In Vivo Implications of Agonist Selective Activation of Delta Opioid Receptor

Delta 阿片受体激动剂选择性激活的体内影响

• 批准号: 8609140
• 负责人: Amynah Amir Ali Pradhan
• 金额: $ 24.9万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-05-15 至 2016-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8609140
• 关键词: Absence of pain sensation; Accounting; Acute; Adult; Adverse effects; Affect; Agonist; Analgesics; Animals; Anxiety; Arrestins; Behavior; Behavioral; Binding; Biological Assay; California; Chronic; Chronic Disease; Clinical; Collaborations; Complex; Convulsions; Core Facility; Coupling; DNA Microarray Chip; Development; Educational process of instructing; Electrophysiology (science); Emotional; Faculty; Fentanyl; G-Protein-Coupled Receptors; GTP-Binding Proteins; Gene Activation; Gene Expression; Genes; Goals; In Vitro; Institution; Ion Channel; Knock-out; Knockout Mice; Laboratories; Learning; Ligands; Ligation; Los Angeles; Mediating; Mediator of activation protein; Mental Depression; Mentors; Modeling; Morphine; Mus; Mutant Strains Mice; Neurons; Opioid; Opioid Receptor; Pain; Pharmacology; Pharmacotherapy; Phase; Phosphoproteins; Positioning Attribute; Postdoctoral Fellow; Primary Cell Cultures; Property; Proteins; RNA Interference; Receptor Activation; Receptor Inhibition; Receptor Signaling; Research; Research Personnel; Role; Seizures; Signal Transduction; Signaling Protein; Site; Spinal Ganglia; Structure; Technical Expertise; Techniques; Therapeutic; Time; Universities; Validation; Work; addiction; arrestin 1; arrestin 2; brain tissue; cell type; chronic pain; delta opioid receptor; desensitization; experience; in vivo; in vivo Model; inflammatory pain; inhibitor/antagonist; interest; knowledge base; novel; receptor; receptor binding; receptor downregulation; receptor internalization; response; skills; trafficking

Project Summary I am currently a senior level postdoctoral fellow with a primary research focus in opioid receptor pharmacology. I intend to transition to a junior faculty position in the next two years, and my current mentored position has been structured to accomplish this goal. I have a wide-range of experience with in vivo models of pain and addiction, as well as extensive experience with techniques to assess receptor binding, function, and localization ex vivo. I am currently working in the laboratory of Dr. Christopher Evans, a prominent opioid researcher, at the University of California Los Angeles (UCLA). During my time at UCLA, I will enhance my skill set by learning new techniques in primary cell culture, electrophysiology, and arrays. My mentor and I have also planned several strategies to facilitate my transition to a faculty position, including budgetary responsibility, teaching, and course work. UCLA is a renowned institution with a number of high profile researchers, and during this mentored phase I will develop important collaborations that I hope to sustain into my independence. In addition, the university also has several state-of-the-art core facilities through which I will enhance my research opportunities. My research focus is on the in vivo consequences of ligand directed signaling at the delta (¿) opioid receptor. Agonists for this receptor are being developed for clinical use, as activation of ¿ receptors relieves pain, and reduces anxiety and depression. However, a major limitation to the clinical use of these compounds is that a subset of ¿ agonists also produce convulsions, the mechanism of which is unknown. This agonist-selective behavior could be due to differential trafficking and signaling of the ¿ receptor, and my recent work shows that the internalizing agonist, SNC80, produces convulsions which are not observed with the non-internalizing agonist, ARM390. ¿-arrestins are major mediators of receptor internalization, and also mediate subsequent receptor signaling. One of the aims of this proposal is to determine the role of ¿-arrestins in SNC80 and ARM390-induced behaviors, using ¿-arrestin 1 and 2 knockout mice. In addition, to characterize the different signaling mechanisms regulating this functional selectivity, the distinct signaling complexes formed by SNC80 and ARM390 will be determined using proximity ligation assays and phosphoprotein arrays. Furthermore, I have found that repeated use of these agonists in an inflammatory pain model also results in differential tolerance. SNC80 produces receptor downregulation and generalized tolerance to all agonist-induced effects. In contrast, ARM390-tolerant animals show intact ¿ receptors, and analgesic tolerance only - the mechanism of which is unknown. A further aim of this application is to characterize this differential tolerance, by looking for changes in receptor-ion channel coupling within the dorsal root ganglia following chronic use. I will also examine the role of ¿-arrestins in these two types of tolerance, and explore possible mechanisms underlying these differences using DNA microarrays. This work has important therapeutic implications, and will enhance our understanding of in vivo opioid receptor trafficking and signaling.

项目摘要 我目前是高级博士后研究员，在阿片类药物接收器中进行了主要研究重点 药理。我打算在接下来的两年中过渡到初级教师职位，而我目前的事很重要 职位已经结构化，以实现这一目标。我在体内模型中有很多经验 疼痛和成瘾，以及具有评估受体结合，功能和功能的技术的丰富经验 本地化。我目前正在著名的阿片类药物克里斯托弗·埃文斯（Christopher Evans）的实验室工作 加州大学洛杉矶分校（UCLA）的研究员。在加州大学洛杉矶分校期间，我将增强我的 通过学习原代细胞培养，电生理学和阵列中的新技术来设定的技能。我的精神和我 还计划了几种策略，以促进我过渡到教师职位，包括预算 责任，教学和课程工作。加州大学洛杉矶分校是一个著名的机构，有许多知名度 研究人员，在此修订阶段，我将建立重要的合作 我的独立性。此外，大学还拥有几个最先进的核心设施 增强我的研究机会。我的研究重点是指向配体的体内后果 在三角洲（）阿片受体的信号传导。该受体的激动剂正在为临床使用开发 接收器的激活减轻疼痛，减轻焦虑和抑郁。但是，对 这些化合物的临床用途是，一部分 - 激动剂也产生抽搐，机制 这是未知的。这种激动剂选择性行为可能是由于运输的差异和信号造成的 接收器和我最近的工作表明，内部化激动剂SNC80产生的抽搐不是 用非内在化激动剂ARM390观察到。 � -arrestins是接收者的主要调解人 内在化，还介导随后的接收器信号传导。该提议的目的之一是 使用� -arrestin 1和2敲除在SNC80和ARM390诱导的行为中的作用 老鼠。此外，要表征调查这种功能选择性的不同信号传导机制 SNC80和ARM390形成的不同信号复合物将使用接近连接分析确定 和磷蛋白阵列。此外，我发现在炎症中反复使用这些激动剂 疼痛模型还会导致差异。 SNC80产生受体下调和广义 对所有激动剂诱导的作用的耐受性。相比之下，ARM390耐受性动物显示完整的接收器，并且 仅镇痛耐受性 - 其机制未知。该应用程序的另一个目的是 通过寻找背侧接收器离子通道耦合的变化来表征这种差异公差 长期使用后的根神经节。我还将研究� -arrestin在这两种类型的宽容中的作用， 并探索使用DNA微阵列的这些差异的可能机制。这项工作有 重要的治疗实施，并将增强我们对体内阿片类药物受体贩运和 信号。