Synaptic Mechanisms of Addiction-Related Behaviors in the Nucleus Accumbens

伏核成瘾相关行为的突触机制

• 批准号: 8637959
• 负责人: Brad Alan Grueter
• 金额: $ 24.9万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-04-01 至 2016-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8637959
• 关键词: Abstinence; Academia; Acute Pain; Address; Affect; Area; Bacterial Artificial Chromosomes; Behavior; Behavioral; Belief; Biological Process; Brain; Cells; Cocaine; Communication; Complex; Development; Disease; Disease model; Doctor of Medicine; Doctor of Philosophy; Dopamine D2 Receptor; Drug Exposure; Drug abuse; Exposure to; Foundations; Future; Glutamate Receptor; Goals; In Vitro; Institutes; Interneurons; Knowledge; Label; Learning; Mental disorders; Mentors; Metabotropic Glutamate Receptors; Minnesota; Modeling; Modification; Molecular; Mus; Names; National Institute of Drug Abuse; Neurons; Nucleus Accumbens; Output; Paper; Patch-Clamp Techniques; Pathway interactions; Pharmaceutical Preparations; Phase; Physiology; Process; Property; Publications; Publishing; Recording of previous events; Recruitment Activity; Research; Rewards; Rhodopsin; Role; Running; Scientist; Slice; Synapses; Synaptic Transmission; Synaptic plasticity; System; Techniques; Training; Transgenic Mice; Transgenic Organisms; United States National Institutes of Health; Viral; Whole-Cell Recordings; addiction; behavior influence; career; cholinergic; chronic pain; cocaine exposure; drug of abuse; effective therapy; expectation; experience; in vivo; interest; learned behavior; light effects; meetings; mesolimbic system; motivated behavior; nerve supply; neural circuit; neuroregulation; optogenetics; post-doctoral training; postsynaptic; promoter; research study; synaptic function; tool; transmission process

Early in my career as a scientist, I decided that communication was the key to all biological processes. This ideology has evolved into a keen interest in the study of synaptic transmission. In the lab of Danny G. Winder, Ph.D. I began my training in synaptic physiology publishing many papers on modulation of excitatory synaptic transmission by metabotropic glutamate receptors and their in vivo recruitment by cocaine. During my postdoctoral training, I have continued to study synaptic transmission in reward related circuitry in the lab of Robert C. Malenka, M.D., Ph.D. Dr. Malenka is a renowned synaptic physiologist and has trained many prominent figures in the synaptic physiology and addiction fields, Antonello Bonci (NIDA), Karl Deisseroth (Stanford), Dan Feldman (UC Berkeley), Pablo Castillo (Albert Einstein), John Isaac (NIH, now Lilly), Anatol Kreitzer (UCSF), Michael Crair (Yale), and Mark Thomas (Univ. Minnesota) to name a few. It is my goal to continue this tradition by obtaining an independent lab in academia studying the role of synaptic transmission in rewarding and aversive behaviors. My area of study differs from Dr. Malenka in that in addition to excitatory transmission, I am also interested in studying inhibitory and neuromodulatory transmission onto NAc interneurons as well as primary output neurons. Eventually, I plan to expand disease models and study effects of aversive states, such as acute and chronic pain, on NAc synaptic circuitry. To do this I am utilizing state of the art techniques and sophisticated yet simple approaches to address these issues in the Nucleus Accumbens (NAc). The ultimate goal of this project is to gain a better understanding of synaptic function in the NAc circuitry and to begin to address how these circuits are recruited to elicit addiction related behaviors. The NAc, as part of the mesolimbic dopamine system, integrates a complex mix of excitatory, inhibitory and modulatory inputs to optimize adaptive motivated behaviors. Dynamic alterations in synaptic transmission within this circuitry are strongly implicated in the development and expression of addictive disorders. The specific aims involve using whole-cell recordings from in vitro slices to define basic properties of NAc neurons and how these are modified by in vivo cocaine exposure. In this proposal, the effects of in vivo cocaine exposure on synaptic properties of NAc output neurons and local microcircuit interneurons (INs) will be delineated utilizing bacterial artificial chromosome (BAC) transgenic marker mice that specifically label direct and indirect pathway medium spiny neurons (MSNs), GABAergic and cholinergic INs. Also, the synaptic properties of three distinct excitatory inputs onto NAc, MSNs and INs will be characterized and the consequences of in vivo cocaine experience on these specific inputs will be determined using virally expressed channel rhodopsin (ChR2). The objectives for the mentored phase of this proposal are: (1) to examine the synaptic properties of direct and indirect MSNs following in vivo cocaine experience, and (2) to determine if specific excitatory synaptic inputs onto these MSNs are differentially altered following in vivo cocaine experience utilizing optogenetic approaches. The independent phase will address: (1) afferent specific basal and drug-induced alterations in excitatory synaptic properties of NAc INs, (2) basal and drug-induced changes in synaptic connectivity between INs and MSNs and (3) behavioral effects of light-induced (ChR2) activation of NAc INs in drug related context, all of which utilize state-of-the-art optogenetic approaches. The results of the proposed experiments will provide a fundamental knowledge of the changes in the synaptic circuitry of the NAc in a pathophysiological state and have implications on future targets for treatment of addiction related behaviors. Additionally, the careful detailed approach of this study provides the foundation for the study of other drugs of abuse and addiction models, as well as additional affirmative disorders associated with maladaptive processes in the NAc. It is my expectation that upon completion of the mentored project I will have the technical and intellectual expertise to successfully run my own independent lab at a respected institute of higher learning. I will have developed to tools to effectively communicate these findings at meetings and in publications. Additionally, it is my belief that upon completion of these projects, I will be able to successfully compete for an R01. Ultimately, this will allow me to continue to contribute to the field of addiction research both by producing quality research and mentoring young scientists.

