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

项目总结 众所周知，压力是引发复发、促进重酒(酒精)消费的一个重要因素， 并产生降低行为灵活性的适应，从而损害对饮酒的控制 并增加复发的易感性。不幸的是，复合体背后的机制和神经回路 压力和饮酒之间的相互作用还没有被很好地理解。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 间歇性乙醇(CIE)暴露选择性地增加依赖(CIE暴露)小鼠的饮酒量。 在目前的资助期间，我们确立了促应激神经肽强啡肽/kappa阿片的作用。 应激性饮酒中的DYN/KOR系统同时，我们使用了OPERANT 条件反射程序证明抗应激神经肽催产素(OT)减少酒精自身 用药和应激诱导的复发样行为。这个研究项目的重点是Dyn/KOR的作用 和OT系统在展示压力相互作用能力的模型中与压力相关的回路中 长期饮酒促进饮酒升级，增强复发行为，并受损 行为灵活性。该项目将采用两种应激源(强迫游泳应激(FSS)和捕食者气味(TMT)) 应激-酒精相互作用的两种模型(CIE-FSS饮酒和应激(TMT)诱导的操纵子 酗酒寻求/饮酒)和两种衡量行为(In)灵活性的指标(偏重酒精的选择行为 以及不顾厌恶坚持饮酒)。在我们之前工作的基础上，展示了一个重要的 DYN/KOR活动在中央杏仁核(CEA)中的作用和飞行员数据显示DYN对CEA的强烈输入 从岛叶皮质(一个对压力和酒精敏感的皮质区域)，研究将检验Dyn/KOR的作用 岛叶皮质(IC)投射至CEA的信号(IC-CeADYN通路)。其他试点数据表明， OT的应激缓冲作用可能是通过CEA中的信号转导来实现的。因此，研究将检查OT 下丘脑(PVN)向CEA的投射活动(PVN-CeAOT途径)。此外，研究还将检查 PVN中Dyn/KOR和OT系统对行为的潜在独特交互作用 压力和酒精相互作用的后果。一般的实验策略将需要使用转基因 小鼠和药理学、化学遗传学和CRISPR/CAS9靶向操作方法 在这些特定神经回路内的特定神经元中的神经肽。总的来说，这些研究的重点是 应激相关神经回路中前应激(Dyn/KOR)和抗应激(OT)系统的适应性 不同的压力源和不同的饮酒模式，从而直接与总体主题框架相联系 财团的成员。最终目标是获得新的知识，这将为开发提供更有效的信息 治疗与压力相关的酗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.