Physiologic Analysis of Two GABAR Gamma2-Subunit Domains

描述(由申请者提供)：本建议书描述了一项为期5年的培训计划，旨在发展神经学和癫痫的学术生涯。这位首席研究员已经在圣路易斯的华盛顿大学完成了神经学住院医师培训，并将于2002年6月在华盛顿大学完成临床癫痫奖学金。然后，他将作为范德比尔特大学医学院癫痫学部的神经学助理教授，扩展他的科学技能。该计划将促进电生理学的指挥，如应用于癫痫。罗伯特·L·麦克唐纳博士将指导首席研究员的科学发展。麦克唐纳博士是电生理学领域公认的领导者。他是神经病学主任，培养了许多K08获奖者、博士后研究员和研究生。此外，与神经科学系教职员工的密切互动将提供额外的科学和职业建议。研究将集中在A型氨基丁酸受体(GABAAR)的生理学和药理学上，GABAAR是中枢神经系统中主要的快速抑制离子通道。GABAAR是几种抗癫痫药物的靶点，与至少两种形式的人类家族性癫痫的点突变有关，并被认为在颞叶癫痫中将具有锌的改变调制。该实验需要构建突变型和嵌合型GABAARs，在培养细胞中表达重组受体，并通过快速给药到大片上和通过分析单通道电流来确定其生理动力学参数。其具体目的包括：1)评估包含人类癫痫中发现的点突变的生理学GABAAR，2)确定变构调节剂对相同GABAAR突变体的影响，3)确定锌的结合结构域，4)确定GABAAR调节剂对锌的抑制作用。范德比尔特大学神经病学系通过将来自不同资源的专业知识整合到定制的课程中，为培训内科科学家提供了理想的环境。这样的环境最大限度地增加了首席研究员建立科学地位并开始成功的独立学术生涯的可能性。

DESCRIPTION (provided by applicant): This proposal describes a 5-year-training program for the development of an academic career in neurology and epilepsy. The principal investigator has completed residency training in neurology at Washington University in St. Louis and will complete a clinical epilepsy fellowship at Washington University in June 2002. He will then expand upon his scientific skills as an Assistant Professor of Neurology in the Epilepsy Division at Vanderbilt University Medical School. This program will promote the command of electro physiology, as applied to epilepsy. Robert L. Macdonald, MD, PhD will mentor the principal investigator's scientific development. Dr. Macdonald is a recognized leader in the field of electro physiology. He is the Chair of Neurology and has trained numerous K08 recipients, post-doctoral fellows and graduate students. In addition, close interaction with faculty in the Department of Neuroscience will provide additional scientific and career advice. Research will focus on the physiology and pharmacology of the gamma amino butyric acid receptor type A (GABAAR), the main fast inhibitory ion channel in the central nervous system. The GABAAR is the target of several anti-epileptic drugs, is associated with point mutations in at least two forms of human familial epilepsy, and is hypothesized will have an altered modulation by zinc in temporal lobe epilepsy. The proposed experiments entail construction of mutant and chimeric GABAARs, expressing the recombinant receptors in cultured cells, and determining their physiological kinetic parameters by rapid-application of drugs to macropatches and by analysis of single channel currents. The Specific Aims include: 1) evaluating the physiology GABAAR containing the point mutations found in human epilepsy, 2) determining the effect of allosteric modulators on the same GABAAR mutants, 3) determining the binding domains of zinc, and 4) determining the effect of GABAAR modulators on zinc inhibition. The Neurology Department at Vanderbilt University provides an ideal setting for training physician-scientists by incorporating expertise from diverse resources into customized programs. Such an environment maximizes the probability that the principal investigator will establish a scientific niche and embark upon a successful independent academic career.