The role of amygdalar estrogen receptors in ethanol reward and binge drinking

 描述（由申请人提供）：酒精使用障碍（AUD）每年影响数百万美国人，造成巨大的个人、社会和经济损失。妇女特别容易受到酒精滥用的负面健康后果的影响，在酒精滥用程度低于男子的情况下，她们遭受神经、心血管和肝脏损害。令人担忧的是，近几十年来，女性人群中AUD的患病率急剧增加。目前的治疗策略主要是基于在雄性动物中进行的研究，对大脑对乙醇反应的性别差异知之甚少。 越来越多的证据表明，雌激素受体（ER）能够增加雌性动物的酒精消费量和酒精体验的主观快乐（或“奖励”）。在初步研究中，我们的实验室已经证明了用卵巢类固醇激素雌二醇治疗的雌性小鼠的乙醇奖励和狂欢样饮酒行为增加。杏仁核是一个在乙醇成瘾动物模型中作为AUD发展介导者的脑区，表达高密度的ER。此外，ER调节多种基因的表达，这些基因已知调节酒精对杏仁核神经传递的影响。因此，本项目的目标是：1）确定经典ER，ERα或ERβ中的哪一个主要负责乙醇奖励和狂欢样饮酒行为的雌激素增强，2）研究雌激素和乙醇对杏仁核基因表达的影响，3）确定ER作用于增强雌性小鼠乙醇相关行为的杏仁核区域。 首先，使用ERα和ERβ特异性激动剂PPT和DPN进行系统治疗，以确定哪种ER介导雌二醇分别在条件性位置偏好（CPP）和黑暗中饮酒（DID）范式中增强乙醇奖励和狂欢样饮酒行为。第二，杏仁核组织将从用ER激动剂处理的雌性小鼠中分离，并在DID范例中进行酒精暴露，实时聚合酶链反应（RT-PCR）分析将测量已知参与乙醇行为反应的子区域中关键基因的表达。第三，在中央和内侧杏仁核（CeA和MeA）中ERα或ERβ表达的RNAi敲低将用于定位ER对自然循环雌性小鼠中乙醇相关行为的影响。 该项目采用成熟的行为和分子技术来研究杏仁核内质网在调节女性大脑对乙醇的反应中的作用。因此，该研究培训计划将为科学理解女性AUD发展做出重要贡献，同时为申请人在乙醇研究中的成功学术生涯做好准备。

 DESCRIPTION (provided by applicant): Alcohol use disorders (AUDs) affect millions of Americans every year, at great personal, social, and economic cost. Women are particularly vulnerable to the negative health consequences of alcohol abuse, suffering neurological, cardiovascular, and liver damage at lower levels of alcohol abuse than men. Alarmingly, recent decades have seen a dramatic increase in the prevalence AUDs among the female population. Current treatment strategies are based on studies conducted predominantly in male animals, and little is known about sex differences in the brain's response to ethanol. A growing body of evidence suggests that estrogen receptors (ERs) are able to increase both the consumption of alcohol and the subjective pleasure (or "reward") of the alcohol experience in female animals. In preliminary studies, our lab has demonstrated increased ethanol reward and binge-like drinking behavior in female mice treated with the ovarian steroid hormone estradiol. The amygdala, a brain region well established as a mediator of AUD development in animal models of ethanol addiction, expresses a high density of ERs. Furthermore, ERs regulate the expression of multiple genes that are known modulate the effects of alcohol on amygdalar neurotransmission. The objectives of this project are therefore: 1) Determine which of the classical ERs, ERα or ERβ, is primarily responsible for estrogenic enhancement of ethanol reward and binge-like drinking behavior, 2) Investigate the effects of estrogen and ethanol on gene expression in the amygdala, and 3) Identify regions of the amygdala where ERs act to enhance ethanol-related behaviors in female mice. First, systemic treatment with the ERα- and ERβ-specific agonists PPT and DPN will be used to determine which ER mediates estradiol's enhancement of ethanol reward and binge-like drinking behavior in the conditioned place preference (CPP) and drinking in the dark (DID) paradigms, respectively. Second, amygdalar tissue will be isolated from female mice treated with ER agonists and subjected to binge ethanol exposure in the DID paradigm, and real-time polymerase chain reaction (RT-PCR) analysis will measure the expression of key genes in subregions known to be involved in behavioral responses to ethanol. Third, RNAi knockdown of ERα or ERβ expression in the central and medial amygdalar nuclei (CeA and MeA) will be used to localize ER effects on ethanol-related behaviors in naturally cycling female mice. The proposed project implements well-established behavioral and molecular techniques to investigate the role of amygdalar ERs in regulating the female brain's response to ethanol. This Research Training Plan will therefore make an important contribution to scientific understanding of AUD development in women while preparing the applicant for a successful academic career in ethanol research.