Defect in hematopoiesis and embryonic lethality at midgestation of Vps13a/Vps13c double knockout mice 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.05.09.653147v1?rss=1"
</link>
<description><![CDATA[
VPS13 is the founding member of a family of proteins that mediate lipid transfer at intracellular membrane contact sites by a bridge-like mechanism. Mammalian genomes comprise 4 VPS13 genes encoding proteins with distinct localizations and function. The gene duplication resulting in VPS13A and VPS13C is the most recent in evolution and, accordingly, these two proteins are the most similar to each other. However, they have distinct subcellular localizations and their loss of function mutations in humans are compatible with life but result in two different age-dependent neurodegenerative diseases, chorea-acanthocytosis and Parkinsons disease, respectively. Thus, it remains unclear whether these two proteins have overlapping functions. Here, we show that while Vps13a KO and Vps13c KO mice are viable, embryonic development of Vps13a/Vps13c double knockout (DKO) mice is arrested at midgestation. Prior to death, DKO embryos were smaller than controls, were anemic and had a smaller liver, the key erythropoietic site at this developmental stage. Further analyses of erythroid precursor cells showed that their differentiation was impaired and that this defect was accompanied by activation of innate immunity as revealed by upregulation of interferon stimulated genes (ISGs). Additionally, the RIG-I and MDA5 components of dsRNA triggered innate immunity were found upregulated in the DKO fetal liver. Activation of innate immunity may result from loss of integrity of the membranes of intracellular organelles, such as mitochondria and autophagic lysosomes, due to the absence of these lipid transport proteins. The surprising and striking synthetic effect resulting for the combined loss of VPS13A and VPS13C suggests that despite of the different localization of these two proteins, the lipid fluxes that they mediate are partially redundant.
]]></description>
<dc:creator>Xu, P.</dc:creator>
<dc:creator>Mancuso, R. I.</dc:creator>
<dc:creator>Leonzino, M.</dc:creator>
<dc:creator>Zeiss, C. J.</dc:creator>
<dc:creator>Krause, D. S.</dc:creator>
<dc:creator>De Camilli, P.</dc:creator>
<dc:date>2025-05-13</dc:date>
<dc:identifier>doi:10.1101/2025.05.09.653147</dc:identifier>
<dc:title><![CDATA[Defect in hematopoiesis and embryonic lethality at midgestation of Vps13a/Vps13c double knockout mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-13</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.05.09.653000v1?rss=1">
<title>
<![CDATA[
Introducing PIGMO, a novel PIGmented MOuse model of Parkinson's disease (V1) 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.05.09.653000v1?rss=1"
</link>
<description><![CDATA[
There is a pressing need for the development, characterization, and standardization of animal models of Parkinsons disease (PD) that properly mimic the cardinal features of this disorder, comprising both the motor phenotype and neuropathological signatures. In the past few years, animal modeling has moved from neurotoxin-based approaches toward viral vectors carrying a given genetic payload of interest. Here, to induce pigmentation of the mouse brain upon systemic delivery, we took advantage of a modified adeno-associated viral vector capsid engineered to bypass the blood-brain barrier and coding for the human tyrosinase gene (AAV9-P31-hTyr). Obtained results revealed an ongoing pigmentation of catecholaminergic centers related to the pathophysiology of PD, such as the substantia nigra pars compacta, ventral tegmental area, and locus coeruleus. Moreover, pigmented dopaminergic neurons exhibited Lewy body-like intracytoplasmic inclusions, a progressive nigrostriatal degeneration, and a time-dependent PD motor phenotype. The bilateral pigmented mouse model of PD generated this way is highly reproducible, does not require stereotaxic surgery for viral vector deliveries, and opens unprecedented possibilities for preclinical testing of therapeutic candidates designed to reduce disease progression rates.
]]></description>
<dc:creator>Chocarro, J.</dc:creator>
<dc:creator>Marana, S.</dc:creator>
<dc:creator>Espelosin, M.</dc:creator>
<dc:creator>Rico, A. J.</dc:creator>
<dc:creator>Ariznabarreta, G.</dc:creator>
<dc:creator>Lorenzo-Ramos, E.</dc:creator>
<dc:creator>Illarduya, M. M.</dc:creator>
<dc:creator>Hernandez-Alcoceba, R.</dc:creator>
<dc:creator>Chillon, M.</dc:creator>
<dc:creator>Vila, M.</dc:creator>
<dc:creator>Kordower, J.</dc:creator>
<dc:creator>Schapira, A. H. V.</dc:creator>
<dc:creator>Garcia-Osta, A.</dc:creator>
<dc:creator>Cuadrado-Tejedor, M.</dc:creator>
<dc:creator>Lanciego, J. L.</dc:creator>
<dc:date>2025-05-14</dc:date>
<dc:identifier>doi:10.1101/2025.05.09.653000</dc:identifier>
<dc:title><![CDATA[Introducing PIGMO, a novel PIGmented MOuse model of Parkinson's disease (V1)]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.05.16.654599v1?rss=1">
<title>
<![CDATA[
End-product inhibition of the LRRK2-counteracting PPM1H phosphatase 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.05.16.654599v1?rss=1"
</link>
<description><![CDATA[
PPM1H phosphatase reverses Parkinsons disease-associated, Leucine Rich Repeat Kinase 2-mediated, Rab GTPase phosphorylation. We showed previously that PPM1H relies on an N-terminal amphipathic helix for Golgi membrane localization and this helix enables PPM1H to associate with liposomes in vitro; binding to highly curved liposomes activates PPM1Hs phosphatase activity. We show here that PPM1H also contains an allosteric binding site for its non-phosphorylated reaction products, Rab8A and Rab10. Microscale thermophoresis revealed that PPM1H binds thio-phosphorylated Rab8A at the active site with a KD of [~]1{micro}M; binding of Rab8A and Rab10 to an alternative site is of similar affinity and is not detected for another LRRK2 substrate, Rab12. Non-phosphorylated Rab8A or Rab10 inhibit PPM1H phosphatase reactions at concentrations consistent with their measured binding affinities and fail to inhibit PPM1H L66R phosphatase reactions. Independent confirmation of non-phosphorylated Rab binding to PPM1H was obtained by sucrose gradient co-flotation of non-phosphorylated Rabs with liposome-bound PPM1H. Finally, Rab8A or Rab10 binding also requires PPM1Hs amphipathic helix, without which the interaction affinity is decreased about 6-fold. These experiments indicate that Golgi associated Rab proteins contribute to the localization of PPM1H and non-phosphorylated Rabs regulate PPM1H phosphatase activity via an allosteric site. Targeting this site could represent a strategy to enhance PPM1H-mediated dephosphorylation of LRRK2 substrates, offering a potential therapeutic approach to counteract LRRK2-driven Parkinsons disease.
]]></description>
<dc:creator>Adhikari, A.</dc:creator>
<dc:creator>Tripathi, A.</dc:creator>
<dc:creator>Chiang, C. Y.</dc:creator>
<dc:creator>Sherpa, P. L.</dc:creator>
<dc:creator>Pfeffer, S. R.</dc:creator>
<dc:date>2025-05-17</dc:date>
<dc:identifier>doi:10.1101/2025.05.16.654599</dc:identifier>
<dc:title><![CDATA[End-product inhibition of the LRRK2-counteracting PPM1H phosphatase]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.05.16.654530v1?rss=1">
<title>
<![CDATA[
Inflamed Microglia like Macrophages in the Central Nervous System of Prodromal Parkinson's Disease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.05.16.654530v1?rss=1"
</link>
<description><![CDATA[
We investigated the role of inflammation in the pathogenesis of prodromal Parkinsons Disease (PD), performing single-cell RNAseq analysis of cerebrospinal fluid (CSF) and blood from 111 individuals, comparing control subjects with early prodromal PD and later PD to patients with multiple sclerosis (MS). Surprisingly, we identified a pleocytosis in the CSF, most pronounced in patients with early PD. Single-cell RNAseq revealed increases in CSF-specific microglia-like macrophages expressing JAK-STAT and TNF signaling signatures in prodromal PD, with a lack of T cell activation in the CSF. The CSF macrophages exhibited similar transcriptional profiles to dural macrophages from human -synuclein-expressing PD model mice. These findings uncover a myeloid-mediated TNF inflammatory process in the CNS of patients with prodromal PD, suggesting a novel pathological mechanism in disease etiology.
]]></description>
<dc:creator>Zhang, L.</dc:creator>
<dc:creator>Yasumizu, Y.</dc:creator>
<dc:creator>Deerhake, M. E.</dc:creator>
<dc:creator>Moon, J.</dc:creator>
<dc:creator>Buitrago-Pocasangre, N.</dc:creator>
<dc:creator>Russo, A.</dc:creator>
<dc:creator>Wang, H.</dc:creator>
<dc:creator>Zhu, B.</dc:creator>
<dc:creator>Seibyl, J. P.</dc:creator>
<dc:creator>Reddy, V.</dc:creator>
<dc:creator>Wang, Q.</dc:creator>
<dc:creator>Spillantini, M. G.</dc:creator>
<dc:creator>Posner, D. A.</dc:creator>
<dc:creator>Clatworthy, M.</dc:creator>
<dc:creator>Sumida, T. S.</dc:creator>
<dc:creator>Longbrake, E. E.</dc:creator>
<dc:creator>Cedarbaum, J. M.</dc:creator>
<dc:creator>Hafler, D. A.</dc:creator>
<dc:date>2025-05-21</dc:date>
<dc:identifier>doi:10.1101/2025.05.16.654530</dc:identifier>
<dc:title><![CDATA[Inflamed Microglia like Macrophages in the Central Nervous System of Prodromal Parkinson's Disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.05.16.654598v1?rss=1">
<title>
<![CDATA[
Repetitive Levodopa Treatment Drives Cell Type-Specific Striatal Adaptations Associated With Progressive Dyskinesia in Parkinsonian Mice 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.05.16.654598v1?rss=1"
</link>
<description><![CDATA[
The use of levodopa to manage Parkinsons disease (PD) symptoms leads to levodopa-induced dyskinesia (LID) and other motor fluctuations, which worsen with disease progression and repeated treatment. Aberrant activity of striatal D1- and D2-expressing medium spiny neurons (D1-/D2-MSNs) underlies LID, but the mechanisms driving its progression remain unclear. Using the 6-OHDA mouse model of PD/LID, we combined in vivo and ex vivo recordings to isolate the effect of repeated treatment in LID worsening and other motor fluctuation-related phenotypes. We found that LID worsening is linked to potentiation of levodopa-evoked responses in both D1-/D2-MSNs, independent of changes in dopamine release or MSN intrinsic excitability. Instead, strengthening of glutamatergic synapses onto D1-MSNs emerged as a key driver. Moreover, we found changes in D2-MSN activity that specifically influenced LID duration, potentially contributing to motor fluctuations, which paralleled a reduction in D2R sensitivity. These findings reveal striatal adaptations contributing to worsening of levodopa-related complications.
]]></description>
<dc:creator>Paz, R. M.</dc:creator>
<dc:creator>Ryan, M. B.</dc:creator>
<dc:creator>Marcott, P. F.</dc:creator>
<dc:creator>Girasole, A. E.</dc:creator>
<dc:creator>Faryean, J.</dc:creator>
<dc:creator>Duong, V.</dc:creator>
<dc:creator>Sridhar, S.</dc:creator>
<dc:creator>Nelson, A. B.</dc:creator>
<dc:date>2025-05-21</dc:date>
<dc:identifier>doi:10.1101/2025.05.16.654598</dc:identifier>
<dc:title><![CDATA[Repetitive Levodopa Treatment Drives Cell Type-Specific Striatal Adaptations Associated With Progressive Dyskinesia in Parkinsonian Mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.05.21.655268v1?rss=1">
<title>
<![CDATA[
Spatio-molecular analysis of primate subthalamus defines anatomical domains relevant to Parkinson disease and neuropsychiatry 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.05.21.655268v1?rss=1"
</link>
<description><![CDATA[
The brain area including and surrounding the subthalamic nucleus (STN) contains several targets for deep brain stimulation (DBS): dorsal STN for the treatment of motor symptoms in Parkinsons disease (PD), anteromedial STN for obsessive compulsive disorder (OCD), zona incerta (ZI) for PD and essential tremor, and medial forebrain bundle (MFB) for OCD and depression. Despite clinical application, molecular and anatomical understanding of this area has remained elusive. To help solving this, we took advantage of our recent data from transcriptomics analysis in mice, and applied fluorescent in situ hybridization (FISH) analysis on macaque and human brain sections. In accordance with a glutamatergic phenotype, we observed VGLUT2 mRNA across the STN, with quantification revealing higher VGLUT2 density in the ventromedial (vm) STN than dorsal STN. Unexpectedly, we identified HTR2C mRNA uniquely localized to vmSTN, while absent in dorsal STN. HTR2C thereby molecularly defines a distinct vmSTN domain, hereafter named vmSTNHTR2C. Based on the role of the serotonin (5-HT) 2C receptor in mood, vmSTNHTR2Cmay spatio-molecularly correspond to the limbic STN. The ventralized HTR2C profile was strongest in anterior STN and coincided with serotonergic innervation. The presence of vmSTNHTR2C was identified also in the human brain, but absent in the mouse. Strong HTR2C anteriorly revealed a new "far-anterior" STN domain. Comparing HTR2C with TAC1, a para-STN marker, para-STN was identified at the anterior STN, partly intermingled with the MFB. Thus, anterior STN shows several unique molecular and anatomical features in the primate brain. Upon analysis of multiple markers, the STN, para-STN and ZI could be divided into several expression-based domains, which we used to construct molecularly supported anatomical maps. This data will inform neuromodulation strategies, such as DBS, and pharmacological treatment approaches.
]]></description>
<dc:creator>Dumas, S.</dc:creator>
<dc:creator>Francois, C.</dc:creator>
<dc:creator>Karachi, C.</dc:creator>
<dc:creator>Mackenzie, A.</dc:creator>
<dc:date>2025-05-21</dc:date>
<dc:identifier>doi:10.1101/2025.05.21.655268</dc:identifier>
<dc:title><![CDATA[Spatio-molecular analysis of primate subthalamus defines anatomical domains relevant to Parkinson disease and neuropsychiatry]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.03.17.641409v1?rss=1">
<title>
<![CDATA[
17q21.31 locus regulates Parkinson's disease relevant pathways through KANSL1 activity 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.03.17.641409v1?rss=1"
</link>
<description><![CDATA[
An inversion polymorphism at the 17q21.31 locus defines the H1 and H2 haplotypes, with the former linked to multiple neurodegenerative disorders, including an increased risk of Parkinsons disease (PD). Although the high linkage disequilibrium at this locus has made it difficult to decipher which gene(s) drive the PD association, there is increasing evidence to support the role of KANSL1 as a risk gene. KANSL1 has been shown to regulate the expression of some PD-associated genes and pathways, likely as part of the histone acetylating non-specific lethal (NSL) complex. Here for the first time, we studied the global effects of 17q21.31 haplotype variation using bulk and single-nuclear RNA-sequencing data from control and PD patient brain. We first analysed differential gene expression across haplotype groups, and then assessed the contribution of KANSL1 by comparing with the results of an siRNA knockdown in neuronal and glial human cell lines. We demonstrated that the PD risk-associated H1 haplotype downregulates autophagy, lysosomal and mitochondrial processes, all of which have already been implicated in PD aetiology. Furthermore, these effects were apparent in both neuronal and glial cell types, and in the case of the latter, appear to be associated with the modulation of innate and adaptive immune responses. Thus, we identify important links between NSL complex activity and PD pathophysiology that can be leveraged for novel therapeutic interventions.
]]></description>
<dc:creator>Hicks, A. R.</dc:creator>
<dc:creator>O'Callaghan, B.</dc:creator>
<dc:creator>Brenton, J. W.</dc:creator>
<dc:creator>Grant-Peters, M.</dc:creator>
<dc:creator>Fairbrother-Browne, A.</dc:creator>
<dc:creator>Rocamora Perez, G.</dc:creator>
<dc:creator>Loh, C. A.</dc:creator>
<dc:creator>Reynolds, R. H.</dc:creator>
<dc:creator>Gustavsson, E. K.</dc:creator>
<dc:creator>Montgomery, K.</dc:creator>
<dc:creator>Tsalenchuk, M.</dc:creator>
<dc:creator>Real, R.</dc:creator>
<dc:creator>Morris, H.</dc:creator>
<dc:creator>Lunnon, K.</dc:creator>
<dc:creator>Marzi, S. J.</dc:creator>
<dc:creator>Jaunmuktane, Z.</dc:creator>
<dc:creator>Hardy, J.</dc:creator>
<dc:creator>Plun-Favreau, H.</dc:creator>
<dc:creator>Ryten, M.</dc:creator>
<dc:date>2025-03-17</dc:date>
<dc:identifier>doi:10.1101/2025.03.17.641409</dc:identifier>
<dc:title><![CDATA[17q21.31 locus regulates Parkinson's disease relevant pathways through KANSL1 activity]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.04.29.651228v1?rss=1">
<title>
<![CDATA[
Single-cell multiomics identifies both shared and unique features of immune dysfunction in the colon, plasma and stool from individuals diagnosed with Parkinson's disease or inflammatory bowel disease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.04.29.651228v1?rss=1"
</link>
<description><![CDATA[
Parkinsons disease (PD) is the fastest growing neurodegenerative disease in the world1. Gastrointestinal (GI) dysfunction can occur decades before motor impairments and in up to 80% of individuals living with PD2,3,4. We investigated peripheral relationships that may underlie mechanisms along the gut-blood axis that contribute to PD pathogenesis. Single-cell multiomic spatial molecular imaging (SMI) of colonic tissue localized inflammatory injury within epithelial cells that appear to be associated with iron mishandling in both inflammatory bowel disease (IBD) and PD biosamples. We found that both the single-cell SMI of RNA and protein revealed parallel cross-modal dysregulation in the gut epithelium, in both IBD and PD biosamples. These data are accompanied by plasma (PD) and stool (IBD) protein depletion of CCL22. Our findings suggest iron mishandling along the gut barrier likely contributes to systemic inflammation, which may be the catalyst that primes circulating immune cells to body-first PD pathogenesis.
]]></description>
<dc:creator>Bolen, M. L.</dc:creator>
<dc:creator>buendia, m.</dc:creator>
<dc:creator>Shi, J.</dc:creator>
<dc:creator>Staley, H.</dc:creator>
<dc:creator>Kachergus, J. M.</dc:creator>
<dc:creator>Efron, P. A.</dc:creator>
<dc:creator>Park, G.</dc:creator>
<dc:creator>Nagpal, R.</dc:creator>
<dc:creator>Xue, Q.-S.</dc:creator>
<dc:creator>McFarland, N. R.</dc:creator>
<dc:creator>Zimmermann, E. M.</dc:creator>
<dc:creator>Forsmark, C. E.</dc:creator>
<dc:creator>Menees, K. B.</dc:creator>
<dc:creator>Salas, A.</dc:creator>
<dc:creator>Thompson, E. A.</dc:creator>
<dc:creator>Tansey, M. G.</dc:creator>
<dc:date>2025-05-02</dc:date>
<dc:identifier>doi:10.1101/2025.04.29.651228</dc:identifier>
<dc:title><![CDATA[Single-cell multiomics identifies both shared and unique features of immune dysfunction in the colon, plasma and stool from individuals diagnosed with Parkinson's disease or inflammatory bowel disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.03.04.641379v1?rss=1">
<title>
<![CDATA[
Molecular and cellular signatures differentiate Parkinson's disease from Parkinson's disease with dementia 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.03.04.641379v1?rss=1"
</link>
<description><![CDATA[
Parkinsons disease (PD) affects millions of people worldwide, and up to 40% of these patients develop dementia, profoundly affecting their quality of life. Whether Parkinsons disease dementia (PDD) simply represents a late stage of PD or constitutes a distinct neurodegenerative process remains unresolved. To clarify this, we generated the largest single nuclear transcriptomic atlas of PD and PDD to date--almost one million nuclei derived from the anterior cingulate cortex and inferior parietal lobule of 64 post-mortem donors. By integrating these data with long-read RNA-seq, we found that the cellular compositions, biological pathways, and molecular profiles diverge substantially between PD and PDD, with limited overlap in differentially expressed genes and pathways. While PD was characterised by widespread upregulation of gene expression programs and robust regional signatures, PDD showed extensive pathway downregulation, loss of cortical regional identity, and significant shifts in transcript usage, including alterations in APP isoforms that may influence pathological amyloid beta accumulation. These findings reveal that PD and PDD represent fundamentally distinct disease states, offering important insights for understanding their underlying mechanisms and will guide the development of targeted therapies and more effective clinical trials.
]]></description>
<dc:creator>Fairbrother-Browne, A.</dc:creator>
<dc:creator>Grant-Peters, M.</dc:creator>
<dc:creator>Brenton, J. W.</dc:creator>
<dc:creator>Nelvagal, H.</dc:creator>
<dc:creator>Reynolds, R. H.</dc:creator>
<dc:creator>Lim, Y. M.</dc:creator>
<dc:creator>Gustavsson, E. K.</dc:creator>
<dc:creator>Macpherson, H.</dc:creator>
<dc:creator>Montgomery, K.</dc:creator>
<dc:creator>Evans, J. R.</dc:creator>
<dc:creator>Hicks, A. R.</dc:creator>
<dc:creator>Chiraki, N.</dc:creator>
<dc:creator>Curless, T.</dc:creator>
<dc:creator>Real, R.</dc:creator>
<dc:creator>Xenakis, T.</dc:creator>
<dc:creator>Houlden, H.</dc:creator>
<dc:creator>Morris, H. R.</dc:creator>
<dc:creator>Gandhi, S.</dc:creator>
<dc:creator>Wood, N. W.</dc:creator>
<dc:creator>Hardy, J.</dc:creator>
<dc:creator>Jaunmuktane, Z.</dc:creator>
<dc:creator>Ryten, M.</dc:creator>
<dc:date>2025-03-04</dc:date>
<dc:identifier>doi:10.1101/2025.03.04.641379</dc:identifier>
<dc:title><![CDATA[Molecular and cellular signatures differentiate Parkinson's disease from Parkinson's disease with dementia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-03-04</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.05.02.651888v1?rss=1">
<title>
<![CDATA[
LRP10 promotes trafficking of progranulin and prosaposin to lysosomes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.05.02.651888v1?rss=1"
</link>
<description><![CDATA[
Mutations in LRP10, a low-density lipoprotein receptor family member, cause familial Parkinsons disease and dementia with Lewy bodies. However, its direct cellular functions remain largely undefined. Using a multidisciplinary approach, our new data shows that LRP10 is required for the efficient trafficking of progranulin and prosaposin to lysosomes. Loss of LRP10 resulted in aberrant Golgi accumulations of progranulin and prosaposin and their reduced abundance in lysosomes. Disease-linked LRP10 missense mutations failed to support this lysosomal trafficking. Moreover, LRP10 KO mice developed striking microgliosis marked by enlarged and hyper-ramified microglia, accompanied by progranulin accumulation in the Golgi. Our results define LRP10 as a positive regulator of progranulin and prosaposin lysosomal protein trafficking and microglia homeostasis and thus shed new light on how its dysfunction may drive neurodegeneration in Parkinsons Disease and dementia with Lewy bodies.
]]></description>
<dc:creator>Filippini, F.</dc:creator>
<dc:creator>Devireddy, S.</dc:creator>
<dc:creator>Ferguson, S.</dc:creator>
<dc:date>2025-05-03</dc:date>
<dc:identifier>doi:10.1101/2025.05.02.651888</dc:identifier>
<dc:title><![CDATA[LRP10 promotes trafficking of progranulin and prosaposin to lysosomes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.05.16.653159v1?rss=1">
<title>
<![CDATA[
Matters arising: Forced Symmetry Artifacts in a Prohibitin Complex Structure 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.05.16.653159v1?rss=1"
</link>
<description><![CDATA[
In a recent publication, Lange et al.1 proposed an 11-spoked wheel model for the mitochondrial prohibitin complex consisting of six PHB1 and five PHB2 subunits built into low resolution density obtained from 11-fold symmetry averaging. The proposed unequal stoichiometry is inherently incompatible with symmetry. The building of an asymmetric molecular model into density obtained by 11-fold symmetry averaging is inherently self-contradictory. This contradiction alone calls the validity of the derived structure into question. Our own data revealed that the complex primarily adopts an asymmetric open conformation in situ. We show that it was the incorrect symmetry imposition on the open conformation that led to the spoked wheel density reported by Lange et al.. Further validation by re-analysis of crosslinking-mass spectrometry data, and a test of in situ template matching all support a larger prohibitin complex architecture. These findings underscore the need for great care in the imposition of symmetry cryo-ET at less than atomistic resolution.
]]></description>
<dc:creator>Herrmann, E.</dc:creator>
<dc:creator>Rose, K. M.</dc:creator>
<dc:creator>Hurley, J. H.</dc:creator>
<dc:date>2025-05-20</dc:date>
<dc:identifier>doi:10.1101/2025.05.16.653159</dc:identifier>
<dc:title><![CDATA[Matters arising: Forced Symmetry Artifacts in a Prohibitin Complex Structure]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-05-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.06.03.657460v1?rss=1">
<title>
<![CDATA[
Parkinson's disease-vulnerable and -resilient dopamine neurons display opposite responses to excitatory input 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.06.03.657460v1?rss=1"
</link>
<description><![CDATA[
Dopamine (DA) neurons of the substantia nigra (SN) are essential for motor control and selectively degenerate in Parkinsons disease (PD). However, DA neurons are molecularly heterogeneous, with some showing greater vulnerability and others resilience. Here, we show that the DA subtype marker Anxa1, identified in mice, labels PD-vulnerable DA neurons in human SN. Using mice, we found that excitatory inputs from subthalamic (STN) and pedunculopontine (PPN) nuclei evoked frequency-dependent excitation in SN GABA neurons, but complex multiphasic DA neuron responses, suggesting heterogeneous DA subtype responses. Indeed, excitatory inputs evoked differential DA responses in striatal subregions, an increase in caudal striatum, but inhibition followed by rebound in dorsolateral striatum. Additionally, PD-resilient Vglut2+ DA neurons were excited by STN/PPN input, while vulnerable Anxa1+ DA neurons were inhibited. These findings demonstrate that DA subtypes are embedded in distinct functional networks, suggesting that some therapeutic interventions may differentially impact vulnerable and resilient DA subtypes.
]]></description>
<dc:creator>Hadjas, L. C.</dc:creator>
<dc:creator>Kollman, G. J.</dc:creator>
<dc:creator>Linderhof, L.</dc:creator>
<dc:creator>Xia, M.</dc:creator>
<dc:creator>Mansur, S.</dc:creator>
<dc:creator>Saint-Pierre, M.</dc:creator>
<dc:creator>Lim, B.</dc:creator>
<dc:creator>Lee, E. B.</dc:creator>
<dc:creator>Cicchetti, F.</dc:creator>
<dc:creator>Awatramani, R.</dc:creator>
<dc:creator>Hollon, N. G.</dc:creator>
<dc:creator>Hnasko, T. S.</dc:creator>
<dc:date>2025-06-07</dc:date>
<dc:identifier>doi:10.1101/2025.06.03.657460</dc:identifier>
<dc:title><![CDATA[Parkinson's disease-vulnerable and -resilient dopamine neurons display opposite responses to excitatory input]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.06.06.657909v1?rss=1">
<title>
<![CDATA[
JIP4 deficiency causes a novel lysosome storage disease arising from impaired cystine efflux 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.06.06.657909v1?rss=1"
</link>
<description><![CDATA[
Lysosomes break down macromolecules, clear cellular waste and recycle nutrients such as cystine. We describe a novel mechanism whereby JIP4 regulates lysosomal cystine storage by controlling the abundance of cystinosin (CTNS), the transporter responsible for lysosomal cystine efflux. To this end, JIP4, previously characterized as a motor adaptor and kinase signaling scaffold, suppresses TMEM55B-dependent ubiquitylation of CTNS. Loss of JIP4 reduces CTNS protein levels, leading to lysosomal cystine accumulation and lysosomal storage defects that phenocopy loss of CTNS in both human cells and the renal proximal tubules of JIP4 knockout mice. These phenotypes mirror cystinosis, the lysosomal storage disease caused by CTNS loss-of-function. Our findings thus reveal a fundamental process that controls the efflux of lysosomal cystine and has relevance to understanding human disease arising from JIP4 mutations.
]]></description>
<dc:creator>Nassar, L. M.</dc:creator>
<dc:creator>Shi, x.</dc:creator>
<dc:creator>Roczniak-Ferguson, A.</dc:creator>
<dc:creator>Shen, H.</dc:creator>
<dc:creator>Ferguson, S. M.</dc:creator>
<dc:date>2025-06-09</dc:date>
<dc:identifier>doi:10.1101/2025.06.06.657909</dc:identifier>
<dc:title><![CDATA[JIP4 deficiency causes a novel lysosome storage disease arising from impaired cystine efflux]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.06.15.659794v1?rss=1">
<title>
<![CDATA[
Generation of knock-in Cre and FlpO mouse lines for precise targeting of striatal projection neurons and dopaminergic neurons 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.06.15.659794v1?rss=1"
</link>
<description><![CDATA[
The basal ganglia and midbrain dopaminergic systems are critical for motor control, reward processing, and reinforcement learning, with dysfunction in these systems implicated in numerous neurodegenerative and neuropsychiatric disorders. To enable precise genetic targeting of key neuronal populations, we generated and characterized five knock-in mouse lines: Drd1-Cre, Adora2a-Cre, Drd1-FlpO, Adora2a-FlpO, and DAT-FlpO. These lines allow for Cre-or FlpO-mediated recombination in dopamine D1 receptor-expressing spiny projection neurons (SPNs), adenosine A2a receptor-expressing SPNs, and dopamine transporter (DAT)-expressing neurons in the midbrain. Histological analyses confirmed recombinase activity in expected brain regions, and whole-cell electrophysiological recordings validated the intrinsic excitability profiles of each neuronal subpopulation. These tools provide high specificity and reliability for studying basal ganglia circuitry and dopaminergic neurons. By enabling targeted manipulations, these openly available knock-in lines will advance research into the neural mechanisms underlying motor control, reward, and neuropsychiatric diseases.
]]></description>
<dc:creator>Albarran, E.</dc:creator>
<dc:creator>Fushiki, A.</dc:creator>
<dc:creator>Nelson, A.</dc:creator>
<dc:creator>Ng, D.</dc:creator>
<dc:creator>Chaimowitz, C.</dc:creator>
<dc:creator>Nikoobakht, L.</dc:creator>
<dc:creator>Sippy, T.</dc:creator>
<dc:creator>Peterka, D. S.</dc:creator>
<dc:creator>Costa, R. M.</dc:creator>
<dc:date>2025-06-15</dc:date>
<dc:identifier>doi:10.1101/2025.06.15.659794</dc:identifier>
<dc:title><![CDATA[Generation of knock-in Cre and FlpO mouse lines for precise targeting of striatal projection neurons and dopaminergic neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-15</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.06.23.661126v1?rss=1">
<title>
<![CDATA[
Bidirectional regulation of glycoprotein nonmetastatic melanoma protein B by β-glucocerebrosidase deficiency in GBA1 isogenic dopaminergic neurons from a patient with Gaucher disease and parkinsonism 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.06.23.661126v1?rss=1"
</link>
<description><![CDATA[
Variants in GBA1 are common genetic risk factors for several synucleinopathies. The increased risk has been attributed to the toxic effects of misfolded glucocerebrosidase (GCase) (gain-of-function), and the accumulation of lipid substrates due to reduced enzyme activity (loss-of-function). To delineate GBA1 pathogenicity, an iPSC line was generated from a patient with both type 1 Gaucher disease (GBA1: N370S/N370S; p.N409S/p.N409S) and Parkinson disease (PD). From this line, we created a reverted wild-type (WT) line and a GBA1 knockout (KO) line to eliminate misfolded GCase and intensify lipid accumulation. N370S/N370S and KO dopaminergic neurons (DANs) exhibited decreasing GCase levels and progressive accumulation of lipid substrates compared to WT DANs. Notably, the expression of GPNMB, whose levels correlate with PD risk, was upregulated by the mild lipid accumulation in N370S/N370S DANs but disrupted in KO DANs. These findings refine the loss-of-function mechanism by associating PD risk levels of GPNMB with lipid levels specific to GBA1 risk variants.
]]></description>
<dc:creator>Chen, C.</dc:creator>
<dc:creator>Ma, C.</dc:creator>
<dc:creator>Sam, R.</dc:creator>
<dc:creator>Lichtenberg, J.</dc:creator>
<dc:creator>Chen, T.</dc:creator>
<dc:creator>Hao, Y.</dc:creator>
<dc:creator>Li, Z.</dc:creator>
<dc:creator>Kowal, I.</dc:creator>
<dc:creator>Andersh, K.</dc:creator>
<dc:creator>Qi, Y. A.</dc:creator>
<dc:creator>Perez, G.</dc:creator>
<dc:creator>Hertz, E.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Williams, D.</dc:creator>
<dc:creator>Henderson, M. J.</dc:creator>
<dc:creator>Park, M.</dc:creator>
<dc:creator>Jiang, X.</dc:creator>
<dc:creator>Jerez, P. A.</dc:creator>
<dc:creator>Blauwendraat, C.</dc:creator>
<dc:creator>Sidransky, E.</dc:creator>
<dc:creator>Chen, Y.</dc:creator>
<dc:date>2025-06-25</dc:date>
<dc:identifier>doi:10.1101/2025.06.23.661126</dc:identifier>
<dc:title><![CDATA[Bidirectional regulation of glycoprotein nonmetastatic melanoma protein B by β-glucocerebrosidase deficiency in GBA1 isogenic dopaminergic neurons from a patient with Gaucher disease and parkinsonism]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.06.24.660984v1?rss=1">
<title>
<![CDATA[
EVscope: A Comprehensive Bioinformatics Pipeline for Accurate and Robust Analysis of Total RNA Sequencing from Extracellular Vesicles 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.06.24.660984v1?rss=1"
</link>
<description><![CDATA[
MotivationExtracellular vesicle (EV) RNA sequencing has emerged as a powerful approach for studying RNA biomarkers and intercellular communication. Nevertheless, the extremely low abundance, fragmented nature and ubiquitous tissue origin of EV RNAs, alongside potential contamination from co-isolated materials, such as free DNA and bacterial RNA, pose substantial analytical challenges. These complexities highlight a pressing need for a standardized, computational workflow that ensures robust quality control and EV RNA characterization.

