Excessive alcohol drinking and CNS regional changes in gene expression

描述（由申请人提供）：本提案的长期目标是更好地了解过度饮酒发展的分子神经生物学事件，并有助于过度饮酒的长期后果。需要验证的总体假设是，在扩展杏仁核（E-AMYG）的离散区域内，涉及神经传递、神经可塑性和细胞内信号通路的基因表达的变化有助于过度饮酒的发展。过量饮酒被定义为血液酒精浓度（BACs）在100-150毫克%范围内，在长期内反复达到。整个假设将通过选择性繁殖的酒精偏好(P)和高酒精饮用（HAD）大鼠进行检验。微穿孔技术将用于获取含有伏隔核壳（ACB-sh）和杏仁核中央核（CeA）的样品。基因表达的变化将使用Affymetrix微阵列来确定。RT-PCR和原位杂交将用于验证微阵列实验的关键发现。过量饮酒范例将被使用为“在黑暗中饮酒多计划访问”（DID-MSA）程序。基因表达的时间变化将在饮酒后和过度饮酒的发展过程中确定。具体目的是确定P和HAD大鼠在过度饮酒之前、开始期间和发展之后ACB-sh和CeA基因表达的变化。慢性酒精暴露和过度饮酒的发展受到多种遗传和环境因素的影响。该项目将提供遗传易感受试者E-AMYG离散区域的重要分子神经生物学信息，可以更全面地描述导致过度饮酒的发展、维持和后果的复杂细胞间和细胞内事件。这些信息对于制定治疗酒精中毒和酒精滥用的药物治疗策略至关重要。

DESCRIPTION (provided by applicant): The long-range goals of this proposal are to better understand the molecular neurobiological events that underlie the development of excessive alcohol drinking, and contribute to the long-range consequences of excessive alcohol drinking. The overall hypothesis to be tested is that changes in the expression of genes involved in neurotransmission, neuroplasticity, and intracellular signaling pathways within discrete regions of the extended amydala (E-AMYG) contribute to the development of excessive alcohol drinking. Excessive drinking is defined as sustainable blood alcohol concentrations (BACs) in the range of 100-150 mg% that are repeatedly attained over a chronic period. The overall hypothesis will be tested using selectively bred alcohol-preferring (P) and high-alcohol-drinking (HAD) rats. Micro-punch techniques will be used to obtain samples containing the nucleus accumbens shell (ACB-sh) and central nucleus of the amygdala (CeA). Changes in gene expression will be determined using Affymetrix microarrays. RT-PCR and in situ hybridization will be used to verify key findings from the microarray experiments. The excessive alcoholdrinking paradigm to be used will be the 'drinking in the dark multiple scheduled access' (DID-MSA) procedure. Time-course changes in gene expression will be determined following a drinking episode, and during the development of excessive alcohol drinking. The specific aims will be designed to determine changes in gene expression within the ACB-sh and CeA of P and HAD rats prior to, during initiation of, and following development of excessive alcohol drinking. The effects of chronic alcohol exposure and the development of excessive alcohol drinking are influenced by multiple genetic and environmental factors. This project will provide important molecular neurobiological information in discrete regions of the E-AMYG of genetically vulnerable subjects that could more comprehensively describe the complex inter- and intracellular events leading to the development, maintenance, and consequences of excessive alcohol drinking. Such information would be critically important for developing pharmacotherapeutic strategies to treat alcoholism and alcohol abuse.