Dock3/Paks对癫痫突触可塑性的调控及异常神经网络形成机制研究

项目名称： Dock3/Paks对癫痫突触可塑性的调控及异常神经网络形成机制研究

项目编号： No.81671289

项目类型： 面上项目

立项/批准年度： 2017

项目学科： 医药、卫生

项目作者： 李杰

作者单位： 新乡医学院

项目金额： 25万元

中文摘要： 癫痫的发病机制尚不完全清楚，神经元之间异常突触联系和病理性神经环路的形成为癫痫的发生提供了解剖学基础。我们最近发现癫痫患者及动物模型脑组织中异常表达Dock3，电生理研究提示Dock3对海马锥体神经元兴奋性有显著影响，且Dock3 shRNA干预影响了海马锥体细胞NMDA受体依赖的突触后电流。为进一步探讨Dock3在癫痫发生中的作用及其机制，本研究拟通过体外细胞培养、离体脑片与动物实验相结合的方法，运用神经电生理、神经行为学及分子生物学等技术研究Dock3对癫痫形成影响，从而在多个层面论证我们的设想。我们提出假说：Dock3通过激活相关下游信号分子Paks/limk1,2影响了肌动蛋白的解聚平衡，与异常神经网络形成有关，且影响突触可塑性改变，参与了癫痫的发生。上述问题为进一步阐明Dock3信号通路在癫痫发生中的分子机制，寻找新的药物治疗靶点和药物提供理论依据。

中文关键词： 难治性癫痫；细胞质分裂蛋白3；p21激活激酶；神经网络

英文摘要： The mechanisms of epilepsy still remain incompletely understood. Abnormal synaptic connection between neurons, pathological reconstruction of neurocircuitry and the reconstruction of the synaptic plasticity provide anatomic basis of epileptogenesis. In our previous study, we found that the expression of Dock3 were significant increased in temporal lobe epilepsy patients and animal epilepsy models compared with normal controls. Electrophysiological studies also showed that Dock3 shRNA treatment resulted in significantly decreased the frequency of action potential and mEPSC in the hippocampal slices. And Dock3 shRNA significantly decreased the average amplitude of NMDA-EPSC without affecting the AMPA current. Dock3 shRNA also impaired the severity of status epilepticus during the acute stage in animal models. But the specific molecular mechanism is unclear. In this study, we use electrophysiological, cell culture, molecular neurobiological and animal behavior observation to explore the mechanisms of Dock3 on the structure and function of hippocampus neuron dendritic spines and abnormal neural network. We hypothesized that Dock3 involved in epileptogenesis process associated with abnormal reconstruction of neurocircuitry through its downstream protein Paks/limk1,2 by affecting the depolymerization balance of actin and microtubule. In this study, we try to prove Dock3 is involved in synaptic plasticity and the formation of abnormal neural network. And to further explore its mechanism by electroneurophysiology and molecular biotechnology in epileptogenesis. This experiment will provide new therapeutic targets for the treatment of epilepsy.

英文关键词： intractable epilepsy;Dock3;Paks;neural network