Molecular genetics of ion channels

可兴奋细胞表面的配基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各种各样的病理疾病，这些疾病可能会影响神经分化并导致神经退化。这项建议涉及多个学科，从分子结构、利用生物物理技术调节离子通道功能、细胞生物学，到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以及电压门控钙通道。这些离子通道为药物干预疾病状态提供了重要的场所，如成瘾、癫痫、创伤或中风导致的神经元死亡，以及神经退行性疾病。正在开发的方法包括：蛋白质结构域的核磁共振表达和分析，细胞系中正常或转基因离子通道的重建以确定其与神经毒素的结合和相互作用，膜片钳分析以检测通道剪接变异体的生物物理性质，通过支架蛋白和蛋白质磷酸化来调节离子通道，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进行靶向离子通道敲除和敲入实验，以进一步揭示特定通道突变的生理后果，并阐明特定通道亚单位组合的生理作用。

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.