ResultsHere, we present EVscope, an open-source bioinformatics pipeline designed specifically for processing EV RNA-seq datasets. EVscope employs an optimized genome-wide expectation-maximization (EM) algorithm that significantly improves multi-mapping read assignment at single-base resolution by effectively leveraging alignment scores (AS) and local read coverage, specifically tailored for fragmented and low-abundance EV RNAs. Notably, EVscope uniquely generates EM-based BigWig files for downstream analysis, a capability currently unavailable in existing EM-based BigWig quantification tools. The pipeline systematically integrates 27 major steps, including quality control, analysis of library structure, contamination assessment, read alignment, read strandedness detection, UMI-based deduplication, RNA quantification, genomic DNA (gDNA) contamination correction, cellular and tissue source inference and visualization with a comprehensive HTML report. EVscope incorporates a comprehensive, updated annotation covering 19 distinct RNA biotypes, encompassing protein-coding genes, lncRNAs, miRNAs, piRNAs, retrotransposons (LINEs, SINEs, ERVs), and additional non-coding RNAs (tRNAs, rRNAs, snoRNAs). Furthermore, it leverages two highly balanced circRNA detection algorithms for robust circular RNA identification. Notably, a downstream module enables the inference of the tissue/cellular origins of EV RNAs using bulk and single-cell RNA-seq reference datasets. EVscope is implemented as a convenient, single-command Bash pipeline leveraging Conda-managed standard software packages and custom scripts, ensuring reproducibility and straightforward deployment.

Availability and implementationCode, documentation, and tutorials are available at GitHub (https://github.com/TheDongLab/EVscope) and archived on Zenodo (https://zenodo.org/records/15577789).
]]></description>
<dc:creator>Zhao, Y.</dc:creator>
<dc:creator>Chintalapudi, H.</dc:creator>
<dc:creator>Xu, Z.</dc:creator>
<dc:creator>Liu, W.</dc:creator>
<dc:creator>Hu, Y.</dc:creator>
<dc:creator>Grassin, E.</dc:creator>
<dc:creator>Song, M.</dc:creator>
<dc:creator>Hong, S.</dc:creator>
<dc:creator>Lee, L. P.</dc:creator>
<dc:creator>Dong, X.</dc:creator>
<dc:date>2025-06-27</dc:date>
<dc:identifier>doi:10.1101/2025.06.24.660984</dc:identifier>
<dc:title><![CDATA[EVscope: A Comprehensive Bioinformatics Pipeline for Accurate and Robust Analysis of Total RNA Sequencing from Extracellular Vesicles]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.06.24.661260v1?rss=1">
<title>
<![CDATA[
TX2P: A Proteogenomic Tool for Comprehensive Transcript Analysis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.06.24.661260v1?rss=1"
</link>
<description><![CDATA[
Long-read RNA sequencing has expanded our understanding of the transcriptome, revealing unannotated transcripts. However, interpreting their functional relevance remains challenging. To address this, we developed TX2P, a user-friendly tool that integrates transcriptomic and proteomic data to link RNA discoveries with protein function. Applied to epilepsy-associated genes, TX2P identified novel protein-coding transcripts, with peptide evidence supporting 17.2% transcripts and 18.0% of unique open reading frames. By seamlessly integrating transcriptomic and proteomic insights, TX2P facilitates transcriptome interpretation.
]]></description>
<dc:creator>Murphy, D.</dc:creator>
<dc:creator>Wood, N. W.</dc:creator>
<dc:creator>Ryten, M.</dc:creator>
<dc:creator>Gustavsson, E. K.</dc:creator>
<dc:date>2025-06-27</dc:date>
<dc:identifier>doi:10.1101/2025.06.24.661260</dc:identifier>
<dc:title><![CDATA[TX2P: A Proteogenomic Tool for Comprehensive Transcript Analysis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-27</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.07.19.665650v1?rss=1">
<title>
<![CDATA[
AutoMorphoTrack: An Automated Python Package for Organelle Morphology, Motility, and Colocalization Analysis in Live-Cell Imaging 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.07.19.665650v1?rss=1"
</link>
<description><![CDATA[
Quantitative imaging of organelle dynamics provides crucial insights into cellular function, state, and organization; however, existing analysis workflows often require advanced coding expertise and multiple software tools. AutoMorphoTrack is an open-source Python toolkit that automates organelle detection, morphology classification, motility tracking, and colocalization from multichannel fluorescence microscopy image stacks. The platform includes adaptive segmentation, organelle trajectory reconstruction, and pixel-level overlap quantification within a unified, reproducible framework that can be executed as an interactive Jupyter notebook, a modular Python package, or through AI-assisted natural-language commands. Each analysis step outputs publication-ready images, time-lapse videos, and standardized quantitative data tables. To complement the main pipeline, an accompanying script--AMTComparison.py--is provided to demonstrate how AutoMorphoTracks outputs can be extended for comparative analysis across individual neurons or experimental conditions. Together, these tools provide an accessible and framework for high-content, reproducible quantification of subcellular morphology, motility, and interactions at single-cell resolution.

