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The study of protein associated with seizure-susceptibility in El mouse brain as a model of epilepsy

作为癫痫模型的El小鼠大脑中与癫痫易感性相关的蛋白质的研究

基本信息

批准号：
63570510
负责人：
KAWAKITA Yukio
金额：
$ 1.34万
依托单位：
Osaka City University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1988
资助国家：
日本
起止时间：
1988 至 1989
项目状态：
已结题

项目摘要

Polysomal messenger RNAs (mRNA) isolated from four parts of brain such as cerebrum, diencephalon, pons and medulla oblongata, and cerebellum were incubated with [^<35>S]-methionine in rabbit reticulocyte lysate system. After the incubation, translation products were analyzed by two dimensional gel electrophoresis. There were twenty proteins, in which protein P of El mice was more labeled in cerebellum than in other parts, but less than that of ddY mice. Comparison of translation products of mRNA from whole brain indicated that labeled protein P significantly decreased immediately after the seizures and after 30 min it restored to normal level. These results suggest that mRNA regulated protein P synthesis is associated with seizure-diathesis.In the synaptic plasma membranes of El mice, ^<32>P incorporation into proteins of molecular weights 200K in cortex and 15OKd in hippocampus was markedly stimulated by combined calcium and calmodulin, whereas it showed a significant inhibition of 20OKd, 10OKd, and 2OKd proteins in amygdala, and 2OKd in hippocampus. The difference of Ca^<2+>-calmodulin kinase II activity between El and ddY mice was remarkable in both hippocampus and amygdala, less pronouns in cortex, and not significant in septa, striatum, thalamus and cerebellum. Thus, hippocampus and amygdala may be responsible for induced-seizure of this animal. Further experiments are need to investigate mRNA species coding for protein P or Ca^<2+>-calmodulin kinase II, and the molecular cloning of cDNA probes being specific relative to such proteins will provide a basis to elucidate the epileptogenesis.

用[^<35>S]-蛋氨酸在兔网织红细胞裂解液系统中孵育从大脑、中脑、脑桥、延髓和小脑等4个部位分离的多体信使rna （mRNA）。孵育后，用二维凝胶电泳分析翻译产物。有20种蛋白，其中El小鼠的P蛋白在小脑的标记量高于其他部位，但低于ddY小鼠。全脑mRNA翻译产物比较表明，标记蛋白P在癫痫发作后立即显著降低，30min后恢复正常。这些结果表明mRNA调控的蛋白P合成与癫痫发作有关。在El小鼠的突触质膜中，钙与钙调蛋白联合作用可显著刺激^<32 b> P掺入皮质和海马的分子量为200K的蛋白中，而杏仁核的20OKd、10OKd和20OKd蛋白以及海马的20OKd蛋白则受到显著抑制。El和ddY小鼠海马和杏仁体Ca^<2+>-钙调蛋白激酶II活性差异显著，皮质代词活性差异显著，中隔、纹状体、丘脑和小脑活性差异不显著。因此，海马和杏仁核可能与该动物的诱导性癫痫发作有关。P蛋白或Ca^<2+>-钙调蛋白激酶II编码的mRNA种类有待进一步的实验研究，对这些蛋白特异性的cDNA探针的分子克隆将为阐明癫痫发生提供基础。