Real-time imaging of μ-opioid receptor signaling, phosphorylation and desensitization (C05)

μ-阿片受体信号传导、磷酸化和脱敏的实时成像 (C05)

• 批准号: 279229656
• 金额: --
• 依托单位: Institut für Pharmakologie und Toxikologie
• 依托单位国家: 德国
• 项目类别: CRC/Transregios
• 财政年份: 2015
• 资助国家: 德国
• 起止时间: 2014-12-31 至 2019-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/279229656?language=en
• 关键词: Real; time; imaging; opioid; receptor

G protein receptor kinases (GRKs) and β-arrestins are key regulators of μ-opioid receptor (MOR) signaling and trafficking. In the previous funding period, we showed that GRK2/3 is recruited more quickly than β-arrestins. β-Arrestin recruitment required GRK2 activity and MOR phosphorylation, but GRK recruitment also depended on phosphorylation sites in the C-terminal tail. Consistent with this, phosphorylation-deficient MORs mediate strongly enhanced acute analgesia in vivo and upon chronic administration analgesic tolerance to fentanyl is abrogated. In this proposal, we will (I) delineate the role of PKC in non-canonical MOR desensitization and tolerance, (II) evaluate the physiological role of unconventional β-arrestin signaling in vivo and in vitro, and (III) utilize advanced imaging techniques to monitor agonist-dependent changes in endogenous MOR receptor organization in living neurons.

G 蛋白受体激酶 (GRK) 和 β-抑制蛋白是 μ-阿片受体 (MOR) 信号传导和运输的关键调节因子。在之前的资助期间，我们表明 GRK2/3 的招募速度比 β-arrestins 更快。 β-Arrestin 招募需要 GRK2 活性和 MOR 磷酸化，但 GRK 招募还取决于 C 末端尾部的磷酸化位点。与此一致的是，磷酸化缺陷的 MOR 介导强烈增强的体内急性镇痛作用，并且长期给药后对芬太尼的镇痛耐受被消除。在本提案中，我们将（I）描述 PKC 在非典型 MOR 脱敏和耐受中的作用，（II）评估非常规 β-arrestin 信号在体内和体外的生理作用，以及（III）利用先进的成像技术监测活体神经元内源性 MOR 受体组织的激动剂依赖性变化。