Molecular genetics of ion channels

可兴奋细胞表面的配体门控和电压调节离子通道介导突触传递和神经分泌。毫不奇怪，离子通道基因的突变会导致多种病理性疾病，这些疾病会影响神经分化并导致神经退行性变。这个建议是多学科的，从分子结构，使用生物物理技术调节离子通道功能，细胞生物学，研究离子通道突变对动物行为的影响。正在研究的通道包括多聚体门控烟碱乙酰胆碱和谷氨酸受体和电压门控钙通道。这些离子通道为药物干预疾病状态提供了重要的位点，所述疾病状态例如成瘾、癫痫和由于创伤或中风以及神经变性疾病引起的神经元死亡。正在开发的方法包括通过NMR表达和分析蛋白质结构域，在细胞系中重建正常或遗传修饰的离子通道以确定它们与神经毒素的结合和相互作用，膜片钳分析以检查通道剪接变体的生物物理特性，通过支架蛋白和蛋白质磷酸化调节离子通道，切片记录和动物行为分析。在此背景下，我们将使用转基因方法的靶向离子通道敲除和敲入实验，以进一步揭示特定的通道突变的生理后果，并阐明特定的通道亚基组合的生理作用。

Ligand-gated and voltage-regulated ion channels on the surface of excitable cells mediate synaptic transmission and neurosecretion. Not surprisingly, mutations of ion channel genes contribute to a wide variety of pathological disorders that can affect neural differentiation and cause neurodegeneration. This proposal is multi-disciplinary ranging from molecular structure, regulation of ion channel function using biophysical techniques, cell biology, to studying the effects of ion channel mutations on animal behavior. The channels being studied include the multimeric gated nicotinic acetylcholine and glutamate receptors and the voltage-gated calcium channels. These ion channels provide important sites for pharmacological intervention in disease status, such as, addiction, epilepsy, and neuronal death due to trauma or stroke, as well as neurodegenerative disease. Approaches being exploited include, expression and analysis of protein domains by NMR, reconstitution of normal or genetically modified ion channels in cell lines to determine their binding and interaction with neurotoxins, patch-clamp analysis to examine the biophysical properties of channel splice variants, modulation ion channels by scaffolding proteins and protein phosphorylation, slice-recording and analysis of animal behavior. Within this context we will use transgenic methods for targeted ion channel knock-out and knock-in experiments to further reveal the physiological consequences of specific channel mutations and to elucidate the physiological roles of specific channel subunits combinations.