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Transcriptional profiling of pre- and post-natal glutamatergic progenitors and nascent neurons by single cell RNA sequencing

ABSTRACT: Purpose: We labeled synchronous cohorts of prenatal and postnatal glutamatergic progenitors to study their lineage relationship and transcriptional specificities. Method and results: We performed scRNA-seq of postnatal glutamatergic progenitors isolated from microdissected pallium at E15.5 or dorsal subventricular zone at P2 in the mouse. Our results highlight transcriptional dysregulation at P2 which includes genes involved in metabolism, differentition and migration.

ORGANISM(S): Mus musculus

PROVIDER: GSE109556 | GEO | 2018/01/25

REPOSITORIES: GEO

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Project description:Throughout postnatal life in mammals, neural stem cells (NSCs) are located in the subventricular zone (SVZ) of the lateral ventricles. The greatest diversity of neuronal and glial lineages they generate occurs during early postnatal life in a region-specific manner. In order to evaluate potential heterogeneity in the NSC pool, we microdissected the dorsal and lateral SVZ at different postnatal ages and isolated NSCs and their immediate progeny based on their expression of Hes5-EGFP/Prominin1 and Ascl1-EGFP, respectively. Whole genome comparative transcriptome analysis revealed transcriptional regulators as major hallmarks that sustain postnatal SVZ regionalization. Manipulation of single genes encoding for locally enriched transcription factors influenced NSC specification indicating that the fate of regionalized postnatal SVZ NSCs can be readily modified . These findings reveal functional heterogeneity of NSCs in the postnatal SVZ and provide targets to recruit region-specific lineages in regenerative contexts. Microarrays of neural stem cells, early progenitors and the tissue from subregions of the subventricular zone were compiled to screen for the full extent of heterogeneity in this region during postnatal life. Spatially distinct regions of the developing forebrain subventricular zone (SVZ) aged at P4, P8 and P11 were microdissected in RNAse free/sterile conditions. Mice expressing Ascl1-EGFP in the SVZ were used to aid accurate microdissection of the dorsal and lateral wall of each of the studied time points as per our previous publications characterizing this method. As well as at the whole microdomain level, additionally, NSCs (Hes5-EGFP+/Prom1+) and early progenitors (Ascl1-EGFP+) from each microdomain were further isolated by FAC sorting methods. This was to provide a comprehensive gene expression analysis at the tissue level and at the cellular level. Generally, 1 litter was used to yield 1 'n' number of replicates. A total of 23 affymetrix analysis were performed.

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Transcriptional profiling of pre- and post-natal glutamatergic progenitors and nascent neurons by single cell RNA sequencing

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