Insular Cortex-BNST neural circuit regulation of chronic alcohol abstinence-induced negative affect

项目总结/摘要 酒精使用障碍（AUD）每年困扰着数百万人及其家庭。澳元经常 与焦虑和抑郁共病，表明饮酒动机的潜在来源。 酗酒者通常将压力源和消极情感状态列为渴望和复发的主要触发因素， 并且疾病状态的严重性与复发易感性相关。健康和经济负担 与AUD相关的研究强调了迫切需要更多的研究来了解这种相互作用， AUD和负性情感障碍之间的联系识别导致这种衰弱的关键神经适应 对于开发更好的诊断工具和治疗酒精中的情感症状至关重要 禁欲越来越多的证据表明，从社会使用到消极的替代驱动的转变 酒精寻求涉及一组统称为扩展杏仁核的大脑结构。床上 终纹核（BNST）是杏仁核延伸网络的一个组成部分，是一个关键节点， 与压力有关的疾病，如焦虑、抑郁和成瘾。BNST在许多方面发挥着重要作用 酒精成瘾的各个方面，包括酗酒和戒断的渴望。BNST与许多 皮质、皮质下、中脑和后脑区域来确定一般情感。我们最近的工作确定了一个 岛叶皮质（insular cortex，BNST）与BNST之间的功能联系。大脑参与了内感受性 意识、认知控制和感觉处理，越来越多的证据表明， 酒精中毒和消极情感障碍我们使用了一个小鼠模型，慢性饮酒，然后强迫 禁欲（CDFA），概述了胰岛素-BNST途径在禁欲诱导的负性 影响，这项建议将大大建立在这一基础证据。我们将联合收割机体内的钙 纤维光度法和离体电生理学与遗传小鼠系和各种复杂的病毒， 遗传技术来分离特定回路的神经元集合。辅导（K99）阶段将提供 纤维光度测定和复杂的病毒遗传操作策略的培训，以确定功能和 接受岛叶输入的BNST神经元的生理状态（Aim 1）和投射到 BNST（目标2）在CDFA的长期禁欲期。独立（R 00）阶段将识别二阶 输入到insulaàBNST神经元，目标是进一步描绘复杂的神经回路调节 禁欲导致的负面影响本研究建议的研究和相关职业发展培训计划 独立奖之路共同提供了一个理想的机制，将申请人过渡到一个 作为一名独立的成瘾神经学家。这些结果将大大促进我们对 慢性酒精滥用后长期戒酒的神经适应。

PROJECT SUMMARY/ABSTRACT Alcohol use disorder (AUD) afflicts millions of individuals and their families each year. AUD is frequently comorbid with anxiety and depression, indicating potential sources for the motivation to consume alcohol. Alcoholics commonly list stressors and negative affective states as leading triggers of cravings and relapse, and severity of the disease state correlates with relapse susceptibility. The health and financial burden associated with AUD highlights the pressing need for more research focused on understanding the interaction between AUD and negative affective disturbances. Identifying key neuroadaptations driving this debilitating disease is essential for developing better diagnostic tools and treatments for affective symptoms in alcohol abstinence. Converging evidence suggests the transition from social use to negative reinforcement-driven alcohol seeking involves a set of brain structures collectively referred to as the extended amygdala. The bed nucleus of the stria terminalis (BNST), a component of the extended amygdala network, is a critical node for stress-related disorders such as anxiety, depression, and addiction. The BNST plays a prominent role in many facets of alcohol addiction including binge drinking and cravings in withdrawal. The BNST interacts with many cortical, subcortical, midbrain, and hindbrain regions to determine general affect. Our recent work identified a functional connection between the insular cortex (insula) and the BNST. The insula is involved in interoceptive awareness, cognitive control, and sensory processing, and mounting evidence suggests a role for the insula in alcoholism and negative affective disturbances. We used a mouse model of chronic drinking followed by forced abstinence (CDFA) to outline a definitive role for the insula-BNST pathway in abstinence-induced negative affect, and this proposal will significantly build on this foundational evidence. We will combine in vivo calcium fiber photometry and ex vivo electrophysiology with genetic mouse lines and a variety of sophisticated viral- genetic techniques to isolate circuit-specific neuronal ensembles. The mentored (K99) phase will provide training in fiber photometry and complex viral-genetic manipulation strategies to determine the functional and physiological state of BNST neurons receiving insular inputs (Aim 1) and insular neurons that project to the BNST (Aim 2) in the protracted abstinence phase of CDFA. The independent (R00) phase will identify 2nd order inputs onto insulaàBNST neurons, with the goal of further delineating the complex neurocircuitry regulating abstinence-induced negative affect. The proposed studies and related career development training plan in this Pathway to Independence Award collectively provide the ideal mechanism to transition the applicant to a career as an independent addiction neuroscientist. The results will significantly advance our understanding of the neural adaptations that occur in protracted abstinence following chronic alcohol abuse.