Study of Molecular Organization of Mouse Cerebellum

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1.By the differential display, microarray, and GeneChip analysis approaches, we explored the gene expression profiles specific to the postnatal developmental stages of mouse cerebellum on a genome-wide basis. In addition, we revealed the spatio-temporal expression patterns of specific genes by RT-PCR and in situ hybridization methods. Integrating all these lines of gene expression information, we have generated the "Cerebellar Development Transcriptome database". By this study, we have succeeded in cloning of many candidates for novel genes responsible for brain development as follows.2.We showed that dendritic clustering and synaptic targeting of Cupidin/Homer, an adaptor protein at postsynaptic density (PSD), coincides those of glutamate receptor-related proteins NR2B and PSD-95 during development of hippocampal neurons, that Cupidin act s as a mobile adaptor in cerebellar granule cells in an activity-dependent manner, and that the Homer family members have differential cellular distributions in developing mouse brains.3.We cloned CAPS2, a paralog of CAPS that regulates Ca2+-dependent exocytosis of secretory granules. CAPS2 was localized to vesicular structures, in which neurotrophins BDNF and NT-3 are included, in parallel fiber terminals of cerebellar granule cells. The overexpression of exogenous CAPS2 in primary cultured granule cells augmented NT-3 release in a depolarization-dependent manner, resulting in promoting survival of Purkinje cells. These data indicate that CAPS2 is a molecule that regulates activity-dependent release of neurotrophins that are indispensable for differentiation and survival of cerebellar neurons.4.We analyzed the structure, function, and brain expression of three genes related to protein phsophorylation (apoptosis activity-related tyrosine kinase AATYK, novel Ser/Thr kinase Ebr, and tyrosine kinase adaptor Cas) and a gene encoded a novel myelin paranodal loop protein.