Graphical Abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=98 SRC="FIGDIR/small/665650v5_ufig1.gif" ALT="Figure 1">
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org.highwire.dtl.DTLVardef@103ecb0org.highwire.dtl.DTLVardef@a81b6forg.highwire.dtl.DTLVardef@a145fcorg.highwire.dtl.DTLVardef@1f9c27c_HPS_FORMAT_FIGEXP  M_FIG C_FIG
]]></description>
<dc:creator>Bayati, A.</dc:creator>
<dc:creator>Schumacher, J. G.</dc:creator>
<dc:creator>Chen, X.</dc:creator>
<dc:date>2025-07-23</dc:date>
<dc:identifier>doi:10.1101/2025.07.19.665650</dc:identifier>
<dc:title><![CDATA[AutoMorphoTrack: An Automated Python Package for Organelle Morphology, Motility, and Colocalization Analysis in Live-Cell Imaging]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-07-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.07.31.667659v1?rss=1">
<title>
<![CDATA[
Elevated brain α-synuclein, phosphorylated-tau, and oxidative stress in mice that survived influenza A pneumonitis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.07.31.667659v1?rss=1"
</link>
<description><![CDATA[
Influenza virus infection increases the incidence of parkinsonism in humans. We have previously shown that allelic variants at the Parkinsons disease (PD)-linked Lrrk2 locus modulate host responses in vivo. Here, we asked whether Lrrk2 kinase activity alters disease outcomes in adult mice that survived a nasally acquired infection. We inoculated mice with the mouse adapted A/Fort Monmouth/1/1947 influenza A virus, serotype H1N1 (LD50, 2x103 plaque forming units), leading to pneumonitis. We found that neither the hyperkinase-active Lrrk2 p.G2019S knock-in mutant nor the kinase-dead Lrrk2 p.D1994S mutant altered the course of an acute H1N1 lung infection. We then probed for long-term effects of H1N1 pneumonitis on brain health by monitoring surviving mice for six weeks post-inoculation. Intriguingly, at this time point, when mice had recovered and showed no detectable viral proteins in the brain, levels of H2O2 and protein nitrotyrosination were significantly elevated in H1N1 survivors vs. mock-treated littermates. In addition, total -synuclein concentrations were increased in an infection-dependent manner but independent of the Lrrk2 genotype. Intriguingly, at the same timepoint, the ratio of phosphorylated-to-total tau (but not total tau itself) was significantly increased in the brains of H1N1-virus exposed Lrrk2 p.G2019S mice compared to wild-type animals. Our collective results demonstrate that a preceding pneumotropic influenza A virus infection can lead to a rise in several neurodegeneration-linked markers in the brains of surviving mice. The increased ratio of phosphorylated-to-total tau in Lrrk2 p.G2019S animals adds to the growing evidence of interactions between specific microbial pathogens and allelic variants at the Lrrk2 locus. The described outcomes in animals that survived an influenza A virus infection may be of relevance to the incidence of neurodegenerative diseases in ageing humans.
]]></description>
<dc:creator>Lunn, M. O.</dc:creator>
<dc:creator>Lengacher, N. A.</dc:creator>
<dc:creator>Hake-Volling, Q.</dc:creator>
<dc:creator>Tomlinson, J. J.</dc:creator>
<dc:creator>aSCENT-PD Investigators,</dc:creator>
<dc:creator>Brown, E. G.</dc:creator>
<dc:creator>Schlossmacher, M. G.</dc:creator>
<dc:date>2025-08-01</dc:date>
<dc:identifier>doi:10.1101/2025.07.31.667659</dc:identifier>
<dc:title><![CDATA[Elevated brain α-synuclein, phosphorylated-tau, and oxidative stress in mice that survived influenza A pneumonitis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-01</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/641233v1?rss=1">
<title>
<![CDATA[
Neuron subtype-specific effector gene expression in the Motor Ganglion of Ciona 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/641233v1?rss=1"
</link>
<description><![CDATA[
The central nervous system of the Ciona larva contains only 177 neurons. The precise regulation of neuron subtype-specific morphogenesis and differentiation observed in during the formation of this minimal connectome offers a unique opportunity to dissect gene regulatory networks underlying chordate neurodevelopment. Here we compare the transcriptomes of two very distinct neuron types in the hindbrain/spinal cord homolog of Ciona, the Motor Ganglion (MG): the Descending decussating neuron (ddN, proposed homolog of Mauthner Cells in vertebrates) and the MG Interneuron 2 (MGIN2). Both types are invariantly represented by a single bilaterally symmetric left/right pair of cells in every larva. Supernumerary ddNs and MGIN2s were generated in synchronized embryos and isolated by fluorescence-activated cell sorting for transcriptome profiling. Differential gene expression analysis revealed ddN- and MGIN2-specific enrichment of a wide range of genes, including many encoding potential "effectors" of subtype-specific morphological and functional traits. More specifically, we identified the upregulation of centrosome-associated, microtubule-stabilizing/bundling proteins and extracellular matrix proteins and axon guidance cues as part of a single intrinsic regulatory program that might underlie the unique polarization of the ddNs, the only descending MG neurons that cross the midline.
]]></description>
<dc:creator>Gibboney, S.</dc:creator>
<dc:creator>Kim, K.</dc:creator>
<dc:creator>Johnson, C. J.</dc:creator>
<dc:creator>Orvis, J.</dc:creator>
<dc:creator>Martinez-Feduchi, P.</dc:creator>
<dc:creator>Lowe, E. K.</dc:creator>
<dc:creator>Sharma, S.</dc:creator>
<dc:creator>Stolfi, A.</dc:creator>
<dc:date>2019-05-17</dc:date>
<dc:identifier>doi:10.1101/641233</dc:identifier>
<dc:title><![CDATA[Neuron subtype-specific effector gene expression in the Motor Ganglion of Ciona]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2019-05-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.06.04.657838v1?rss=1">
<title>
<![CDATA[
Genetic modulation of immune gene co-expression in the aged mouse hippocampus by the Apbb1ip locus 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.06.04.657838v1?rss=1"
</link>
<description><![CDATA[
Ageing is a major risk factor for many neurodegenerative diseases, and the hippocampus is particularly vulnerable to the effects of ageing. To define the transcriptomic changes related to ageing and the impact of genetic variation, we analysed hippocampal gene expression data generated from a genetically diverse panel of inbred mouse strains belonging to the BXD Family. We applied a combination of differential expression, differential correlation, and weighted gene co-expression network analyses, followed by genetic mapping to delineate age-associated transcriptomic patterns and genetic modulators. This study revealed an upregulation in immune response and microglial genes. We identified a key age-associated co-expression module enriched in immune related genes, and quantitative trait locus mapping of this module uncovered a genetic regulatory locus in which Apbb1ip was the primary candidate gene. Additionally, gene level differential correlation analysis identified a substantial restructuring of the hippocampal transcriptome during ageing. Notably, Ywhab, an important signalling chaperone displayed altered gene expression correlations with >70 genes, between young and aged mice. These results provide novel insights into transcriptional dynamics of hippocampal ageing and identified the immune gene Apbb1ip as a potential modulator of immune response and microglial gene upregulation with implications for neurodegenerative disease pathogenesis.

HighlightsO_LIComplementary gene expression analyses indicated an upregulation of immune responses in the ageing hippocampus of a genetically diverse mouse cohort, likely driven by microglia subtypes.
C_LIO_LIA systems genetics approach revealed the Apbb1ip locus as a modulator of immune gene co-expression, highlighting a novel candidate regulator of age-associated neuroimmune dynamics.
C_LIO_LISignalling chaperone Ywhab gene expression was highly differentially correlated with other transcripts during ageing which may underlie widespread age-associated molecular perturbations in the hippocampus.
C_LI
]]></description>
<dc:creator>Tomkins, J. E.</dc:creator>
<dc:creator>Methi, A.</dc:creator>
<dc:creator>Bouland, G.</dc:creator>
<dc:creator>Doludda, B.</dc:creator>
<dc:creator>Murphy, K.</dc:creator>
<dc:creator>Miller, J. A.</dc:creator>
<dc:creator>Overall, R. W.</dc:creator>
<dc:creator>Mozhui, K.</dc:creator>
<dc:date>2025-06-08</dc:date>
<dc:identifier>doi:10.1101/2025.06.04.657838</dc:identifier>
<dc:title><![CDATA[Genetic modulation of immune gene co-expression in the aged mouse hippocampus by the Apbb1ip locus]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.06.13.659385v1?rss=1">
<title>
<![CDATA[
Patch deconvolution for Fourier light-field microscopy 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.06.13.659385v1?rss=1"
</link>
<description><![CDATA[
Imaging flow cytometry using Fourier light-field microscopy enables high-throughput three-dimensional cellular imaging, capable of capturing thousands of events per second. However, volumetric reconstruction speed remains orders of magnitude slower than the acquisition speed. The current state of art uses Richardson-Lucy algorithm, restricted to just 5-10 reconstructed per second with GPU acceleration. This limitation hinders real-time applications such as cell sorting and thus has bottlenecked the widespread adoption of 3D imaging flow cytometry. We introduce patch deconvolution, an optimisation compatible with the Richardson-Lucy framework that significantly accelerates convergence, achieving over 100-200 reconstructions per second on standard GPUs, a 20-40 fold improvement over Richardson-Lucy. Validated on both simulated and experimental datasets, patch deconvolution achieves reconstruction quality comparable to Richardson-Lucy in both static and flow data. This supports rapid cell sorting based on spatial features and enables advanced applications, such as detecting rare spatial events in large cell populations, which would otherwise be indistinguishable in traditional flow cytometry.
]]></description>
<dc:creator>Fu, B.</dc:creator>
<dc:creator>Jones, C. L.</dc:creator>
<dc:creator>Heraghty, D.</dc:creator>
<dc:creator>Yang, S.</dc:creator>
<dc:creator>O'Brien-Ball, C.</dc:creator>
<dc:creator>Junghans, V.</dc:creator>
<dc:creator>Yang, H.</dc:creator>
<dc:creator>Knowles, T.</dc:creator>
<dc:creator>Weiss, L. E.</dc:creator>
<dc:creator>Fernandes, R. A.</dc:creator>
<dc:creator>Lee, S.</dc:creator>
<dc:date>2025-06-17</dc:date>
<dc:identifier>doi:10.1101/2025.06.13.659385</dc:identifier>
<dc:title><![CDATA[Patch deconvolution for Fourier light-field microscopy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.06.18.660098v1?rss=1">
<title>
<![CDATA[
Development of a protocol utilizing single-cell analysis for the differentiation of human iPSCs into SOX6+ midbrain dopaminergic neurons. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.06.18.660098v1?rss=1"
</link>
<description><![CDATA[
Parkinsons disease (PD) is characterized by the progressive loss of midbrain dopaminergic (mDA) neurons, leading to severe debilitating motor impairment. Recent single-cell analyses have revealed that specific mDA subpopulations, such as SOX6 AGTR1 neurons, are most vulnerable to degeneration. Current human pluripotent stem cell differentiation protocols fail to selectively generate these subtypes. Here, we describe a multiomic-guided strategy that enriches SOX6+ mDA neurons by combining enhancing Sonic Hedgehog agonism with prolonged Wnt activation. This approach accelerates floor plate specification, increases expression of mDA developmental markers, and substantially increases the yield of SOX6 mature mDA neurons compared to prior attempts. Following intrastriatal transplantation into hemiparkinsonian mice, these cells restored motor function in approximately 4 months and generated grafts containing SOX6 A9-like neurons. Our work thus establishes a reproducible differentiation platform for generating the PD-susceptible mDA subtype, providing a foundation for precision disease modelling and subtype-targeted cell replacement therapies.
]]></description>
<dc:creator>Lyu, G.</dc:creator>
<dc:creator>Garcia Swinburn, R.</dc:creator>
<dc:creator>Kreutzmann, J. C.</dc:creator>
<dc:creator>Xiong, A.</dc:creator>
<dc:creator>Kojima, R.</dc:creator>
<dc:creator>Abaurre, C.</dc:creator>
<dc:creator>Tremolanti, C.</dc:creator>
<dc:creator>Gellhaar, S.</dc:creator>
<dc:creator>Uhlen, P.</dc:creator>
<dc:creator>Svenningsson, P.</dc:creator>
<dc:creator>Castelo Branco, G.</dc:creator>
<dc:creator>Dagliyan, O.</dc:creator>
<dc:creator>Salto, C.</dc:creator>
<dc:creator>Arenas, E.</dc:creator>
<dc:date>2025-06-22</dc:date>
<dc:identifier>doi:10.1101/2025.06.18.660098</dc:identifier>
<dc:title><![CDATA[Development of a protocol utilizing single-cell analysis for the differentiation of human iPSCs into SOX6+ midbrain dopaminergic neurons.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-06-22</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.08.16.670665v1?rss=1">
<title>
<![CDATA[
A lipid transfer-dependent feedback loop activates ATG9A compartments in autophagy initiation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.08.16.670665v1?rss=1"
</link>
<description><![CDATA[
Autophagy degrades cellular material by sequestering it in autophagosomes, which form de novo from precursors called phagophores. Phagophore assembly and expansion require ATG9A-positive seed compartments, the lipid transfer protein ATG2A, and the class III phosphatidylinositol 3-phosphate kinase complex I (PI3KC3-C1). PI3KC3-C1 synthesizes phosphatidylinositol 3-phosphate (PI3P), a key lipid that drives downstream processes for phagophore expansion, including ATG8 lipidation. We find that ATG9A compartments contain only traces of phosphatidylinositol (PI), likely insufficient for efficient PI3P production or recruitment of PI3P-binding effectors. Nevertheless, ATG2A is recruited to these compartments and mediates lipid transfer, including PI, into them. Remarkably, even without detectable PI3P, ATG9A compartments can support ATG8 lipidation, and ATG8 proteins themselves enhance ATG2A-mediated lipid transfer. In cells, ATG2A is essential for the appearance of PI3P on ATG9A compartments. Together, these findings support a model in which a lipid transfer-driven feedback loop, rather than pre-existing PI content, is required to activate ATG9A compartments for phagophore expansion.

TeaserA feedback loop driven by lipid transfer activates ATG9A compartments for autophagosome biogenesis.
]]></description>
<dc:creator>Holzer, E.</dc:creator>
<dc:creator>Sawa-Makarska, J.</dc:creator>
<dc:creator>Bernklau, D.</dc:creator>
<dc:creator>Romanov, J.</dc:creator>
<dc:creator>Schuschnig, M.</dc:creator>
<dc:creator>Martens, S.</dc:creator>
<dc:date>2025-08-18</dc:date>
<dc:identifier>doi:10.1101/2025.08.16.670665</dc:identifier>
<dc:title><![CDATA[A lipid transfer-dependent feedback loop activates ATG9A compartments in autophagy initiation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-18</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.08.19.671144v1?rss=1">
<title>
<![CDATA[
Neuromodulator control of energy reserves in dopaminergic neurons 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.08.19.671144v1?rss=1"
</link>
<description><![CDATA[
The brain is a metabolically vulnerable organ as neurons have both high resting metabolic rates and the need for local rapid conversion of carbon sources to ATP during activity. Midbrain dopamine neurons are thought to be particularly vulnerable to metabolic perturbations, as a subset of these are the first to undergo degeneration in Parkinsons disease (PD), a neurodegenerative disorder long suspected to be in part driven by deficits in mid-brain bioenergetics (1). In skeletal muscle, energy homeostasis under varying demands is achieved in part by its ability to rely on glycogen as a fuel store, whose conversion to ATP is under hormonal regulatory control. In neurons however the absence of easily observable glycogen granules has cast doubt on whether this fuel store is operational, even though brain neurons express the key regulatory enzymes associated with building or burning glycogen (2). We show here that that in primary mid brain dopaminergic neurons, glycogen availability is under the control of dopamine auto receptors (D2R), such that dopamine itself provides a signal to store glycogen. We find that when glycogen stores are present, they provide remarkable resilience to dopamine nerve terminal function under extreme hypometabolic conditions, but loss of this dopamine derived signal, or impairment of access to glycogen, makes them hypersensitive to fuel deprivation. These data show that neurons can use an extracellular cue to regulate local metabolism and suggest that loss of dopamine secretion might make dopamine neurons particularly subject to neurodegeneration driven by metabolic stress.

Significance StatementThis work demonstrates that a neurons metabolic resilience is actively shaped by local neuromodulation, providing a potential explanation for why different neuronal subtypes exhibit unequal vulnerability to metabolic stressors and why dopamine neurons become more vulnerable when they lose the capacity to release dopamine.
]]></description>
<dc:creator>Pulido, C.</dc:creator>
<dc:creator>Gentry, M. S.</dc:creator>
<dc:creator>Ryan, T. A.</dc:creator>
<dc:date>2025-08-19</dc:date>
<dc:identifier>doi:10.1101/2025.08.19.671144</dc:identifier>
<dc:title><![CDATA[Neuromodulator control of energy reserves in dopaminergic neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.08.19.670819v1?rss=1">
<title>
<![CDATA[
α-Synuclein driven cell susceptibility in Parkinson's disease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.08.19.670819v1?rss=1"
</link>
<description><![CDATA[
Early cellular events in Parkinsons disease (PD) remain elusive. While aggregation of -synuclein (Syn) into Lewy bodies marks advanced pathology, smaller Syn oligomers have been implicated in prodromal stages. Here we map Syn oligomers at single-particle resolution in post-mortem brain tissue from Braak stage 3/4 PD cases and matched controls. Quantitative imaging of 9,882 neurons across four regions captured over 112 million Syn oligomers. Mean intracellular -Syn burden was unchanged between groups, but PD samples contained a higher fraction of neurons whose oligomer load exceeded a specific aggregation threshold. We term these aggregation-susceptible cells (ASCs). ASC enrichment in vulnerable regions supports a population-level model in which early pathology arises from a stochastic shift in cellular composition rather than altered Syn aggregation kinetics. This human-tissue, large-scale dataset provides a quantitative framework for detecting ASCs and for testing population-level interventions in PD and related proteinopathies.
]]></description>
<dc:creator>Breiter, J. C.</dc:creator>
<dc:creator>Beckwith, J. S.</dc:creator>
<dc:creator>Brock, E. E.</dc:creator>
<dc:creator>Lachica, J.</dc:creator>
<dc:creator>Toomey, C. E.</dc:creator>
<dc:creator>Fu, B.</dc:creator>
<dc:creator>Ryten, M.</dc:creator>
<dc:creator>Weiss, L. E.</dc:creator>
<dc:creator>Wood, N. W.</dc:creator>
<dc:creator>Gandhi, S.</dc:creator>
<dc:creator>Vendruscolo, M.</dc:creator>
<dc:creator>Lee, S. F.</dc:creator>
<dc:date>2025-08-23</dc:date>
<dc:identifier>doi:10.1101/2025.08.19.670819</dc:identifier>
<dc:title><![CDATA[α-Synuclein driven cell susceptibility in Parkinson's disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.08.24.671754v1?rss=1">
<title>
<![CDATA[
Integrated Representations of Threat and Controllability in the Lateral Frontal Pole 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.08.24.671754v1?rss=1"
</link>
<description><![CDATA[
Emotional processing is ubiquitous in everyday life, informing goal pursuit not only in response to current demands, but also in anticipation of future outcomes. Lateral prefrontal (LPFC) function supports cognitive control, and emerging evidence suggests a unique role for its anterior-most region--the lateral frontal pole (FPl)--in integrating putatively amygdala-originated emotion signals with goal information. However, whether these organizational properties of LPFC are expressed during the anticipation of future threat remains unknown. Here, we used FIR modeling and pattern similarity analysis to examine dynamic engagement and representational properties of distinct LPFC regions during threat anticipation requiring goal-directed action. Healthy participants (n=67) were scanned during a threat-of-shock paradigm consisting of a prolonged (18s) countdown to possible shock administration. Threat unpleasantness and controllability were manipulated orthogonally: in controllable trials, participants could avoid an unpleasant or mild shock by making a successful time-sensitive response; in uncontrollable trials, shocks were administered regardless of performance. LPFC robustly coded for anticipated threat unpleasantness, with FPl showing the strongest modulation by threat unpleasantness and controllability relative to caudal and mid-LPFC regions. Caudal and mid-LPFC maintained independent representations of threat unpleasantness and controllability. In contrast, FPl held conjunctive threat-and-controllability representations, which were associated with successful motor performance following anticipation of unpleasant shocks. Stronger conjunctive FPl representations were also associated with greater inverse amygdala-FPl coupling. Together, these findings provide insight into LPFC organization under naturalistic emotional challenges and highlight a key role for FPl in integrating affective and control-related information during threat anticipation to support goal-directed action.

