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Neural Stem/Progenitor Cells Derived from the Embryonic Dorsal Telencephalon of D6/GFP Mice Differentiate Primarily into Neurons After Transplantation into a Cortical Lesion

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Abstract

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.

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Acknowledgments

The authors would like to thank to Dr. Z. Kozmik and Dr. O. Machon for their generous gift of D6/GFP mice. This study was supported by the following grants: 305/09/0717 and 309/08/H079 from the Grant Agency of the Czech Republic, AVOZ50390512 from the Academy of Sciences of the Czech Republic, LC554 and 1M0538 from the Ministry of Education, Youth and Sports of the Czech Republic and GAUK62/2006/C/2. LF from the Grant Agency of Charles University.

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  1. Laboratory of Neurobiology, Department of Cellular Neurophysiology, Institute of Experimental Medicine ASCR v.v.i., Videnska 1083, 142 20, Prague 4, Czech Republic

    Iva Prajerova, Pavel Honsa, Alexandr Chvatal & Miroslava Anderova

  2. Second Medical Faculty, Department of Neuroscience, Charles University, V Uvalu 84, 150 06, Prague 5, Czech Republic

    Iva Prajerova, Pavel Honsa & Alexandr Chvatal

  3. Second Medical Faculty, Center for Cell Therapy and Tissue Repair, Charles University, V Uvalu 84, 150 06, Prague 5, Czech Republic

    Alexandr Chvatal & Miroslava Anderova

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Prajerova, I., Honsa, P., Chvatal, A. et al. Neural Stem/Progenitor Cells Derived from the Embryonic Dorsal Telencephalon of D6/GFP Mice Differentiate Primarily into Neurons After Transplantation into a Cortical Lesion. Cell Mol Neurobiol 30, 199–218 (2010). https://doi.org/10.1007/s10571-009-9443-x

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