在我作为科学家的职业生涯早期，我认为沟通是所有生物过程的关键。这 意识形态已经演变成对突触传递研究的浓厚兴趣。在Danny G.温德 博士我开始学习突触生理学，发表了许多关于兴奋性突触调节的论文。 通过代谢型谷氨酸受体的传递和它们通过可卡因的体内募集。在我 在博士后培训期间，我继续在实验室研究奖赏相关回路中的突触传递， Robert C. Malenka，医学博士，博士Malenka博士是一位著名的突触生理学家， 突触生理学和成瘾领域的杰出人物，Antonello Bonci（NIDA），Karl Deisseroth （斯坦福大学）、丹·费尔德曼（加州大学伯克利分校）、巴勃罗·卡斯蒂略（阿尔伯特·爱因斯坦）、约翰·艾萨克（NIH，现为礼来公司）、阿纳托尔 Kreitzer（加州大学旧金山分校），Michael Crair（耶鲁大学）和Mark托马斯（明尼苏达大学）仅举几例。我的目标是 通过在学术界获得一个独立的实验室来继续这一传统，研究突触传递的作用 奖励行为和厌恶行为。我的研究领域与Malenka博士的不同之处在于，除了兴奋性 传递，我也有兴趣研究抑制和神经调节传递到NAc 中间神经元以及初级输出神经元。最终，我计划扩展疾病模型并研究其影响 NAc突触回路上的厌恶状态，例如急性和慢性疼痛。为了做到这一点，我利用国家的 艺术技术和复杂而简单的方法来解决这些问题的核心 Accumbens（NAc）。 本项目的最终目标是更好地了解NAc回路中的突触功能， 开始解决这些回路是如何被招募来引发成瘾相关行为的。NAC作为 中脑边缘多巴胺系统整合了兴奋性、抑制性和调节性输入的复杂混合物， 优化适应性动机行为。在这个回路中，突触传递的动态变化是 与成瘾性疾病的发展和表达密切相关。具体目标包括使用 来自体外切片的全细胞记录，以定义NAc神经元的基本特性以及这些特性是如何被修改的 体内可卡因暴露。在这个建议中，体内可卡因暴露对突触特性的影响， NAc输出神经元和局部微回路中间神经元（IN）将利用细菌人工神经细胞模型来描绘。 染色体（BAC）转基因标记小鼠，其特异性标记直接和间接途径培养基多刺 神经元（MSNs），GABA能和胆碱能神经元。此外，三种不同的兴奋性神经元的突触特性 将对NAc、MSN和IN的输入进行表征，并将体内可卡因经验对 这些特异性输入将使用病毒表达的通道视紫红质（ChR2）来确定。 本建议的指导阶段的目标是：（1）检查直接和 间接MSN后，在体内可卡因的经验，和（2）以确定是否特定的兴奋性突触输入 在这些MSN上，在利用光遗传学方法的体内可卡因经历之后，这些MSN被差异地改变。 独立阶段将解决：（1）传入特定的基础和药物诱导的兴奋性改变 NAc INs的突触特性，（2）INs之间突触连接的基础和药物诱导的变化， MSNs和（3）药物相关背景下光诱导（ChR2）激活NAc INs的行为效应，所有 其利用最先进的光遗传学方法。所提出的实验的结果将提供一个 病理生理状态下NAc突触回路变化的基本知识， 对未来治疗成瘾相关行为的目标有影响。此外，细心的 本研究的详细方法为其他药物滥用和成瘾的研究提供了基础 模型，以及额外的肯定性障碍与适应不良的过程中NAC。 这是我的期望，在指导项目完成后，我将有技术和智力 专业知识，成功地运行我自己的独立实验室在一个受人尊敬的高等学府。我将不得不 开发工具，以便在会议上和出版物中有效地传播这些调查结果。此外，还 我相信在完成这些项目后，我将能够成功地竞争R01。最后， 这将使我能够继续为成瘾研究领域做出贡献， 和指导年轻科学家。