Significance StatementAnticipating emotionally-charged events--such as a painful outcome we may or may not be able to avoid--requires integrating emotion and control to guide behavior. However, the neural mechanisms through which emotional states influence goal-directed behavior in naturalistic, anticipatory emotional contexts remain unclear. Using a threat-of-shock paradigm and multivariate analyses we show that the anterior-most region of the lateral prefrontal cortex (LPFC)--the lateral frontal pole (FPl)--uniquely integrates information about the emotional unpleasantness and controllability of a future event, and that the strength of this integrated signal predicts better behavioral performance. These findings extend models of LPFC function to naturalistic, emotional contexts, and highlight the FPl as a key node for translating emotional and behavioral-control information into adaptive action.
]]></description>
<dc:creator>Stasiak, J. E.</dc:creator>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Dundon, N. M.</dc:creator>
<dc:creator>Rizor, E. J.</dc:creator>
<dc:creator>Villanueva, C. M.</dc:creator>
<dc:creator>Barandon, P. L.</dc:creator>
<dc:creator>Grafton, S. T.</dc:creator>
<dc:creator>Lapate, R. C.</dc:creator>
<dc:date>2025-08-24</dc:date>
<dc:identifier>doi:10.1101/2025.08.24.671754</dc:identifier>
<dc:title><![CDATA[Integrated Representations of Threat and Controllability in the Lateral Frontal Pole]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.08.25.671921v1?rss=1">
<title>
<![CDATA[
Microglial activation and alpha-synuclein oligomers drive the early inflammatory phase of Parkinson's disease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.08.25.671921v1?rss=1"
</link>
<description><![CDATA[
Parkinsons disease (PD) is characterised by insoluble -synuclein (Syn) aggregates in Lewy bodies (LBs) within the substantia nigra, with cortical pathology appearing as the disease progresses. Late-stage LB deposition, cellular stress, and neuronal loss obscure disease-driving events, we therefore performed multi-regional transcriptomic and aggregate profiling in early-midstage PD brains (Braak 3-4), where cortical regions are pathologically unaffected. We report neuroimmune activation as an early PD feature, characterised by the expansion of a high-SNCA-expressing microglial state. This robust immune signature occurs prior to LB formation, but is associated with oligomeric Syn within cortical microglia. In hiPSC-derived microglia, both endogenous Syn oligomerisation, and exogenous oligomer uptake, trigger transcriptional reprogramming, characterised by interferon-driven inflammation, antigen presentation, and mitochondrial suppression, closely mirroring the early PD brain. These findings describe mechanisms by which Syn oligomerisation potently initiates early neuroinflammation, highlighting a critical interplay between proteinopathy and immune activation at the earliest stages of disease.
]]></description>
<dc:creator>Evans, J. R.</dc:creator>
<dc:creator>Grant-Peters, M.</dc:creator>
<dc:creator>Beckwith, J. S.</dc:creator>
<dc:creator>Toomey, C. E.</dc:creator>
<dc:creator>Popovic, R.</dc:creator>
<dc:creator>Breiter, J. C.</dc:creator>
<dc:creator>Brenton, J. W.</dc:creator>
<dc:creator>Fairbrother-Browne, A.</dc:creator>
<dc:creator>Strohbuecker, S.</dc:creator>
<dc:creator>Lachica, J.</dc:creator>
<dc:creator>Rodriguez-Lopez, M.</dc:creator>
<dc:creator>Brock, E. E.</dc:creator>
<dc:creator>Fu, B.</dc:creator>
<dc:creator>Nahidiazar, L.</dc:creator>
<dc:creator>Lopez-Garcia, P.</dc:creator>
<dc:creator>Ferguson, R.</dc:creator>
<dc:creator>Saleeb, R. S.</dc:creator>
<dc:creator>Lucas-Clarke, H.</dc:creator>
<dc:creator>Penverne, A.</dc:creator>
<dc:creator>D'Sa, K.</dc:creator>
<dc:creator>Pang, C. W.</dc:creator>
<dc:creator>Horrocks, M. H.</dc:creator>
<dc:creator>Vendruscolo, M.</dc:creator>
<dc:creator>Wood, N.</dc:creator>
<dc:creator>Lee, S. F.</dc:creator>
<dc:creator>Ryten, M.</dc:creator>
<dc:creator>Gandhi, S.</dc:creator>
<dc:date>2025-08-28</dc:date>
<dc:identifier>doi:10.1101/2025.08.25.671921</dc:identifier>
<dc:title><![CDATA[Microglial activation and alpha-synuclein oligomers drive the early inflammatory phase of Parkinson's disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.08.28.672006v1?rss=1">
<title>
<![CDATA[
Leucine-rich repeat kinase 2 impairs the release sites of Parkinson's disease vulnerable dopamine axons 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.08.28.672006v1?rss=1"
</link>
<description><![CDATA[
The end-stage pathology of Parkinsons disease (PD) involves the loss of dopamine-producing neurons in the substantia nigra pars compacta (SNc). However, synaptic deregulation of these neurons begins much earlier. Understanding the mechanisms behind synaptic deficits is crucial for early therapeutic intervention, yet these remain largely unknown. In the SNc, different dopamine neuron subtypes show varying susceptibility patterns to PD, complicating our understanding. This study uses intersectional genetic mouse models to uncover synaptic perturbations in vulnerable dopamine neurons, focusing on the LRRK2 kinase, a protein closely linked to PD. Through a combination of immunofluorescence and advanced proximity labeling methods, we found higher LRRK2 expression in the most vulnerable dopamine neuron subclusters. High-resolution imaging revealed that pathogenic LRRK2 disrupts release sites in vulnerable dopamine axons, leading to decreased in vivo evoked striatal dopamine release in mice with LRRK2 mutations. Proteomic and biochemical analyses indicate that mutant LRRK2 increases the phosphorylation of RAB3 proteins, reducing their interactions with RIM1/2 effector proteins and impacting their synaptic functions. Overall, this research highlights the cell-autonomous dysfunctions caused by mutant LRRK2 in the neurons that are primarily affected by the disease. It also provides a framework for therapeutic strategies for early nigrostriatal synaptic deficits in PD.
]]></description>
<dc:creator>Chen, C.</dc:creator>
<dc:creator>He, Q.</dc:creator>
<dc:creator>Tombesi, G.</dc:creator>
<dc:creator>Napier, E.</dc:creator>
<dc:creator>Jaconelli, M.</dc:creator>
<dc:creator>Moreno-Ramos, O.</dc:creator>
<dc:creator>Serio, H.</dc:creator>
<dc:creator>Naaldijk, Y.</dc:creator>
<dc:creator>Promes, V.</dc:creator>
<dc:creator>Schneeweis, A.</dc:creator>
<dc:creator>Quinn, K.</dc:creator>
<dc:creator>Nasios, C.</dc:creator>
<dc:creator>Greggio, E.</dc:creator>
<dc:creator>Kozorovitskiy, Y.</dc:creator>
<dc:creator>Arango, D.</dc:creator>
<dc:creator>Khan, A. R.</dc:creator>
<dc:creator>Alessi, D. R.</dc:creator>
<dc:creator>Dombeck, D. A.</dc:creator>
<dc:creator>Hilfiker, S.</dc:creator>
<dc:creator>Awatramani, R.</dc:creator>
<dc:creator>Parisiadou, L.</dc:creator>
<dc:date>2025-08-28</dc:date>
<dc:identifier>doi:10.1101/2025.08.28.672006</dc:identifier>
<dc:title><![CDATA[Leucine-rich repeat kinase 2 impairs the release sites of Parkinson's disease vulnerable dopamine axons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.09.04.673943v1?rss=1">
<title>
<![CDATA[
TDP-43 loss of function drives aberrant splicing in Parkinson's disease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.09.04.673943v1?rss=1"
</link>
<description><![CDATA[
Introductory paragraphWhile mRNA splicing dysregulation is a well-established contributor to neurodegeneration in disorders such as amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD), its role in Parkinsons disease (PD) remains underexplored. Here, we analyse transcriptomic data from >500 post-mortem human brain samples from individuals with and without PD to show that splicing alterations are frequently detected. Differentially spliced genes were significantly more enriched for those causally-implicated in both PD and ALS than genes that were differentially expressed. Furthermore, we observed a strong association between these splicing alterations and dysfunction of the RNA-binding protein (RBP), TAR DNA-binding protein 43 (TDP-43). Strikingly, genes and exon junctions affected by TDP-43 knockdown overlapped significantly with those dysregulated across brain regions in PD. In brains from individuals with the LRRK2 c.6055G>A (p.G2019S) mutation, the most common genetic cause of PD, we also observed significant enrichment of TDP-43-dependent splicing changes. This finding was corroborated in human pluripotent stem cell-derived midbrain dopaminergic neurons and a LRRK2 p.G2019S knock-in mouse model, where reduced nuclear TDP-43 levels evidenced the well-recognised loss-of-function mechanism contributing to splicing dysregulation. By leveraging our RNA-based analyses we predicted TDP-43-dependent novel peptide sequences and validated their existence within human LRRK2 mutation mDNs, while also demonstrating an overall loss of protein and mRNA expression in mis-spliced genes. Collectively, our findings reveal that PD is marked by extensive splicing dysregulation dependent on TDP-43, making TDP-43 a promising new therapeutic target in PD.
]]></description>
<dc:creator>Brenton, J. W.</dc:creator>
<dc:creator>Follett, J.</dc:creator>
<dc:creator>Nirujogi, R.</dc:creator>
<dc:creator>Toomey, C. E.</dc:creator>
<dc:creator>Lopez-Garcia, P.</dc:creator>
<dc:creator>Evans, J. R.</dc:creator>
<dc:creator>Lee, Y. J.</dc:creator>
<dc:creator>Syed, K. M.</dc:creator>
<dc:creator>Rocamora Perez, G.</dc:creator>
<dc:creator>Fairbrother-Browne, A.</dc:creator>
<dc:creator>D'Sa, K.</dc:creator>
<dc:creator>Grant-Peters, M.</dc:creator>
<dc:creator>Lachica, J.</dc:creator>
<dc:creator>Hicks, A. R.</dc:creator>
<dc:creator>Wagen, A. Z.</dc:creator>
<dc:creator>O'Callaghan, B.</dc:creator>
<dc:creator>Macpherson, H.</dc:creator>
<dc:creator>Montgomery, K.-a.</dc:creator>
<dc:creator>Busquets, O.</dc:creator>
<dc:creator>Reynolds, R. H.</dc:creator>
<dc:creator>Garcia Ruiz, S.</dc:creator>
<dc:creator>Cao, T.</dc:creator>
<dc:creator>Chen, Z.</dc:creator>
<dc:creator>Plun-Favreau, H.</dc:creator>
<dc:creator>Wong, P. C.</dc:creator>
<dc:creator>Farrer, M.</dc:creator>
<dc:creator>Lashley, T.</dc:creator>
<dc:creator>Soldner, F.</dc:creator>
<dc:creator>Hockemeyer, D.</dc:creator>
<dc:creator>Alessi, D.</dc:creator>
<dc:creator>Wood, N. W.</dc:creator>
<dc:creator>Hardy, J.</dc:creator>
<dc:creator>Rio, D. C.</dc:creator>
<dc:creator>Jaunmuktane, Z.</dc:creator>
<dc:creator>Gustavsson, E. K.</dc:creator>
<dc:creator>Gandhi, S.</dc:creator>
<dc:creator>Ryten, M.</dc:creator>
<dc:date>2025-09-09</dc:date>
<dc:identifier>doi:10.1101/2025.09.04.673943</dc:identifier>
<dc:title><![CDATA[TDP-43 loss of function drives aberrant splicing in Parkinson's disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.09.18.677192v1?rss=1">
<title>
<![CDATA[
Faecalibacterium prausnitzii, depleted in the Parkinson's disease microbiome, improves motor deficits in α-synuclein overexpressing mice 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.09.18.677192v1?rss=1"
</link>
<description><![CDATA[
Gut microbiome composition is altered in Parkinsons disease (PD), a neurodegenerative disorder characterized by motor dysfunction and frequently accompanied by gastrointestinal (GI) symptoms. Notably, microbial taxa with anti-inflammatory properties are consistently depleted in PD patients compared to controls. To explore whether specific gut bacteria may be disease-protective, we assembled a microbial consortium of 8 human-associated taxa that are reduced in individuals with PD across multiple cohorts and geographies. Treatment of -synuclein overexpressing (Thy1-ASO) mice, an animal model of PD, with this consortium improved motor and GI deficits. A single bacterial species from this consortium, Faecalibacterium prausnitzii, was sufficient to correct gut microbiome deviations in Thy1-ASO mice, induce anti-inflammatory immune responses, and promote protective colonic gene expression profiles. Accordingly, oral treatment with F. prausnitzii robustly ameliorated motor and GI symptoms and reduced -synuclein aggregates in the brain. These findings support the emerging hypothesis of functional contributions by the microbiome to PD and embolden development of potential probiotic therapies.
]]></description>
<dc:creator>Moiseyenko, A.</dc:creator>
<dc:creator>Antonello, G.</dc:creator>
<dc:creator>Schonhoff, A. M.</dc:creator>
<dc:creator>Boktor, J. C.</dc:creator>
<dc:creator>Long, K.</dc:creator>
<dc:creator>Dirks, B.</dc:creator>
<dc:creator>Oguienko, A. D.</dc:creator>
<dc:creator>Winnett, A. V.</dc:creator>
<dc:creator>Simpson, P.</dc:creator>
<dc:creator>Daeizadeh, D.</dc:creator>
<dc:creator>Ismagilov, R. F.</dc:creator>
<dc:creator>Krajmalnik-Brown, R.</dc:creator>
<dc:creator>Segata, N.</dc:creator>
<dc:creator>Waldron, L. D.</dc:creator>
<dc:creator>Mazmanian, S. K.</dc:creator>
<dc:date>2025-09-21</dc:date>
<dc:identifier>doi:10.1101/2025.09.18.677192</dc:identifier>
<dc:title><![CDATA[Faecalibacterium prausnitzii, depleted in the Parkinson's disease microbiome, improves motor deficits in α-synuclein overexpressing mice]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.10.01.679724v1?rss=1">
<title>
<![CDATA[
Alternate Splicing Directs PMCA2 to Lysosomes and is Linked to Neurodegeneration 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.10.01.679724v1?rss=1"
</link>
<description><![CDATA[
Plasma membrane calcium ATPases (PMCAs) are believed to function exclusively at the plasma membrane where they expel calcium from the cytosol. We have unexpectedly identified a splice variant-dependent localisation of the PMCA isoform PMCA2 to the lysosome, where it forms an evolutionarily conserved complex with NPC1, the lysosomal membrane protein defective in the rare lysosomal storage disease Niemann-Pick disease type C (NPC). This interaction is required for lysosomal Ca2+ homeostasis and implicates PMCA2 as a mediator of Ca2+ uptake into lysosomes. Disruption of the NPC1-PMCA2 complex contributes to the pathophysiology of both Niemann-Pick disease type C and Parkinsons disease, revealing an unrecognised intracellular function for PMCA2 and a shared mechanism linking lysosomal Ca2+ and lipid regulation in neurodegeneration.
]]></description>
<dc:creator>Fernandez-Suarez, M. E.</dc:creator>
<dc:creator>Bush, R.</dc:creator>
<dc:creator>Brenton, J. W.</dc:creator>
<dc:creator>Pereira, G.</dc:creator>
<dc:creator>Grant-Peters, M.</dc:creator>
<dc:creator>Reynolds, R. H.</dc:creator>
<dc:creator>te Vruchte, D.</dc:creator>
<dc:creator>Shepherd, D.</dc:creator>
<dc:creator>Weng, Y.</dc:creator>
<dc:creator>Artaza-Fernandez, E.</dc:creator>
<dc:creator>Lis, P.</dc:creator>
<dc:creator>Sanchez-Pulido, L.</dc:creator>
<dc:creator>Morgan, A.</dc:creator>
<dc:creator>Davis, L.</dc:creator>
<dc:creator>Gomez-Coronado, D.</dc:creator>
<dc:creator>Eden, E. R.</dc:creator>
<dc:creator>Alessi, D. R.</dc:creator>
<dc:creator>Ponting, C.</dc:creator>
<dc:creator>Galione, A.</dc:creator>
<dc:creator>Newstead, S.</dc:creator>
<dc:creator>Hardy, J.</dc:creator>
<dc:creator>Patel, S.</dc:creator>
<dc:creator>Ryten, M.</dc:creator>
<dc:creator>Platt, F. M.</dc:creator>
<dc:date>2025-10-02</dc:date>
<dc:identifier>doi:10.1101/2025.10.01.679724</dc:identifier>
<dc:title><![CDATA[Alternate Splicing Directs PMCA2 to Lysosomes and is Linked to Neurodegeneration]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.10.06.680693v1?rss=1">
<title>
<![CDATA[
Structural remodeling of the mitochondrial protein biogenesis machinery under proteostatic stress 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.10.06.680693v1?rss=1"
</link>
<description><![CDATA[
Cells have evolved organelle-specific responses to maintain protein homeostasis (proteostasis). During proteostatic stress, mitochondria downregulate translation and enhance protein folding, yet the underlying mechanisms remain poorly defined. Here, we employed cryo-electron tomography to observe the structural consequences of mitochondrial proteostatic stress within human cells. We detected protein aggregates within the mitochondrial matrix, accompanied by a marked remodeling of cristae architecture. Concomitantly, the number of mitochondrial ribosome complexes was significantly reduced. Mitochondrial Hsp60 (mHsp60), a key protein folding machine, underwent major conformational changes to favor complexes with its co-chaperone mHsp10. We visualized the interactions of mHsp60 with native substrate proteins, and determined in vitro mHsp60 cryo- EM structures enabling nucleotide state assignment of the in situ structures. These data converge on a model of the mHsp60 functional cycle and its essential role in mitochondrial proteostasis. More broadly, our findings reveal structural mechanisms governing mitochondrial protein biosynthesis and their remodeling under proteostatic stress.
]]></description>
<dc:creator>Ehses, K. L.</dc:creator>
<dc:creator>Lopez-Alonso, J. P.</dc:creator>
<dc:creator>Antico, O.</dc:creator>
<dc:creator>Azem, A.</dc:creator>
<dc:creator>Muqit, M.</dc:creator>
<dc:creator>Ubarretxena-Belandia, I.</dc:creator>
<dc:creator>Fernandez-Busnadiego, R.</dc:creator>
<dc:date>2025-10-06</dc:date>
<dc:identifier>doi:10.1101/2025.10.06.680693</dc:identifier>
<dc:title><![CDATA[Structural remodeling of the mitochondrial protein biogenesis machinery under proteostatic stress]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-06</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.10.08.681047v1?rss=1">
<title>
<![CDATA[
Proteome Landscapes Decode Organelle Vulnerabilities in cortical and dopaminergic-like induced neurons Across Lysosomal Storage Disorders 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.10.08.681047v1?rss=1"
</link>
<description><![CDATA[
Lysosomes maintain cellular homeostasis by degrading proteins delivered via endocytosis and autophagy and recycling building blocks for organelle biogenesis. Lysosomal Storage Disorders (LSDs) comprise a broad group of diseases affecting lysosomal degradation, ion flux, and lipid catabolism. Within this group, sphingolipidoses genes involved in glycosphingolipid breakdown are known (GBA1) or candidate (SMPD1, ASAH1) risk factors for Parkinsons Disease, though disease mechanisms remain unclear. Using our previously reported LSD mutant proteomic landscape in HeLa cells, we observed pronounced variability in endolysosomal proteome signatures among sphingolipid pathway mutants, with ASAH1-/- cells showing altered lysosomal lipid composition, impaired endocytic trafficking, and disrupted ultrastructure by cryo- electron tomography. To extend these findings in a more physiologic context, we generated a human embryonic stem (ES) cell library comprising 23 LSD gene knockouts and profiled proteomic changes during differentiation into cortical and midbrain dopaminergic neurons over a 7 to 10 week period. LSD mutants exhibited lineage-specific alterations in organellar proteomes, revealing diverse vulnerabilities. Notably, GBA1-/- and ASAH1-/- dopaminergic neurons showed disruptions in synaptic and mitochondrial compartments, correlating with impaired dopaminergic neuronal firing and disrupted presynaptic protein localization. This LSD mutant toolkit and associated proteomic landscape provides a resource for defining molecular signatures of LSD gene loss and highlights convergence of lysosomal dysfunction, synaptic integrity, and mitochondrial health as potential links between sphingolipidoses and PD risk.
]]></description>
<dc:creator>Kraus, F.</dc:creator>
<dc:creator>He, Y.</dc:creator>
<dc:creator>Jiang, Y.</dc:creator>
<dc:creator>Li, D.</dc:creator>
<dc:creator>Ambaw, Y. A.</dc:creator>
<dc:creator>Gasparoli, F. M.</dc:creator>
