The Alpha-1 GABAA Receptor Regulates Alcohol-Drinking Behaviors

Alpha-1 GABAA 受体调节饮酒行为

• 批准号: 7595244
• 负责人: Harry L June
• 金额: $ 17.81万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-04-01 至 2011-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7595244
• 关键词: Alcohol consumption; Alcohols; Behavior; Bilateral; Breeding; Cannulas; Control Locus; Ethanol; Fostering; Globus Pallidus; Goals; HSV vector; Immunoblotting; Immunohistochemistry; In Situ; In Situ Hybridization; In Vitro; Infusion procedures; Lead; Ligands; Mediating; Methodology; Molecular Biology Techniques; Mutant Strains Mice; Play; Property; Psychological reinforcement; RNA Interference; Rat-1; Regulation; Research; Research Personnel; Reverse Transcriptase Polymerase Chain Reaction; Rewards; Role; Simplexvirus; Small Interfering RNA; Specificity; Sucrose; Techniques; Testing; Viral Vector; Virus; drinking behavior; in vivo; motivated behavior; neurobehavioral; novel; putamen; receptor; reinforcer; success; vector; zolpidem

DESCRIPTION (provided by applicant): It is well established that the rewarding properties of ethanol [EtOH] are mediated in part by GABAA-receptor mechanisms; however, only recently has research demonstrated that the a1 receptor subunit, particularly those within the ventral pallidum [VP], may be the critical GABAergic receptor regulating EtOH reinforcement. The primary objective of this proposal will be to further evaluate the role of the GABAA a1-containing receptor subunit in regulating EtOH-seeking behaviors by employing a combination of neuropsychopharmacological and molecular biology techniques. To accomplish this goal, the selectively-bred high alcohol-drinking [HAD-1] rats will be used. The primary hypothesis to be tested is that a selective inhibition of the GABAA a1 receptor containing subunit within the VP of HAD-1 rats will lead to selective reductions in EtOH-motivated behavior, with little or no effect on sucrose-motivated behaviors. The inhibition of the GABAA a1 subunit will be accomplished by constructing a herpes simplex virus [HSV] vector, which will utilize the siRNA sequence specific, posttranscriptional gene silencing mechanism. This vector will then be infused directly into the VP of HAD-1 rats via bilateral guide cannulae. It is hypothesized that a selective reduction in EtOH-maintained responding will be observed following infusion of the vector-mediated siRNA amplicon into the VP. Specifically, reinforcer and neuroanatomical specificity will be expected, as neither suppression on sucrose maintained responding following infusion of the active virus in the VP, nor suppression on alcohol responding infusion of the active virus into the neuroanatomical control locus [e.g., caudate putamen] will be expected. In subsequent studies, a combination of in situ reverse transcriptase-PCR, immunohistochemistry, and Western analyses [immunoblotting] will be used to determine the success of the HSV-siRNA viral vector manipulations. These studies should extend our understanding of the role of the GABAA a1 receptor subtype in the regulation of alcohol-drinking behaviors.

描述（由申请人提供）：众所周知，乙醇[etoH]的奖励性能部分由GABAA受体机制介导；然而，直到最近研究表明，A1受体亚基，尤其是腹侧粒子[VP]中的A1受体亚基可能是调节ETOH增强的关键GABA能受体。该提案的主要目的是进一步评估含GABAA A1受体亚基在调节寻求EtOH寻求行为中的作用，通过采用神经心理药物学和分子生物学技术的组合。为了实现这一目标，将使用有选择的高饮酒[HAD-1]大鼠。要测试的主要假设是，在HAD-1大鼠VP中，对含有亚基的GABAA A1受体的选择性抑制作用将导致EtOH动机行为的选择性减少，而对糖果动机行为的影响很小或没有影响。 GABAA A1亚基的抑制作用将通过构造单纯疱疹病毒[HSV]载体来实现，该载体将利用特定于转录后的siRNA序列，后期的后基因沉默机制。然后，该矢量将通过双侧指南插管直接注入HAD-1大鼠的VP。假设在向载体介导的siRNA扩增子输注到VP之后，将观察到ETOH维护响应的选择性减少。具体而言，预计将预期增强和神经解剖学特异性，因为在VP中注入活性病毒后，蔗糖均未抑制反应，也不会抑制活性病毒注入神经植物控制基因座中的酒精反应[例如，预计Caudate Putamen]。在随后的研究中，将使用原位逆转录酶-PCR，免疫组织化学和西方分析[免疫印迹]的组合来确定HSV-SIRNA病毒载体操纵的成功。这些研究应扩展我们对GABAA A1受体亚型在调节酒精饮用行为中的作用的理解。