ETHANOL MECHANISMS IN GABAAR GENE TARGETED MICE

描述：(改编自《调查员摘要》)：焦点 这个项目继续是转基因解剖的推定 乙醇的分子靶点解释神经生理学基础 行动。中心假设是A型伽马-氨基丁酸 酸受体(GABA-A受体)是该机制的关键组成部分 对酒精的行为反应有贡献的行为。 已经/或将会产生具有特定改变的突变体 与GABA的神经化学反应密切相关的基因。这个 正在/将对老鼠进行一系列测试，这些测试跨越了 分子、细胞和行为层面；这种多层面的方法 将允许正确地将响应归因于乙醇的位置 行动。具体地说，无处不在缺乏测试版的老鼠品系 GABA-A受体的Gamma 2L或Delta亚基已经 产生的；电生理和行为反应 动物对乙醇的反应正在调查中。两个新的鼠标品系正在开发中 创建(并将被分析)包含组织特异性敲除的 蛋白激酶C的伽马亚型；这个基因已经被证明 对酒精的行为反应至关重要。战略上的 这种有条件淘汰法的固有优势是 避免了基因改变的不良发育影响， 这项技术使人们能够绘制出神经解剖位置(S) 与观察到的行为表型有关。另外三部小说 正在创建并将分析鼠标线；这些线是：(A) 普遍存在的GABA-A受体α1亚单位基因敲除，(B) 表达Ala(291)Trp点突变的转基因 GABA-A受体的亚单位；研究人员已经证明 从电生理角度来看，这种突变可以消除乙醇的增强作用 GABA作用；相同Ala(291)Trp突变的敲门突变 它取代了内源性的alpha1基因。总而言之，这些研究 提供一种综合的方法来了解乙醇是如何发挥作用的 它对中枢神经系统的影响。

DESCRIPTION: (Adapted from the Investigator's Abstract): The focus of this project continues to be transgenic dissection of the putative molecular targets of ethanol to explain the neurophysiological basis of action. The central hypothesis is that the type A gamma-aminobutyric acid receptor (GABA-A receptor) is a critical component of the mechanism of action that contributes to the behavioral response to ethanol. Mutants have been/ or will be created that harbor specific alterations in genes intimately involved in the neurochemical response to GABA. The mice are being/ will be analyzed with a battery of tests that span the molecular, cellular and behavioral levels; such a multi-level approach will allow responses to be properly attributed to a site of ethanol action. Specifically, mouse lines that ubiquitously lack either the beta 3, gamma 2L or delta subunits of the GABA-A receptor have already been produced; the electrophysiologic and behavioral responses of these animals to ethanol is being investigated. Two new mouse lines are being created (and will be analyzed) that harbor tissue-specific knockouts of the gamma isoform of protein kinase C; this locus has been demonstrated to be critical for behavioral responses to ethanol. The strategic advantages inherent in this conditional knockout approach are that untoward developmental effects of the genetic alteration are avoided, and the technique allows one to map the neuroanatomic location(s) that contribute to the behavioral phenotype observed. Three additional novel mouse lines are being created and will be analyzed; these are: (a) a ubiquitous knockout of the alpha1 subunit of the GABA-A receptor, (b) a transgene that expresses an Ala (291) Trp point mutation in the alpha1 subunit of the GABA-A receptor; the investigators have demonstrated electrophysiologically that this mutation abolishes ethanol enhancement of GABA action; a knockin mutation of the same Ala (291) Trp mutation that replaces the endogenous alpha1 gene. Together, these studies provide an integrated approach toward understanding how ethanol exerts its effects on the central nervous system.