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

从大脑、间脑、脑桥和延髓、小脑等4个脑组织中提取多聚体信使RNA（polysomemessenger RNA，mRNA），<35>在兔网织红细胞裂解物体系中与[^ S]-蛋氨酸孵育。孵育后，通过二维凝胶电泳分析翻译产物。共检测到20种蛋白质，其中P蛋白在El鼠小脑中的标记率高于其他部位，而在ddY鼠小脑中的标记率低于其他部位。全脑mRNA翻译产物的比较显示，癫痫发作后即刻标记蛋白P显著降低，30 min后恢复至正常水平。在E1小鼠突触质膜中，Ca ~（2+）和Ca ~（2+）联合作用可<32>显著促进皮质200 K和海马15 OKd蛋白的P ~（2+）掺入，而抑制杏仁核20 OKd、10 OKd和2 OKd蛋白及海马2 OKd蛋白的P ~（2+）掺入。E1和ddY小鼠海马和杏仁核Ca^&lt;2+&gt;-钙调蛋白激酶II活性差异显著，皮质代词较少，隔、纹状体、丘脑和小脑差异不显著。因此，海马和杏仁核可能是导致该动物癫痫发作的原因。需要进一步的实验来研究编码蛋白P或Ca^&lt;2+&gt;-钙调蛋白激酶II的mRNA种类，并且分子克隆与这些蛋白相关的特异性cDNA探针将为阐明癫痫的发生提供基础。

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.