BK通道和TRPV1通道在小鼠海马神经元中的相互作用机制

Ion channels are fundamental elements for maintaining cell membrane potential and neuron excitability. For understanding the role of ion channels in physiological and pathological processes and discovering ion channel related drugs, it is significant to study the interaction between ion channels in native cells. Large conductance voltage and calcium activated potassium (BK) channel is key membrane protein for coupling of cellular calcium and electric signal; transient receptor potential cation channel vanilloid 1 (TRPV1) is non selective calcium channel. Heterologous expressed BK and TRPV1 channel in HEK cells form functionally coupled complex, but it’s unknown whether BK and TRPV1 channel could also interact with each other in central neurons. BK channel mutant D434G is associated with human syndrome of generalized epilepsy and paroxysmal dyskinesia; the activation of TRPV1 channel induces epilepsy; whether the interaction between BK and TRPV1 channel changes in central neuron under epilepsy is unclear. By using patch clamp, fluorescent immunocytochemistry, coimmunoprecipitation and other biochemical methods, the interaction between BK and TRPV1 channel in hippocampus neuron from normal and epileptic mouse will be studied, and the interaction between wild type BK and TRPV1 channel and between mutant D434G and TRPV1 channel will be compared. The prospective results will provide new clue for understanding the underlying mechanism of epilepsy and designing new anti-epilepsy drugs.

离子通道是维持细胞膜静息电位、保证神经元电兴奋性的基础蛋白质。研究天然细胞中离子通道之间的相互作用对于理解离子通道在正常生理和病理条件下的功能及研发相关药物具有重要意义。电压和钙激活的大电导钾通道（BK）是神经元细胞膜上偶联钙信号和电信号的关键蛋白之一，瞬态电压感受器阳离子通道子类V成员1（TRPV1）是高度表达在中枢神经元上的非选择性钙离子通道。本项目拟通过膜片钳、免疫荧光和免疫共沉淀等方法来研究BK和TRPV1通道在正常和癫痫小鼠海马神经元中的相互作用；并通过磷酸钙转染法将野生型和癫痫突变BK（D434G）分别转染到BK通道缺失小鼠海马神经元中来比较两者和内源性TRPV1通道之间的相互作用，从而进一步阐明BK通道和TRPV1通道之间的相互作用变化与癫痫的相关性。预期研究成果将为揭示新的癫痫病发病机制及研发新的抗癫痫药物提供一定的理论基础。