Functional and Epigenetic Consequences of Paternal Alcohol Exposure

项目摘要/摘要 酒精使用障碍(AUD)是一种高度遗传的疾病，在美国影响着数百万人。 尽管AUD的遗传率估计为~50%，但遗传变异仅占不到 患AUD的风险为1%。最近的证据表明，改变基因的表观遗传因素 在不影响潜在DNA序列的情况下，表达可能解释了一些缺失的遗传性 澳元。研究最充分的表观遗传机制是发生在启动子内的dna甲基化。 它主要与基因抑制有关，但并不总是与基因抑制有关。人与人 临床前研究报告称，酒精会改变男性生殖细胞的DNA甲基化状态。在小鼠身上工作 研究表明，这些变化可以遗传给第一代后代，而父亲的酒精 暴露会影响后代的饮酒行为和对酒精的行为敏感性。然而，没有一项研究 研究了父亲酒精暴露对子代操作酒精自我给药的影响。因此， 拟议项目的总体目标是使用Wistar大鼠来检查酒精自我给药行为 以及酒精依赖男性后代DNA甲基化水平的变化。要做到这一点，男性Wistar 将大鼠暴露在酒精蒸气中超过6周，以诱导酒精依赖。在酒精暴露后， 雄性将在8周内不受干扰(大鼠精子发生的一个周期)，然后与酒精交配 天真的雌性。这些配对的后代将接受自我管理行为的测试和评估 成年期DNA甲基化的变化。提议的项目的第一个目标是确定父系 酒精暴露改变男性和男性操作性酒精的获得和维持自我给药 雌性后代。此外，第二个目标将确定精子中的DNA甲基化水平是否发生了变化 如果这些变化在后代的大脑和组织中保持不变。结果将进一步 了解父亲先入为主的酒精暴露的长期后果，并可能澄清 新的遗传行为和/或生物标志物，可用于开发新的或改进现有的预防措施 以及治疗澳元病的策略。

Project Summary/Abstract Alcohol use disorder (AUD) is a highly heritable disease affecting millions of people in the United States. Although the heritability of AUD has been estimated to be ~50%, genetic variants only account for less than 1% of the risk for developing AUD. Recent evidence suggests that epigenetic factors, which alter gene expression without affecting the underlying DNA sequence, may explain some of the missing heritability of AUD. The most well studied epigenetic mechanism is DNA methylation occurring at sites within the promoter regions of genes, and it is mostly, although not always, associated with gene repression. Human and preclinical studies report that alcohol can alter the DNA methylation profile in male germ cells. Work in mice has shown that these changes can be transmitted to first generation progeny and that paternal alcohol exposure affects offspring's alcohol drinking behaviors and behavioral sensitivity to alcohol. However, no study has examined the effects of paternal alcohol exposure on operant alcohol self-administration in offspring. Thus, the overall goal of the proposed project is to use Wistar rats to examine alcohol self-administration behaviors and alterations to DNA methylation levels in the offspring of alcohol-dependent males. To do this, male Wistar rats will be exposed to alcohol vapor over 6 weeks to induce alcohol dependence. After alcohol exposure, males will be left undisturbed for 8 weeks (one cycle of spermatogenesis in rats) and then mated with alcohol naïve females. Offspring from these mating pairs will be tested on self-administration behaviors and assessed for DNA methylation changes in adulthood. The first aim of the proposed project is to identify whether paternal alcohol exposure changes acquisition and maintenance of operant alcohol self-administration in male and female offspring. Additionally, the second aim will determine if DNA methylation levels are altered in the sperm of alcohol sires, and if these changes are maintained in brain and tissue of offspring. The results will further knowledge as to the long term consequences of paternal preconception alcohol exposure and may elucidate novel inherited behaviors and/or biomarkers that can be used to develop new, or refine existing, preventive and therapeutic strategies for AUD.