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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要我目前是一名高级博士后研究员，主要研究方向是阿片受体我打算在未来两年内过渡到初级教员职位，目前的导师是我。我的职位就是为了实现这一目标而设计的，我在体内模型方面拥有广泛的经验。疼痛和成瘾，以及评估受体结合、功能和技术的丰富经验我目前在 Christopher Evans 博士的实验室工作，他是一位著名的阿片类药物。加州大学洛杉矶分校 (UCLA) 的研究员在 UCLA 期间，我将提高我的水平。我和我的导师通过学习原代细胞培养、电生理学和阵列方面的新技术来掌握技能。还有一些计划策略来促进我过渡到教职职位，包括预算加州大学洛杉矶分校是一所著名的机构，拥有许多知名人士。研究人员，在这个指导阶段，我将开展重要的合作，我希望将其维持下去此外，大学还拥有一些最先进的核心设施，我可以通过这些设施。增加我的研究机会。我的研究重点是配体定向的体内后果。 δ (¿) 阿片受体的信号传导正在开发用于临床用途，如激活 ¿然而，受体可以缓解疼痛，并减少焦虑和抑郁。这些化合物的临床用途是 ¿激动剂也会产生惊厥，其机制这是未知的。这种激动剂选择性行为可能是由于 ¿ 的不同运输和信号传导所致。受体，我最近的工作表明，内化激动剂 SNC80 会产生抽搐，而抽搐则不会使用非内化激动剂 ARM390 观察到。 -抑制素是受体的主要介质内化，并介导随后的受体信号传导该提案的目的之一是确定 ¿ 的角色-SNC80 和 ARM390 诱导行为中的抑制，使用 ¿ -arrestin 1 和 2 敲除此外，为了表征调节这种功能选择性的不同信号机制，由 SNC80 和 ARM390 形成的不同信号复合物将使用邻近连接测定来确定此外，我发现在炎症中重复使用这些激动剂。疼痛模型还导致 SNC80 产生受体下调和普遍耐受。相比之下，ARM390 耐受的动物表现出完整的 ¿受体，和仅止痛耐受——其机制尚不清楚。本申请的进一步目的是。通过寻找背侧受体-离子通道耦合的变化来表征这种差异耐受性长期使用后我还将检查根神经节的作用。 -抑制这两种类型的耐受性，并利用 DNA 微阵列探索这些差异背后的可能机制。重要的治疗意义，并将增强我们对体内阿片受体贩运和发信号。