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Ethanol Remodeling of Striatal Microcircuits

纹状体微电路的乙醇重塑

基本信息

批准号：
8902749
负责人：
BRIAN NEIL MATHUR
金额：
$ 22.35万
依托单位：
UNIVERSITY OF MARYLAND BALTIMORE
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-09-02 至 2016-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8902749
关键词：
Acute Alcohol abuse Alcohol consumption Alcohol dependence Alcoholism Animal Behavior Axon Basal Ganglia Behavior Behavioral Biological Assay Brain Brain region CNR1 gene Cannabinoids Chronic Collaborations Control Groups Corpus striatum structure Data Depressed mood Educational process of instructing Endocannabinoids Ethanol Ethanol dependence Foundations Future Generations Grant Habits Human Image Immunohistochemistry In Vitro Interneurons Knowledge Laboratories Learning Long-Term Depression Mass Spectrum Analysis Mediating Mentors Mentorship Methods Modeling Molecular Mus National Institute on Alcohol Abuse and Alcoholism Neurons Neurosciences Research Operative Surgical Procedures Output Pathway interactions Phase Presynaptic Terminals Property Proteins Psychological reinforcement Public Health Research Research Proposals Resources Rewards Role Signal Transduction Site Slice Synapses Synaptic plasticity System Techniques Testing Toxin Tracer Training Veronica Water Work Writing alcohol effect alcohol exposure alcohol research anandamide authority behavior measurement behavioral pharmacology brain circuitry cell type collaborative environment drinking drinking behavior effective therapy endogenous cannabinoid system experience in vivo innovation interdisciplinary approach millisecond motor control neurotransmitter release novel therapeutics optogenetics post-doctoral training presynaptic response responsible research conduct skills synaptic function tool transmission process

项目摘要

DESCRIPTION (provided by applicant): The research proposal investigates how ethanol exposure induces brain circuit remodeling to result in persistent ethanol consumption. This project will focus on a brain region that strongly controls habits, the striatum. Habit formation i dependent upon endocannabinoid signaling through the cannabinoid type 1 receptor (CB1), a presynaptic terminal protein that mediates lasting forms of synaptic plasticity. The majority of striatal CB1 is localized to inhibitory microcircuit terminals, which potently control the functionl output of this brain region. Interestingly, several lines of evidence now indicate a direct effect f ethanol on the endocannabinoid system. Despite the relevance of striatal control of habits to alcoholism, a paucity of data exists in this regard. This proposal seeks to understand how acute and chronic ethanol exposure remodels the striatal inhibitory microcircuitry to promote habitual ethanol consumption. This will be investigated through three specific experimental aims. 1) Determine the acute effects of ethanol on GABAergic transmission at specific striatal inhibitory microcircuit synapses. This aim will be accomplished by exposing acute mouse brain slices to ethanol and optogenetically assessing effects on the strength and plasticity of specific striatal inhibitory microcircuit synapses. The molecular mechanism underlying these changes will be examined by assessing the interaction between ethanol and the endocannabinoid system. 2) Assess the chronic effects of ethanol drinking in the dark (DID) on GABAergic transmission at synapses of the striatal inhibitory microcircuitry. The DID paradigm models long-term, binge-like ethanol consumption present in the human condition. To examine DID-induced lasting changes to striatal inhibitory microcircuit synapses of mice, brain slices will be made and the ethanol-induced changes to efficacy and plasticity of specific microcircuit synapses will be assessed. We will test how these maladaptive homeostatic responses of the inhibitory microcircuitry arise from ethanol-induced perturbation of the endocannabinoid system. 3) Determine the contribution of the striatal inhibitory microcircuitry to DID-induced increased ethanol consumption using in vivo optogenetics. To accomplish this aim, a specific cell type comprising the inhibitory microcircuitry will be reversibly inhibited on-line, with millisecond precision, durng drinking behavior in the DID paradigm using an optogenetics approach in vivo. The contribution of this manipulation during DID to post-DID ethanol consumatory behavior will be assessed using a two-bottle (ethanol and water) choice assay. The results of these studies should provide significant advances in our knowledge of how the reinforcing effects of ethanol are mediated by brain circuit-specific alterations. In my graduate and postdoctoral training, I gained the necessary training in neuroanatomical and electrophysiological techniques necessary to effectively execute this research proposal. In my graduate work under Dr. Ariel Y. Deutch, I trained extensively in neuroanatomical and behavioral pharmacology techniques including stereotaxic microinfusion of neuronal tract tracers and toxins, imaging mass spectrometry, immunohistochemistry, and behavioral measures of motor control. Through this training, I gained an expert ability to conceptualize brain circuitry. In my postdoctoral training under Dr. David M. Lovinger, I have become proficient in electrophysiological analyses assessing synaptic function. Moreover, I have pioneered the use of optogenetics in the lab, aided by my stereotaxic surgery abilities, for determining the contribution of synapse-specific changes to circuit remodeling. During the mentored phase of the proposal, research will be conducted at NIAAA in the laboratory of Dr. David M. Lovinger, my mentor. Dr. Andrew Holmes will serve as my co-mentor. Dr. Lovinger is an authority in ethanol and synaptic plasticity research and has extensive experience in in vitro and in vivo electrophysiological recording. Dr. Holmes, a behavioral neuroscience and ethanol research expert, will oversee my continued training in assessing animal behavior. With the combined collaboration of Dr. Veronica Alvarez and Dr. Geroge Kunos, both at NIAAA, all experimental tools and research guidance necessary for executing the research proposal will be readily accessible. Finally, the resources available at NIAAA for developing my teaching, mentorship, managerial, grant writing, and responsible conduct of research skills create a supportive environment in which to readily accomplish my proposed training plan.