<dc:creator>Paulo, J. A.</dc:creator>
<dc:creator>Walther, T. C.</dc:creator>
<dc:creator>Farese, R.</dc:creator>
<dc:creator>Gygi, S. P.</dc:creator>
<dc:creator>Wilfling, F.</dc:creator>
<dc:creator>Harper, J. W.</dc:creator>
<dc:date>2025-10-08</dc:date>
<dc:identifier>doi:10.1101/2025.10.08.681047</dc:identifier>
<dc:title><![CDATA[Proteome Landscapes Decode Organelle Vulnerabilities in cortical and dopaminergic-like induced neurons Across Lysosomal Storage Disorders]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.02.13.638174v1?rss=1">
<title>
<![CDATA[
Robust analytical methods for bis(monoacylglycero)phosphate profiling in health and disease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.02.13.638174v1?rss=1"
</link>
<description><![CDATA[
Bis(monoacylglycero)phosphate (BMP), a distinct anionic phospholipid predominantly found in late endosomes and lysosomes, plays a pivotal role in supporting lysosomal functions and maintaining metabolic homeostasis. Its impaired function is associated with an array of disorders, notably neurodegenerative diseases. However, the identification and quantitation of BMP remains difficult due to its structural similarity to isomer phosphatidylglycerol (PG), thus necessitating robust analytical methods for accurate and reliable BMP profiling. In this study, we present comprehensive liquid chromatography - tandem mass spectrometry (MS2) methodologies for the precise and systematic analysis of BMP species in biological samples. We detail LC/MS methods for both an untargeted Orbitrap mass spectrometer and a targeted triple quadrupole (QQQ) mass spectrometer. We utilize differences in polarity and structure to annotate BMPs and PGs based on retention time and positive mode MS2 fragmentation patterns, respectively. Further, we propose a new approach for overcoming common challenges in BMP profiling by leveraging the newly discovered biochemical function of CLN5 as the BMP synthase. Since genetic ablation of CLN5 leads to specific depletion of BMPs but not PGs, we use lipid extracts from CLN5 knockout (KO) and wild-type (WT) cells as biological standards to confidently annotate BMPs as targets with significantly low BMP Identification Index (BMPII), defined as BMPII = CLN5 KO / WT. We additionally propose the BMP enrichment score (BMPES) as a secondary validation metric, defined as lysosomal abundance of BMP / whole-cell abundance. Altogether, this approach constitutes a robust method for BMP profiling and annotation, furthering research into health and disease.
]]></description>
<dc:creator>Dong, W.</dc:creator>
<dc:creator>Nyame, K.</dc:creator>
<dc:creator>Rawat, E. S.</dc:creator>
<dc:creator>Medoh, U. N.</dc:creator>
<dc:creator>Xiong, J.</dc:creator>
<dc:creator>Bonin, C. C.</dc:creator>
<dc:creator>Alsohybe, H. N.</dc:creator>
<dc:creator>Liu, H. Y.</dc:creator>
<dc:creator>Gomes, S.</dc:creator>
<dc:creator>Hsieh, T.</dc:creator>
<dc:creator>Arnold, M.</dc:creator>
<dc:creator>Hsieh, F.</dc:creator>
<dc:creator>Sammler, E.</dc:creator>
<dc:creator>Abu-Remaileh, M.</dc:creator>
<dc:date>2025-02-17</dc:date>
<dc:identifier>doi:10.1101/2025.02.13.638174</dc:identifier>
<dc:title><![CDATA[Robust analytical methods for bis(monoacylglycero)phosphate profiling in health and disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-02-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.08.19.670962v1?rss=1">
<title>
<![CDATA[
Structural basis for binding of RILPL1 to TMEM55B reveals a lysosomal platform for adaptor assembly through a conserved TBM motif 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.08.19.670962v1?rss=1"
</link>
<description><![CDATA[
Inherited mutations in VPS35 and the kinase LRRK2 lead to hyperphosphorylation of Rab GTPases and promote the formation of phospho-Rab signalling complexes. A subset of RH2 domain-containing proteins from the RILP-homology family, including RILP, RILPL1, RILPL2, JIP3, and JIP4 are Rab effectors that recognize the LRRK2-phosphorylated switch 2 threonine of phospho-Rab8A and phospho-Rab10. More recently, phospho-Rabs have been found on lysosomal membranes within multi-protein assemblies involving TMEM55B and RILPL1. TMEM55B is a 284-residue lysosomal membrane protein with no homology to known proteins. It comprises a 218-residue cytosolic N-terminal region and two predicted transmembrane -helices. Residues 80- 160, which face the cytosol, mediate binding to a C-terminal motif of RILPL1, formed after RILPL1 associates with phospho-Rab8A. Here, we report the crystal structures of TMEM55B alone and in complex with a C-terminal RILPL1 peptide, encompassing the TMEM55B interaction region, which we define as the TMEM55B Binding Motif (TBM). The cytosolic domain of TMEM55B adopts a rigid architecture of two tandem RING-like domains, each forming a Zn{superscript 2}-stabilized 40-residue {beta}-sandwich. TBM binding is mediated primarily by backbone hydrogen bonding and anchored by two glutamate residues from RILPL1. These findings support a model in which RILPL1 is recruited to phospho-Rab8A-positive lysosomes prior to TMEM55B engagement. Further co-immunoprecipitation and mutational analyses indicate that TMEM55B forms complexes independently of phospho-Rabs with proteins containing a conserved TBM, like that of RILPL1, including JIP3, JIP4, OCRL, WDR81, and TBC1D9B. Together, these findings uncover previously unrecognized regulatory networks associated with TMEM55B and lysosomal function and suggest that TMEM55B serves as a central hub for adaptor recruitment at the lysosomal membrane.
]]></description>
<dc:creator>Waschbuesch, D.</dc:creator>
<dc:creator>Pal, P.</dc:creator>
<dc:creator>Nirujogi, R. S.</dc:creator>
<dc:creator>Cavin, M.</dc:creator>
<dc:creator>Singh, J.</dc:creator>
<dc:creator>Alessi, D. R.</dc:creator>
<dc:creator>Khan, A. R.</dc:creator>
<dc:date>2025-08-24</dc:date>
<dc:identifier>doi:10.1101/2025.08.19.670962</dc:identifier>
<dc:title><![CDATA[Structural basis for binding of RILPL1 to TMEM55B reveals a lysosomal platform for adaptor assembly through a conserved TBM motif]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-24</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.08.25.672135v1?rss=1">
<title>
<![CDATA[
LRRK2G2019S acts as a dominant interfering mutant in the context of iron overload 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.08.25.672135v1?rss=1"
</link>
<description><![CDATA[
A key pathological feature of Parkinsons Disease (PD) is the loss of neuromelanin, an iron chelator within the dopaminergic neurons, which results in iron toxicity thought to result in selective neuronal vulnerability. This implicated iron handling pathways as an early target of research in PD. Given the critical role of PD-related activating mutations in LRRK2 (leucine-rich repeat protein kinase 2) within membrane trafficking pathways we examined the impact of mutant LRRK2G2019S on iron homeostasis within a model macrophage cell line known to have high iron capacity. Proteomics analysis revealed a dysregulation of iron-related proteins in steady state with highly elevated levels of ferritin light chain and a reduction of ferritin heavy chain. LRRK2 mutant cells showed efficient ferritinophagy upon iron chelation, but upon iron overload there was a near complete block in the degradation of the ferritinophagy adaptor NCOA4. Surprisingly, NCOA4 levels were not rescued upon inhibition of the LRRK2 kinase activity in iron overload conditions, nor was the phosphorylation of substrate Rab GTPases. We therefore generated a CRISPR mutation to delete the kinase domain of LRRK2 and express only the Rab-binding armadillo repeat domain. Although the kinase domain was deleted, the truncation mutant of LRRK2 showed strong Rab8 phosphorylation in conditions of iron overload, similar to LRRK2G2019S cells, with the phosphorylated Rab8 accumulating at the plasma membrane. These data indicate that the G2019S mutation acts as a kinase independent, dominant-interfering mutant specifically in conditions of iron overload. Together, our data implicate LRRK2 as a key regulator of iron homeostasis and point to the need for an increased focus on the mechanisms of iron dysregulation in PD.
]]></description>
<dc:creator>Goldman, A.</dc:creator>
<dc:creator>Nguyen, M.</dc:creator>
<dc:creator>Lanoix, J.</dc:creator>
<dc:creator>Fahmy, A.</dc:creator>
<dc:creator>Zhong Xu, Y.</dc:creator>
<dc:creator>Schurr, E.</dc:creator>
<dc:creator>Thibault, P.</dc:creator>
<dc:creator>Desjardins, M.</dc:creator>
<dc:creator>McBride, H.</dc:creator>
<dc:date>2025-08-25</dc:date>
<dc:identifier>doi:10.1101/2025.08.25.672135</dc:identifier>
<dc:title><![CDATA[LRRK2G2019S acts as a dominant interfering mutant in the context of iron overload]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-08-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.09.03.673956v1?rss=1">
<title>
<![CDATA[
Activation of transposable elements is linked to a region- and cell-type-specific interferon response in Parkinson's disease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.09.03.673956v1?rss=1"
</link>
<description><![CDATA[
Parkinsons disease (PD) is a common age-related neurodegenerative disorder involving a neuroinflammatory response, the cause of which remains unclear. Transposable elements (TE) have been linked to inflammatory states, but their potential role in PD has not been explored. Using bulk- and single nuclei RNAseq of postmortem brain tissue from four brain regions, we studied TE transcriptional activation and its correlation with neuroinflammation in PD. Over a thousand TE loci, including LINE-1s and ERVs, were highly expressed in a cell-type and region-specific manner in the human brain. Increased TE expression was found in microglia and neurons in the substantia nigra and putamen in the PD brains, but not amygdala or prefrontal cortex, compared to age-matched control tissue. This TE activation correlated with innate immune transcriptional responses, characterized by the expression of interferon-related and viral response genes, in the same brain regions. The link between an interferon response and TE activation was mechanistically confirmed using human pluripotent stem cell-derived microglia and neurons. Our findings provide a unique insight into TE transcription in the PD brain and suggests that TEs play a role in chronic neuroinflammatory processes and the progression of this neurodegenerative disorder.
]]></description>
<dc:creator>Garza, R.</dc:creator>
<dc:creator>Adami, A.</dc:creator>
<dc:creator>Thiruvalluvan, A.</dc:creator>
<dc:creator>Wijesinghe, S.</dc:creator>
<dc:creator>Curle, A. J.</dc:creator>
<dc:creator>Tam, O.</dc:creator>
<dc:creator>Forcier, T.</dc:creator>
<dc:creator>Lagka, D. A.</dc:creator>
<dc:creator>Kazakou, N. L.</dc:creator>
<dc:creator>Atacho, D. A. M.</dc:creator>
<dc:creator>Sharma, Y.</dc:creator>
<dc:creator>Jonsson, M. E.</dc:creator>
<dc:creator>Horvath, V.</dc:creator>
<dc:creator>Bermudez, S.</dc:creator>
<dc:creator>Johansson, J. G.</dc:creator>
<dc:creator>Rainbow, D. B.</dc:creator>
<dc:creator>Castilla-Vallmanya, L.</dc:creator>
<dc:creator>Jones, J. L.</dc:creator>
<dc:creator>Quaegebeur, A.</dc:creator>
<dc:creator>Gale Hammell, M.</dc:creator>
<dc:creator>Kirkeby, A.</dc:creator>
<dc:creator>Barker, R. A.</dc:creator>
<dc:creator>Jakobsson, J.</dc:creator>
<dc:date>2025-09-07</dc:date>
<dc:identifier>doi:10.1101/2025.09.03.673956</dc:identifier>
<dc:title><![CDATA[Activation of transposable elements is linked to a region- and cell-type-specific interferon response in Parkinson's disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.09.03.674114v1?rss=1">
<title>
<![CDATA[
Proteome-wide impact of LRRK1 and 2 inhibitors on protein interactions and phosphorylation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.09.03.674114v1?rss=1"
</link>
<description><![CDATA[
The Leucine-rich repeat kinases 1 and 2 (LRRK1 and 2) are large, multidomain proteins and closely related members of the Roco protein family. They share a high similarity in domain structure and are both phosphorylate members of the Rab GTPase family. However, despite these similarities, there are substantial differences between the two kinases. While mutations to LRRK1 are only implicated in rare cases of osteopetrosis, LRRK2 is associated with multiple diseases, most prominently with familial and sporadic forms of Parkinsons disease, where pathogenic LRRK2 is associated with an increased kinase activity. While LRRK2 has received major attention from the research community, LRRK1 has been largely understudied. In this work, we employ proximity labelling mass spectrometry in combination with quantitative phosphoproteomics in a model cell line to obtain the cellular interactomes of LRRK1 and LRRK2 and corresponding phosphorylation sites. We then use this dataset to characterize the impact of small molecules targeting both LRRK1 and 2. We identify phosphorylation sites across the proteome that are impacted by these inhibitors and identify novel candidate substrates for LRRK2, including MICALL2. Taken together our data provide a powerful resource for future studies on the cellular role and function of LRRK proteins and their potential use as therapeutic targets.
]]></description>
<dc:creator>Jansen, J.</dc:creator>
<dc:creator>Wendel, M.</dc:creator>
<dc:creator>Raig, N. D.</dc:creator>
<dc:creator>Kraus, T. V.</dc:creator>
<dc:creator>Surridge, K. J.</dc:creator>
<dc:creator>Mahesula, S.</dc:creator>
<dc:creator>Richter-Muller, N.</dc:creator>
<dc:creator>Mathea, S.</dc:creator>
<dc:creator>Reck-Peterson, S. L.</dc:creator>
<dc:creator>Knapp, S.</dc:creator>
<dc:creator>Stengel, F.</dc:creator>
<dc:date>2025-09-07</dc:date>
<dc:identifier>doi:10.1101/2025.09.03.674114</dc:identifier>
<dc:title><![CDATA[Proteome-wide impact of LRRK1 and 2 inhibitors on protein interactions and phosphorylation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.09.09.675138v1?rss=1">
<title>
<![CDATA[
A molecular atlas of cell-type specific signatures in the parkinsonian striatum. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.09.09.675138v1?rss=1"
</link>
<description><![CDATA[
The progressive degeneration of dopaminergic projections to the striatum is a key disease mechanism in Parkinsons disease (PD). To define the cellular landscape in the parkinsonian striatum, we mapped the cell-type specific transcriptional landscape in early and progressive PD mouse models and in human PD stages. Our analyses revealed substantial transcriptomic changes across both neuronal and glial populations, with astrocytes and oligodendrocytes exhibiting distinct disease-associated gene expression profiles. Notably, progressive dopamine depletion uncovered differential neuronal vulnerability, identifying eccentric striatal projection neurons (SPNs) and Chst9-expressing direct-pathway SPNs as among the most resilient subtypes in both species. This cross-species resource establishes a comprehensive framework for investigating cell-state dynamics in the parkinsonian striatum and uncovers selectively vulnerable and resistant cell types that can inspire new therapeutic strategies.
]]></description>
<dc:creator>Graziano, M.</dc:creator>
<dc:creator>Mantas, I.</dc:creator>
<dc:creator>Masarapu, Y.</dc:creator>
<dc:creator>Frapard, S.</dc:creator>
<dc:creator>Garza, R.</dc:creator>
<dc:creator>Adami, A.</dc:creator>
<dc:creator>Fazal, S.</dc:creator>
<dc:creator>Quaegebeur, A.</dc:creator>
<dc:creator>Barker, R.</dc:creator>
<dc:creator>Jakobsson, J.</dc:creator>
<dc:creator>Giacomello, S.</dc:creator>
<dc:creator>Meletis, K.</dc:creator>
<dc:date>2025-09-10</dc:date>
<dc:identifier>doi:10.1101/2025.09.09.675138</dc:identifier>
<dc:title><![CDATA[A molecular atlas of cell-type specific signatures in the parkinsonian striatum.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-09-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.10.13.682089v1?rss=1">
<title>
<![CDATA[
The SNCA A53T mutation sensitizes human neurons and microglia to ferroptosis 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.10.13.682089v1?rss=1"
</link>
<description><![CDATA[
The major pathological hallmarks of sporadic and familial forms of Parkinsons disease (PD) are the targeted and progressive loss of midbrain dopaminergic neurons (mDA), associated with systemic iron accumulation, -synuclein (syn) accumulation and aggregation, and lipid peroxidation amongst other reactive oxygen species (ROS) generation. Therapeutic strategies aimed towards dopamine restoration, syn removal and iron chelation have provided symptomatic relief but failed to prevent or slow disease progression. This is in part due to the lack of understanding of the exact pathways leading to neuronal death in PD. In this study, we investigate ferroptosis, a unique cell death mechanism sharing multiple features with PD pathology, as a relevant pathway with implications in disease pathogenesis. We identified an enrichment of ferroptosis genes dysregulated throughout PD postmortem brain samples and several neuronal and glial PD models. Using CRISPR/Cas9 technology, we generated a rapid iPSC-derived synucleinopathy neuronal model harbouring the SNCA A53T mutation and report increased ROS generation, reduced levels of antioxidant glutathione (GSH), impaired mitophagy and a heightened vulnerability to ferroptosis-induced lipid peroxidation and cell death. Critically, inhibition of the key lipid peroxidation enzyme and driver of ferroptosis, 15-lipoxygenase (15-LO), rescued synucleinopathy associated pathologies and prevented pathological syn oligomerisation in SNCA A53T neurons. Furthermore, we report enhanced microglial ferroptosis susceptibility in models of synucleinopathy. In summary, we highlight a new mechanism by which the familial PD-associated SNCA A53T mutation causes cell death and propose 15-LO inhibition as a tractable therapeutic opportunity in PD.
]]></description>
<dc:creator>Mahoney-Sanchez, L.</dc:creator>
<dc:creator>Clarke-Lucas, H.</dc:creator>
<dc:creator>Penverne, A.</dc:creator>
<dc:creator>Evans, J. R.</dc:creator>
<dc:creator>D'Sa, K.</dc:creator>
<dc:creator>Strohbuecker, S.</dc:creator>
<dc:creator>Lopex Garcia, P.</dc:creator>
<dc:creator>Cosker, K.</dc:creator>
<dc:creator>Soltic, D.</dc:creator>
<dc:creator>O'Callaghan, B. J.</dc:creator>
<dc:creator>Griffiths, A.</dc:creator>
<dc:creator>Pintchovski, S. A.</dc:creator>
<dc:creator>Plun-Favreau, H.</dc:creator>
<dc:creator>Hallqvist, J.</dc:creator>
<dc:creator>Mills, K.</dc:creator>
<dc:creator>Gandhi, S.</dc:creator>
<dc:date>2025-10-14</dc:date>
<dc:identifier>doi:10.1101/2025.10.13.682089</dc:identifier>
<dc:title><![CDATA[The SNCA A53T mutation sensitizes human neurons and microglia to ferroptosis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-14</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.10.24.684366v1?rss=1">
<title>
<![CDATA[
A framework for efficient CRISPRi-mediated silencing of retrotransposons in human pluripotent stem cells 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.10.24.684366v1?rss=1"
</link>
<description><![CDATA[
This protocol describes a workflow for transcriptional silencing of transposable elements (TEs) in human induced pluripotent stem cells (hiPSCs). It illustrates how to design gRNAs to target TE families or unique TE loci and how to validate the efficiency and specificity of large-scale CRISPRi-based silencing using a multiome approach that combines bulk RNA sequencing, CUT&RUN epigenetic profiling, and proteomics. This unique framework optimizes performance and interpretation of in vitro functional studies based on transcriptional manipulation of TEs in hiPSC models.

