ETHANOL MECHANISMS IN GABAAR GENE TARGETED MICE

描述：（改编自研究者摘要）： 这个项目继续是转基因解剖假定的 乙醇的分子靶点来解释神经生理学基础， 行动上核心假设是A型γ-氨基丁酸 酸受体（GABA-A受体）是该机制的关键组成部分 对乙醇的行为反应。 突变体已经或将要产生， 与GABA的神经化学反应密切相关的基因。的 小鼠正在/将被分析与电池的测试，跨越 分子、细胞和行为水平;这种多层次的方法 将允许响应适当地归因于乙醇的位置 行动上具体来说，小鼠品系普遍缺乏β 3，GABA-A受体的γ 2L或δ亚基已经被发现。 产生;这些电生理和行为反应 动物对乙醇的耐受性正在研究中。两条新的鼠标线正在 创建（并将进行分析），其具有组织特异性敲除 蛋白激酶C的γ亚型;该位点已被证明 对乙醇的行为反应至关重要。战略 这种条件敲除方法的固有优点是， 避免了遗传改变对发育的不利影响， 并且该技术允许绘制神经解剖位置， 有助于观察到的行为表型。3新小说 正在创建鼠标线，并将进行分析;这些是：（a）a GABA-A受体α 1亚基的普遍敲除，（B） 在α 1中表达Ala（291）Trp点突变的转基因， GABA-A受体的亚基;研究人员已经证明， 在电生理学上，该突变消除了乙醇增强 GABA作用;相同Ala（291）Trp突变的敲入突变 替代了内源性的α 1基因这些研究一起 提供一个综合的方法来了解乙醇如何发挥 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.