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导致过度饮酒发展的分子神经生物学事件，并导致过度饮酒的长期后果。待检验的总体假设是，在扩展杏仁核（E-AMYG）的离散区域内参与神经传递、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量饮酒的发展。过量饮酒被定义为持续血液酒精浓度（BAC）在100-150 mg%的范围内，在慢性期内反复达到。将使用选择性繁殖的酒精偏好（P）和高酒精饮用（HAD）大鼠来检验总体假设。将使用微穿孔技术获得含有杏仁核壳（ACB-sh）和杏仁核中央核（CeA）的样本。 基因表达的变化将使用Affytron微阵列测定。RT-PCR和原位杂交将用于验证微阵列实验的关键发现。将使用的过度饮酒范例是“在黑暗中饮酒多个预定访问”（DID-MSA）程序。基因表达的时程变化将在饮酒事件后和过度饮酒的发展过程中确定。具体的目的将被设计为确定在ACB-sh和CeA的P和HAD大鼠的基因表达的变化之前，期间开始的，和之后的发展过度饮酒。慢性酒精暴露的影响和过度饮酒的发展受到多种遗传和环境因素的影响。该项目将提供重要的分子神经生物学信息的离散区域的E-AMYG的遗传易感的科目，可以更全面地描述复杂的内部和细胞内的事件，导致发展，维护，和后果的过度饮酒。 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.