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The Role of Protein Phosphorylation and dephosphorylation in Neuronal Signal Transduction

蛋白质磷酸化和去磷酸化在神经信号转导中的作用

基本信息

批准号：
02454137
负责人：
TANAKA Chikako
金额：
$ 4.48万
依托单位：
Kobe University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1990
资助国家：
日本
起止时间：
1990 至 1991
项目状态：
已结题

项目摘要

Recently, protein phosphorylation and dephosphorylation are thought to be a central event in cellular signal transduction. Large amount of protein kinases and protein phosphatases are found in the central nervous system, suggesting that these enzymes are involved in the neuronal signal transduction. In this fiscal year, we obtained following findings on the protein kinase C and calcineurin.1) Developmental change in the distribution of protein kinase C subspecies in rat brain was examined. alpha and beta I subspecies were found when rat was born, on the other hand, beta II and gamma subspecies appeared one to two weeks after the birth when the synaptogenesis occurred.2) Immunohistochemical study on the localization of four protein kinase C subspecies and neurotransmitters in striate-nigral system was performed. Dopaminergic neurons in the substantia nigra and cholinergic neurons in the striatum contained alpha subspecies, and GABAergic neurons in the striatum contained beta -I subspecies.3) Distribution of calcineurin A subtypes were examined using in situ hybridization technique and immunohistochemistry in rat brain. The activation mechanism of calcineurin was studied usign calmodulin antagonists.4) GABA_B receptor was expressed in Xenopus oocyte and its regulation by protein kinase C was demonstrated.5) cDNA cloning of testis-specific calcineurin B-like protein and cDNA cloning of yeast homolog of calcineurin B were performed.

最近，蛋白质磷酸化和去磷酸化被认为是细胞信号转导中的中心事件。中枢神经系统中存在大量的蛋白激酶和蛋白磷酸酶，表明这些酶参与神经元信号转导。在本财年，我们获得了以下关于蛋白激酶C和钙调磷酸酶的发现。1)检查了大鼠脑中蛋白激酶C亚种分布的发育变化。 α和βI亚种在大鼠出生时就已发现，而βII和γ亚种则在出生后1-2周突触发生时出现。2)对四种蛋白激酶C亚种和神经递质在纹状体-黑质系统中的定位进行免疫组织化学研究。黑质多巴胺能神经元和纹状体胆碱能神经元含有α亚种，纹状体GABA能神经元含有β-I亚种。3)采用原位杂交技术和免疫组化技术检测大鼠脑内钙调神经磷酸酶A亚型的分布。利用钙调蛋白拮抗剂研究了钙调磷酸酶的激活机制。4)在非洲爪蟾卵母细胞中表达了GABA_B受体，并证实了其受蛋白激酶C的调节。5)进行了睾丸特异性钙调磷酸酶B样蛋白的cDNA克隆和钙调磷酸酶B的酵母同源物的cDNA克隆。