Neural Substrates of Binge Drinking

酒精滥用每年造成约8.8万人死亡，每年造成约2500亿美元的损失 美国。根据美国国家酒精研究所的定义，酗酒占这些成本的75%左右 滥用和酗酒，如在2小时内喝了4-5杯酒和/或血液酒精含量达到80毫克/分升 期间(Sack等人，2010年)。在美国，酗酒正在增加，在男性和女性中都非常普遍 现役军人和退伍军人，是酒精使用障碍的强烈预测因素(AUD；han et 等人，2017年；Stahre等人，2009年；Gowin等人，2017年)。超过40%的美国退伍军人一生 酒精使用障碍史(Fuerlein等人，2016年)。因此，酒精对健康、社会和 经济问题，特别是退伍军人事务部内部的问题。 个体对第一次饮酒经历的反应与他们患上 AUD，包括成人早期经常酗酒(King等人，2011年；McCarty等人，2004年)。辨认大脑 这种危险饮酒模式背后的回路是了解脆弱性的重要第一步 澳元的影响因素。我们最近的研究确认了伏隔核(NAC)核心在类似狂欢中的关键作用 使用DID饮酒(在黑暗中饮酒；一种模拟小鼠暴饮式饮酒的范例)和 化学遗传学[即DREADDS(专门由特制药物激活的特制受体)]。基于电路的 对动物的研究与患有AUD的人类有很强的相关性。患有AUD的抗药性男性 诊断显示，随着NAC的脑深部刺激，饮酒、渴求和复发率减少(皮尔斯 和瓦索勒，2013年)。NAC核心接收来自多个地区的预测，并对许多地区很重要 行为。为了进一步了解狂饮的回路，我们将确定对NAC的神经投射 在高饮酒C57BL/6J小鼠DID过程中参与的核心。我们会注射逆行标记， RAAVretro-GFP，进入NAC核心，将小鼠暴露于乙醇、糖精或水中，然后 定量检测GFP阳性神经元的c-Fos免疫反应。拟议的工作将是第一个确定 在狂饮期间，NAC核心电路在男性和女性中都使用。我们假设 在乙醇注射过程中，几个NAC投射脑区将被激活，包括中央和基底外侧 杏仁核、大脑皮层、脑岛、腹侧被盖区和腹侧海马体。 我们的初步数据支持测试来自中央核的未被充分研究的投射的作用 在酗酒中，杏仁核(CEA)到NAC核心。众所周知，这两个区域都与酒精有关 饮酒，但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 向NAC输入(通过化学遗传学和双重病毒投射靶向)，以确定该投射是否 调节狂欢般的饮酒。CEA神经元含有许多神经肽和调节剂；因此，我们将 用免疫组织化学方法确定这些投射的性质。基于文献和研究结果 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)。我们会 伏隔体内注射促肾上腺皮质激素释放因子(CRF)以确定它是否改变了狂欢样 喝酒。然后，我们将测试CRF受体的拮抗是否可以阻断刺激CEA-A的作用。 DID上的&gt；NAC核心预测(通过化学遗传学)。我们假设CEA的化学生成刺激- NAC预测将减少饮酒，NAC内部核心CRF将减少酗酒，NAC内部CRF将减少酗酒 拮抗剂将阻止通过CEA-&NAC核心投射的化学生成刺激而出现的饮酒减少。 该提案将使用基于行为、药理学和电路的方法来更好地 了解酗酒的神经基础。我们将确定对NAC核心的神经元投射重要 用于调节高饮酒C57BL/6J小鼠品系暴饮性饮酒。我们假设CEA 到NAC核心的突起对于调节狂饮很重要，并且与狂饮有关。

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.