进行性肌阵挛性癫痫（PME）是一种少见的癫痫综合征，有关PME突变基因的谱系研究尚不完善。我们前期家系研究发现新型突变CACNA1Ac.6975_6976insCAG 的存在且位于CDS区，影响氨基酸编码功能，但是否参与PME发病机制尚不明确。为此，本项目拟采取基因克隆及转染技术培养突变型神经元样细胞PC12，建立突变型基因鼠模型；通过分生及流式细胞学，对神经细胞膜P/Q型钙离子通道Cav2.1的alpha-1A亚单位及抑制性神经递质GABAA-R亚单位定量分析，观察细胞周期；膜片钳技术对电压依赖性钙离子通道进行测量；荧免技术对突变鼠脑组织钙离子相关蛋白Ryanodine 受体、TH、Calretinin定量测定；AAV光遗传学技术在体蓝光诱发行为学改变。探讨CACNA1A 基因CDS区新型突变点对P/Q型钙离子通道Cav2.1、抑制性GABAA受体功能的影响及在PME发病机制中的作用。

The manifestation of the epilepsy syndrome is complex and the accurate diagnosis usually depends on gene detection methods. Progressive Myoclonic Epilepsy (PME) is a rare epilepsy syndrome with insufficient spectrum of gene mutation. We have recently found that a novel mutation of CACNA1A c.6975_6976insCAG in a PME family. The involvement of CACNA1A in the pathogenesis of PME, however, merits further investigation. We aim to test the function of alpha-1A submit of the P/Q-type Cav2.1channel,GABAARγ2 submit of the inhibitory GABAAR, and make an analysis of the distribution of mutant CACNA1A, quantify the expression of the Ryanodine receptor, TH, Calretinin, by using gene clone, gene transfection technique, patch-clamp, molecular biology and immunofluorescence, as well as flow cytometry. Then observe the behavioral changes induced by blue light, by AAV optogenetics technique. We will also cultivate the mutant neuronal cells and further establish the model of mutant rats. Our project may clarify whether the novel mutation in the CDS region of CACNA1A has an effect on the P/Q-type Cav2.1 channel or has involved in the pathogenesis of the PME, which on the long run may help screen new drug targets.