For complete details on the use and execution of this protocol, please refer to Adami et al. (2025)1.

Graphical abstract

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=197 SRC="FIGDIR/small/684366v2_ufig1.gif" ALT="Figure 1">
View larger version (48K):
org.highwire.dtl.DTLVardef@154560aorg.highwire.dtl.DTLVardef@1dce192org.highwire.dtl.DTLVardef@5d7097org.highwire.dtl.DTLVardef@8aea84_HPS_FORMAT_FIGEXP  M_FIG C_FIG
]]></description>
<dc:creator>Adami, A.</dc:creator>
<dc:creator>Garza, R.</dc:creator>
<dc:creator>Dorazehi, F.</dc:creator>
<dc:creator>Douse, C. H.</dc:creator>
<dc:creator>Jakobsson, J.</dc:creator>
<dc:date>2025-10-28</dc:date>
<dc:identifier>doi:10.1101/2025.10.24.684366</dc:identifier>
<dc:title><![CDATA[A framework for efficient CRISPRi-mediated silencing of retrotransposons in human pluripotent stem cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-28</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.10.29.685288v1?rss=1">
<title>
<![CDATA[
Extracellular space diffusion modelling identifies distinct functional advantages of archetypical glutamatergic and GABAergic synapse geometries 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.10.29.685288v1?rss=1"
</link>
<description><![CDATA[
The brain extracellular space (ECS) is a convoluted compartment of nano- and microscale interconnected ducts. A key step in signaling between neural cells is diffusion of signaling molecules through the ECS, yet, signaling is generally considered solely from the stance of cells and their properties. Where ECS diffusion is addressed, this is commonly done using volume-averaging techniques blind to individual signaling events and ECS geometry. We hypothesized that ECS geometry can shape local diffusion and thereby tune signaling arising from point-sources. To access the scale of individual transmitter release events and synapse geometries, we developed a computational diffusion model, DifFlux, based on super-resolved images of hippocampal ECS in live mouse brain slices and combined this with single molecule Monte Carlo diffusion simulations. Our approach allows us to simulate diffusion of molecules of our choosing in true live ECS geometries. We asked how the ECS shapes local diffusion in dense neuropil and along larger cellular processes in CA1 stratum radiatum. We observed local diffusional anisotropy and directionality imposed by ECS geometry. Further, we identified distinct functional advantages of dendritic spine and somatodendritic synapse ECS geometries, shedding light on the longstanding conundrum of why glutamatergic and GABAergic synapses are so conspicuously morphologically different. Our modelling broadly identifies ECS structure as a direct modulator of extrasynaptic signaling that can operate in parallel to conventional regulation mechanisms.
]]></description>
<dc:creator>Gimenez, P.</dc:creator>
<dc:creator>Shakya, R. T.</dc:creator>
<dc:creator>Santamaria, F.</dc:creator>
<dc:creator>Tonnesen, J.</dc:creator>
<dc:date>2025-10-29</dc:date>
<dc:identifier>doi:10.1101/2025.10.29.685288</dc:identifier>
<dc:title><![CDATA[Extracellular space diffusion modelling identifies distinct functional advantages of archetypical glutamatergic and GABAergic synapse geometries]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-10-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.11.07.687251v1?rss=1">
<title>
<![CDATA[
Reconstitution of multistep recruitment of ULK1 to membranes in autophagy 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.11.07.687251v1?rss=1"
</link>
<description><![CDATA[
The ULK1 complex (ULK1C) and the class III phosphatidylinositol 3-kinase complex I (PI3KC3-C1) act together to initiate autophagy. Human ULK1C consists of ULK1 itself, FIP200, and the HORMA domain heterodimer ATG13:ATG101. PI3P generated by PI3KC3-C1 is essential to recruit and stabilize ULK1C on membranes for ULK1 to phosphorylate its membrane-associated substrates in autophagy induction, even though ULK1C subunits do not contain any PI3P-binding domains. Here we show that the ATG13:ATG101 dimer forms a tight complex with the PI3P-binding protein WIPI3, as well as with WIPI2. Bound to WIPI2-3, ATG13:ATG101 aligns with the membrane to insert its Trp-Phe (WF) finger into the membrane. Molecular dynamics simulations show that alignment of WIPIs and the ATG101 WF finger cooperatively stabilizes the complex on membranes, explaining the essential role of the WF residues in autophagy. Biochemical reconstitution and a cell-based assay show that WIPI3:ATG13 engagement is required for ATG16L1 phosphorylation by ULK1, ATG13 puncta formation, and bulk autophagic flux. We further showed that a kinase domain (KD)-proximal PVP motif within the ULK1 IDR docks onto the surface of the ATG13:ATG101 HORMA dimer and used molecular modeling to show how the ULK1 KD is brought close to the membrane surface. Biochemical reconstitution and cell-based assays show that the PVP motif is essential for in vitro ULK1 phosphorylation of ATG16L1 and important for starvation-induced autophagy and BNIP3/NIX-dependent mitophagy. These data establish a stepwise pathway for recruitment of the ULK1 KD to the vicinity of the membrane surface downstream of PI3KC3-C1.
]]></description>
<dc:creator>Duan, Y.</dc:creator>
<dc:creator>Lu, Y.</dc:creator>
<dc:creator>Paul, S.</dc:creator>
<dc:creator>Betz, J.</dc:creator>
<dc:creator>Wilhelm, L. P.</dc:creator>
<dc:creator>Cook, A. S. I.</dc:creator>
<dc:creator>Ren, X.</dc:creator>
<dc:creator>Adriaenssens, E.</dc:creator>
<dc:creator>Martens, S.</dc:creator>
<dc:creator>Ganley, I.</dc:creator>
<dc:creator>Hummer, G.</dc:creator>
<dc:creator>Hurley, J. H.</dc:creator>
<dc:date>2025-11-09</dc:date>
<dc:identifier>doi:10.1101/2025.11.07.687251</dc:identifier>
<dc:title><![CDATA[Reconstitution of multistep recruitment of ULK1 to membranes in autophagy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.11.10.687702v1?rss=1">
<title>
<![CDATA[
Insights into the regulation of VPS13 family bridge-like lipid transfer proteins from the structure of VPS13C. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.11.10.687702v1?rss=1"
</link>
<description><![CDATA[
Bridge-like lipid transfer proteins (BLTPs) play central roles in redistributing lipids from their primary site of synthesis in the endoplasmic reticulum to other organelles. They comprise bridge-domains spanning between organelles at contact sites that allow lipids to transit the cytosol between adjacent membranes. The assembly of BLTPs into complexes with adaptor proteins enables their lipid transfer ability. To address the mechanisms underlying assembly and regulation of BLTP complexes, we used cryo-EM to resolve the structure of one such BLTP, the Parkinsons protein VPS13C, at near-atomic resolution. The structure identifies a lipid-transfer-nonpermissive conformation, where the built-in C-terminal VAB adaptor module blocks the end of the lipid transfer bridge, interfering with lipid delivery. We also identify calmodulin, central to calcium signaling, as a VPS13 partner, suggesting calcium regulation of VPS13 function. Altogether, this structure of intact VPS13C serves as starting point to understand its regulation and, more broadly, that of other BLTPs.
]]></description>
<dc:creator>Li, D.</dc:creator>
<dc:creator>Wang, X.</dc:creator>
<dc:creator>Hu, B.</dc:creator>
<dc:creator>Hao, H.</dc:creator>
<dc:creator>Hamill, S.</dc:creator>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Chen, G.</dc:creator>
<dc:creator>De Camilli, P.</dc:creator>
<dc:creator>Reinisch, K.</dc:creator>
<dc:date>2025-11-11</dc:date>
<dc:identifier>doi:10.1101/2025.11.10.687702</dc:identifier>
<dc:title><![CDATA[Insights into the regulation of VPS13 family bridge-like lipid transfer proteins from the structure of VPS13C.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.11.16.688672v1?rss=1">
<title>
<![CDATA[
Pathway and gates for ATG2A-mediated lipid transport in autophagy 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.11.16.688672v1?rss=1"
</link>
<description><![CDATA[
Autophagy is a complex process in which eukaryotic cells degrade cytosolic components by delivering them to lysosomes via double-membrane autophagosomes. The lipid transfer protein ATG2A plays a crucial role in autophagosome formation by tethering the phagophore membrane to the ER and delivering a significant fraction of the required lipids. The mechanism by which ATG2A shuttles lipids from one membrane to the other, however, remains elusive. Here, we combine structural predictions, molecular dynamics simulations and in vitro lipid transfer assays to gain mechanistic insights into ATG2A-mediated lipid transport. Using this integrative approach, we characterize the contact sites of the protein with donor and acceptor membranes. Our simulations capture multiple events of lipid uptake and delivery from and to the bound membrane. Conformational rearrangements of N-terminal amphipathic helices emerge as a critical factor for facile lipid transport. With this insight, we designed an ATG2A mutant that transfers lipids three times faster than the wild type in vitro. In complex with ATG9A, ATG2A forms a bridge between two parallel membranes at 12 nm separation. Overall, our findings suggest that ATG2A is a lipid transporter gated at the N-terminus by blocking helices that, upon release, act as additional membrane tethers.
]]></description>
<dc:creator>Cabezudo, A. C.</dc:creator>
<dc:creator>Holzer, E.</dc:creator>
<dc:creator>Martens, S.</dc:creator>
<dc:creator>Hummer, G.</dc:creator>
<dc:date>2025-11-16</dc:date>
<dc:identifier>doi:10.1101/2025.11.16.688672</dc:identifier>
<dc:title><![CDATA[Pathway and gates for ATG2A-mediated lipid transport in autophagy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-16</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.11.16.688724v1?rss=1">
<title>
<![CDATA[
A Multimodal Atlas Reveals the Anatomical Distribution of Medium Spiny Neuron Subtypes and a Novel RGS6+ Population in the Primate Striatum 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.11.16.688724v1?rss=1"
</link>
<description><![CDATA[
The primate striatum and its principal neuron type, the medium spiny neuron (MSN), integrate cortical and subcortical signals related to movement, cognition, and emotion. These signals are processed through cell type specific circuits traditionally defined by MSN dopamine receptor expression. However, classification by dopamine receptor type alone fails to fully specify MSN diversity and falls short of capturing the functional complexity of the striatum. Here, we combined single-nucleus multi-omic sequencing and high-plex spatial transcriptomics to build a comprehensive atlas of MSNs in the macaque striatum. Using multi-omic sequencing, we profiled MSNs across four anatomically and functionally defined territories, and we mapped these subtypes back into their anatomical context by integrating the multi-omic data with [~]5.4 million spatially resolved cells sampled across the full rostral-caudal and dorsal-ventral extent of the striatum. This approach revealed two previously undocumented ventral striatum (VS) subtypes, D1-VS-RGS6 and D2-VS-RGS6, which are molecularly distinct from known ventral striatal MSNs yet share core limbic features. We also uncovered gradients in matrix-compartment cell types along the rostral-caudal axis. Finally, by integrating MSN subtype-specific transcriptomes and ATAC-seq-derived regulatory annotations with human GWAS data, we demonstrate strong, cell-type-specific enrichment of polygenic risk for Parkinsons disease, substance use disorders, and psychiatric and cognitive traits, including a striking association of D2-VS-RGS6 with schizophrenia and bipolar disorder. Together, this multimodal atlas provides a foundation for linking primate striatal cell types to circuit function and disease mechanisms.

HIGHTLIGHTSO_LIMultimodal analysis of NHP striatum reveals heterogeneous cell type distribution
C_LIO_LITwo previously uncharacterized MSN subtypes in the ventral striatum express RGS6
C_LIO_LIVentral striatum cell types exhibit similar characteristics across the Rostro-Caudal axis
C_LIO_LINHP cell types show strong, cell type specific associations to genomic disease predictors
C_LI
]]></description>
<dc:creator>Abdelhady, G.</dc:creator>
<dc:creator>Brull, O. R.</dc:creator>
<dc:creator>He, J.</dc:creator>
<dc:creator>Lin, M. K.</dc:creator>
<dc:creator>Galvan, A.</dc:creator>
<dc:creator>Pfenning, A. R.</dc:creator>
<dc:creator>Bostan, A. C.</dc:creator>
<dc:creator>Stauffer, W. R.</dc:creator>
<dc:date>2025-11-17</dc:date>
<dc:identifier>doi:10.1101/2025.11.16.688724</dc:identifier>
<dc:title><![CDATA[A Multimodal Atlas Reveals the Anatomical Distribution of Medium Spiny Neuron Subtypes and a Novel RGS6+ Population in the Primate Striatum]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.11.19.689251v1?rss=1">
<title>
<![CDATA[
LRRK2 integrates Rab and GABARAP interactions to sense and respond to distinct lysosomal stresses 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.11.19.689251v1?rss=1"
</link>
<description><![CDATA[
Increased activity of leucine-rich repeat kinase 2 (LRRK2) is an important risk factor for Parkinsons disease. LRRK2 localizes to lysosomal membranes, and changes in lysosome physiology are emerging as key regulators of its activation, yet the mechanisms by which distinct perturbations engage this kinase remain unclear. Analysis of osmotic and membrane-integrity challenges revealed that LRRK2 integrates multiple upstream cues through parallel interactions with Rab GTPases and GABARAP. Manipulations that caused lysosome enlargement, including inhibition of PIKfyve, showed that osmotic swelling leads to the accumulation of multiple Rabs on lysosomes and Rab-dependent LRRK2 activation independently of GABARAP. In contrast, under conditions of lysosome deacidification, CASM-dependent lipidation of GABARAP creates a platform that cooperates with Rabs in LRRK2 activation. These findings demonstrate how LRRK2 interprets perturbations of lysosome function through a combination of Rab- and GABARAP-dependent mechanisms, providing a framework for understanding both normal physiological regulation and pathological dysregulation in Parkinsons disease.

