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Neurogenesis and gliogenesis after ischemic brain injury in EGFP/GFAP mice
Dlouhá, Veronika ; Anděrová, Miroslava (advisor) ; Vargová, Lýdia (referee)
Focal ischemia induces enhancement of neurogenesis/gliogenesis in the subventricular zone (SVZ) of the lateral ventricle and it also leads to glial scar formation in the vicinity of the ischemic lesion. The gliotic scar is mainly formed by reactive astrocytes that express glial fibrilarly acidic protein (GFAP), nevertheless this protein is also expressed in adult multipotent neural stem cells (NSCs). Therefore, we have used the strain of transgenic mice (GFAP/EGFP mice), in which the enhanced green fluorescent protein (EGFP) is expressed under human GFAP promoter in astrocytes as well as in NSCs, thus allowing us an immediate vizualization of these cells, and to estimate the effect of ischemic injury on their fate during proliferation and differentiation in vitro. Focal ischemia was induced by the occlusion of the middle cerebral artery (MCAO) and 3 days post injury, an immunohistochemical analysis was carried out. Furthermore, the cell isolation from SVZ and the region of gliotic scar was performed, followed by their cultivation under proliferative conditions (as neurospheres) and their differentiation for 7-10 days. The differentiation potential of these cells was studied using immunocytochemical analyses and patch clamp technique was employed to estimate their membrane properties. Based on increased...
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