GABAa-R Alpha4 Subunit in Ethanol-Related Behaviors

说明(申请人提供)：A型伽马氨基丁酸 受体(GABAA-R)被广泛认为是血管紧张素转换酶的重要组成部分。 乙醇的作用机理。然而，个人的贡献 GABAA-R亚基对乙醇相关行为的研究还不够充分 地址。GABAA-R的Alpha4亚基被明确牵连 在调节各种实验动物的行为过度兴奋方面 情况包括对酒精的依赖和戒断。对.的使用 先进的转基因和基因打靶技术现在允许假说 以明确的方式进行测试，使用精确的分子工具 整个动物的背景。通过使用Alternate测试各种小鼠模型 α4在一组细胞上表达的区域和时间模式 分子、细胞和行为分析，我们有机会 阐明α4亚单位对临床相关性的贡献 与酒精相关的行为。我们相信这些研究提供了一个完整的 我们的目标是了解乙醇如何在人体内发挥作用 中枢神经系统。为了实现我们的目标，我们的具体目标如下 建议：A1.创建并鉴定转基因小鼠品系 过表达四环素调节的GABAA-Rα4亚单位。A2.创建和 描述了携带普遍存在的基因敲除的基因工程小鼠的特征 GABAA-R的α4亚基。A3.创造和表征新型转基因 指导可诱导的神经元特异性基因重组的小鼠品系。A4. 建立并鉴定可诱导的、神经元特异性的GABAα4基因敲除 老鼠线。A5.用一组分析这些基因操纵的动物 评估基本生理、行为和表型改变的测试 乙醇诱导的行为。

DESCRIPTION (provided by applicant): The gamma-aminobutyric acid type A receptor (GABAA-R) has been extensively implicated as a key component of the mechanism of action of ethanol. However, the contribution of individual subunits of the GABAA-R to ethanol-related behaviors has not been adequately addressed. The alpha4 subunit of the GABAA-R has specifically been implicated in modulating behavioral hyperexcitibility in a variety of experimental conditions including ethanol dependence and withdrawal. The use of sophisticated transgenic and gene targeting technology now allows hypotheses to be tested in an unambiguous manner, using precise molecular tools within the context of the whole animal. By testing a variety of mouse models with altered regional and temporal patterns of expression of alpha4 on a battery of molecular, cellular, and behavioral assays, we have the opportunity to elucidate the contribution of the alpha4 subunit to clinically relevant ethanol-related behaviors. We believe these studies provide an integrated approach toward our goal of understanding how ethanol exerts its effects in the central nervous system. To accomplish our goal, the following specific aims are proposed: A1. Create and characterize a transgenic mouse line that overexpresses a tetracycline regulated GABAA-R alpha4 subunit. A2. Create and characterize genetically engineered mice that harbor a ubiquitous knockout of the alpha4 subunit of the GABAA-R. A3. Create and characterize novel transgenic mouse lines that direct inducible, neuron-specific gene recombination. A4. Create and characterize an inducible, neuron-specific GABAAR alpha4 knockout mouse line. A5. Analyze these genetically manipulated animals with a battery of tests to assess basic physiology, behavior, and phenotypic alterations in ethanol-induced behaviors.