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Diverse gene expression of diencephalon astrocytes reveals proliferation regulated by Smad4 - scRNA-seq II

ABSTRACT: Diverse gene expression of diencephalon astrocytes reveals proliferation regulated by Smad4 - scRNA-seq II

PROVIDER: PRJNA736322 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR14766504.fastq.gz	Fastqsanger.gz
	SRR14766505.fastq.gz	Fastqsanger.gz

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Diverse gene expression of diencephalon astrocytes reveals proliferation regulated by Smad4 - scRNA-seq II

Project description:Astrocytes are crucial for neural network function throughout the brain and also show region-specific gene expression differences. To explore this at the single cell level, we isolated cells by ACSA2 MACS from the diencephalon. Single cell RNA-seq revealed two super clusters of cells expressing astrocyte markers, one of which expressed many ependymal markers. Fluorescent labelling of ependymal cells allowed removing them during dissection. The remaining astrocytes and astrocyte-like cells show considerable heterogeneity with some subsets or gene expression states showing more wide-spread and others more restricted mapping of gene expression throughout the forebrain. Surprisingly, diencephalic astrocytes express proliferation regulating genes. Immunostaining for proiliferation markers, DNA-base incorporation and clonal analysis revealed ongoing low level astrocytogenesis even in 8 months old mice. Comparing gene expression of astrocytes in diencephalon and cortex grey matter identified Smad4 as a key regulator of astrocyte in vivo proliferation and in vitro neurosphere formation. Thus, astrocyte diversity is seemingly partitioned in wide-spread and region-specific subsets and reveals the novel concept of adult astrocytogenesis in the diencephalon in a Smad4-dependent manner.

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Proliferation and neural stem cell potential of diencephalic astrocytes revealed by genome-wide expression is SMAd4 dependent – RNAseq

Project description:Neural stem cells (NSCs) have astroglial hallmarks, while astrocytes in the intact adult mammalian brain parenchyma are thought to be postmitotic and lack NSC hallmarks that are activated only after injury. Analysis of genome-wide expression at population and single cell level in astrocytes from the diencephalon (DIE) and the cerebral cortex Grey grey Matter matter (CTX GM) revealed cell cycle associated gene expression, that we verified in situ by PCNA immunostaining and incorporation of the thymidine analog EdU. Strikingly, clonal analysis using GLASTCreERT2/confetti mice showed that only diencephalic astrocytes give rise to daughter cells in multi-cellular clones. In addition, subsets of astrocytes only from the diencephalon form multipotent, self-renewing neurospheres in vitro. Given the differential expression levels of genes related to Smad mediated transcriptional regulation between CTX GM and DIE astrocytes, we deleted the common mediator Smad4 in adult astrocytes. This abrogates proliferation of diencephalic astrocytes in vivo and their neurosphere formation in vitro. This work therefore identified a novel astrocyte subtype with NSC hallmarks in the diencephalon and implicates ongoing astrocytogenesis as a region-specific hallmark in the adult brain.

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Proliferation and neural stem cell potential of diencephalic astrocytes revealed by genome-wide expression is SMAd4 dependent – scRNA-seq

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Proliferation and neural stem cell potential of diencephalic astrocytes revealed by genome-wide expression is SMAd4 dependent

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Proliferation and neural stem cell potential of diencephalic astrocytes revealed by genome-wide expression is SMAd4 dependent – RNAseq

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