D6 is a promoter/enhancer of the mDach1 gene that is involved in the development of the neocortex and hippocampus. It is expressed by proliferating neural stem/progenitor cells (NSPCs) of the cortex at early stages of neurogenesis. The differentiation potential of NSPCs isolated from embryonic day 12 mouse embryos, in which the expression of green fluorescent protein (GFP) is driven by the D6 promoter/enhancer, has been studied in vitro and after transplantation into the intact adult rat brain as well as into the site of a photochemical lesion. The electrophysiological properties of D6/GFP-derived cells were studied using the whole-cell patch-clamp technique, and immunohistochemical analyses were carried out. D6/GFP-derived neurospheres expressed markers of radial glia and gave rise predominantly to immature neurons and GFAP-positive cells during in vitro differentiation. One week after transplantation into the intact brain or into the site of a photochemical lesion, transplanted cells expressed only neuronal markers. D6/GFP-derived neurons were characterised by the expression of tetrodotoxin-sensitive Na(+)-currents and K (A)- and K (DR) currents sensitive to 4-aminopyridine. They were able to fire repetitive action potentials and responded to the application of GABA. Our results indicate that after transplantation into the site of a photochemical lesion, D6/GFP-derived NSPCs survive and differentiate into neurons, and their membrane properties are comparable to those transplanted into the non-injured cortex. Therefore, region-specific D6/GFP-derived NSPCs represent a promising tool for studying neurogenesis and cell replacement in a damaged cellular environment.

• MeSH
• bikukulin metabolismus   MeSH
• biologické markery metabolismus   MeSH
• blokátory draslíkových kanálů metabolismus   MeSH
• buněčná diferenciace fyziologie   MeSH
• embryo savčí anatomie a histologie   fyziologie   MeSH
• GABA antagonisté metabolismus   MeSH
• GABA metabolismus   MeSH
• kmenové buňky cytologie   fyziologie   MeSH
• krysa rodu rattus MeSH
• metoda terčíkového zámku MeSH
• myši MeSH
• neurony cytologie   fyziologie   MeSH
• promotorové oblasti (genetika) MeSH
• rekombinantní fúzní proteiny genetika   metabolismus   MeSH
• telencefalon cytologie   patologie   fyziologie   MeSH
• transplantace kmenových buněk MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Neural progenitors of the mouse forebrain can be propagated in vitro as neurospheres in the presence of bFGF and EGF. However, less is understood whether regional characteristics or developmental stage properties of these cells are maintained in neurosphere cultures. Here we show that the original cell fate is lost in neurosphere cultures. We isolated neural progenitors from the dorsal telencephalon of D6-GFP mice and cultured them in vitro. The expression profile was specifically changed in cultured cells in just three passages. Markers of the dorsal forebrain were downregulated and several ventrally-expressed genes were induced. The altered gene expression led to a profound phenotypic change of cultured cells. D6-GFP positive cortical progenitors produce excitatory neurons in the cortex and few astrocytes in vivo but after culture in vitro, these cells differentiate into many astrocytes and also oligodendrocytes and inhibitory neurons. Wnt signaling in cultured neurospheres was downregulated in the same manner as other dorsal markers but dominant active Wnt signaling slowed down the loss of the dorsal identity in neurospheres.

• MeSH
• astrocyty cytologie   metabolismus   MeSH
• buněčná diferenciace fyziologie   MeSH
• buněčné sféroidy cytologie   MeSH
• buněčný rodokmen fyziologie   MeSH
• financování organizované MeSH
• genetické markery MeSH
• interneurony cytologie   metabolismus   MeSH
• kmenové buňky cytologie   metabolismus   MeSH
• mozková kůra cytologie   embryologie   metabolismus   MeSH
• myši transgenní MeSH
• myši MeSH
• neuroglie cytologie   metabolismus   MeSH
• neurony cytologie   metabolismus   MeSH
• oligodendroglie cytologie   metabolismus   MeSH
• promotorové oblasti (genetika) imunologie   MeSH
• rekombinantní fúzní proteiny genetika   MeSH
• telencefalon cytologie   embryologie   metabolismus   MeSH
• vývojová regulace genové exprese genetika   MeSH
• zelené fluorescenční proteiny genetika   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH