Glycine transporter-1 regulates the proliferation of neural stem/progenitor cells derived from the embryonic mouse hippocampus

Abstract Generation of olfactory bulb (OB) interneurons requires neural stem/progenitor cell specification, proliferation, differentiation, and young interneuron migration and maturation. Here, we show that the homeobox transcription factors Dlx1/2 are central and essential components in the transcriptional code for generating OB interneurons. In Dlx1/2 constitutive null mutants, the differentiation of GSX2+ and ASCL1+ neural stem/progenitor cells in the dorsal lateral ganglionic eminence is blocked, resulting in a failure of OB interneuron generation. In Dlx1/2 conditional mutants (hGFAP-Cre; Dlx1/2F/− mice), GSX2+ and ASCL1+ neural stem/progenitor cells in the postnatal subventricular zone also fail to differentiate into OB interneurons. In contrast, overexpression of Dlx1&2 in embryonic mouse cortex led to ectopic production of OB-like interneurons that expressed Gad1, Sp8, Sp9, Arx, Pbx3, Etv1, Tshz1, and Prokr2. Pax6 mutants generate cortical ectopia with OB-like interneurons, but do not do so in compound Pax6; Dlx1/2 mutants. We propose that DLX1/2 promote OB interneuron development mainly through activating the expression of Sp8/9, which further promote Tshz1 and Prokr2 expression. Based on this study, in combination with earlier ones, we propose a transcriptional network for the process of OB interneuron development.

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Structurally Distinct LewisX Glycans Distinguish Subpopulations of Neural Stem/Progenitor Cells

Journal of Biological Chemistry ◽

10.1074/jbc.m110.201095 ◽

2011 ◽

Vol 286 (18) ◽

pp. 16321-16331 ◽

Cited By ~ 29

Author(s):

Eva Hennen ◽

Tim Czopka ◽

Andreas Faissner

Keyword(s):

Progenitor Cells ◽

Carbohydrate Binding ◽

Neural Cells ◽

Stem Cell Markers ◽

Binding Assays ◽

Embryonic Mouse ◽

Isolation And Characterization ◽

Neural Stem Progenitor Cells

There is increasing evidence that the stem and progenitor cell population that builds the central nervous system is very heterogeneous. Stem cell markers with the potential to divide this cell pool into subpopulations with distinct characteristics are sparse. We were looking for new cell type-specific antigens to further subdivide the progenitor pool. Here, we introduce the novel monoclonal antibody clone 5750. We show that it specifically labels cell surfaces of neural stem and progenitor cells. When 5750-expressing cells were isolated by fluorescence-activated cell sorting from embryonic mouse brains, the sorted population showed increased neurosphere forming capacity and multipotency. Neurospheres generated from 5750-positive cells could self-renew and remained multipotent even after prolonged passaging. Carbohydrate binding assays revealed that the 5750 antibody specifically binds to LewisX-related carbohydrates. Interestingly, we found that the LewisX epitope recognized by clone 5750 differs from those detected by other anti-LewisX antibody clones like 487LeX, SSEA-1LeX, and MMALeX. Our data further reveal that individual anti-LewisX clones can be successfully used to label and deplete different subpopulations of neural cells in vivo and in vitro. In conclusion, we present a new tool for the isolation and characterization of neural subpopulations and provide insights into the complexity of cell surface glycosylation.

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A protocol for isolation of fetal neural progenitor cells from mouse hippocampus

Cell and Organ Transplantology ◽

10.22494/cot.v2i2.32 ◽

2014 ◽

Vol 2 (2) ◽

pp. 155-157

Author(s):

O. Tsupykov

Keyword(s):

Progenitor Cells ◽

Molecular Mechanisms ◽

Neural Progenitor Cells ◽

Neural Progenitor ◽

Homogeneous Population ◽

Mouse Hippocampus ◽

Neural Stem Progenitor Cells ◽

Percoll Density Gradient ◽

Detailed Protocol

Culture of neural stem/progenitor cells are widely used to study the characteristics of these cells under controlled conditions in vitro as well as to study the cellular and molecular mechanisms of CNS diseases and develop strategies for their treatment.This paper provides a detailed protocol to isolate of fetal (E17-18) neural progenitor cells (NPCs) of mouse hippocampus. The technique is based on the use of centrifugation of hippocampal cells suspension in Percoll density gradient to obtain purified NPCs fractions. The cells are cultured in serum-free medium in a monolayer, which creates conditions for more equitable access of FGF-2 to the cells. This method provides a homogeneous population of undifferentiated progenitors from fetal mouse hippocampus.

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Neural Stem/Progenitor Cells Promote Endothelial Cell Morphogenesis and Protect Endothelial Cells against Ischemia via HIF-1α-Regulated VEGF Signaling

Journal of Cerebral Blood Flow & Metabolism ◽

10.1038/jcbfm.2008.38 ◽

2008 ◽

Vol 28 (9) ◽

pp. 1530-1542 ◽

Cited By ~ 65

Author(s):

Tamara Roitbak ◽

Lu Li ◽

Lee Anna Cunningham

Keyword(s):

Endothelial Cell ◽

Progenitor Cells ◽

Adult Brain ◽

Serum Starvation ◽

Protective Effects ◽

Embryonic Mouse ◽

Vegf Signaling ◽

Intracerebral Transplantation ◽

Neural Stem Progenitor Cells

Vascular cells provide a neural stem/progenitor cell (NSPC) niche that regulates expansion and differentiation of NSPCs within the germinal zones of the embryonic and adult brain under both physiologic and pathologic conditions. Here, we examined the NSPC—endothelial cell (NSPC/EC) interaction under conditions of ischemia, both in vitro and after intracerebral transplantation. In culture, embryonic mouse NSPCs supported capillary morphogenesis and protected ECs from cell death induced by serum starvation or by transient oxygen and glucose deprivation (OGD). Neural stem/progenitor cells constitutively expressed hypoxia-inducible factor 1α (HIF-1α) transcription factor and vascular endothelial growth factor (VEGF), both of which were increased approximately twofold after the exposure of NSPCs to OGD. The protective effects of NSPCs on ECs under conditions of serum starvation and hypoxia were blocked by pharmacological inhibitors of VEGF signaling, SU1498 and Flt-1-Fc. After intracerebral transplantation, NSPCs continued to express HIF-1α and VEGF, and promoted microvascular density after focal ischemia. These studies support a role for NSPCs in stabilization of vasculature during ischemia, mediated via HIF-1α-VEGF signaling pathways, and suggest therapeutic application of NSPCs to promote revascularization and repair after brain injury.

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