Physiological and Molecular Mechanisms of Mu Opioid Receptors in Motivation and Affect

Mu阿片受体在动机和情感中的生理和分子机制

• 批准号: 10570885
• 负责人: Daniel Charles Castro
• 金额: $ 24.9万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-01 至 2025-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10570885
• 关键词: Affect; Affective; Animals; Area; Arrestins; Behavior; Behavioral; Behavioral Paradigm; Brain; Budgets; CRISPR/Cas technology; Calcium Signaling; Cessation of life; Chimera organism; Classification; Committee Members; Consumption; Development; Eating; Enkephalins; Ensure; Faculty; Fiber; Food; Food deprivation (experimental); GTP-Binding Proteins; Genetic; Goals; Image; Incentives; Intake; Interdisciplinary Study; Intervention; Knockout Mice; Knowledge; Lead; Leadership; Learning; Measures; Medial; Mediating; Mentors; Mentorship; Microscopy; Modeling; Molecular; Motivation; Mus; Naloxone; Negative Valence; Neurons; Nucleus Accumbens; Opioid; Output; Pathway interactions; Persons; Pharmaceutical Preparations; Pharmacology; Phase; Phosphorylation; Photometry; Photons; Physiological; Physiology; Population; Positioning Attribute; Positive Valence; Postdoctoral Fellow; Presynaptic Terminals; Prosencephalon; Proteins; Quinine; Reaction; Receptor Signaling; Research; Research Personnel; Rewards; Rhodopsin; Role; Scientific Societies; Serine; Shapes; Signal Pathway; Signal Transduction; Signaling Protein; Site; Structure; Substance abuse problem; Sucrose; System; Taste Perception; Technical Expertise; Testing; Therapeutic; Thirst; Time; Training; United States; Viral; Viral Vector; Water; addiction; beta-arrestin; calcium indicator; career; cell type; conditional knockout; dorsal raphe nucleus; drug reward; endogenous opioids; experimental study; food consumption; genetic approach; hedonic; illicit opioid; imaging approach; in vivo; in vivo imaging; interest; knowledgebase; light gated; loss of function; meetings; microendoscopy; motivated behavior; mouse model; mu opioid receptors; neural circuit; neurochemistry; neuromechanism; novel strategies; novel therapeutics; opioid overdose; opioid use disorder; optogenetics; orofacial; pharmacologic; postsynaptic; prescription opioid abuse; presynaptic; prevent; programs; receptor; receptor function; recruit; response; skills; symposium; taste stimuli; training opportunity

Project Summary: The primary goal of this training proposal is to understand, with mechanistic granularity, how mu opioid receptors (MORs) modulate motivated `wanting' versus affective `liking' in nucleus accumbens (NAc). During the proposed K99 training period, I will be trained in two in vivo physiology/imaging approaches (fiber photometry and 1-photon microscopy) and on intracellular/molecular effector systems. Additionally, I will actively participate in professional/career training opportunities and have frequent meetings with my mentoring committee to prepare to apply and succeed in an independent faculty position. The first research aim of my proposal seeks to understand the temporal dynamics and effects of endogenous MOR signaling in the DRNEnkNAc pathway. My previous postdoctoral research, using pharmacology, genetics, and optogenetics/chemogenetics, has identified the terminals of an enkephalinergic dorsal raphe nucleus projection to NAc as the site of MOR action for modulating food intake behaviors. To better understand how MOR signaling affects this pathway, I will use two in vivo imaging approaches, fiber photometry and microendoscopy, to determine how endogenous MOR signals shape neural circuit activity during motivated behaviors (Aim 1, K99). Additionally, because the roles of specific intracellular signaling cascades have not been well defined in brain reward circuits, I will manipulate G-protein and beta-arrestin signaling pathways to determine how each of them contributes to behavior (Aim 2, K99). During the K99 phase, I will also be preparing to lead my own research lab. I will actively participate in scientific society leadership positions (chairing symposia, steering committees), learn about how to direct a lab (budgeting and administrative tasks), and continue to further my scholarly knowledgebase (planned interactions with my mentor and committee members). For the R00 “independent” phase of my proposal, I propose to build on the mentored training above to examine what mechanisms and circuits underlie MOR-mediated affective `liking' in NAc (Aim 3). These studies are of interest because growing evidence suggests that though NAc MORs can robustly modulate both motivation and affect, they likely do so via different neural mechanisms. Understanding how these biopsychological systems are dissociable, even within a specific neurochemical class, has important implications for developing novel therapeutic drugs that are efficacious without also having a propensity for abuse. I will selectively disrupt or restore MOR function using multiple genetic mouse models and viral vector interventions, as well as record endogenous neuronal responses in vivo using microendoscopic approaches. I will perform these experiments while testing mice on the affective test reactivity (TR) test, which classifies and quantifies innate orofacial reactions to taste stimuli and has been used effectively for decades to measure the affective value of taste stimuli. The knowledge gained from these studies and mentorship opportunities will greatly facilitate the development of my own research program, preparing me for the next phase of my career.

项目概要： 本培训计划的主要目标是了解，与机械粒度，如何μ阿片受体 （MORs）调节有动机的“想要”与情感的“喜欢”在丘脑核（NAc）。在拟议 K99培训期间，我将接受两种体内生理学/成像方法（光纤测光和单光子 显微镜）和细胞内/分子效应器系统。此外，我将积极参与 专业/职业培训机会，并经常与我的指导委员会开会， 申请并在独立教师职位上取得成功。我的建议的第一个研究目标是 了解DRNEnk β NAc通路中内源性莫尔信号传导的时间动力学和影响。我 以前的博士后研究，使用药理学，遗传学，光遗传学/化学遗传学，已经确定 脑啡肽能中缝背核的末梢投射到NAc作为莫尔作用的部位， 调节食物摄入行为。为了更好地理解莫尔信号如何影响这一通路，我将使用两个 体内成像方法，纤维光度法和显微内窥镜，以确定内源性莫尔信号 在动机行为过程中塑造神经回路活动（目标1，K99）。此外，由于特定的角色 细胞内信号级联在大脑奖赏回路中还没有很好的定义，我将操纵G蛋白 和β-arrestin信号通路，以确定它们中的每一个如何对行为做出贡献（Aim 2，K99）。期间 在K99阶段，我也将准备领导我自己的研究实验室。我将积极参与科学社会 领导职位（主持研讨会，指导委员会），学习如何指导实验室（预算和 行政任务），并继续推进我的学术知识基础（计划与我的导师互动 和委员会成员）。对于我的建议的R 00“独立”阶段，我建议建立在指导的基础上， 上述训练，以检查在NAc中MOR介导的情感“喜欢”的基础是什么机制和回路（Aim （3）第三章。这些研究很有意义，因为越来越多的证据表明，尽管NAc MORs可以强烈地 调节动机和情感，它们可能通过不同的神经机制来实现。了解这些 生物心理系统是可分离的，即使在一个特定的神经化学类，有重要的影响 用于开发新的治疗药物，这些药物是有效的，但也没有滥用的倾向。我会 使用多种遗传小鼠模型和病毒载体干预选择性地破坏或恢复莫尔功能， 以及使用显微内窥镜方法记录体内内源性神经元反应。我必坚定 这些实验，同时测试小鼠的情感测试反应（TR）测试，分类和量化 先天口面反应的味觉刺激，并已有效地使用了几十年来衡量情感 味觉刺激值。从这些学习和指导机会中获得的知识将大大 促进我自己的研究计划的发展，为我职业生涯的下一阶段做好准备。