Alcohol Actions-Molecular Targets on Brain Proteins

酒精作用——大脑蛋白质的分子靶标

• 批准号: 8631877
• 负责人: Robert A Harris
• 金额: $ 47.1万
• 依托单位: UNIVERSITY OF TEXAS AT AUSTIN
• 依托单位国家: 美国
• 财政年份: 1983
• 资助国家: 美国
• 起止时间: 1983-09-29 至 2019-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8631877
• 关键词: Affect; Alcohol consumption; Alcohol dependence; Alcoholism; Alcohols; Aminobutyric Acids; Attention; Behavior; Behavioral; Binding; Brain; Brain region; Chloride Channels; Collaborations; Communication; Consumption; Dissection; Ethanol; Funding; Gated Ion Channel; Genetic; Glycine; Glycine Receptors; Goals; Human; In Vitro; Individual; Injection of therapeutic agent; Intoxication; Ion Channel; Knock-in Mouse; Knock-out; Knockout Mice; Laboratories; Ligands; Link; Molecular; Molecular Target; Mus; Mutant Strains Mice; Mutation; N-Methylaspartate; Neurons; Neurotransmitter Receptor; Pathway interactions; Pharmaceutical Preparations; Pharmacotherapy; Physical Dependence; Potassium Channel; Preparation; Proteins; Publications; RNA Interference; Recombinants; Resistance; Rewards; Rodent; Role; Site; Structure; Synapses; Taurine; Techniques; Technology; Time; Transcription Coactivator; alcohol abuse pharmacotherapy; alcohol behavior; alcohol effect; alcohol response; brain pathway; cost; in vivo; innovation; interest; molecular site; mutant; new technology; novel; nuclease; public health relevance; receptor

ABSTRACT This is a renewal of ongoing R01 funding to define molecular sites of alcohol action in brain and to link alcohol effects on these receptors with specific behaviors such as consumption, reward/aversion, intoxication and physical dependence. During the current period of funding we were successful in providing molecular and even atomic level of analysis of alcohol sites on pentameric ligand-gated (pLGIC) ion channels, including the first crystallographic structure of alcohols bound within a channel protein. We also succeeded in constructing mutant mice with GABAa receptor subunits which are resistant to alcohol modulation allowing us to link specific GABAa receptors with discrete behavioral actions of alcohol. We are in the middle of the fourth year of this project period and to date have 30 publications with 7 more submitted or in preparation. Although we and others have made considerable progress, the molecular sites of alcohol action in the brain are not completely defined. Emerging evidence from human and rodent genetics, as well as recombinant receptors, indicates that several targets which have received only limited attention may be important sites of alcohol action in brain. We propose to define the role of two groups of ligand-gated ion channels, the glycine-activated chloride channels (GlyR) and GABAaRs formed from ¿ subunits (¿ GABAaRs) in actions of alcohol at the molecular, electrophysiological and behavioral levels. A strength of this proposal is the combined use of knock-out (KO) and knock-in (KI) mice allowing us to define the importance of the presence of a subunit (KO) and the importance of direct alcohol action on the protein (KI) for behavioral actions of alcohol. An innovative aspect is use of a new technology (transcription activator- like effector nuclease; TALEN) to be employed by our collaborator, Dr. Gregg Homanics, that markedly reduces the time and cost required for construction of mutant mice. In addition to our behavioral studies, mutant mice will be used by our collaborator, Dr. Neil Harrison for electrophysiological studies of glycinergic function in accumbal regions. Several GlyRs and ¿ GABAaRs are genetically linked with human alcohol dependence and understanding their role in alcohol actions may provide new targets for pharmacotherapies of alcohol abuse and alcoholism.

抽象的 这是对正在进行的 R01 资助的更新，该资助旨在定义大脑中酒精作用的分子位点并将 酒精对这些受体的特定行为产生影响，例如消费、奖赏/厌恶、 中毒和身体依赖性。在当前的融资期间，我们成功地 提供五聚体配体门控 (pLGIC) 上醇位点的分子甚至原子水平分析 离子通道，包括通道蛋白内结合的醇的第一个晶体结构。我们 还成功构建了具有GABAa受体亚基的耐酒精突变小鼠 调节使我们能够将特定的 GABAa 受体与酒精的离散行为作用联系起来。我们 正处于该项目期间的第四年中期，迄今为止已有 30 种出版物，其中还有 7 种 已提交或正在准备中。 尽管我们和其他人已经取得了相当大的进展，但酒精作用的分子位点 大脑尚未完全定义。来自人类和啮齿动物遗传学的新证据，以及 重组受体，表明仅受到有限关注的几个目标可能是 酒精在大脑中作用的重要部位。我们建议定义两组配体门控离子的作用 通道、甘氨酸激活的氯离子通道 (GlyR) 和由 ¿ 亚基 (¿ GABAaRs）在分子、电生理和行为水平上影响酒精的作用。实力为 该提案结合使用了敲除（KO）和敲入（KI）小鼠，使我们能够定义 亚基 (KO) 存在的重要性以及酒精对蛋白质 (KI) 直接作用的重要性 对于酒精的行为行为。一个创新的方面是使用新技术（转录激活剂- 如效应核酸酶； TALEN）由我们的合作者 Gregg Homanics 博士雇用，这显着 减少了构建突变小鼠所需的时间和成本。除了我们的行为研究之外， 我们的合作者 Neil Harrison 博士将使用突变小鼠进行电生理学研究 伏隔区的甘氨酸功能。一些 GlyR 和 ¿ GABAaR 与以下基因相关： 人类酒精依赖和了解其在酒精行为中的作用可能为以下方面提供新的目标： 酒精滥用和酒精中毒的药物疗法。