描述（由申请人提供）：本研究计划探讨乙醇暴露如何诱导脑回路重塑，从而导致持续的乙醇消耗。这个项目将把重点放在一个强烈控制习惯的大脑区域——纹状体上。习惯的形成依赖于内源性大麻素通过大麻素1型受体（CB1）发出的信号，大麻素1型受体是一种介导突触可塑性持久形式的突触前末端蛋白。纹状体CB1大部分定位于抑制性微电路终端，有效控制该脑区功能输出。有趣的是，现在有几条线索表明乙醇对内源性大麻素系统有直接影响。尽管纹状体控制习惯与酗酒有关，但在这方面缺乏数据。该建议旨在了解急性和慢性乙醇暴露如何重塑纹状体抑制微电路，以促进习惯性乙醇消费。这将通过三个具体的实验目的进行调查。1)确定乙醇对特定纹状体抑制性微回路突触gaba能传递的急性影响。这一目标将通过将急性小鼠脑切片暴露于乙醇中并光遗传学评估对特定纹状体抑制性微回路突触的强度和可塑性的影响来实现。这些变化的分子机制将通过评估乙醇和内源性大麻素系统之间的相互作用来研究。2)评估夜间饮酒（DID）对纹状体抑制性微回路突触gaba能传递的慢性影响。DID模式模拟了人类长期的、酗酒式的乙醇消费。为了检验did诱导的纹状体抑制性微回路突触的持久变化，我们将制作脑切片，并评估乙醇诱导的特定微回路突触的功效和可塑性的变化。我们将测试这些抑制微电路的不适应稳态反应是如何从乙醇诱导的内源性大麻素系统的扰动中产生的。3)利用体内光遗传学技术确定纹状体抑制微电路对did诱导的乙醇消耗增加的贡献。为了实现这一目标，一种包含抑制微电路的特定细胞类型将在体内使用光遗传学方法，在DID范式的饮酒行为期间，以毫秒精度在线可逆地抑制。在DID期间，这种操作对DID后乙醇消耗行为的贡献将使用两瓶（乙醇和水）选择试验进行评估。这些研究的结果应该为我们了解乙醇的强化作用是如何通过脑回路特异性改变介导的提供了重要的进展。在我的研究生和博士后训练中，我获得了有效执行本研究计划所需的神经解剖学和电生理技术的必要训练。在Ariel Y. Deutch博士的指导下，我在神经解剖学和行为药理学技术方面接受了广泛的培训，包括神经束示踪剂和毒素的立体定向微输注、成像质谱、免疫组织化学和运动控制的行为测量。通过这次训练，我获得了将大脑回路概念化的专业能力。在David M. Lovinger博士的博士后培训中，我精通电生理分析，评估突触功能。此外，在我立体定向手术能力的帮助下，我率先在实验室中使用光遗传学来确定突触特异性变化对电路重塑的贡献。在提案的指导阶段，研究将在NIAAA在我的导师David M. Lovinger博士的实验室进行。安德鲁·霍姆斯博士将担任我的共同导师。Lovinger博士是乙醇和突触可塑性研究方面的权威，在体外和体内电生理记录方面拥有丰富的经验。霍尔姆斯博士是行为神经科学和乙醇研究专家，他将监督我继续接受评估动物行为的培训。在NIAAA的Veronica Alvarez博士和george Kunos博士的联合合作下，执行研究计划所需的所有实验工具和研究指导都将随时可用。最后，NIAAA提供的资源可以提高我的教学、指导、管理、拨款写作和负责任的研究技能，为我提供了一个支持性的环境，使我很容易完成我提出的培训计划。