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The study on crosstalk failure in the nervous system.

神经系统串扰失效的研究。

基本信息

批准号：
06044166
负责人：
HATASE Osamu
金额：
$ 4.29万
依托单位：
The Kagawa Medical School
依托单位国家：
日本
项目类别：
Grant-in-Aid for international Scientific Research
财政年份：
1994
资助国家：
日本
起止时间：
1994 至 1995
项目状态：
已结题

项目摘要

The involvement of calmodulin dependent protein kinases (CaM-kinases) and cyclin-dependent kinase 5 (Cdk 5) in epilepsy and Alzheimer's disease was studied.Among CaM-kinases known, CaM-kinase II has been demonstrated to play an important role in the plasticity of neuronal cells. We investigatd the possible involvement of CaM-kinase I and CaM-kinase IV in the neuronal plasticity during epileptogenesis. Amygdaloid-kindled rat brain was used for immunohistochemical study, Western blot analysis, Northern blot analysis and in situ hybridization study.CaM-kinases I and IV were present in the control normal rat brain. Significantly incerased levels of both kinases were detected in the kindled cerebrum, while no obvious change was detected in cerebellum. Cdk 5 showed almost constant expression level in all stages of epileptogenesis. On the other hand, p35, a brain specific activator of Cdk 5, showed a significant increase at stage 3 where new synapse formation is most frequently observed. The level of p35 in other stages (stages 1,2,4, and 5) remained low. When the activity of Cdk 5/p35 was measured, the highest activity was detected at stage 3.Tau protein has been known to be heavily phosphorylated in Alzheimer's brain and was found in paired helical tangles. We therefore checked if Tau was phosphorylated during epileptogenesis. Western blot analysis using an antibody against phosphorylated-Tau (p-Tau) demonstrated the high level of p-Tau presented at stage 3, while a lot less level of p-Tau in other stages. Western blot analysis using an antibody against nonphosphorylated-Tau showed a mirror image of that of p-Tau. We also demonstrated that both p-Tau and Cdk 5/p35 colocalized in Alzheimer's deposit.These results together indicated that CaM-kinases and Cdk 5/p35 might play an pathogenic role in epileptogenesis and Alzheimer's disease.

研究了钙调蛋白依赖性蛋白激酶（CaM-kinases）和细胞周期蛋白依赖性激酶5（Cdk 5）在癫痫和Alzheimer病中的作用，其中CaM-kinases Ⅱ在神经细胞的可塑性中起重要作用。本实验证实了钙调蛋白激酶I和钙调蛋白激酶IV在癫痫发生过程中可能参与了神经元的可塑性。用杏仁核点燃大鼠脑组织进行免疫组织化学、Western印迹、北方印迹和原位杂交研究，正常对照大鼠脑组织中有钙调蛋白激酶I和IV。在点燃的大脑中，两种激酶的水平均显著升高，而小脑中无明显变化。Cdk 5在癫痫发生的各个阶段都有恒定的表达。另一方面，Cdk 5的脑特异性激活剂p35在最常观察到新突触形成的阶段3显示出显著增加。p35在其他阶段（第1、2、4、5期）仍保持低水平。当测量Cdk 5/p35的活性时，在第3阶段检测到最高活性。已知Tau蛋白在阿尔茨海默氏症的脑中被严重磷酸化，并且在成对的螺旋缠结中被发现。因此，我们检查了Tau在癫痫发生过程中是否被磷酸化。使用针对磷酸化-Tau（p-Tau）的抗体的蛋白质印迹分析表明在第3阶段呈现高水平的p-Tau，而在其他阶段中p-Tau的水平低得多。使用针对非磷酸化-Tau的抗体的蛋白质印迹分析显示了p-Tau的镜像。我们还发现p-Tau和Cdk 5/p35共定位于Alzheimer's存款中，这些结果共同表明CaM激酶和Cdk 5/p35可能在癫痫和Alzheimer's病的发病中起作用。