Significance StatementThis study reveals how LRRK2 integrates lysosomal stress signals through coordinated interactions with Rab GTPases and GABARAP. Osmotic swelling drives strong Rab-dependent activation, whereas deacidification requires CASM-mediated GABARAP lipidation as a scaffold for LRRK2 activation at lysosomes. These results define how LRRK2 activation at lysosomes is tuned across physiological and pathogenic contexts.
]]></description>
<dc:creator>Clegg, D.</dc:creator>
<dc:creator>Bentley-DeSousa, A.</dc:creator>
<dc:creator>Roczniak-Ferguson, A.</dc:creator>
<dc:creator>Ferguson, S. M.</dc:creator>
<dc:date>2025-11-19</dc:date>
<dc:identifier>doi:10.1101/2025.11.19.689251</dc:identifier>
<dc:title><![CDATA[LRRK2 integrates Rab and GABARAP interactions to sense and respond to distinct lysosomal stresses]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.12.01.691607v1?rss=1">
<title>
<![CDATA[
Elucidating an anterior cingulate circuit for self-initiated actions and rescue of Parkinsonian akinesia 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.12.01.691607v1?rss=1"
</link>
<description><![CDATA[
Dopamine (DA) depletion is known to result in Parkinsonian symptoms such as the inability to initiate movements (akinesia). While Parkinsonian akinesia is traditionally associated with reduced DA signaling in the striatum, the contribution of cortical regions that also receive DA projections and project to the striatum remains unclear. Here, we identify a previously unexplored cortical circuit involving D1-like dopamine receptor-expressing neurons in the anterior cingulate cortex (ACC) that is critical for initiating goal-directed movements. We find that a selective activation of ACC-D1+ neurons can flexibly drive targeted movement and locomotion even in akinetic mice after dopamine depletion or receptor antagonism. These findings uncover a cortical mechanism for movement initiation and offer promising new therapeutic targets for treating Parkinsonian akinesia.
]]></description>
<dc:creator>Ulloa Severino, F.</dc:creator>
<dc:creator>Lu, B.</dc:creator>
<dc:creator>Kim, J.</dc:creator>
<dc:creator>Friedman, A.</dc:creator>
<dc:creator>Roshchina, M.</dc:creator>
<dc:creator>Johnson, S.</dc:creator>
<dc:creator>Bakhurin, K.</dc:creator>
<dc:creator>Eroglu, C.</dc:creator>
<dc:creator>Yin, H.</dc:creator>
<dc:date>2025-12-03</dc:date>
<dc:identifier>doi:10.64898/2025.12.01.691607</dc:identifier>
<dc:title><![CDATA[Elucidating an anterior cingulate circuit for self-initiated actions and rescue of Parkinsonian akinesia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-03</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.12.03.692128v1?rss=1">
<title>
<![CDATA[
Striatal ensembles specify and control granular forelimb actions 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.12.03.692128v1?rss=1"
</link>
<description><![CDATA[
The ability of the brain to control specific fine actions is crucial for survival. The striatum is a critical brain center for both movement and learning, and its dysfunction underlies numerous movement disorders1-9. Whereas activity in the striatum has been classically viewed as invigorating5,10-13 and reinforcing movements12-20, recent studies suggest that striatal activity encodes specific movements21-24. However, it is not known how granular this activity is, and if it indeed controls specific ongoing movements. We designed a task where mice performed two minimally-different forelimb actions, consisting of a push or pull isometric force on an immobile joystick, and imaged the activity of medium spiny neurons (MSNs) in the dorsolateral striatum using 2-photon microscopy. We observed that striatal activity encoded both the preparation and execution of specific actions, even when those actions were not reinforced. Furthermore, both populations of D1 and D2-MSNs - classically viewed as promoting versus inhibiting movement2,3,25 - equally encoded action identity. We developed a closed-loop system to model and stimulate action-specific neural ensembles deep in the brain, using holographic optogenetics through a GRIN lens. Stimulation of action-specific ensembles of both D1- and D2-MSNs increased the force of ongoing actions, but only when the ensemble stimulated was congruent with the ongoing action. These results reveal that specific ensembles of both D1- and D2-MSNs causally control specific ongoing actions, as granular as different muscle co-contractions of the same forelimb. Such granularity provides a mechanistic framework for understanding how striatal dysfunction can produce highly specific movement impairments in Huntingtons disease2,9 and dystonia2,6.
]]></description>
<dc:creator>Rodrigues-Vaz, I.</dc:creator>
<dc:creator>Athalye, V. R.</dc:creator>
<dc:creator>Peterka, D. S.</dc:creator>
<dc:creator>Costa, R. M.</dc:creator>
<dc:date>2025-12-08</dc:date>
<dc:identifier>doi:10.64898/2025.12.03.692128</dc:identifier>
<dc:title><![CDATA[Striatal ensembles specify and control granular forelimb actions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.12.19.695584v1?rss=1">
<title>
<![CDATA[
Excitatory nicotinic signaling drives action potential bursting in dopaminergic axons 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.12.19.695584v1?rss=1"
</link>
<description><![CDATA[
Nicotinic acetylcholine receptors (nAChRs) facilitate striatal dopamine transmission but also suppress dopamine release during high-frequency stimulation, suggesting they act as as low pass filters of dopamine release. Because axonal excitability is key a determinant of transmission, we combined axonal recordings and calcium imaging to define the physiological conditions under which nAChRs bidirectionally control dopaminergic axons. Activation of cholinergic interneuron (CINs) recruited nAChRs to enhance dopaminergic axon signals under moderate activation but suppressed signals after strong high-frequency stimulation. Axonal recordings revealed that single-pulse striatal stimulation triggered a rapid ([~]125 Hz) burst of 2-3 nAChR-driven spikes in dopaminergic axons followed by a brief refractory period that inhibited further axon spiking. In sum, we show that nAChRs mainly enhance local excitability of striatal dopaminergic axons but also trigger axonal bursting that suppresses axonal excitability. This mechanism expands the computational power of dopaminergic axons and explains the apparent nAChR-mediated low-pass filtering of dopamine release.
]]></description>
<dc:creator>Kramer, P. F.</dc:creator>
<dc:creator>Yanez, A.</dc:creator>
<dc:creator>Clever, F.</dc:creator>
<dc:creator>Zhang, R.</dc:creator>
<dc:creator>Khaliq, Z. M.</dc:creator>
<dc:date>2025-12-21</dc:date>
<dc:identifier>doi:10.64898/2025.12.19.695584</dc:identifier>
<dc:title><![CDATA[Excitatory nicotinic signaling drives action potential bursting in dopaminergic axons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.12.21.695842v1?rss=1">
<title>
<![CDATA[
Incentive valence differentially engages open- and closed-loop basal ganglia circuits during movement initiation 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.12.21.695842v1?rss=1"
</link>
<description><![CDATA[
Incentives modulate voluntary movement, yet the circuitry channeling these signals into motor output remains unclear. Classical models emphasize a closed-loop circuit (CLC) linking dorsal putamen (PUTd) with motor cortex, but this pathway is anatomically segregated from affective processing regions. Anatomical and clinical evidence point to an alternative: an open-loop circuit (OLC) from ventral putamen (PUTv) that may route affective signals to motor cortex. Here, we conducted two experiments to test whether a functional OLC exists in humans and whether it is differentially engaged by incentive conditions. First, in 7 T resting-state fMRI (multi-echo), PUTv showed robust functional connectivity with both affective and motor regions, including the cingulate motor area (CMA), even after accounting for PUTd variance. This connectivity pattern supports the plausibility of an independent pathway linking affective basal ganglia regions to the motor cortex. Second, in 3 T task fMRI (incentivized reaching), jackpot (high-reward) and robber (high-loss avoidance) incentive conditions produced distinct behavioral and neural signatures. Jackpot produced a speed-accuracy trade-off, with faster movement initiation but more false starts. Neurally, this coincided with engagement (BOLD responses relevant for initiation speed) being reduced in CLC nodes but not in OLC. Robber, in contrast, eliminated engagement in both OLC and CLC nodes, instead recruiting stopping-related regions (e.g., STN), consistent with an avoidance phenomenology. Together, these findings support a versatile architecture for movement initiation that flexibly engages distinct cortico-subcortical circuits depending on incentive phenomenology, and offer a candidate mechanism through which affective salience and valence modulate voluntary movement.

Significance StatementAffective signals profoundly influence movement, yet the mechanisms linking motivationally relevant contexts with motor behavior remain unclear. Combining ultra-high field (7 T) connectomics with task-based (3 T) neuroimaging, we provide the first systems-level evidence in humans for such a mechanism: a ventral putamen-centered open-loop circuit (OLC) connecting affective and motor areas, operating alongside the canonical dorsal putamen-centered closed-loop sensorimotor circuit (CLC). Critically, the phenomenological quality of incentive (how it is construed as reward versus threat) rather than magnitude alone, likely determines which circuit dominates during movement initiation. These findings help to explain paradoxical kinesia in Parkinsons disease, where affective contexts can bypass degraded sensorimotor circuits, and establish foundations for context-based therapeutic interventions.
]]></description>
<dc:creator>Dundon, N. M.</dc:creator>
<dc:creator>Rizor, E. J.</dc:creator>
<dc:creator>Stasiak, J.</dc:creator>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Li, T.</dc:creator>
<dc:creator>Sabugo, K.</dc:creator>
<dc:creator>Villanueva, C.</dc:creator>
<dc:creator>Barandon, P.</dc:creator>
<dc:creator>Babenko, V.</dc:creator>
<dc:creator>Beverly-Aylwin, R.</dc:creator>
<dc:creator>Stump, A.</dc:creator>
<dc:creator>Santander, T.</dc:creator>
<dc:creator>Bostan, A. C.</dc:creator>
<dc:creator>Lapate, R. C.</dc:creator>
<dc:creator>Grafton, S. T.</dc:creator>
<dc:date>2025-12-23</dc:date>
<dc:identifier>doi:10.64898/2025.12.21.695842</dc:identifier>
<dc:title><![CDATA[Incentive valence differentially engages open- and closed-loop basal ganglia circuits during movement initiation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-23</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.12.29.696920v1?rss=1">
<title>
<![CDATA[
Cardiac-sympathetic state predicts action restraint, gated by demonstrated agency 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.12.29.696920v1?rss=1"
</link>
<description><![CDATA[
Withholding action until the appropriate moment is a core challenge of motivated behavior. Using beat-to-beat cardiac contractility during an incentivized reaching task, we show that cardiac-sympathetic outflow predicts action restraint. Under high-reward conditions that induce a speed-accuracy tradeoff, reduced contractility at the time of instruction preceded premature responses (false starts). Under high-loss-avoidance conditions, elevated pre-movement contractility predicted slower, more controlled initiation, but only among participants with above-median task success. These findings suggest cardiac-sympathetic engagement does not simply serve mobilization but flexibly supports context-appropriate action regulation, with recruitment for restraint gated by demonstrated agency.
]]></description>
<dc:creator>Dundon, N. M.</dc:creator>
<dc:creator>Rizor, E. J.</dc:creator>
<dc:creator>Stasiak, J.</dc:creator>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Babenko, V.</dc:creator>
<dc:creator>Lapate, R. C.</dc:creator>
<dc:creator>Grafton, S. T.</dc:creator>
<dc:date>2025-12-31</dc:date>
<dc:identifier>doi:10.64898/2025.12.29.696920</dc:identifier>
<dc:title><![CDATA[Cardiac-sympathetic state predicts action restraint, gated by demonstrated agency]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-12-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.12.31.697225v1?rss=1">
<title>
<![CDATA[
System-wide profiling reveals metabolic alterations in a G2019S LRRK2 knockin mouse model of Parkinson's disease: systemic depletion of pyrimidine nucleosides 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.12.31.697225v1?rss=1"
</link>
<description><![CDATA[
BackgroundThe G2019S mutation in LRRK2 is the most common genetic cause of familial Parkinsons disease (PD) and is associated with increased susceptibility to sporadic PD, which likely arises from the interplay of genetic predisposition, environmental exposure, and aging. Metabolic syndrome (MetS) has been implicated as an independent risk factor for PD, but the potential interaction between the G2019S LRRK2 mutation and metabolic stress in disease pathogenesis remains unclear.

MethodsWe employed long-term high-fat diet (HFD) feeding to induce metabolic syndrome in aged LRRK2 mutant mice, followed by system-wide characterization using multi-omic approaches, including metabolomic profiling, proteomics, bulk RNA sequencing, and single-nucleus RNA sequencing.

ResultsWe find that thymidine and deoxyuridine levels are consistently reduced across tissues in G2019S LRRK2 knockin mice at baseline, accompanied by increased hepatic expression of thymidine phosphorylase (TP). HFD exposure further unmasks disruptions in purine and energy metabolism in the brain and lungs of G2019S LRRK2 knockin mice, with astrocytes and oligodendrocytes in the ventral midbrain exhibiting the most pronounced impairment in oxidative phosphorylation transcriptional pathways.

ConclusionsOur findings demonstrate that pre-existing metabolic syndrome unmasks widespread disruptions in systemic nucleotide and energy metabolism and exacerbates mitochondrial dysfunction in G2019S LRRK2 knockin mice. This conditional "two-hit" phenotype underscores the critical role of environmental factors, such as diet, in revealing metabolic vulnerabilities associated with PD-linked genetic backgrounds. Our study provides novel insight into potential metabolic targets or pathways for therapeutic intervention in individuals at-risk for developing PD.
]]></description>
<dc:creator>Ma, Y.</dc:creator>
<dc:creator>Erb, M. L.</dc:creator>
<dc:creator>Sipple, K.</dc:creator>
<dc:creator>Offerman, A.</dc:creator>
<dc:creator>Moore, D. J.</dc:creator>
<dc:date>2026-01-02</dc:date>
<dc:identifier>doi:10.64898/2025.12.31.697225</dc:identifier>
<dc:title><![CDATA[System-wide profiling reveals metabolic alterations in a G2019S LRRK2 knockin mouse model of Parkinson's disease: systemic depletion of pyrimidine nucleosides]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-02</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.01.07.694713v1?rss=1">
<title>
<![CDATA[
Myeloid PINK1 represses mtDNA release and immune signaling that impacts neuronal pathology in patient-derived idiopathic PD models 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.01.07.694713v1?rss=1"
</link>
<description><![CDATA[
Parkinsons disease (PD) is a neurodegenerative disorder marked by the development of cardinal motor deficits preceded by a protracted prodromal period of non-motor symptoms often involving the gastrointestinal (GI) tract. There is an emerging consensus that both the peripheral immune system and local neuroinflammation play key roles in the etiology of PD. We previously demonstrated a critical function for the Parkinsons related proteins PINK1 and Parkin as repressors of the innate to adaptive immune response in cultured cells and mouse models of infection. However, it remained unclear whether these processes were conserved in patient-derived models, and precisely how immune signaling may ultimately drive the death of dopaminergic neurons. Here we show that GI infection of PINK1 knockout (KO) mice triggered acute neurodegeneration which was evident early in the enteric nervous system. Treating wild type enteric or dopaminergic neurons with conditioned medium from immune-stimulated PINK1 KO macrophages was sufficient to promote neuronal disruption in both mouse and human neurons in vitro. Within immune-activated macrophages, we reveal that loss of PINK1 led to an enhanced release of mitochondrial DNA (mtDNA) within mitochondrial derived vesicles, leading to the activation of cGAS/STING pathways. These changes were seen in both mouse/human in vitro models and in PD patient-derived primary macrophages. Notably, pharmacological modulation using a PINK1 activator with high therapeutic potential attenuated pro-inflammatory profiles elicited by the mtDNA-dependent STING/NF-{kappa}B pathway in idiopathic patient-derived macrophages. Ultimately, our study lays the foundation for understanding PINK1-related peripheral macrophage mechanisms in idiopathic PD and provides a target for further development to treat the disease at early stages.

Graphical abstractPINK1-related immune mechanisms of Parkinsons disease and associations with early neurodegenerative events.



O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=120 SRC="FIGDIR/small/694713v1_ufig1.gif" ALT="Figure 1">
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org.highwire.dtl.DTLVardef@6f6488org.highwire.dtl.DTLVardef@13139aaorg.highwire.dtl.DTLVardef@c0d4d1org.highwire.dtl.DTLVardef@1d41e25_HPS_FORMAT_FIGEXP  M_FIG C_FIG
]]></description>
<dc:creator>Recinto, S. J.</dc:creator>
<dc:creator>MacDonald, A.</dc:creator>
<dc:creator>Premachandran, S.</dc:creator>
<dc:creator>Liu, L.</dc:creator>
<dc:creator>Bayati, A.</dc:creator>
<dc:creator>Rodriguez, L.</dc:creator>
<dc:creator>Nguyen, M.</dc:creator>
<dc:creator>Petit, F.</dc:creator>
<dc:creator>Mukherjee, S.</dc:creator>
<dc:creator>Larmanjat, J.</dc:creator>
<dc:creator>Allot, A.</dc:creator>
<dc:creator>Yaqubi, M.</dc:creator>
<dc:creator>McPherson, P. S.</dc:creator>
<dc:creator>Durcan, T. M.</dc:creator>
<dc:creator>Gruenheid, S.</dc:creator>
<dc:creator>Trudeau, L.-E.</dc:creator>
<dc:creator>Drouin-Ouellet, J.</dc:creator>
<dc:creator>McBride, H. M.</dc:creator>
<dc:creator>Stratton, J. A.</dc:creator>
<dc:date>2026-01-08</dc:date>
<dc:identifier>doi:10.64898/2026.01.07.694713</dc:identifier>
<dc:title><![CDATA[Myeloid PINK1 represses mtDNA release and immune signaling that impacts neuronal pathology in patient-derived idiopathic PD models]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.01.12.699117v1?rss=1">
<title>
<![CDATA[
Location and mapping of the human rostromedial tegmental nucleus and associated midbrain inhibitory nuclei regulating dopamine neurons 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.01.12.699117v1?rss=1"
</link>
<description><![CDATA[
BackgroundAnimal experiments reveal distinct GABAergic cell clusters within the dopaminergic midbrain regions in the rostromedial tegmental nucleus (RMTg) and retrorubral fields (RRF) that have yet to be clearly defined in humans. These neurons send prominent inhibitory projections to dopaminergic neurons in the substantia nigra and ventral tegmental area that impact motor, reward and threat processing.

ObjectiveTo identify GABergic RMTg and RRF cell clusters in the human brain.

MethodsTen {micro}m formalin-fixed paraffin-embedded transverse human brainstem sections from ten control cases were obtained from the Sydney Brain Bank and immunohistochemistry for GABA and tyrosine hydroxylase was used to determine the location and cell size of RMTg and RRF GABAergic neurons. These cell populations were further mapped in transverse 50{micro}m thick cresyl violet stained serial midbrain sections (every 750{micro}m) from previously published controls (Halliday et al. 1990a).

ResultsGABAergic neurons were cytoarchitecturally distinct, with the largest GABAergic neurons in the RRF, followed by RMTg neurons which were larger than GABAergic neurons in the interpedunclular nucleus (Kruskal-Wallis test, p<0.0001). RMTg and RRF GABAergic neurons first appear in caudal transverse midbrain sections approximately 38mm above the obex. RMTg moves rostrally and medially from underneath the decussation of the superior cerebellar peduncle to just lateral to the interpeduncular nucleus. The RRF cluster also moves rostrally and medially to the parabrachial pigmented nucleus (PBP) just under the red nucleus.

ConclusionThe GABAergic neurons in RMTg and RRF/PBP that modulate dopamine neuronal excitability have distinct morphologies in humans. Identifying these inhibitory neurons is key to evaluating their role in neurodegenerative diseases.
]]></description>
<dc:creator>Filimontseva, A.</dc:creator>
<dc:creator>Fu, Y.</dc:creator>
<dc:creator>Halliday, G.</dc:creator>
<dc:date>2026-01-12</dc:date>
<dc:identifier>doi:10.64898/2026.01.12.699117</dc:identifier>
<dc:title><![CDATA[Location and mapping of the human rostromedial tegmental nucleus and associated midbrain inhibitory nuclei regulating dopamine neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-12</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.01.16.699530v1?rss=1">
<title>
<![CDATA[
In vivo Proximity & Spatial Proteomics with CRISPR Screening Identify STXBP1 as a Protective Modifier of a-synuclein Toxicity in Dopamine Neurons. 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.01.16.699530v1?rss=1"
</link>
<description><![CDATA[
Parkinsons disease (PD) is a disease of adults involving the loss of dopaminergic neurons after a long, asymptomatic, prodromal period. -synuclein, LRRK2, and VPS35 are linked to familial PD, however, how these mutations predispose dopamine neurons to death during the early prodromal phases remains unclear. Here, we used in vivo native proximity proteomics (iBioID) and dopaminergic neuron-specific subcellular proteomics across multiple PD models to uncover early alterations preceding neuronal loss. Our analyses identified convergent disruptions in synaptic protein abundance, indicating that presynaptic trafficking defects are early events in PD pathogenesis. Using a targeted CRISPR-based genetic screen in dopamine neurons, we demonstrated that mimicking this misregulation of STXBP1 amplifies vulnerability to -synuclein, implicating it as a previously underappreciated toxicity buffering factor. These findings highlight convergent mechanisms that sensitize dopamine neuronal degeneration and that presynaptic vesicle SNARE-complex proteins could serve as key targets for disease-modifying therapies in PD and related neurodegenerative disorders.

