Long-term alcohol drinking alters stress engagement of BNST circuit elements

长期饮酒会改变 BNST 电路元件的压力参与

• 批准号: 10021530
• 负责人: Lara Stephanie Hwa
• 金额: $ 12.13万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-20 至 2020-12-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10021530
• 关键词: Acute; Affect; Alcohol consumption; Alcohol dependence; Alcohol withdrawal syndrome; Alcohols; American; Amygdaloid structure; Anxiety; Area; Atlases; Automobile Driving; Behavior; Behavior Control; Behavioral; Biological Assay; Brain; Brain Diseases; Brain region; Cells; Chronic; Complex; Computer Analysis; Coping Behavior; Data; Development; Disease; Dynorphins; Educational process of instructing; Elements; Exposure to; Foxes; Future; Genetic Recombination; Goals; Heavy Drinking; Image; Individual; Institution; K-Series Research Career Programs; Knowledge; Laboratories; Lead; Light; Maps; Measures; Mediating; Mentors; Methods; Microscopy; Modernization; Mus; Neurons; Neuropeptide Receptor; Neuropeptides; Neurosciences; North Carolina; Odors; Opioid Antagonist; Pathologic; Pathway interactions; Phase; Phenocopy; Phenotype; Physiology; Population; Prevention; Process; Recording of previous events; Relapse; Research; Role; Signal Transduction; Source; Stress; Stress and Coping; Structure of terminal stria nuclei of preoptic region; Synapses; Synaptic Transmission; System; Techniques; Testing; Time; Tracer; Training; Treatment Cost; Universities; Viral; Virus; Visualization; addiction; alcohol exposure; alcohol use disorder; base; behavioral response; biological adaptation to stress; brain circuitry; brain tissue; chronic alcohol ingestion; coping; designer receptors exclusively activated by designer drugs; drinking; environmental stressor; experimental study; glutamatergic signaling; improved; in vivo; kappa opioid receptors; knock-down; neural circuit; new therapeutic target; novel; optogenetics; patch clamp; preclinical study; problem drinker; programs; recruit; relating to nervous system; response; skills; social; social defeat; societal costs; stress management; stressor; tool

Project Summary Alcohol use disorder is a chronically relapsing brain disease that is often precipitated by stress. The source of the stress can either be external, such as an environmental stressor acting upon the affected individual, and/or internal, with endogenous signals challenging the system for more alcohol. This K99/R00 application aims to study the neural circuits modulating long-term, intermittent alcohol (IA) drinking and stress. The dynorphin (DYN)/kappa opioid receptor neuropeptide system has been shown to be involved in the dysphoric phases of alcohol dependence in preclinical studies. The K99 portion of this proposal will involve more extensive training using cell-specific optogenetics and chemogenetics to influence alcohol-affected behavior. Specifically, this proposal tests the control of stress coping behavior in response to a predator odor during protracted withdrawal from alcohol in the bed nucleus of the stria terminalis (BNST), a brain area that has DYN adaptations following chronic IA drinking. Additional training will be gained during the second K99 phase using a discovery-based approach to identify whole brain circuit mapping after escalated alcohol drinking and stress. To probe which BNST inputs are recruited in an unbiased manner, an inducible cre-dependent retrograde virus will be used in fos-cre (Targeted Recombination of Active Populations, TRAP2) mice to identify activated inputs to the BNST that are time-locked to acute predator odor stress; these inputs may be differentially modulated by a history of IA drinking. Next, the TRAPed BNST circuitry will be identified through the iDISCO+ whole brain tissue clearing and immunolabeling method using light sheet microscopy and computational analysis aligned with the Allen Brain Atlas. Further, whole cell patch clamp recordings of DYN cells will be performed to describe the synaptic alterations in the stressed, alcohol-exposed BNST-projecting pathways. These K99 experiments will yield newly identified whole brain neural systems recruited after alcohol and stress as future avenues for independent studies for the R00 phase. The R00 studies will assess the contributions of distinct circuits using both in vivo optogenetics and a multiplexed DREADD approach to inhibit specific BNST projections. This research would thereby enhance our understanding of the functional circuitry of the brain and inform the development of targeted treatments for stress and alcohol disorders. The postdoctoral candidate, Dr. Lara Hwa, will use the K99/R00 career development award to master these modern neuroscience techniques at the University of North Carolina with her qualified mentoring team to become an expert neuroscientist in alcohol and stress interactions. She hopes to lead a strong research and teaching program at an R1 academic institution, poised to probe the complex relationship of alcohol and stress underlying pathological behavior through advanced neuroscience techniques.

项目摘要 酒精使用障碍是一种长期复发的脑疾病，通常由压力引起。的来源 压力可以是外部的，例如作用于受影响个体的环境压力源和/或 内源性信号挑战该系统的更多酒精。此K99/R00应用程序的目的是 研究神经回路调节长期，间歇性酒精（IA）饮酒和压力。 Dynorphin （Dyn）/Kappa阿片受体神经肽系统已显示出参与 临床前研究中的酒精依赖性。该提案的K99部分将涉及更广泛的培训 使用细胞特异性的光遗传学和化学遗传学来影响受酒精影响的行为。具体来说，这是 提案测试响应于长期戒断期间捕食者气味的压力应对行为的控制 从床核中的酒精（BNST）的床核（BNST），一个大脑区域，具有DYN适应的大脑区域 慢性IA喝。在第二个K99阶段将使用基于发现的额外培训获得额外的培训 识别饮酒和压力升级后识别全脑电路映射的方法。探测哪个 BNST输入以公正的方式募集，可诱导的Cre依赖性逆行病毒用于 FOS-CRE（有效群体的靶向重组，陷阱2）小鼠以识别激活的输入到BNST 被锁定到急性捕食者气味应激；这些输入可能是通过历史记录来差异的 ia喝。接下来，将通过IDISCO+全脑组织清除识别带状的BNST电路 使用轻度显微镜和与艾伦对齐的计算分析的免疫标记方法 脑图集。此外，将执行DYN细胞的全细胞贴片夹记录以描述突触 压力，暴露于酒精的BNST项目的改变。这些K99实验将产生 在酒精和压力后，新确定的整个大脑神经系统是未来的途径 R00阶段的独立研究。 R00研究将评估使用不同电路的贡献 体内光遗传学和多重恐惧的方法都可以抑制特定的BNST投影。这 研究将增强我们对大脑功能电路的理解，并告知 开发针对压力和酒精疾病的有针对性治疗方法。博士后候选人Lara博士 HWA，将使用K99/R00职业发展奖来掌握这些现代神经科学技术 北卡罗来纳大学与她合格的指导团队成为酒精的专家神经科学家 和压力相互作用。她希望在R1学术中领导强大的研究和教学计划 机构，准备探究酒精与压力的复杂关系 通过先进的神经科学技术。