Transgenerational effect of gestational ethanol exposure

 描述(由申请人提供)：产前酒精暴露已被证明增加了在以后的生活中使用和滥用乙醇的可能性。一个有待回答的问题是，它也能代代相传吗？我们的数据表明，在青春期，低/中度酒精暴露会导致乙醇摄入量增加，对乙醇催眠作用的敏感度降低，不仅在子宫中暴露于乙醇的幼鼠中，在其后代中也是如此。我们的中心假设是酒精使用/滥用倾向的遗传S是跨代遗传的，并与特定大脑区域的DNA甲基化变化有关，已知这些区域与酒精奖励正确关系有关。本提案的变革性目标是确定这些脑区中涉及乙醇编程的特定基因以及涉及乙醇暴露表型跨代传递的谱系。该项目使用产前酒精暴露的动物模型来研究在子宫中直接接触酒精的后代及其后代中酒精滥用风险增加的传递。具体目标1将确定母系和父系在孕期酒精暴露对三代后代的酒精敏感性和摄入量影响中的贡献。这将通过测试产前暴露的动物和饲养产前暴露(F1)于1)乙醇、2)水或3)所有可能的组合(9；见方法中的表)的雄性和雌性大鼠来实现，以确定涉及传播的血统。我们的假设，即遗传是通过父系血统，将通过检查酒精摄入量在出生后14天(PD)和青春期，以及敏感性的催眠作用(丧失翻正反射试验)在PD 42。具体目标2将使用全基因组硫酸氢测序技术，确定怀孕期间酒精暴露对极端谱系(受影响的谱系和对照)F1和F2基因组范围DNA甲基化的影响。这项工作完成后所获得的结果将使我们能够指导未来的工作，针对涉及乙醇暴露表型传递的靶基因，并更好地理解酒精滥用风险增加的跨代遗传机制。

 DESCRIPTION (provided by applicant): Prenatal alcohol exposure has been shown to increase the probability of use and abuse of ethanol later in life. One question remaining to be answered is, can it also be transmitted across generations? Our data suggest that low/moderate ethanol exposure results in increased ethanol intake and reduced sensitivity to ethanol's hypnotic effects during puberty not only in those infant rats exposed to ethanol in the womb but also in their offspring. Our central hypothesis is that inheritance of ethanol use/abuse liability s transgenerational and associated with variations in DNA methylation in specific brain regions known to be involved in alcohols rewarding proper- ties. The transformative objective of the present proposal is to identify the specific genes in these brain areas that are involved in ethanol programming as well as the lineage involved in transgenerational transmission of the ethanol-exposed phenotype. This project uses an animal model of prenatal ethanol exposure to investigate the transmission of increased liability for ethanol abuse in the offspring directly exposed to alcohol in the womb and their progeny. Specific Aim 1 will determine the contribution of maternal and paternal lineages in the effect of gestational ethanol exposure on ethanol sensitivity and consumption in three generation of off- spring. This will be achieved through testing prenatally exposed animals and breeding male and female rats prenatally exposed (F1) to 1) ethanol, 2) water, or 3) non-manipulated in all possible combinations (9; see table in approach) to determine the lineage involved in transmission. Our hypothesis, that inheritance is through paternal lineage, will be tested by examining ethanol intake at postnatal day (PD) 14 and in adolescence, and sensitivity to the hypnotic effects (loss of righting reflex test) of ethanl at PD 42. Specific Aim 2 will identify the effect of ethanol exposure during gestation on genome-wide DNA methylation of F1 and F2 of the extreme lineage (affected lineage and control), using a whole-genome bisulfate sequencing technique. Results obtained after completion of this work will allow us to direct future work towards target genes involved in the transmission of ethanol-exposed phenotype, and to better understand the mechanisms involved in transgenerational inheritance of increased risk for ethanol abuse.