Functional analysis of neuronal migration-associated genes in murine brains by in utero gene transfer techniques with GFP reporter genes.

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Prenatal exposure to ionizing radiation of low doses in rodents induces deceleration of neuronal migration during cortical histogenesis, and results in disorganized cortical architecture in mature brain. We have attempted to elucidate genes which show changing patterns of expression after exposure to X-rays in fetal mouse brains, by applying two molecular biological techniques, i.e., the restriction landmark cDNA scanning (RLCS) method and the suppression subtractive PCR method.Approximately 13,000 cDNA species have been scanned and it turned out that more than twenty genes among the genes scanned are differentially expressed between X-irradiated embryos and non-irradiated ones. One of the genes showing up-regulation is Rab6A which is known to be associated with vesicle transport from trans-Golgi network. In addition, RabKIFL and GAPCenA genes were up-regulated. We then established a gene transfection method in which EGFP-tagged proteins of interest can be expressed in cultured cells to produce dominant negative effects. Based on these results we have applied an in utero electroporation technique to introduce genes of interest tagged with EGFP to embryonic mouse brains and this study is currently under way.