In Vivo Implications of Agonist Selective Activation of Delta Opioid Receptor

项目摘要 我目前是高级博士后研究员，主要研究阿片受体。 药理学。我打算在接下来的两年内过渡到初级教员的职位，而我目前的导师 为了实现这一目标，已经安排了职位。我对活体模型有广泛的经验 疼痛和成瘾，以及评估受体结合、功能和 体外定位。我目前在著名阿片类药物克里斯托弗·埃文斯博士的实验室工作 加州大学洛杉矶分校(UCLA)研究员。在加州大学洛杉矶分校期间，我将加强我的 通过学习原代细胞培养、电生理学和阵列方面的新技术而获得的技能。我和我的导师 我还计划了几项战略，以促进我向教员职位的过渡，包括预算 责任、教学和课程工作。加州大学洛杉矶分校是一所著名的大学，拥有许多高调的 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 我的独立性。此外，该大学还拥有几个最先进的核心设施，我将通过这些设施 增加我的研究机会。我的研究重点是配体导向的体内后果。 在阿片受体的信号。这种受体的激动剂正在开发用于临床，如 受体的激活可以缓解疼痛，减少焦虑和抑郁。然而，对 这些化合物的临床应用是，激动剂的一部分也会引起惊厥，其机制是 这是个未知数。这种选择激动剂的行为可能是由于不同的贩运和信号传递。 受体，而我最近的工作表明，内化激动剂SNC80会产生惊厥，而不是 用非内在化激动剂ARM390观察。阻滞素是受体的主要介体 内化，也参与随后的受体信号传递。这项提议的目的之一是 使用α-arrestin 1和2基因敲除来确定？-arrestins在SNC80和ARM390诱导的行为中的作用 老鼠。此外，为了表征调节这种功能选择性的不同信号机制， SNC80和ARM390形成的不同信号复合体将使用邻近连接分析来确定 和磷蛋白阵列。此外，我发现反复使用这些激动剂在炎症性疾病中 疼痛模型也会导致差异耐受。SNC80产生受体下调和全身化 对所有激动剂诱导的影响耐受。相比之下，对ARM390耐受的动物表现出完整的受体，并且 仅限止痛耐受-其机制尚不清楚。此应用程序的另一个目标是 通过寻找背侧受体-离子通道偶联的变化来表征这种差异耐受性 长期使用后的根神经节。我还将研究逮捕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。这项工作有 重要的治疗意义，并将加强我们对体内阿片受体贩运和 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.