Proteomic Dissection of Withdrawal-Induced Excessive Drinking

描述（由申请人提供）：长期暴露于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、饮酒失控和复发，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扩展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，遗传差异和/或神经适应在这个电路是负责的个体差异的脆弱性过度饮酒。我们建议使用定量蛋白质组学来剖析分子机制有助于行为表型的差异和过量饮酒（基线饮酒和戒断诱导饮酒）：1）腺苷酸环化酶7（AC 7）转基因动物-使用戒断诱导饮酒（WID）范例，AC 7基因拷贝数的变化产生饮酒表型的变化，以及2）高酒精偏好（HAP）/低酒精偏好（HAP）动物-通过2瓶选择（2BC）范式选择性饲养自由选择酒精消费差异的动物-所选基因型（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， 富集脑组分的定量分析，2）确定基线的总体差异 两种动物模型[AC 7转基因模型（AC 7 转基因对野生型）和HAP/HAP选择性育种模型（HAP对HAP）]，和3）在WID-2BC期间的选定时间点，全局比较动物之间蛋白质表达的纵向变化（AC 7转基因对野生型和HAP对HAP），以鉴定有助于差异和过度饮酒行为的蛋白质。

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.