海馬スライス培養における内在的ニューロン新生

抄録

It has been found that neurogenesis occurs in the dentate granule cell layer (GCL) of the mammalian hippocampus throughout life, but how a newly generated cell differentiates into a neuron and is integrated into hippocampal circuitry remains to be elucidated. We have previously reported that intrinsic neurogensis occurs spontaneously in the organotypic slice culture of a rat hippocampus using 5-bromodeoxyuridine labeling method. In the present study, the newly divided cells and their descendents were labeled in the living slices with the enhanced green fluorescent protein (EGFP) through retrovirus vector transduction. We found that (1) the EGFP-labeled cells increased in number in the first two weeks and (2) the EGFP-labeled cells consisted of variety of phenotypes of both early and late stages of differentiation in four weeks after inoculation: about 25% NeuN-positive cells with appearances of neurons in the GCL, 25% immature neurons immunoreactive with Tuj1, approximately 30% GFAP-positive cells and 30% nestin-positive cells. These results suggest that the slice cultures intrisically retain neurogenic abilities for their cultivation period. Our slice culture system would provide a good model of animal experiments to follow up the newly divided cells for a certain long period. [Jpn J Physiol 54 Suppl:S228 (2004)]