The analyzes of the alteration and activity of protein kinase C in the epileptic mouse brain.

[结果]1。ptz诱导惊厥后的改变(1)PKC活性癫痫发作后10 min，与对照组和强直惊厥小鼠相比，大脑皮层胞质PKC活性显著升高。另一方面，惊厥后皮质膜部分无明显变化。ptz引起的惊厥对海马、纹状体和丘脑/下丘脑的胞质和膜组分PKC活性无显著影响。Western blot分析PKC同工酶惊厥后10 min，胞浆部分80-K PKC β含量升高，膜部分80-K PKC β含量下降。在强直性惊厥期间和惊厥期后10分钟细胞质部分检测到50-K PkC β。PKC α的表达与PKC β相似。而PKC γ、δ和zeta的表达在惊厥后无明显变化。(3)钙蛋白酶活性惊厥后胞浆和膜部分皮质钙蛋白酶活性无明显变化。EL小鼠癫痫发作后的变化(1)PKC同工酶表达的RT-PCR分析PKC α mRNA表达在发作间期较轻，癫痫发作后逐渐降低。发作间期PKC β水平较低，发作后逐渐升高。PKC γ在癫痫发作间期丰富，在癫痫发作后30 min达到峰值，在癫痫发作后120 min恢复到基础水平。间隔期PKC δ的变化不大，在癫痫发作后30min达到峰值。【讨论】ptz诱发癫痫发作后，大脑皮层PKC α和β从膜转移到胞质。EL小鼠癫痫发作后PKC同工酶的变化与ptz诱导的癫痫发作后不同。有必要通过northern和western blot分析进一步检查。

[RESULTS]1.The alteration after PTZ-induced convulsion(1)PKC activityThe cytosolic PKC activity in the cerebral cortex significantly increased 10 min after seizure, compared with that of the control and tonic-convulsive mouse. On the other hand, that in the cortical membrane fraction after convulsion didn't alter significantly. PTZ-induced convulsion didn't considerably change the PKC activity of the cytosolic and membrane fractions in hippocapmus, striatum and thalamus/hypothalamus.Western blot analyzes of PKC isozymesThe content of 80-K PKC beta in the cytosolic fraction increased, whereas that in the membrane fraction decreased at 10 min after convulsion. The 50-K PkC beta was detected in the cytosolic fraction during the tonic convulsion and 10 min after the convulsive phase. The expression of PKC alpha was similar to that of PKC beta. However the expressions of PKC gamma, delta, and zeta didn't alter after convulsions.(3)Calpain activityThe cortical Calpain activities in the cytosolic and membrane fraction didn't change significantly after convulsions.2.The alteration after EL mouse's seizure(1)RT-PCR analysis of PKC isozymes expressionThe mRNA expression of PKC alpha was slight at the interictal period and decreased gradually after the seizure. That of PKC beta was slight at the interictal period but increased gradually after the seizure. That of PKC gamma was abundant at the interictal period, reached a peak at 30 min after seizures and return the basal level at 120 min. That of PKC delta was little at the interictal period and peaked at 30 min after the seizure.[DISCUSSIONS]PKC alpha and beta translocated from membrane to cytosol in the cerebral cortex after PTZ-induced seizures. The alterations of PKC isozymes after seizures of EL mouse were different from those after PTZ-induced seizures. Further examinations are necessary by northern and western blot analyzes.