NeuroMolecular consequences of adolescent binge drinking

描述(由申请人提供)：青少年酗酒是一个日益严重的国家健康问题。卫生与公众服务部：物质滥用和精神健康服务管理局(2005年)的最新数据显示，90%以上的年轻人饮酒遵循“狂欢模式”，即在两小时内摄入足够的酒精，使血液中的酒精浓度超过0.08%的法定上限。此外，他们报告说，在调查前一个月，分别有大约2%、8%、20%和50%的12岁、14岁、16岁和18岁的青少年有酗酒行为。青春期是以神经元结构和功能的形成性变化为标志的；因此，在这一脆弱时期滥用酒精可能会导致永久性的神经损伤和成人行为缺陷。这些拟议研究的总体目标和具体目标将1)阐明青春期酒精暴露改变大脑基因表达的特定机制途径，2)确定这些改变如何持续到成年，以及3)确定它们如何通过表观遗传传递给下一代。这些实验将利用青少年“酗酒”的动物模型(大鼠)和神经细胞培养研究来确定酒精对青少年大脑发育的影响。目的1利用蛋白质组学技术，包括双向荧光差示凝胶电泳法和电喷雾电离(MSN)质谱仪，鉴定酒精暴露后与糖皮质激素受体(GR)相关的特异性蛋白质。此外，功能分析将确定这些蛋白质：蛋白质相互作用如何改变负责调节应激反应的GR目标基因。目的2使用技术来量化和鉴定青少年酒精暴露诱导的应激相关基因DNA甲基化模式的表观遗传学变化。最后，Aim 3将研究青春期接触酒精的动物的后代，以确定与青少年酗酒有关的表观遗传变化传递给后代的程度。所有这些目标的总体研究策略都将使用尖端的蛋白质组和表观遗传学技术，以及经典的社会行为任务，以获得对酒精如何在青少年和成人的压力反应中产生长期、可能是永久性的变化的广泛机制的理解。这些拟议的研究属于NIH任务的更大范围，目的是了解酒精滥用如何导致中枢神经系统的功能变化，从而对人类健康产生不利影响。此外，这些研究符合国家酒精中毒和酒精滥用研究所(NIAAA)的五年战略计划，即对酒精滥用采取“寿命视角”，并确定酒精滥用如何根据特定的生物生命阶段产生不同的生理结果。

DESCRIPTION (provided by applicant): Adolescent alcohol abuse is a growing national health concern. Recent data from the Department of Health and Human Services: Substance Abuse and Mental Health Services Administration (2005) showed that over 90% of alcohol consumption by youth follows a "binge pattern", defined as consuming enough alcohol in a 2- hour time span to raise the blood alcohol concentrations above the legal limit of 0.08%. Moreover, they report that approximately 2, 8, 20, and 50% of teenagers aged 12, 14, 16, and 18 years, respectively, engaged in binge drinking behavior in the month prior to the survey. The adolescent period is marked by formative changes in neuronal structure and function; therefore, alcohol abuse during this vulnerable time period can result in permanent neurological damage and adult behavioral deficits. The overall goals and specific aims of these proposed studies will 1) elucidate the specific mechanistic pathways by which alcohol exposure during adolescence modifies gene expression in the brain, 2) determine how these modifications persist through adulthood, and 3) ascertain how they are transferred by epigenetic inheritance to the next generation. The experiments will utilize both animal models (rat) of adolescent "binge drinking" and neuronal cell culture studies to determine the effects of alcohol on the developing adolescent brain. Aim 1 employs proteomic techniques, including 2-D fluorescence difference gel electrophoresis combined with electrospray ionization (MSn) mass spectrometry, to identify specific proteins that associate with the glucocorticoid receptor (GR) after alcohol exposure. In addition, functional assays will determine how these protein:protein interactions alter GR target genes responsible for regulating the stress response. Aim 2 employs techniques to quantify and identify epigenetic changes in the DNA methylation patterns of stress-related genes that are induced by adolescent alcohol exposure. Finally, Aim 3 will examine the offspring of animals exposed to alcohol during adolescence to determine the extent that the epigenetic changes associated with adolescent alcohol abuse are transferred to the offspring. The general research strategy for all of the aims will employ both cutting-edge proteomic and epigenetic techniques, along with classical social behavior tasks, to gain a broad mechanistic understanding of how alcohol can produce long-term, possibly permanent changes in the adolescent and adult stress response. These proposed studies fall within the larger scope of the NIH mission to understand how alcohol abuse produces functional changes in the central nervous system to adversely affect human health. Furthermore, these studies conform to the 5-year strategic plan of the National Institutes of Alcoholism and Alcohol Abuse (NIAAA) to take a "lifespan perspective" on alcohol abuse and determine how alcohol abuse can produce different physiological outcomes dependent upon specific biological life stages.