Opioid Receptor Signaling: selective mechanism of regulation & receptor crosstalk

主要研究者/项目负责人：Evans，Christopher J.，博士，组分II 组分II标题：阿片受体信号传导：选择性机制 调节和受体相互作用 摘要： 第二部分将研究μ阿片受体激动剂触发差异信号传导的机制， 受体运输在过去的资助期间，我们开发了小鼠背根神经节（DRG） 研究野生型和突变小鼠中天然和病毒表达的阿片受体的原代培养物 (such如在组分I中产生的那些和可通过突变小鼠核心获得的其它物质）。使用 真实的时间PCR，基因阵列，流式细胞术和共聚焦显微镜使我们能够充分表征 调节的mRNA的表达以及蛋白质的表达、分布和状态， 磷酸化使用这些技术结合电生理记录，我们有 开发了吗啡和DAMGO独特的运输和信号特征的证据， 一种主要的调节性田鼠抑制素亚型。根据这些初步数据，第二部分将： 表征最初用DAMGO和吗啡观察到的差异μ受体介导的信号传导 并随后定义其他临床上重要的内源性激动剂的信号传导谱; 2）分析 通过α 2a和δ阿片受体的受体串扰对μ受体运输的贡献 3）测试β-抑制蛋白和c-src的细胞靶向在细胞内的表达和信号传导中至关重要的假设。 确定阿片样物质激动剂信号传导谱和μ阿片样物质受体的运输。 这些目标将为理解通过μ阿片类药物的激动剂导向信号传导提供基础 受体的目的是确定激动剂的性质，可以区分临床上有用的， 阿片类药物的副作用。

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.