Role of nucleus accumbens core in ethanol reward and binge-like drinking: Focus on sex as a biological variable

项目摘要 酗酒是酒精使用障碍(AUD)的危险因素，而酒精使用和AUD 诊断在男性中比在女性中更普遍，这一差距已经大大缩小。有希望的治疗方法 使用啮齿动物模型已经确定了治疗AUD的候选药物；然而，很少有研究做到 将性别作为生物变量(SABV)。增加我们对神经回路中SABV的理解 这是酗酒的基础，可能会导致更多量身定做的干预措施。伏隔核核心 澳大利亚全国反兴奋剂机构(NACC)拥有不同的传入联系，这是其参与澳元所有阶段的关键基础。在治疗中 抵抗AUD，NAcc脑深部刺激的男性患者减少了酒精渴求，并促进了 减少酒精摄入量或完全戒酒，强调这一大脑区域与翻译性AUD的相关性 研究。在啮齿动物中，饮用乙醇(Etoh)会增加NAC中c-Fos(神经元活动的生物标志物)， 而NAC的操作(通过病变、DBS或化学遗传学)改变了乙醇的饮用。削弱NAcc 在雄性DBA/2J小鼠中防止获得乙醇条件性位置偏爱(CPP)，这是一种测量 对乙醇的回报效应的敏感性。由于啮齿动物NAcc中存在一些已知的性别差异， 无论是从根本上还是针对Etoh，都必须在测试SABV重要性的研究中解决SABV问题 乙醇奖励和澳元中的NACC。乙醇饮酒(DID)的摄入量存在明显的性别差异 在对NAcc进行化学发生操作后，抑制减少了C57BL/6J(B6)男性的摄入量，但 增加女性的摄入量。相反，刺激NAcc会减少女性的乙醇摄入量，而不是 在男性身上可以看到这种影响。这些发现表明，NAcc操作可能会改变对奖励的敏感性 乙醇的作用是性别依赖的，如果是这样，这可能是化学遗传诱导的变化的基础 在饮酒方面。总体而言，这项建议解决了NAcc和SABV在B6小鼠体内的重要性 狂欢/醉酒的几个方面-1)改变NAcc活动如何影响 醉酒剂量的乙醇和2)确定NAcc电路在狂欢的饮酒。 目的1测试NAcc的化学操作是否改变了乙醇的条件性奖赏效应 使用乙醇条件性位置偏好(CPP)任务。AIM 2使用全脑成像来识别区域 在DID期间参与(通过分析c-Fos表达)，结合使用病毒逆行示踪剂 给NAcc注射。这将允许对共同标记的c-Fos和表达GFP的NAcc进行量化 用于确定在此行为期间哪些NAcc电路处于工作状态的输入。将两性都包括在内，使我们能够 确定c-fos诱导是否存在差异，以及对NAcc的投射是否存在差异 在DID期间进行差异化处理。拟议的研究将极大地提高我们对1)如何 NACC的活动可能会影响性别特异性的Etoh的奖励效应，以及2)全脑和NAcc特异性 在男性和女性中，像酒精一样狂饮的环路。

Project Summary Binge alcohol drinking is a risk factor for Alcohol Use Disorder (AUD), and while alcohol use and AUD diagnoses are more prevalent in men than women, this gap has drastically narrowed. Promising therapeutic candidates for the treatment of AUD have been identified using rodent models; however, few studies have addressed sex as a biological variable (SABV). Increasing our understanding of SABV in the neural circuitry that underlies binge-like drinking could lead to more tailored interventions. The nucleus accumbens core (NAcc) has diverse afferent connections that underlie its crucial involvement in all stages of AUD. In treatment resistant male patients with AUD, NAcc deep-brain stimulation reduced alcohol craving and promoted either lower alcohol intake or total abstinence, underscoring the relevance of this brain region in translational AUD research. In rodents, ethanol (EtOH) drinking increases c-Fos (a biomarker for neuronal activity) in the NAc, and manipulations of the NAc (via lesion, DBS, or chemogenetics) changes EtOH drinking. Lesioning the NAcc in male DBA/2J mice prevents acquisition of EtOH conditioned place preference (CPP), a measure of sensitivity to the rewarding effects of EtOH. Since there are some known sex-differences in the rodent NAcc, both basally and in response to EtOH, it is imperative to address SABV in studies testing the importance of the NAcc in EtOH reward and AUD. There are apparent sex differences in EtOH Drinking-in-the-Dark (DID) intake following chemogenetic manipulation of the NAcc, where inhibition reduces intake in C57BL/6J (B6) males but increases intake in females. Conversely, stimulation of the NAcc reduces DID EtOH intake in females, with no effect seen in males. These findings suggest that NAcc manipulation may alter sensitivity to the rewarding effects of EtOH in a sex-dependent manner, and if so, this could underlie chemogenetically-induced changes in drinking. Overall, this proposal addresses the importance of the NAcc and SABV in B6 mice during 2 facets of binge/intoxication – 1) how changing NAcc activity impacts the positive subjective effects of an intoxicating dose of EtOH and 2) determining the NAcc circuitry engaged during binge-like drinking. Aim 1 tests whether chemogenetic manipulation of the NAcc alters the conditioned rewarding effects of EtOH using an EtOH conditioned place preference (CPP) task. Aim 2 uses whole-brain imaging to identify regions engaged during DID (through analysis of c-Fos expression), in combination with use of a viral retrograde tracer administered into the NAcc. This will allow for quantification of co-labeled c-Fos and GFP-expressing NAcc inputs to determine which NAcc circuits are engaged during this behavior. Inclusion of both sexes allows us to determine whether there are differences in c-fos induction, and whether projections to the NAcc are differentially engaged during DID. The proposed studies will critically improve our understanding of 1) how NAcc activity may sex-specifically impact the rewarding effects of EtOH, and 2) whole brain and NAcc specific circuits engaged by binge-like EtOH drinking in males and females.