Ventral pallidal circuitry in alcohol seeking and reinstatement by stress

寻找酒精和压力恢复的腹侧苍白球回路

• 批准号: 9385558
• 负责人: Jocelyn M Richard
• 金额: $ 13.88万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2018-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9385558
• 关键词: Abstinence; Alcohol abuse; Alcohol consumption; Alcohol dependence; Alcohols; Amygdaloid structure; Anatomy; Area; Automobile Driving; Award; Behavior; Behavioral; Brain; Calcium; Cells; Characteristics; Chronic; Chronic stress; Cocaine; Collection; Cues; Data; Dependence; Dopamine D1 Receptor; Dopamine D2 Receptor; Electrophysiology (science); Enterobacteria phage P1 Cre recombinase; Environment; Exhibits; Extinction (Psychology); Fiber; Fluorescence; Future; Globus Pallidus; Grant; Histologic; Image; Implant; Injection of therapeutic agent; Laboratories; Learning; Length; Measurement; Measures; Medial; Methods; Modeling; Neurons; Nucleus Accumbens; Opsin; Optics; Organ; Pathway interactions; Pattern; Pharmacology; Phase; Photometry; Play; Population; Prefrontal Cortex; Rattus; Relapse; Resolution; Rewards; Role; Solvents; Specificity; Stress; System; Testing; Three-Dimensional Imaging; Training; Transgenic Organisms; Viral; Virus; Withdrawal; Yohimbine; acute stress; alcohol availability; alcohol cue; alcohol exposure; alcohol relapse; alcohol response; alcohol seeking behavior; awake; base; behavioral response; calcium indicator; dependence relapse; experimental study; interest; neural circuit; neural correlate; novel; optical fiber; optogenetics; relating to nervous system; response; selective expression; sensor; skills; stressor; three dimensional structure; vapor

Project Summary Acute or chronic stress contributes to both escalation of alcohol use and relapse to alcohol seeking after abstinence. Stressors are hypothesized to contribute to relapse in part by promoting responses to cues in the environment that have been previously associated with alcohol availability, yet the brain mechanisms of this interaction between cues and stress are poorly understood. An area of the brain implicated in both stress- and cue-induced reward seeking is the ventral pallidum (VP). I have previously found that VP neurons encode the vigor of cue-elicited reward seeking, and my preliminary results suggest that VP neurons encode reinstatement of reward seeking by the pharmacological stressor, yohimbine. In the current proposal, I aim to confirm and expand my findings regarding VP encoding of reinstatement by stress, using a behavioral stressor, and to further dissect the neural circuit mechanisms by which VP neurons contribute to alcohol seeking. Here, in the K99 phase, I will examine VP encoding of cued alcohol seeking and reinstatement induced by stress in dependent versus non-dependent alcohol seeking rats. This aim will provide me with new training in models of alcohol dependence and stress reinstatement. I will then examine encoding of alcohol seeking behavior in inputs to VP from the prefrontal cortex (PFC) and basolateral amygdala (BLA), using a novel method for projection specific measurement of activity. This method, fiber photometry, uses viral-based expression of a fluorescent calcium indicator paired with optical measurement of fluorescence. Training in this skill will allow me to measure projection-specific neural activity in the proposed aims and future projects. Further, this aim will provide critical information regarding the timing of activity in BLA and PFC inputs during alcohol seeking, to be tested in the R00 phase. I will also assess the collateralization of these neurons, comparing two methods: traditional sectioning and immunolabeling-enabled three-dimensional imaging of solvent-cleared organs (iDISCO). This method provides unprecedented access to the three-dimensional structure of immunolabeled cells, and the opportunity to obtain both large-scale and high resolution information simultaneously, crucial for planned experiments in my future laboratory. In the R00 phase I will test the functional role of temporally precise activity in BLA and PFC inputs to VP during cued alcohol seeking and reinstatement of alcohol seeking, using optogenetics. I will also measure activity in genetically-defined populations of neurons projecting from the nucleus accumbens (NAc), which include both D1- and D2-dopamine receptor dominant neurons. Using transgenic rats I will selectively express calcium indicators in either D1- or D2-dominant neurons projecting to VP. These experiments will provide novel data regarding the differential activity of D1 and D2 neurons projecting to VP and will provide an important framework for future experiments and grants. These experiments will characterize the neural circuit mechanisms by which VP neurons drive alcohol seeking, and how they differ across different states of dependence and following stress.

项目概要 急性或慢性压力会导致饮酒增加和酗酒复发 节制。据推测，压力源部分通过促进对暗示的反应而导致复发。 以前认为环境与酒精的可用性有关，但这种情况的大脑机制 人们对线索和压力之间的相互作用知之甚少。大脑中与压力和压力有关的区域 线索诱导的奖励寻求是腹侧苍白球（VP）。我之前发现 VP 神经元编码 线索引发的奖励寻求的活力，我的初步结果表明 VP 神经元编码恢复 药理应激源育亨宾寻求奖励的过程。在当前的提案中，我的目的是确认并 使用行为压力源扩展我关于压力恢复的 VP 编码的发现，并进一步 剖析 VP 神经元促进酒精寻找的神经回路机制。 在这里，在 K99 阶段，我将检查提示酒精寻求和恢复诱导的 VP 编码 依赖与非依赖酒精寻求大鼠的压力。这个目标将为我提供新的训练 酒精依赖和压力恢复模型。然后我将检查酒精寻求的编码 使用一种新方法研究前额皮质 (PFC) 和基底外侧杏仁核 (BLA) 对 VP 输入的行为 用于活动的投影特定测量。这种方法，即光纤光度测定法，使用基于病毒的表达 荧光钙指示剂与荧光光学测量配对。这项技能的培训将使我 测量拟议目标和未来项目中特定于投影的神经活动。此外，这一目标将 提供有关寻酒期间 BLA 和 PFC 输入的活动时间的关键信息，以便 在R00阶段进行了测试。我还将评估这些神经元的抵押，比较两种方法： 溶剂清除器官的传统切片和免疫标记三维成像 （iDISCO）。该方法为免疫标记的三维结构提供了前所未有的途径 细胞，以及同时获得大规模和高分辨率信息的机会，这对于 计划在我未来的实验室进行实验。 在 R00 阶段，我将测试 VP 的 BLA 和 PFC 输入中时间精确活动的功能作用 在提示饮酒和恢复饮酒过程中，使用光遗传学。我也会测量活动 在从伏隔核 (NAc) 投射的基因定义的神经元群体中，其中包括 D1-和D2-多巴胺受体优势神经元。使用转基因大鼠我将选择性地表达钙 D1 或 D2 主导神经元中的指标投射到 VP。这些实验将提供新的数据 关于投射到 VP 的 D1 和 D2 神经元的差异活动，将提供一个重要的框架 用于未来的实验和资助。这些实验将表征神经回路机制 VP 神经元驱动酒精寻找，以及它们在不同的依赖状态和压力后的差异。