Role of calmodulin kinases on the regulation of Ca^<2+> signaling of central nervous systems

钙调蛋白激酶对中枢神经系统Ca^2信号传导调节的作用

基本信息

批准号：
10470484
负责人：
YAMAUCHI Takashi
金额：
$ 8.51万
依托单位：
The University of Tokushima
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 2001
项目状态：
已结题

项目摘要

Neuronal Ca^<2+> /calmodulin-dependent protein kinase II (CaM kinase II) regulates important neuronal functions, including neurotransmitter synthesis and neurotransmitter release, modulation of ion channel activity, synaptic plasticity, and gene expression. The following results were obtained in this study; 1. when CaM kinase II was introduced in neuronal cells, neurite outgrowth was stimulated, indicating that CaM kinase II played a role stimulation of neurite outgrowth; 2. Ca^<2+> independent activity of the kinase autophosphorylated at Thr-286 involved in neurite outgrowth; 3. more than 28 protein substrates of CaM kinase II were found in the postsynaptic density (PSD), and almost all of these proteins were identified by protein sequencing and mass spectrometry; 4. when the developmental change of the kinase was investigated during the neural differentiation of cultured cells, it was found that CaM kinase II was upregulated during the neuronal differentiation; 5. cell type distinctive changes of splicing pattern of δ isoform were found not only during development of rat brain tissue, but also during differentiation of cultured neuronal cells, indicating that alternatively spliced variants of δ isoform of CaM kinase II were expressed during neural differentiation; 6. when we used the deletion mutants for β-specific insertions to explore the difference between α and β CaM kinase II in enzymatic properties, it was found that β specific insertion of β CaM kinase II played an important role in the cellular distribution of the kinase; 7 The gene encoding the β isoform of rat Ca^<2+>/calmodulin-dependent protein kinase II was cloned, and its structure and exon-intron organization were determined; 8. When the promoter activity of the gene was analyzed using nueronal and non-neuronal cells, neuronal cell type-specific promoter activity was found in the 5'-upstream region of the gene.

神经元Ca^<2+> /钙调蛋白依赖性蛋白激酶II（CAM激酶II）调节重要的神经元功能，包括神经递质合成和神经递质释放，离子通道活性的调节，突触可变性和基因表达。在本研究中获得了以下结果。 1。当在神经元细胞中引入CAM激酶II时，刺激了神经蛋白的出生，表明CAM激酶II发挥了神经元出现的作用模拟。 2。Ca^<2+>在THR-286中涉及神经元出生时的激酶自磷酸化的独立活性； 3。在突触后密度（PSD）中发现了28多个CAM激酶II的蛋白质底物，几乎所有这些蛋白质都是通过蛋白测序和质谱法鉴定的。 4。当在培养细胞的神经元分化过程中研究了激酶的发育变化时，发现在神经元分化过程中CAM激酶II已更新。 5。在大鼠脑组织的发育过程中，在分化培养的神经元细胞的分化过程中，发现了δ同工型的剪接模式的独特变化，表明在神经元分化过程中表达了CAM激酶II的δ同工型的剪接变体； 6。当我们将缺失突变体用于β特异性插入来探索酶特性中α和βCAM激酶II之间的差异时，发现β的特异性插入βCAM激酶II在激酶的细胞分布中起重要作用。 7将编码大鼠Ca^<2+>/钙调蛋白依赖性蛋白激酶II的β同工型的基因克隆，并确定其结构和外显子内组织； 8。当使用Nueronal和非神经元细胞分析基因的启动子活性时，在基因的5'上游区域中发现了神经元细胞类型特异性启动子活性。