Early binge drinking and gene regulation

描述(由申请人提供)：INIA的这一新组成部分的总体目标是了解过度饮酒的神经生物学。流行病学数据表明，早期饮酒，尤其是过度饮酒，是最终导致酒精中毒的一个主要风险因素。我们的假设是，基因表达的先天差异和对酒精反应的差异，以及在狂欢饮酒范式中大量接触酒精的特殊性，都有助于过度饮酒的发展。我们假设酒精暴露将影响DNA甲基化，这将对基因表达和剪接产生持久影响，从而增加长期酗酒的易感性。所有这三个特定的目标将紧密联系在一起，通过在相同组织上以协调的方式检查基因表达和剪接、DNA甲基化和microRNAs，以便我们可以阐明酒精暴露发挥影响的机制。我们的策略是研究来自酒精偏好不同品系的酒精天真动物，找出容易过度饮酒的预先存在的遗传差异，并研究出生后第30天至58天反复酗酒的动物，以确定这如何影响基因表达和DNA甲基化，以及差异是否持续。我们将在反复暴饮暴食结束时以及30天后分析暴露和未暴露的动物之间的差异，看看哪些差异仍然存在。我们将关注关键的INIA区域：伏隔核壳(NAC-Shell)、杏仁中央核(CEA)和腹侧被盖区(VTA)；我们有来自成年男性的微阵列数据，显示这些区域的基因表达存在许多显著差异。在全球研究中发现的差异，以及从人类研究中挑选的候选基因，将进行测试，以确定它们是否在顺转录病毒中受到控制。我们将研究同一组织中的基因表达、剪接、DNA甲基化和microRNAs，以便对酒精可能导致过度饮酒的影响有一个完整的、系统的了解。这些机制研究不能在人类身上进行，因为我们既不能控制酒精暴露，也不能接触到脑组织。出于这个原因，另一个翻译部分将是将乙醇对大脑关键区域的影响与可接触的组织血液中的影响进行比较，以期进行潜在的人体研究。

DESCRIPTION (provided by applicant): The overall aim of this new component for INIA is to understand the neurobiology of excessive alcohol consumption. Epidemiological data demonstrate that early initiation of drinking, and particularly excessive drinking, is a major risk factor for eventual alcoholism. Our hypothesis is that both innate differences in gene expression and differences in response to alcohol, particularity to heavy exposure to alcohol in a binge drinking paradigm, contribute to the development of excessive alcohol drinking. We hypothesize that alcohol exposure will affect DNA methylation which will have lasting effects on gene expression and splicing in a way that increases long-term vulnerability to excessive alcohol drinking. All three Specific Aims will be closely linked, by examining gene expression and splicing, DNA methylation, and microRNAs in a coordinated way on the same tissues, so that we can elucidate mechanisms by which alcohol exposure exerts its effects. Our strategy is to study both alcohol-naive animals from lines selected for difference in alcohol preference, to find pre-existing genetic differences that predispose to excessive drinking, and also to study animals that have experienced repeated binge drinking from postnatal day 30 to 58 to determine how that affects gene expression and DNA methylation, and whether the differences persist. We will analyze differences between exposed and unexposed animals at the end of the repeated binges and also 30 days later to see which differences persist. We will focus on key INIA regions: the shell of the nucleus accumbens (Nac-shell), the central nucleus of the amygdala (CeA), and the ventral tegmental area (VTA); we have microarray data from adult males that show many significant differences in gene expression in these regions. Differences found in global studies, and selected candidate genes from human studies, will be tested to determine whether they are controlled in cis. We will study gene expression, splicing, DNA methylation and microRNAs in the same tissues, to allow an integrated, systems-wide understanding of the effects of alcohol that might underlie excessive alcohol consumption. These mechanistic studies cannot be carried out in humans because we can neither control alcohol exposure nor access brain tissue. For that reason, an additional translational component will be to compare the effects of ethanol on key brain regions with those in an accessible tissue, blood, looking forward to potential human studies.