Prefrontal cortical microcircuit mechanisms for reciprocal interactions between arousal and ethanol consumption

项目总结/摘要 酒精使用障碍（AUD）对个人、经济和社会造成重大损失。鉴于其高寿命 因此，迫切需要了解AUD病理生理学基础的神经生物学机制 开发有效的治疗策略。应激相关的神经内分泌功能障碍和自主觉醒 与酒精相关的行为高度相关。反复使用酒精会增加性唤起。 此外，寻求治疗的AUD患者的较高唤醒水平与较高的复发率相关。这些 研究结果表明，唤醒和饮酒之间存在相互作用，例如，较高的基础唤醒 促进饮酒，这进一步加剧了唤醒。理解神经生物学的挑战 介导觉醒和饮酒之间相互作用的机制是缺乏临床前动物模型， 允许量化饮酒以及唤醒和神经元活动的纵向测量。我们 通过为头部固定的小鼠设计一种自愿的乙醇消耗模式， 用于神经元活动记录的光子钙成像和用于测量唤醒的瞳孔测量。我们的初步 实验表明，前额叶皮层的前扣带皮层（ACC）分区有力地增加了 兴奋基础唤醒水平和唤醒相关ACC活性与乙醇量相关 消费，表明ACC有助于唤醒调制饮酒。大脑皮层的活动 由局部中间神经元的抑制作用形成。表达血管活性肠肽（VIP）的中间神经元 特别重要，因为它们抑制其他中间神经元，导致ACC的去抑制性兴奋。 假设ACC VIP神经元是唤醒和乙醇之间相互作用的关键节点 消费在目标1和2中，我们检验了VIP神经元介导的去抑制增加唤醒的假设 乙醇消费。在目标3中，我们将确定重复的乙醇消耗是否会增加VIP介导的 抑制解除，导致过度兴奋，进一步促进饮酒。我们将使用一个 结合先进的方法，包括双光子钙成像，光遗传学， 机器视觉辅助闭环光遗传学和离体切片电生理学。所有这些 机制研究将确定VIP神经元如何促进饮酒并受到饮酒的影响， 它们作为调节由异常唤醒驱动的饮酒的关键治疗靶点。

Project Summary/Abstract Alcohol use disorder (AUD) exacts a major personal, economical, and societal toll. Given its high lifetime prevalence, there is an urgent need to understand neurobiological mechanisms underlying AUD pathophysiology to develop effective therapeutic strategies. Dysfunction of stress-related neuroendocrine and autonomic arousal pathways are highly associated with alcohol-related behaviors. Repeated alcohol use increases arousal. Moreover, higher arousal levels in treatment-seeking AUD patients correlates with higher rates of relapse. These findings suggest reciprocal interactions between arousal and alcohol use, such that higher basal arousal promotes alcohol drinking, which further exacerbates arousal. A challenge in understanding the neurobiological mechanisms mediating interactions between arousal and drinking is the lack of preclinical animal models that allow quantification of drinking together with longitudinal measurements of arousal and neuronal activity. We addressed this by designing a voluntary ethanol consumption paradigm for head-fixed mice combined with two- photon calcium imaging for neuronal activity recordings and pupillometry for measuring arousal. Our preliminary experiments show that the anterior cingulate cortex (ACC) subdivision of the prefrontal cortex potently increases arousal. Basal levels of arousal and arousal-related ACC activity are correlated with the amount of ethanol consumption, suggesting that the ACC contributes to arousal modulation of drinking. Cortical activity is critically shaped by inhibition from local interneurons. The vasoactive intestinal polypeptide (VIP) expressing interneurons are particularly important as they inhibit other interneurons, leading to disinhibitory excitation of the ACC. We hypothesize that ACC VIP neurons are a key node for reciprocal interactions between arousal and ethanol consumption. In Aims 1 and 2, we test the hypothesis that VIP neuron-mediated disinhibition increases arousal and ethanol consumption. In Aim 3, we will determine if repeated ethanol consumption increases VIP-mediated disinhibition, leading to hyperarousal that further promotes drinking. We will test these hypotheses using a combination of advanced approaches including multicolor two-photon calcium imaging, multicolor optogenetics, machine vision-assisted closed-loop optogenetics, and ex-vivo slice electrophysiology. Together, these mechanistic studies will determine how VIP neurons both contribute to and are affected by drinking, establishing them as a key therapeutic target for modulating drinking driven by aberrant arousal.