Comparative analysis of ependymal cells from the lateral ventricular wall and the spinal cord of adult mice
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ABSTRACT: In contrast to ependymal cells in the lateral ventricle wall (LVW), spinal cord (SC) ependymal cells possess certain neural stem cell characteristics. Isolated CD133+/CD24+/CD45-/CD34- ependymal cells from the SC displayed in vitro self renewal and differentiation capacity, whereas those from the LVW did not. The molecular basis of this difference is unknown. In this study, antibodies against multiple surface markers were applied to isolate SC and LVW ependymal cells which allowed a direct comparison of their in vitro behavior and gene expression profile. cRNA samples of three independent biological replicates of CD133+/CD24+/CD45-/CD34- LVW ependymal cells, CD133+/CD24+/CD45-/CD34- SC ependymal cells, CD133+/CD24-/CD45-/CD34- radial glial cells, and SC ependymal cell-derived NSPs were hybridized onto Illumina MouseWG-6 v1.1 gene expression arrays
Project description:In contrast to ependymal cells in the lateral ventricle wall (LVW), spinal cord (SC) ependymal cells possess certain neural stem cell characteristics. Isolated CD133+/CD24+/CD45-/CD34- ependymal cells from the SC displayed in vitro self renewal and differentiation capacity, whereas those from the LVW did not. The molecular basis of this difference is unknown. In this study, antibodies against multiple surface markers were applied to isolate SC and LVW ependymal cells which allowed a direct comparison of their in vitro behavior and gene expression profile.
Project description:Using CD133 as a pan-ependymal cell marker, we wished to understand whether CD133+ DNGR-1 traced cells constituted a distinct subset of ependymal cells by comparing these at the single cell level with CD133+ non-traced cells purified from spinal cords of DNGR-1 lineage tracer mice.
Project description:Through single cell transcriptome analysis, we uncovered molecular signatures of CD133+/GFAP- ependymal (E) cells, CD133+/GFAP+ neural stem (B) cells, Dlx2+ neuroblasts (A cells), and Sox10+ oligodendrocyte progenitors (O cells) in the adult mouse forebrain neurogenic zone. prominent hub genes of the gene network unique to ependymal CD133+/GFAP- quiescent cells are enriched for receptors of angiogenic factors and immune-responsive genes. Administration of VEGF activated CD133+ ependymal stem cells lining not only the lateral, but also the 4th ventricles, and together with bFGF, elicited subsequent neural lineage differentiation and migration.
Project description:Through single cell transcriptome analysis, we uncovered molecular signatures of CD133+/GFAP- ependymal (E) cells, CD133+/GFAP+ neural stem (B) cells, Dlx2+ neuroblasts (A cells), and Sox10+ oligodendrocyte progenitors (O cells) in the adult mouse forebrain neurogenic zone. prominent hub genes of the gene network unique to ependymal CD133+/GFAP- quiescent cells are enriched for receptors of angiogenic factors and immune-responsive genes. Administration of VEGF activated CD133+ ependymal stem cells lining not only the lateral, but also the 4th ventricles, and together with bFGF, elicited subsequent neural lineage differentiation and migration. Examination of 28 single cells and 4 populations of 10 cells from adult mouse forebrain neurogenic zone.
Project description:We performed single-cell (sc)RNA-Seq on male donors FACS-sorted “primitive” (CD34+ CD133+ CD90+) and “progenitor” (CD34+ CD133+ CD90-) HSPC cells treated with IL2RG ZFN, High Specificity (HS), (Low Specificity (LS) or control RNP in a ratio 1 to 1. We also included HS RNP and the cognate repair template containing a GFP expression cassette delivered by AAV6 and sorted for targeted integration (HS/AAV6 GFP+ vs. HS/AAV6 GFP-)
Project description:Total RNA was isolated from GFAP::GFP+CD133+EGFR-CD24- (quiescent neural stem cells, qNSCs), GFAP::GFP+CD133+EGFR+CD24- (activated neural stem cells, aNSCs) and GFAP::GFP+CD133- EGFR+CD24- (transit amplifying cells, TACs) cells from the adult mouse ventricular-subventricular zone (V-SVZ) (GFAP::GFP mice, Jackson Mice Stock number 003257).
Project description:The spinal cord neural stem cell potential is contained within the ependymal cells lining the central canal. Ependymal cells are, however, heterogeneous and we know little about what this reflects. To gain new insights into ependymal cell heterogeneity, we microdissected the ependymal cell layer from the thoracic spinal cord of 4 FOXJ1-EGFP transgenic mice (2.5-to-3-month old). After after dissociating the tissue into a cell suspension, we sorted single GFP-positive ependymal cells into lysis plates. cDNA synthesis was performed using Smart-seq2 technology.
Project description:After injury, mammalian spinal cords develop scars to confine the lesion and prevent further damage. However, excessive scarring can hinder neural regeneration and functional recovery. These competing actions underscore the importance of developing therapeutic strategies to dynamically modulate scar progression. Previous research on scarring has primarily focused on astrocytes, but recent evidence has suggested that ependymal cells also participate. Ependymal cells normally form the epithelial layer encasing the central canal, but they undergo massive proliferation and differentiation into astroglia following certain injuries, becoming a core scar component. However, the mechanisms regulating ependymal proliferation in vivo remain unclear. Here we uncover an endogenous κ-opioid signalling pathway that controls ependymal proliferation. Specifically, we detect expression of the κ-opioid receptor, OPRK1, in a functionally under-characterized cell type known as cerebrospinal fluid-contacting neuron (CSF-cN). We also discover a neighbouring cell population that expresses the cognate ligand prodynorphin (PDYN). Whereas κ-opioids are typically considered inhibitory, they excite CSF-cNs to inhibit ependymal proliferation. Systemic administration of a κ-antagonist enhances ependymal proliferation in uninjured spinal cords in a CSF-cN-dependent manner. Moreover, a κ-agonist impairs ependymal proliferation, scar formation and motor function following injury. Together, our data suggest a paracrine signalling pathway in which PDYN+ cells tonically release κ-opioids to stimulate CSF-cNs and suppress ependymal proliferation, revealing an endogenous mechanism and potential pharmacological strategy for modulating scarring after spinal cord injury.
Project description:Ependymomas are glial tumors that share morphologic similarities with ependymal cells. Here we laser microdissected ependymoma cells of 15 infratentorial tumors and generated gene expression profiles. These profiles were compared to those of 7 laser microdissected ependymal tissues. 10 000 laser microdissected cells of each of the 15 different infratentorial ependymomas and seven ependymal autopsy tissues were lysed and gene expression profiles were generated. Expression profiles of ependymomas were compared to those of ependymal cells.
Project description:Microarray and miRNA analysis of CD133(+), CD34(+)CD133(-) and CD133(-)CD34(-) cells The goal of the experiment was the comparison of expression levels of mRNA and miRNA in CD133(+) and CD34(+)CD133(-) cells