Neural Substrates of Binge Drinking

酒精滥用每年造成约88，000人死亡，每年造成约2500亿美元的损失。 美方美国国家酒精研究所对酗酒的定义是， 滥用和酒精中毒：在2小时内饮用4-5杯饮料和/或血液酒精水平>80 mg/dL 时期（Sacks等人，2010年）。美国酗酒现象正在增加，在男性和女性中都非常普遍 现役军人和退伍军人，并且是酒精使用障碍的强有力预测因素（AUD; Han et 例如，2017; Stahre等人，2009; Gowin等人，2017年）。超过40%的美国退伍军人拥有一辈子 酒精使用障碍史（Fuerlein等，2016）。因此，酒精对健康、社会和环境造成巨大负担。 经济问题，特别是在退伍军人事务部。 个人对酒精的第一次体验的反应与他们患上糖尿病的风险有关。 AUD，包括频繁的早期成人狂饮（King等人，2011; McCarty等人，2004年）。识别大脑 这种危险的饮酒模式背后的神经回路是了解脆弱性的重要的第一步 澳元的因素。我们最近的研究确定了一个关键作用的核神经元（NAc）的核心，在暴食样 使用DID（在黑暗中饮酒;在小鼠中模拟狂欢式饮酒的范例）饮酒， 化学遗传学[即DREADDs（设计者受体仅由设计者药物激活）]。基于电路 动物研究与患有AUD的人类密切相关。AUD耐药男性 诊断显示减少饮酒，渴望，和复发率与脑深部刺激的NAc（皮尔斯 and Vassoler，2013）. NAc核心接收来自几个地区的预测，对许多地区来说很重要。 行为。为了进一步了解狂饮的神经回路，我们将确定NAc的神经投射。 在高饮酒C57 BL/6 J小鼠中，在DID期间参与的核心。我们会注射逆行标记， 将rAAV逆转录-GFP导入NAc核心，将小鼠暴露于乙醇、糖精或水DID程序，然后 定量GFP阳性神经元中的c-Fos免疫反应性。拟议的工作将首先确定 NAc核心回路在男性和女性酗酒期间都参与其中。我们假设 在乙醇DID期间，几个NAc投射脑区将参与，包括中央和基底外侧 杏仁核、前边缘皮质和海马、腹侧被盖区和腹侧海马。 我们的初步数据支持测试的作用，一个欠研究的投影从中央核， 杏仁核（CeA）到NAc核心。众所周知，这两个地区都与酒精有关 然而，人们对这种投射在饮酒中的性质和作用知之甚少。我们将操纵CeA 输入到NAc（通过化学遗传学和双重病毒投射靶向），以确定该投射是否 调节狂饮CeA神经元含有许多神经肽和调节剂;因此，我们将 使用免疫组织化学鉴定这些投射的性质。根据文献和结果， 我们的初步研究，我们集中在肽类神经递质，促肾上腺皮质激素释放因子（CRF）。我们将 给予促肾上腺皮质激素释放因子（CRF），以确定它是否改变了暴食样 喝酒然后，我们将测试CRF受体的拮抗作用是否可以阻断刺激CeA的作用。 > DID的NAc核心预测（通过化学遗传学）。我们假设CeA-的化学遗传刺激 >NAc预测将减少饮酒，NAc核心CRF内将减少酗酒，NAc CRF内将减少酗酒。 拮抗剂将阻断用CeA->NAc核心投射的化学发生刺激所观察到的饮酒减少。 该提案将使用行为，药理学和基于电路的方法，以更好地 了解酗酒的神经基础我们将确定神经元投射到NAc核心重要 用于调节高饮酒量C57 BL/6 J小鼠品系中的暴饮暴食。我们假设CeA 投射到NAc核心对于调节酗酒是重要的，并且参与酗酒。

Alcohol misuse is responsible for ~88,000 deaths annually and exerts an annual cost of ~$250 billion in the US. Binge drinking, which accounts for ~75% of these costs, is defined by the National Institute on Alcohol Abuse and Alcoholism as consuming 4-5 drinks and/or achieving a blood alcohol level >80 mg/dL within a 2 hr period (Sacks et al., 2010). Binge drinking is increasing in the US, is highly prevalent in both male and female active military duty personnel and veterans, and is a strong predictor of an alcohol use disorder (AUD; Han et al., 2017; Stahre et al., 2009; Gowin et al., 2017). More than 40% of US military veterans have a lifetime history of alcohol use disorder (Fuerlein et al., 2016). Thus, alcohol exerts a large burden on health, social, and economic problems, especially within the Department of Veterans Affairs. How individuals respond to their first experiences with alcohol are related to their risk for developing an AUD, including frequent early adult binge drinking (King et al., 2011; McCarty et al., 2004). Identifying the brain circuitry that underlies this dangerous pattern of drinking is an important first step in learning about vulnerability factors for AUD. Our recent studies identified a key role for the nucleus accumbens (NAc) core in binge-like drinking using DID (Drinking in the Dark; a paradigm which models binge-like drinking in mice) and chemogenetics [i.e. DREADDs (designer receptors exclusively activated by designer drugs)]. Circuitry-based studies in animals are powerfully relevant for humans with AUD. Treatment-resistant males with an AUD diagnosis exhibit reduced drinking, craving, and rates of relapse with deep brain stimulation of the NAc (Pierce and Vassoler, 2013). The NAc core receives projections from several regions and is important for many behaviors. To further understand the circuitry of binge drinking, we will identify neural projections to the NAc core that are engaged during DID in high drinking C57BL/6J mice. We will inject the retrograde marker, rAAVretro-GFP, into the NAc core, expose mice to ethanol, saccharin, or water DID procedures, and then quantify c-Fos immunoreactivity in GFP positive neurons. The proposed work will be the first to identify the NAc core circuitry engaged during binge drinking using both in males and females. We hypothesize that several NAc projecting brain regions will be engaged during ethanol DID, including the central and basolateral amygdala, prelimbic cortex, and insula, ventral tegmental area, and ventral hippocampus. Our preliminary data support testing the role of an understudied projection from the central nucleus of the amygdala (CeA) to the NAc core in binge drinking. Both regions are well known to be involved in alcohol drinking, yet very little is known about the nature and role of this projection in drinking. We will manipulate CeA inputs to the NAc (via chemogenetics and dual virus projection targeting) to determine whether this projection modulates binge-like drinking. CeA neurons contain many neuropeptides and modulators; therefore, we will identify the nature of these projections using immunohistochemistry. Based on the literature and the results of our preliminary studies, we focus on the peptide neurotransmitter, corticotropin releasing factor (CRF). We will administer corticotropin releasing factor (CRF) intra-accumbens to determine whether it alters binge-like drinking. We will then test whether antagonism of CRF receptors can block the effects of stimulating CeA- >NAc core projections (via chemogenetics) on DID. We hypothesize that chemogenetic stimulation of CeA- >NAc projections will reduce drinking, intra-NAc core CRF will reduce binge-like drinking, and intra-NAc CRF antagonists will block reductions in drinking seen with chemogenetic stimulation of CeA->NAc core projections. This proposal will use behavioral, pharmacological, and circuitry based approaches to better understand the neural substrates of binge drinking. We will identify neuronal projections to NAc core important for regulating binge-like drinking in the high drinking C57BL/6J mouse strain. We hypothesize that CeA projections to the NAc core are important for regulating binge drinking and are engaged by binge drinking.