Proteomic Dissection of Withdrawal-Induced Excessive Drinking

描述(申请人提供)：长期接触酒精会导致神经适应现象，包括耐受性、敏感化、依赖、戒断、饮酒失控和复发，这些都会导致过度饮酒。INIA(酒精中毒综合神经科学倡议)联盟的目标是确定与杏仁核及其连接相关的大脑奖赏回路中发生的分子、细胞和行为神经适应。据推测，该回路中的遗传差异和/或神经适应是导致个体对过度饮酒易感性的差异的原因。我们建议使用定量蛋白质组学在两个动物模型中分析导致差异饮酒和过度饮酒(基线饮酒和戒断诱导饮酒)行为表型的分子机制：1)腺酰环化酶7(AC7)转基因动物-AC7基因拷贝数的变化导致使用戒断诱导饮酒(WID)范式的饮酒表型变化；2)高酒精偏好(HAP)/低酒精偏好(LAP)动物-通过2瓶-选择(2BC)范式选择性地培育出针对自由选择饮酒差异的动物-所选的基因型(多个基因的变化)有助于基线饮酒。我们的目标是1)开发和优化蛋白质组方法学 对丰富的脑部分进行定量分析，2)确定基线的全球差异 两种动物模型所选品系之间的蛋白质表达[AC7转基因模型(AC7 3)建立WID-2BC期间特定时间点动物间蛋白质表达的纵向比较[AC7转基因与野生型以及HAP与LAP]，以找出导致差异饮酒行为及过度饮酒行为的相关蛋白质。

DESCRIPTION (provided by applicant): Chronic exposure to alcohol results in neuroadaptive phenomena, including tolerance, sensitization, dependence, withdrawal, loss of control of drinking, and relapse that contribute to the development of excessive alcohol consumption. The goal of the INIA (Integrative Neuroscience Initiative on Alcoholism) Consortium is to identify the molecular, cellular, and behavioral neuroadaptations that occur in the brain reward circuits associated with the extended amygdala and its connections. It is hypothesized that genetic differences and/or neuroadaptations in this circuitry are responsible for the individual differences in vulnerability to the excessive consumption of alcohol. We propose to use quantitative proteomics to dissect the molecular mechanisms contributing to the behavioral phenotype of differential and excessive drinking (both baseline drinking and withdrawal induced drinking) in two animal models: 1) Adenylyl Cyclase 7 (AC7) transgenic animals - changes in the copy number of the AC7 gene produces changes in drinking phenotype using the Withdrawal Induced Drinking (WID) paradigm and 2) High Alcohol Preference (HAP)/Low Alcohol Preference (LAP) animals - animals selectively bred for differences in free-choice alcohol consumption by the 2 Bottle-Choice (2BC) paradigm - the selected genotype (changes in multiple genes) contributes to differential baseline drinking. Our goals are 1) to develop and optimize proteomic methodology for the quantitative analysis of enriched brain fractions, 2) to identify global differences in baseline protein expression between selected lines of the two animal models [AC7 transgenic model (AC7 transgenic versus wildtype) and HAP/LAP selective breeding model (HAP versus LAP)], and 3) to globally compare longitudinal changes in protein expression between animals (AC7 transgenic versus wildtype and HAP versus LAP) at selected time points during WID-2BC to identify proteins that contribute to the differential and excessive drinking behaviors.