Proteomic Dissection of Withdrawal-Induced Excessive Drinking

描述（由申请人提供）：长期接触酒精会导致神经适应性现象，包括耐受性、致敏性、依赖性、戒断、失去对饮酒的控制和复发，从而导致过度饮酒的发展。INIA（酒精中毒综合神经科学倡议）联盟的目标是确定与扩展的杏仁核及其连接相关的大脑奖励回路中发生的分子、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量饮酒易感性差异的原因。我们建议在两种动物模型中使用定量蛋白质组学来剖析导致差异和过度饮酒（基线饮酒和戒断性饮酒）行为表型的分子机制：1)腺苷酸环化酶7 （AC7）转基因动物-使用戒断诱导饮酒（WID）范式，AC7基因拷贝数的变化产生饮酒表型的变化；2)高酒精偏好(HAP)/低酒精偏好（LAP）动物-通过2瓶选择（2BC）范式选择性饲养的动物自由选择酒精消费量的差异-选择的基因型（多个基因的变化）有助于差异基线饮酒。我们的目标是1)开发和优化蛋白质组学方法

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.