05-08-08(18:09:58)
Authors:
Prajerova I. 1,2
Anderova M. 1,3
Honsa P. 1
Machon O. 4
and Chvatal A. 1-3
Institutions:
1 Department of Cellular Neurophysiology - Laboratory of Neurobiology, Institute of Experimental Medicine, ASCR;
2 Department of Neuroscience, 2nd Med. Faculty, Charles University;
3 Center for Cell Therapy and Tissue Repair, Charles University;
4 Department of Transcriptional Regulation, Institute of Molecular Genetics, ASCR; Prague, Czech Republic
Title of abstract : Immunohistochemical and electrophysiological analysis of D6/GFP-neural stem/progenitor cells during in vitro differentiation and after transplantation into the injured rat brain.
Abstract text:
D6 is a promoter/enhancer of mDach1 gene, which is involved in the development of neocortex including ventricular zone and hippocampus, and is expressed in the proliferating neural stem/progenitor cells of the cortex (Machon et al., 2002). Embryonic neural stem/progenitor cells were isolated from E12 mouse embryos of D6-GFP transgenic mice, in which the expression of GFP is driven by D6 promoter/enhancer. The differentiation potential of these cells was first characterized in vitro. Electrophysiological and immunohistochemical analysis revealed two distinct cell populations. Large flat cells forming an underlying layer expressed GFAP and/or nestin. Smaller cells with multiple long processes expressed neuronal markers beta-III tubulin, MAP-2 or DCX. These cells with an average membrane potential of -54 mV, a membrane resistance of 2326 MOhms and a membrane capacitance of 9.8 pF displayed voltage-dependent A-type K+-channels, delayed outwardly rectifying K+-channels and Na+-channel, which was blocked by TTX. To study their survival and differentiation potential in vivo, D6-GFP neural stem/progenitor cells were transplanted into the intact tissue or into the site of the photochemical lesion (a model of thrombotic stroke). One week after the transplantation the cells survived and expressed markers of mature neurons (NeuN-, NF68-, beta-III tubulin- and MAP2- positive cells). Based on these data, D6/GFP-cells could provide a suitable tool for studying cell survival, migration and differentiation under pathological conditions.
Supported by GACR305/06/1316, AVOZ50390512, LC554, 1M0538 and GAUK62/2006/C/2.LF.
