不同功能状态KCNA2突变通过影响神经网络E/I平衡导致癫痫脑病的机制研究

神经网络兴奋性/抑制性（excitatory/inhibitory，E/I）失衡是导致癫痫脑病的重要原因。Kv1.2通道由KCNA2编码，通过介导D型钾电流调控神经网络的兴奋性。目前已知KCNA2功能缺失型，缺失合并获得型及获得型突变均可导致癫痫、认知障碍和共济失调等，但由于大脑神经网络联系的复杂性，目前的研究不能解释KCNA2三种功能状态突变均可致病的机制。本研究首先收集KCNA2基因突变患者信息，探讨基因型与表型的关系；随后构建三种不同功能状态的点突变小鼠及兴奋性或抑制性神经元KCNA2基因条件性敲除小鼠，通过膜片钳、电镜、qPCR、Western Blots、免疫荧光、激光共聚焦及行为学测试等方法分别从细胞-神经环路-整体水平探讨KCNA2基因突变对不同神经元类型及神经网络的影响，阐明KCNA2相关癫痫的复杂致病机制，以期为儿童KCNA2相关癫痫脑病的诊断和治疗提供理论依据。

Excitatory/inhibitory imbalance in neural networks plays a critical role in epileptic encephalopathies. Kv1.2 channels, encoded by KCNA2, contribute to the regulation of excitability in neural networks by mediating D-type currents. Current studies have reported that KCNA2 mutations could cause only loss-of-function (LOF), loss and gain of function (LOF/GOF), and only gain of function (GOF) of Kv1.2 channels. Surprisingly, all of the three types of KCNA2 functional changes can cause epilepsy. However, since the associations of neuronal network in brain are complicated, the underlying mechanisms still remain to be elucidated. Herein, we dedicate this study to exploring the possible mechanisms underlying KCNA2 related epilepsy. Firstly, we will collect more clinical information about patients with KCNA2 mutation to identify the associations between genotypes and phenotypes. Then, we will construct transgenic animal models including KCNA2 site-mutant mouse (LOF, GOF/LOF and GOF mutation) and conditional knockout mouse (targeted at excitatory or inhibitory neurons), which may help to investigate the effect of KCNA2 mutations on different type of neurons and neuronal networks by means of patch-clamp, qPCR, electron microscope, western blots, immunostaining, confocal microscope and behavior test. The research will be beneficial to explain the phenotypes of patients with KCNA2 mutations so as to provide theoretical foundation for diagnosis and treatment.