Proteomic Dissection of Withdrawal-Induced Excessive Drinking

描述(由申请人提供)：酒精中毒综合神经科学倡议(INIA)-WEST联盟的首要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度饮酒易感性的差异的原因。因此，我们的主要关注点之一是识别其表达受酒精调控的基因，以及与任何给定的过度饮酒模式有关的基因。Wu UOL是INIA-West联盟的18个U01之一，专注于对来自具有良好特征的饮酒动物模型的脑部分进行差异蛋白质组分析，目的是确定与行为表型相关的基因靶标。最近，该联盟的集体努力已经集中在51个基因靶点上。我们建议扩大WU U01家长资助的范围，包括蛋白质组服务部分(称为INIA-WEST定量蛋白质组学核心)，以满足INIA-WEST的分析蛋白质组学需求。具体地说，将为最近选择的51个INIA-WEST基因靶点开发有针对性的蛋白质组分析(“质谱学WESTN”)，并将用于在INIA-WEST研究人员产生的大脑样本中对这些靶点进行量化。目前，INIA-WEST没有指定的蛋白质组学核心资源，也没有对INIA-WEST样本进行蛋白质组学分析的机制。通过Wu UOL母公司拨款申请补充资金将为INIA-West提供一个集中的服务部分，以集中分析INIA-West基因目标蛋白的量化，并促进对多种过度饮酒动物模型和范例的比较分析。 公共卫生相关性：对选定的基因目标蛋白进行高通量多路测量，将加快理解导致过度饮酒的脆弱性的分子差异。

DESCRIPTION (provided by applicant): The overarching goal of the Integrative Neuroscience Initiative on Alcoholism (INIA)-West 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. Therefore, one of our main focuses is the identification of genes whose expression is regulated by alcohol and which are responsible for any given model of excessive alcohol consumption. The Wu UOl is one 18 U01s of the INIA-West Consortium and is focused on the differential proteomic analysis of brain fractions enriched from well-characterized animal models of drinking for the purpose of identifying gene targets associated with the behavioral phenotype. Recently, the collective efforts of the Consortium have converged on 51 gene targets. We propose to expand the scope of the Wu U01 parent grant to include a proteomic service component (to be referred to as the INIA-West Quantitative Proteomics Core) to meet the analytical proteomic needs of INIA-West. Specifically, targeted proteomic assays ("mass spectrometry Westerns") will be developed for each of the recently selected 51 INIA-West gene targets and will be used to quantify these targets in brain samples generated from the INIA-West investigators. Currently, INIA-West has no designated Proteomics Core Resource and no mechanism for the proteomic analyses of INIA-West samples. This request for supplementary funds through the Wu UOl parent grant will provide a centralized service component to INIA-West to focus analytical efforts on the quantification of INIA-West gene target proteins and facilitate comparative analyses across the multiple animal models and paradigms of excessive drinking. PUBLIC HEALTH RELEVANCE: High-throughput multiplexed measurements of selected gene target proteins will expedite the understanding of molecular differences underlying the vulnerability to excessive alcohol consumption.