Opioid Receptor Signaling: selective mechanism of regulation & receptor crosstalk

首席研究员/项目主任：埃文斯，克里斯托弗·J，博士，组成部分二 第二部分标题：阿片受体信号转导：选择性机制 调节和受体串扰 摘要： 组件II将研究MU阿片激动剂触发差异信号和 受体交易。在过去的资助期间，我们培育了小鼠背根神经节(DRG)。 原代培养研究野生型和突变型小鼠的天然和病毒表达的阿片受体 (例如在组件I中生成的那些和通过Mutant Mouse Core提供的其他那些)。它的用途 实时聚合酶链式反应、基因阵列、流式细胞仪和共聚焦显微镜使我们能够充分表征 受调控的mRNAs的表达以及蛋白的表达、分布和状态 磷酸化。将这些技术与电生理记录相结合，我们有 发现了吗啡和DAMGO独特的贩运和信号特征的证据 作为一种主要的arrestin亚型的调控田鼠。根据这些初步数据，构成部分二将：1) DAMGO和吗啡最初观察到的Mu受体介导的差异信号的特征 并随后定义了其他临床上重要的和内源性激动剂的信号特征；2)分析 通过α2a和β阿片受体的受体串扰在Mu受体转运中的作用 和信号传递；3)测试假设，即β-抑制素和c-src的细胞靶向在 确定阿片激动剂的信号特征和Mu阿片受体的贩运。 总之，这些目标将为理解通过Mu阿片类药物的激动剂导向信号提供基础。 受体，目的是识别激动剂的特性，这些特性可能区分临床上有用的 阿片类药物的有害影响。

Principal Investigator/Program Director: Evans, Christopher J., Ph.D., Component II Component II title: Opioid Receptor Signaling: selective mechanisms of regulation and receptor cross-talk ABSTRACT: Component II will study the mechanisms by which mu opioid agonists can trigger differential signaling and receptor trafficking. During the past funding period we have developed mouse dorsal root ganglia (DRG) primary cultures to study both native and virally expressed opioid receptors in wild-type and mutant mice (such as those generated in Component I and others available through the Mutant Mouse Core). The use of real time PCR, gene arrays, flow cytometry and confocal microscopy has enabled us to fully characterize the expression of regulated mRNAs as well as protein expression, distribution, and states of phosphorylation. Using these techniques in combination with electrophysiological recording, we have developed evidence for unique trafficking and signaling profiles for morphine and DAMGO with implication for a major regulatory Vole of arrestin isoforms. Based upon these preliminary data Component II will: 1) Characterize the differential mu-receptor-mediated signaling initially observed with DAMGO and morphine and subsequently define signaling profiles of other clinically important and endogenous agonists; 2) Analyze the contributions of receptor cross-talk via alpha 2a and delta opioid receptors on mu-receptor trafficking and signaling; 3) Test the hypothesis that beta-arrestins, and the cellular targeting of c-src are critical in determining both the opioid agonist signaling-profiles and trafficking of mu opioid receptors. Together these aims will provide a foundation for understanding agonist-directed signaling via mu opioid receptors with the goal of identifying agonist properties that may differentiate the clinically useful from the detrimental effects of opioid drugs.