In Vivo Implications of Agonist Selective Activation of Delta Opioid Receptor

项目摘要 我目前是一名高级博士后研究员，主要研究阿片受体 药理学我打算在未来两年内过渡到初级教师职位，我目前的导师是 我们的立场是为实现这一目标而建立的。我有一个广泛的经验，在体内模型， 疼痛和成瘾，以及评估受体结合，功能和 离体定位。我目前在克里斯托弗·埃文斯博士的实验室工作， 加州洛杉矶大学（UCLA）的研究员。在加州大学洛杉矶分校期间，我将提高我的 通过学习原代细胞培养，电生理学和阵列的新技术来掌握技能。我和我的导师 我还计划了几个策略，以促进我过渡到教师的立场，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， 研究人员，在这个指导阶段，我将发展重要的合作，我希望能维持到 我的独立此外，大学还拥有几个最先进的核心设施，通过这些设施，我将 增加我的研究机会。我的研究重点是在体内的后果配体定向 在δ阿片受体上的信号传导。该受体的激动剂正在开发用于临床用途， 受体的激活可以缓解疼痛，减少焦虑和抑郁。然而，一个主要的限制， 这些化合物的临床用途是，一个子集的激动剂也产生惊厥， 这是未知的。这种激动剂选择性行为可能是由于不同的运输和信号的？ 受体，我最近的工作表明，内化激动剂，SNC 80，产生抽搐，而不是 用非内化激动剂ARM 390观察到。抑制蛋白是受体的主要介质 内化，并且还介导随后的受体信号传导。这项建议的目的之一是 使用<$-arrestin 1和2敲除，确定<$-arrestins在SNC 80和ARM 390诱导的行为中的作用 小鼠此外，为了表征调节这种功能选择性的不同信号传导机制， SNC 80和ARM 390形成的不同信号传导复合物将使用邻位连接测定来确定 和磷蛋白阵列。此外，我还发现，反复使用这些激动剂， 疼痛模型也导致不同的耐受。SNC 80产生受体下调和全身性 对所有激动剂诱导的作用的耐受性。相比之下，ARM 390耐受动物显示出完整的受体， 仅镇痛耐受-其机制尚不清楚。本申请的另一个目的是 通过寻找背侧皮层内受体-离子通道耦合的变化来表征这种差异耐受性。 根神经节慢性使用。我还将研究抑制蛋白在这两种耐受中的作用， 并使用DNA微阵列探索这些差异背后的可能机制。这项工作 重要的治疗意义，并将提高我们对体内阿片受体贩运的理解， 发信号。

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.