Study of Molecular Organization of Mouse Cerebellum

小鼠小脑分子组织的研究

基本信息

批准号：
13308047
负责人：
FURUICHI Teiichi
金额：
$ 29.7万
依托单位：
The Institute of Physical and Chemical Research (RIKEN)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2003
项目状态：
已结题

项目摘要

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.

1.通过差异显示，微阵列和祖先分析方法，我们探索了小鼠小脑在全基因组的基础上特有的基因表达谱。此外，我们通过RT-PCR和原位杂交方法揭示了特定基因的时空表达模式。整合了所有这些基因表达信息，我们生成了“小脑发展转录组数据库”。通过这项研究，我们成功地将许多候选者克隆为负责大脑发育的新基因的克隆。2。我们表明，树突状聚类和突触靶标丘比丁素/荷马是一种突触后密度（PSD）的衔接蛋白（PSD）的衔接蛋白，重合谷氨酸受体受体蛋白NRRSNR2B和PSD-95在PSD-95中，该蛋白在PSD-95中，该蛋白质在upipd-95中，upipd-95 hippipt as sipd-95小脑颗粒细胞中的移动适配器以活性依赖性的方式，并且荷马家族成员在发育中的小鼠大脑中具有差异性细胞分布。3。我们克隆的CAPS2（克隆CAPS2），这是一种调节Ca2+依赖性颗粒的cap的caps2。 CAPS2位于囊泡结构中，其中包括神经营养蛋白BDNF和NT-3，在小脑颗粒细胞的平行纤维末端中包括。原代培养的颗粒细胞中外源CAPS2的过表达以去极化依赖性方式增强NT-3释放，从而促进了Purkinje细胞的存活。这些数据表明CAPS2是一种分子，可调节神经营养蛋白的活性依赖性释放，这对于小脑神经元的分化和存活是必不可少的。适配器cas）和一个基因编码了一种新型的髓磷酸偏曲loop蛋白。