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Spatially and temporally distinct patterns of expression for VPS10P domain receptors in human cerebral organoids.

ABSTRACT: Spatially and temporally distinct patterns of expression for VPS10P domain receptors in human cerebral organoids.

PROVIDER: PRJNA976668 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR24746235_1.fastq.gz	Fastqsanger.gz
	SRR24746235_2.fastq.gz	Fastqsanger.gz
	SRR24746236_1.fastq.gz	Fastqsanger.gz
	SRR24746236_2.fastq.gz	Fastqsanger.gz

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Spatially and temporally distinct patterns of expression for VPS10P domain receptors in human cerebral organoids

Project description:Spatially and temporally distinct patterns of expression for VPS10P domain receptors in human cerebral organoids

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Spatially and temporally distinct patterns of expression for VPS10P domain receptors in human cerebral organoids

Project description:Vacuolar protein sorting 10 protein (VPS10P) domain receptors are a unique class of intracellular sorting receptors that emerge as major risk factors associated with psychiatric and neurodegenerative diseases, including bipolar disorders, autism, schizophrenia, as well as Alzheimer’ disease and frontotemporal dementia. Yet, the lack of suitable experimental models to study receptor functions in the human brain have hampered elucidation of receptor actions in brain disease. Here, we have adapted protocols to the study of VPS10P domain receptors in human cerebral organoids and applied these to the detailed characterization of receptor expression during development and in mature tissue, including single-cell RNA sequencing. These studies uncovered spatial and temporal patterns of expression unique to individual receptor species in the human brain. While SORL1 expression is abundant in stem cells and SORCS1 peaks at onset of neurogenesis, SORT1 and SORCS2 show increasing expression coinciding with maturation of neuronal and non-neuronal cell types, arguing for distinct functions in development versus the adult brain. In neurons, subcellular localization also distinguishes between two types of receptors, either mainly localized to the cell soma (SORL1 and SORT1) or also to neuronal projections (SORCS1 and SORCS2), suggesting divergent functions in protein sorting between Golgi and the endo-lysosomal system or along axonal and dendritic tracks. Taken together, our findings provide an important resource concerning temporal, spatial, and subcellular patterns of VPS10P domain receptor expression in cerebral organoids for further elucidation of receptor (dys)functions causative of behavioral and cognitive defects of the human brain.

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DIURNAL RHYTHMS SPATIALLY AND TEMPORALLY ORGANIZE AUTOPHAGY

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A method for spatially and temporally resolved protein network interrogation in living cells

Project description:Cells operate through protein interaction networks organized in space and in time, but current methods to interrogate such biology typically resolve only one of these dimensions. Here, we describe a method to resolve both dimensions simultaneously using proximity labeling mediated by engineered ascorbic acid peroxidase (APEX). APEX has been used to label organelle proteomes with high temporal resolution, but its spatial resolution is generally thought to be limited because of its larger labeling radius. We describe an analytical pipeline, based on quantitative proteomics using spatial references, which overcomes this barrier. As proof-of-principle, we apply the methodology to interrogate protein interaction networks of G protein coupled receptors both temporally and spatially. Using this approach, we identify known binding partners as well as novel receptor regulators. Validating the methodology as a discovery tool, we demonstrate that two of these components drive ubiquitin-linked down-regulation of opioid receptors.

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