1/8: INIA Stress and Chronic Alcohol Interactions: Role of Dynorphin/KOR and Oxytocin Systems in Stress-Enhanced Alcohol Drinking, Relapse, and Impaired Behavioral Flexibility

项目摘要 已知压力是触发复发、促进重度酒精（乙醇）消耗、 并产生适应性，降低行为灵活性，从而损害对饮酒的控制 和增加复发的可能性。不幸的是，复杂的机制和神经回路 压力和饮酒之间相互作用还不太清楚。作为INIAstress联盟的一部分， 我们开发了一种小鼠模型，在该模型中，反复短暂的强迫游泳应激（FSS）暴露与慢性应激相互作用。 间歇性乙醇（CIE）暴露以选择性地增强依赖性（CIE暴露）小鼠的饮酒。 在目前的资助期间，我们建立了前应激神经肽强啡肽/κ阿片样物质的作用， 受体（DYN/KOR）系统在应激性饮酒中的作用。与此同时，我们使用操作性 条件反射程序，以证明抗应激神经肽催产素（OT）减少酒精自我- 给药和应激诱导的复发样行为。本研究项目的重点是DYN/KOR的作用 和OT系统在模型中的压力相关电路证明压力的能力，以相互作用， 慢性酒精促进饮酒升级，增强复发样行为， 行为灵活性该项目将采用两种压力源（强迫游泳压力（FSS）和捕食者气味（TMT）） 暴露），两种模型的压力-酒精相互作用（CIE-FSS饮酒和压力（TMT）诱导的操作 酒精复吸寻求/饮酒），以及两种行为（不）灵活性（酒精偏见选择行为 饮酒，虽有不敬之意。在我们先前工作的基础上， DYN/KOR活性在中央杏仁核（CeA）中的作用，试验数据显示强DYN输入到CeA 从岛叶皮层（一个对压力和酒精敏感的皮层区域），研究将检查DYN/KOR的作用。 岛叶皮质（IC）投射到CeA的信号传导（IC-CeADYN通路）。其他试点数据显示， OT的应激缓冲作用可能是通过CeA中的信号传导介导的。因此，研究将检查OT 下丘脑（PVN）投射到CeA（PVN-CeAOT途径）的活性。此外，研究将审查 PVN中DYN/KOR和OT系统对行为的潜在独特交互作用 压力-酒精相互作用的后果。一般的实验策略将需要利用转基因 小鼠和药理学，化学遗传学和CRISPR/Cas9方法来靶向操纵这些 这些特定神经回路中的特定神经元中的神经肽。这些研究集中在 应激相关神经回路中的促应激（DYN/KOR）和抗应激（OT）系统的适应， 不同的压力源和不同的饮酒模式，从而直接与整体主题框架相结合 的财团。最终目标是获得新的知识， 治疗与压力有关的过度饮酒和复发的干预措施。

PROJECT SUMMARY Stress is known to be a significant factor in triggering relapse, promoting heavy alcohol (ethanol) consumption, and producing adaptations that reduce behavioral flexibility, thereby compromising control over alcohol drinking and elevating susceptibility to relapse. Unfortunately, mechanisms and neurocircuitry underlying the complex interaction between stress and alcohol drinking are not well understood. As part of the INIAstress Consortium, we developed a mouse model in which repeated brief forced swim stress (FSS) exposure interacts with chronic intermittent ethanol (CIE) exposure to selectively enhance alcohol drinking in dependent (CIE-exposed) mice. During the current funding period we established a role for the pro-stress neuropeptide dynorphin/kappa opioid receptor (DYN/KOR) system in stress-enhanced alcohol drinking. At the same time, we used operant conditioning procedures to demonstrate that the anti-stress neuropeptide oxytocin (OT) reduced alcohol self- administration and stress-induced relapse-like behavior. This research project focuses on the role of DYN/KOR and OT systems within stress-relevant circuits in models demonstrating the ability of stress to interact with chronic alcohol in promoting escalation of alcohol drinking, enhanced relapse-like behavior, and impaired behavioral flexibility. The project will employ two stressors (forced swim stress (FSS) and predator odor (TMT) exposure), two models of stress-alcohol interactions (CIE-FSS Drinking and stress (TMT)-induced operant alcohol relapse seeking/drinking), and two measures of behavioral (in)flexibility (alcohol-biased choice behavior and persistence of alcohol drinking despite aversion). Building on our previous work demonstrating an important role for DYN/KOR activity in the central amygdala (CeA) and pilot data showing strong DYN input to the CeA from the insular cortex (a stress and alcohol sensitive cortical area), studies will examine the role of DYN/KOR signaling in insular cortical (IC) projections to the CeA (IC-CeADYN pathway). Other pilot data suggest that the stress-buffering effects of OT may be mediated by signaling in the CeA. Thus, studies will examine OT activity in hypothalamic (PVN) projections to the CeA (PVN-CeAOT pathway). Further, studies will examine the potential unique interactive effects of the DYN/KOR and OT systems in the PVN on behavioral consequences of stress-alcohol interactions. The general experimental strategy will entail utilizing transgenic mice and pharmacological, chemogenetic, and CRISPR/Cas9 approaches to target manipulation of these neuropeptides in specific neurons within these specific neurocircuits. Collectively, these studies focus on adaptations in pro-stress (DYN/KOR) and anti-stress (OT) systems within stress-relevant neurocircuits using different stressors and different drinking models, thereby directly interfacing with the overall thematic framework of the consortium. The ultimate goal is to gain new knowledge that will inform development of more effective interventions for treating stress-related excessive alcohol drinking and relapse.