HighlightsO_LIIn vivo native-BioID mapping of multiple Parkinsons disease (PD) protein interactomes revealed a convergent presynaptic network.
C_LIO_LIiBioID analysis on mutant PD proteins (-synuclein A30P, LRRK2 G2019S, VPS35 D620N) uncovered mutation-specific shifts in local proximity networks, notably in endocytic and vesicle recycling pathways.
C_LIO_LISpatial proteomics (iBioCoFrac) of dopamine neurons in vivo identified functional modules with reduced levels of key synaptic proteins in PD models.
C_LIO_LIComparative proteomics using iBioCoFrac revealed synaptic vesicle regulation as a primary site of molecular convergence and early molecular signatures in dopamine neurons across multiple PD mouse models.
C_LIO_LIAn in vivo CRISPR screen pinpointed the presynaptic protein Stxbp1/Munc18-1 as an -synuclein toxicity modifier in dopaminergic neurons.
C_LI
]]></description>
<dc:creator>Shonai, D.</dc:creator>
<dc:creator>Kent, J.</dc:creator>
<dc:creator>Okafor, A.</dc:creator>
<dc:creator>Gao, Y.</dc:creator>
<dc:creator>Parameswaran, P.</dc:creator>
<dc:creator>Bustamante, E.</dc:creator>
<dc:creator>Ramani, B.</dc:creator>
<dc:creator>Diao, Y.</dc:creator>
<dc:creator>Kampmann, M.</dc:creator>
<dc:creator>Soderblom, E. J.</dc:creator>
<dc:creator>Soderling, S. H.</dc:creator>
<dc:date>2026-01-17</dc:date>
<dc:identifier>doi:10.64898/2026.01.16.699530</dc:identifier>
<dc:title><![CDATA[In vivo Proximity & Spatial Proteomics with CRISPR Screening Identify STXBP1 as a Protective Modifier of a-synuclein Toxicity in Dopamine Neurons.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-17</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.01.17.700056v1?rss=1">
<title>
<![CDATA[
Cathepsin-dependent amyloid formation drives mechanical rupture of lysosomal membranes 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.01.17.700056v1?rss=1"
</link>
<description><![CDATA[
Lysosomal membrane integrity is essential for cellular homeostasis, and its failure drives lysosomal storage disorders (LSD) and neurodegeneration. The dipeptide L-leucyl-L-leucine methyl ester (LLOMe) is widely used to model lysosomal damage, yet its mechanism remains poorly understood. The prevailing view holds that LLOMe polymerizes into membrane-permeabilizing peptide chains within the lysosomal lumen. Using cryo-electron tomography in cultured cells and primary neurons, we visualized the structural basis of LLOMe-induced lysosomal damage. We reveal that LLOMe forms amyloid structures within lysosomes that directly interact with and rupture the limiting membrane through mechanical stress. In vitro reconstitution confirms this amyloid-mediated mechanism. These findings establish a structural paradigm for lysosomal membrane disruption and provide insights into how disease-relevant protein aggregates, implicated in neurodegeneration and LSD, may compromise lysosomal integrity.
]]></description>
<dc:creator>Li, D.</dc:creator>
<dc:creator>Zhang, W.</dc:creator>
<dc:creator>Medina, M.</dc:creator>
<dc:creator>Stuke, J. F. M.</dc:creator>
<dc:creator>Schwarz, A.</dc:creator>
<dc:creator>Brill, J.</dc:creator>
<dc:creator>Brenner, J.</dc:creator>
<dc:creator>Kraus, F.</dc:creator>
<dc:creator>Ohlerich, S.</dc:creator>
<dc:creator>Lizarrondo, J.</dc:creator>
<dc:creator>Pflaum, J.</dc:creator>
<dc:creator>Grass, J. H.</dc:creator>
<dc:creator>Soltow, L.-M.</dc:creator>
<dc:creator>Hammerschmid, D.</dc:creator>
<dc:creator>Weber, N.</dc:creator>
<dc:creator>Welsch, S.</dc:creator>
<dc:creator>Langer, J.</dc:creator>
<dc:creator>Windbergs, M.</dc:creator>
<dc:creator>Harper, W. J.</dc:creator>
<dc:creator>Schuman, E.</dc:creator>
<dc:creator>Hummer, G.</dc:creator>
<dc:creator>Grotjahn, D. A.</dc:creator>
<dc:creator>Wilfling, F.</dc:creator>
<dc:date>2026-01-19</dc:date>
<dc:identifier>doi:10.64898/2026.01.17.700056</dc:identifier>
<dc:title><![CDATA[Cathepsin-dependent amyloid formation drives mechanical rupture of lysosomal membranes]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-19</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.01.20.700614v1?rss=1">
<title>
<![CDATA[
An anatomical hotspot for striatal dopamine-acetylcholine interactions during reward and movement 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.01.20.700614v1?rss=1"
</link>
<description><![CDATA[
Dopamine (DA) and acetylcholine (ACh) are key neuromodulators that regulate striatal circuits underlying movement and reinforcement learning. Evidence suggests that DA and ACh systems interact, but where and how interactions are expressed across striatal regions in behaviorally relevant release dynamics remains unknown. We applied micro-fiber arrays to simultaneously measure striatum-wide DA and ACh in behaving mice, revealing an anatomical organization in which DA-ACh anti-correlations were concentrated in a hotspot in the anterior dorsolateral striatum (aDLS). Anti-correlations resulted from temporally coincident pairs of spontaneous and event-locked transient peak and dip events occurring in a DA[-&gt;]ACh sequence. The aDLS localized hotspot was consistently expressed within distinct signals associated with unpredicted rewards, learned and extinguished Pavlovian cues, and locomotion initiation and invigoration phases, for which we revealed novel, opposing DA-ACh dynamics. Optogenetic activation of DA neurons selectively suppressed spontaneous ACh release within the aDLS hotspot, and ex vivo recordings revealed enhanced D2-mediated ACh inhibition in aDLS relative to ventral regions, suggesting a mechanistic basis for this spatial specificity. These findings demonstrate that DA-ACh interactions during behavior are spatially organized, rather than uniformly conserved, and shape behaviorally relevant dynamics to regulate region-specific functions in learning and movement control.
]]></description>
<dc:creator>Bouabid, S.</dc:creator>
<dc:creator>Vu, M.-A. T.</dc:creator>
<dc:creator>Noggle, C.</dc:creator>
<dc:creator>Vietti-Michelina, S.</dc:creator>
<dc:creator>Brimblecombe, K.</dc:creator>
<dc:creator>Platt, N.</dc:creator>
<dc:creator>Zhang, L.</dc:creator>
<dc:creator>Joshi, A.</dc:creator>
<dc:creator>Cragg, S.</dc:creator>
<dc:creator>Howe, M. W.</dc:creator>
<dc:date>2026-01-21</dc:date>
<dc:identifier>doi:10.64898/2026.01.20.700614</dc:identifier>
<dc:title><![CDATA[An anatomical hotspot for striatal dopamine-acetylcholine interactions during reward and movement]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-21</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2025.11.06.687049v1?rss=1">
<title>
<![CDATA[
Parkinsons disease modeling in regenerative spiny mice (Acomys dimidiatus) captures key disease-relevant behavioral, histological, and molecular signatures 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2025.11.06.687049v1?rss=1"
</link>
<description><![CDATA[
Parkinsons disease (PD) is a multifactorial neurodegenerative disorder that has been modeled extensively in animals, primarily rodents, but also in non-human primates and non-mammalian organisms. However, no single animal model fully recapitulates the hallmarks of PD pathology. Here, we extend this work by modeling PD for the first time in the spiny mouse (Acomys dimidiatus), a mammal notable for its robust regeneration of multiple tissues. We show that the nigrostriatal pathway of A. dimidiatus is vulnerable to both acute 6-hydroxydopamine (6-OHDA) toxicity and chronic -synuclein (Syn) preformed fibril-induced aggregation. Mouse Syn PFFs produced widespread pS129-positive Syn inclusions across multiple brain regions, mirroring a key pathological hallmark of PD. Compared to C57BL/6J mice, A. dimidiatus exhibited more pronounced behavioral impairments, greater nigrostriatal degeneration, and higher pS129-Syn inclusion burden within substantia nigra pars compacta (SNpc) neurons. To probe the molecular underpinnings behind the vulnerability, we performed single-cell spatial proteomics, which revealed extensive proteomic alterations in dopaminergic neurons associated with Syn aggregation. Multiple proteins were dysregulated in A. dimidiatus, including those involved in proteasomal function, mitochondrial pathways, and oxidative stress regulation, which are processes commonly implicated in PD. Notably, proteomic analysis identified heightened astrocytic activation in the SNpc, which we validated histologically, suggesting a distinct glial response compared to mice. Together, these findings expand our understanding of PD-relevant pathophysiology across species and establish A. dimidiatus as a model for studying mechanisms of neurodegeneration.
]]></description>
<dc:creator>Dutta, S.</dc:creator>
<dc:creator>Pang, M.</dc:creator>
<dc:creator>Donahue, R. R.</dc:creator>
<dc:creator>Chou, T.-F.</dc:creator>
<dc:creator>Seifert, A. W.</dc:creator>
<dc:creator>Gradinaru, V.</dc:creator>
<dc:date>2025-11-08</dc:date>
<dc:identifier>doi:10.1101/2025.11.06.687049</dc:identifier>
<dc:title><![CDATA[Parkinsons disease modeling in regenerative spiny mice (Acomys dimidiatus) captures key disease-relevant behavioral, histological, and molecular signatures]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2025-11-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.01.28.702423v1?rss=1">
<title>
<![CDATA[
Machine learning-based image analysis of Parkinson's disease iPS-derived neurons predicts genotype and reveals mitochondria-lysosome abnormalities 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.01.28.702423v1?rss=1"
</link>
<description><![CDATA[
Mitochondrial and lysosomal dysfunction are central features of Parkinsons disease (PD) across major genetic forms including PRKN, SNCA, and LRRK2. We applied cell morphomics, a machine-learning-based framework combining high-content imaging with quantitative feature extraction, to analyse mitochondrial and lysosomal morphology at single-cell resolution in iPS cell-derived cortical neurons from PD patients and healthy controls (13 lines total). Supervised machine-learning models distinguished PD neurons from controls with high accuracy (AUC = 0.87) and reliably separated individual genotypes. Feature importance and attribution analysis revealed genotype-specific organelle biases, with mitochondrial features dominating classification in PRKN neurons, balanced mitochondrial and lysosomal contributions in SNCA neurons, and a greater lysosomal contribution in LRRK2 neurons. Multi-class models retained strong performance, and findings were reproduced across two independent laboratories using different dyes and imaging conditions. These results demonstrate that morphomics provides a robust and scalable framework to quantify genotype-specific organelle abnormalities in PD neurons and supports its application for cellular stratification and biomarker discovery.
]]></description>
<dc:creator>Li, Y.</dc:creator>
<dc:creator>Powell, M.</dc:creator>
<dc:creator>Chedid, J.</dc:creator>
<dc:creator>Sutharsan, R.</dc:creator>
<dc:creator>Garrido, A. L.</dc:creator>
<dc:creator>Abu-Bonsrah, D.</dc:creator>
<dc:creator>Pavan, C.</dc:creator>
<dc:creator>Fraser, T.</dc:creator>
<dc:creator>Ovchinnikov, D.</dc:creator>
<dc:creator>Zhong, M.</dc:creator>
<dc:creator>Davis, R.</dc:creator>
<dc:creator>Strbenac, D.</dc:creator>
<dc:creator>Johnston, J. A.</dc:creator>
<dc:creator>Thompson, L. H.</dc:creator>
<dc:creator>Kirik, D.</dc:creator>
<dc:creator>Parish, C. L.</dc:creator>
<dc:creator>Halliday, G. M.</dc:creator>
<dc:creator>Sue, C. M.</dc:creator>
<dc:creator>Dzamko, N.</dc:creator>
<dc:creator>Wali, G.</dc:creator>
<dc:date>2026-01-29</dc:date>
<dc:identifier>doi:10.64898/2026.01.28.702423</dc:identifier>
<dc:title><![CDATA[Machine learning-based image analysis of Parkinson's disease iPS-derived neurons predicts genotype and reveals mitochondria-lysosome abnormalities]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-01-29</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.02.19.706842v1?rss=1">
<title>
<![CDATA[
Distinct amygdalar pathways to frontopolar cortex and medial temporal lobe support temporal memory for emotional events 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.02.19.706842v1?rss=1"
</link>
<description><![CDATA[
Growing evidence suggests that emotion shapes temporal aspects of memory--such as remembering when an event occurred--yet the neural bases of these effects remain unclear. Prior work indicates that temporal-context representations in memory are supported by the function of medial temporal lobe (MTL) and prefrontal regions with privileged access to amygdala-originated affective signals, which may mediate emotional influences on temporal memory--including hippocampus, perirhinal cortex, and lateral frontopolar cortex (FPl). To test whether these candidate pathways shape temporal memory for emotional events, participants encoded trial-unique negative and positive images in the scanner, followed by a surprise temporal memory task. We found that more negative stimuli were associated with greater amygdala activation and lower temporal-memory error for when emotional images occurred. Replicating and extending prior work, higher stimulus-evoked multivoxel pattern similarity in the hippocampus (between trials) and FPl (within and between trials) was associated with higher temporal memory errors. Notably, greater amygdala responding to emotional images predicted higher between-trial pattern similarity in the hippocampus, but lower within- and between-trial similarity in FPl. Amygdala engagement was also correlated with greater perirhinal activation, which in turn predicted stronger item memory and a recency bias in temporal memory estimates. Collectively, these findings reveal distinct--and opposing--modulatory effects of amygdala-FPl and amygdala-MTL pathways on temporal memory, with the potential to sharpen and blur memory for when emotional events occurred.

Significance statementEmotional events influence how we remember time. Prior work implicates function of the lateral frontal pole (FPl) and medial temporal lobe (MTL) regions--including hippocampus and perirhinal cortex--in temporal memory. Yet, whether and how these areas support temporal memory for emotional events remains unclear. Using trial-unique emotional images, we found that negative-emotional valence was associated with stronger amygdala activation and temporal-memory accuracy. Higher pattern similarity in hippocampus and FPl predicted worse temporal memory. Notably, amygdala activation predicted higher between-trial hippocampal pattern similarity but lower within-trial FPl similarity. Perirhinal engagement, which correlated with amygdala activation and item memory, predicted a recency bias in temporal-memory judgments. Together, these findings reveal distinct amygdala-MTL and amygdala-prefrontal pathways through which emotion shapes temporal memory.
]]></description>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Stasiak, J. S.</dc:creator>
<dc:creator>Dundon, N. M.</dc:creator>
<dc:creator>Rizor, E.</dc:creator>
<dc:creator>Barandon, P. L.</dc:creator>
<dc:creator>Villanueva, C. M.</dc:creator>
<dc:creator>Grafton, S. T.</dc:creator>
<dc:creator>Lapate, R. C.</dc:creator>
<dc:date>2026-02-20</dc:date>
<dc:identifier>doi:10.64898/2026.02.19.706842</dc:identifier>
<dc:title><![CDATA[Distinct amygdalar pathways to frontopolar cortex and medial temporal lobe support temporal memory for emotional events]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-20</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.02.25.708014v1?rss=1">
<title>
<![CDATA[
Selective loss of Primary Cilia and Neurotrophic Signaling in G51D alpha-Synuclein Mice Highlights a Common Pathway to Parkinsons Disease 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.02.25.708014v1?rss=1"
</link>
<description><![CDATA[
Parkinsons disease is characterized by dopaminergic neuron loss and accumulation of -synuclein aggregates in the brain. G51D -synuclein knock-in mice provide a genetically and clinically relevant model of disease, exhibiting early olfactory deficits, age-dependent motor impairment, and progressive phospho--synuclein accumulation. In multiple Parkinsons disease models, striatal cholinergic and parvalbumin interneurons, as well as astrocytes, lose primary cilia and the neurotrophic signaling needed to sustain dopaminergic neurons. We show here that G51D--synuclein mice share these phenotypes. Phospho-Ser129 -synuclein accumulation correlates with cilia loss in cholinergic interneurons but not in medium spiny neurons that accumulate higher phospho--synuclein levels. In the piriform cortex, parvalbumin neurons lose primary cilia and downregulate Neurturin, potentially contributing to olfactory dysfunction. Within the peripheral olfactory epithelium, horizontal basal cells lose cilia, whereas multi-ciliated olfactory sensory neuron cilia remain intact. These findings reveal convergent cellular vulnerabilities across Parkinsons disease models and highlight a pathogenic role for impaired ciliary signaling.

TeaserLoss of primary cilia may contribute to dopamine neuron loss in both inherited and common Parkinsons disease.
]]></description>
<dc:creator>Lin, Y.-E.</dc:creator>
<dc:creator>Jaimon, E.</dc:creator>
<dc:creator>Kim, Y.</dc:creator>
<dc:creator>Loftman, A.</dc:creator>
<dc:creator>Vijayakumaran, A.</dc:creator>
<dc:creator>Belfort, B. D.</dc:creator>
<dc:creator>Chiang, C. Y.</dc:creator>
<dc:creator>Arenkiel, B. R.</dc:creator>
<dc:creator>Zoghbi, H. Y. Y.</dc:creator>
<dc:creator>Pfeffer, S. R.</dc:creator>
<dc:date>2026-02-25</dc:date>
<dc:identifier>doi:10.64898/2026.02.25.708014</dc:identifier>
<dc:title><![CDATA[Selective loss of Primary Cilia and Neurotrophic Signaling in G51D alpha-Synuclein Mice Highlights a Common Pathway to Parkinsons Disease]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-25</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://biorxiv.org/cgi/content/short/2026.02.23.707545v1?rss=1">
<title>
<![CDATA[
Neural Control of Autonomic Arousal During Threat Anticipation Revealed by High-Resolution Cardiac Contractility 
]]>
</title>
<link>
https://biorxiv.org/cgi/content/short/2026.02.23.707545v1?rss=1"
</link>
<description><![CDATA[
The sympathetic nervous system prepares the organism for adaptive action by shaping physiological, affective, and behavioral responses to environmental demands. Yet, how sympathetic signals dynamically couple with neural systems supporting emotional experience and behavior remains poorly understood, in part because common indices such as skin conductance responses lack sufficient temporal resolution to track these dynamics. Here, we evaluated trans-radial electrical bioimpedance velocimetry (TREV), a non-invasive measure of beat-to-beat cardiac contractility, and compared it with skin conductance responses during threat anticipation and simultaneous fMRI. Participants (n=60) completed a threat-of-shock paradigm requiring goal-directed action. Cardiac contractility increased during threat anticipation, covaried with skin conductance responses, and independently predicted self-reported emotional intensity. Critically, threat-related increases in contractility--but not skin conductance--tracked threat-related modulation of activation in dorsomedial prefrontal cortex, posterior parietal cortex, and cerebellum, with contractility-modulated cerebellar activation predicting faster motor responses under threat. These findings establish TREV-derived cardiac contractility as a physiological signal linking sympathetic drive with neural responding, emotional experience, and adaptive behavioral mobilization during emotion-guided action.
]]></description>
<dc:creator>Stasiak, J. E.</dc:creator>
<dc:creator>Wang, J.</dc:creator>
<dc:creator>Dundon, N. M.</dc:creator>
<dc:creator>Rizor, E. J.</dc:creator>
<dc:creator>Barandon, P. L.</dc:creator>
<dc:creator>Villanueva, C. M.</dc:creator>
<dc:creator>Babenko, V.</dc:creator>
<dc:creator>Li, T. L.</dc:creator>
<dc:creator>Sabugo, K. M.</dc:creator>
<dc:creator>Grafton, S. T.</dc:creator>
<dc:creator>Lapate, R. C.</dc:creator>
<dc:date>2026-02-26</dc:date>
<dc:identifier>doi:10.64898/2026.02.23.707545</dc:identifier>
<dc:title><![CDATA[Neural Control of Autonomic Arousal During Threat Anticipation Revealed by High-Resolution Cardiac Contractility]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory Press</dc:publisher>
<prism:publicationDate>2026-02-26</prism:publicationDate>
<prism:section></prism:section>
</item>
</rdf:RDF>
