Alcohol/Stress Effects on Kappa Opioid Receptors in the Amygdala and Accumbens

总结 压力是一个众所周知的因素，促进酗酒和复发饮酒的个人与酒精使用 障碍（AUD）和避免酒精戒断的负面情感症状（例如焦虑和 抑郁症）也被认为在复发中起关键作用。此外，负面情绪障碍可能 这些疾病的相互作用可能会加剧两者的症状。一 脑应激系统的一个组成部分是由神经肽强啡肽和它的目标，κ 阿片受体（KOR），并且在暴露于慢性酒精和/或压力后增加。我们将使用一个 慢性间歇性乙醇（CIE）暴露与慢性强迫游泳应激或 小鼠单次长时间应激，以了解KOR在适应不良神经生物学中的作用 应激/乙醇相互作用引起的变化。我们以前的工作表明， 慢性乙醇可显著上调中脑核强啡肽/KOR系统 在小鼠CIE模型和长期暴露模型中， 长期自愿饮用乙醇的猴模型。此外，在小鼠中，我们发现全身给予 KOR拮抗剂减少焦虑/强迫行为（埋大理石）和戒断诱导的过度 喝酒多巴胺（DA）参与对压力的负性情感反应，并且已知DA终末 KOR活动的目标。因此，我们将采用多管齐下的方法来研究KOR对DA的监管 调节饮酒动机的NAc和调节饮酒动机的基底外侧杏仁核（BLA）中的信号传导。 调节压力/焦虑相关的行为，使用来自这些区域的KOR的条件性病毒敲低， 微透析以评估暴露于CIE和/或应激后的基础多巴胺和强啡肽水平 范例和荧光传感器dLight来查询这些动物中的实时多巴胺信号， 对时间锁定行为的反应。本提案的总体目标是界定知识产权代表在发展议程中的作用 慢性酒精和/或压力暴露后NAc和BLA中的信号传导，以及这些适应性变化是如何发生的。 在压力反应性边缘电路中，负责与退缩相关的负面影响的节点可以 用于治疗AUD。

SUMMARY Stress is a well-known factor in promoting heavy drinking and relapse to drinking in individuals with alcohol use disorder (AUD), and avoidance of the negative affective symptoms of ethanol withdrawal (e.g. anxiety and depression) is also believed to play a critical role in relapse. Further, disorders of negative affect may predispose individuals to AUD and the interaction of these disorders may exacerbate symptoms of both. One component of the brain stress system is comprised of the neuropeptide dynorphin and its target, the kappa opioid receptors (KOR), and it is augmented following exposure to chronic alcohol and/or stress. We will use a mouse model of chronic intermittent ethanol (CIE) exposure combined with either chronic forced swim stress or mouse single prolonged stress to understand the role that KORs play in the maladaptive neurobiological changes induced by stress/ethanol interactions. Our previous work revealed that the function of the dynorphin/KOR system in the nucleus accumbens (NAc) was robustly up-regulated by chronic ethanol exposure and withdrawal in a time/exposure dependent manner in both the CIE model in mice and in a long- term voluntary ethanol drinking model in monkeys. In addition, in mice we found that systemic administration of a KOR antagonist reduced anxiety/compulsive behaviors (marble burying) and withdrawal-induced excessive drinking. Dopamine (DA) is involved in negative affective responses to stress, and DA terminals are known targets of KOR activity. Therefore, we will use a multipronged approach to examine KOR regulation of DA signaling in the NAc, which regulates motivation to drink alcohol, and the basolateral amygdala (BLA), which regulates stress/anxiety-related behaviors, using conditional viral knockdown of KORs from these regions, microdialysis to assess basal dopamine and dynorphin levels following exposure to CIE and/or stress paradigms, and the fluorescent sensor dLight to query real-time dopamine signaling in these animals in response to time-locked behaviors. The overall goal of this proposal is to define the role of KORs on DA signaling in the NAc and BLA following exposure to chronic alcohol and/or stress, and how these adaptations in stress-responsive limbic circuitry nodes that are responsible for withdrawal-associated negative affect can be targeted for the treatment of AUD.