FUNCTIONAL REGULATION OF OPIOID SIGNALLING

阿片类信号传导的功能调节

• 批准号: 8025976
• 负责人: EVA V VARGA
• 金额: $ 16.29万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8025976
• 关键词: Adenylate Cyclase; Afferent Neurons; Agonist; Analgesics; BDKRB2 gene; Binding Sites; Bradykinin Receptor; Capsaicin; Cell Line; Cells; Chronic; Cyclic AMP; Cyclic AMP-Dependent Protein Kinases; Development; Dinoprostone; Dynamin 2; Dynorphin A; Dynorphins; Event; Forskolin; Future; Hand; Human; Hyperalgesia; In Vitro; Investigation; Lead; Mediating; Modeling; Molecular; Morphine; Neonatal; Neuromodulator; Neuronal Plasticity; Neurons; Neurotransmitters; Nociception; Opioid; Opioid Analgesics; Opioid Peptide; Opioid Receptor; Pain; Pathway interactions; Physiological; Play; Preparation; Process; Protein Kinase; Protein Kinase Inhibitors; Proteins; Proto-Oncogene Proteins c-raf; Rattus; Receptor Signaling; Recombinants; Regulation; Role; Sensory; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Site; Spinal; Spinal Cord; Stimulus; Testing; Up-Regulation; Work; allodynia; analog; biphalin; central pain; chronic pain; endogenous opioids; in vitro testing; inhibitor/antagonist; neurotransmitter release; novel; protein kinase inhibitor; receptor; receptor coupling; research study; trafficking

Sustained morphine treatment was shown to increase the concentration of excitatory Gs protein-coupled neuromodulators (such as PGE2 and dynorphin) and augment pain neurotransmitter release in the spinal cord. In Project C we will investigate the role of cAMP-regulated signaling pathways in the regulation of pain neurotransmitter (CGRP) release from cultured neonatal rat primary sensory (DRG) neurons by PGE2 and a non-opioid fragment of spinal dynorphin, dyn2-13. In addition, since earlier we have shown that sustained morphine treatment leads to a Raf-1 -mediated sensitization of adenylyl cyclase(s) (AC superactivation) towards excitatory agents in recombinant cells, in Project C we also will investigate the physiological role of Raf-1-mediated AC superactivation in the sensitization of basal and/or capsaicin-evoked CGRP release from sensory neurons after sustained morphine-treatment. We hypothesize that Raf-1-mediated AC superactivation sensitizes primary sensory neurons to Gs protein-coupled neuromodulators leading to augmented basal and/or evoked CGRP release upon sustained morphine treatment. To evaluate this hypothesis we shall I. investigate the role of Raf-1 in the sensitization of cAMP formation in cultured neonatal rat DRG neurons toward the Gs protein-coupled excitatory neuromodulators, PGE2 and dyn2-13; II. test the role of cAMP, cAMP-dependent protein kinase (PKA) and Raf-1 in the regulation of basal and/or capsaicinevoked CGRP release by PGE2 and dyn2-13 in cultured neonatal rat DRG neurons before and after sustained morphine treatment; and III. study the effect of selected novel compounds - prepared in the Synthetic Core and Project A - on cAMP concentration and basal and capsaicin-evoked CGRP release in cultured neonatal rat DRG neurons, before and after sustained opioid agonist treatment.

持续的吗啡治疗被证明可以增加兴奋性 Gs 蛋白偶联的浓度 神经调节剂（如 PGE2 和强啡肽）并增强脊髓中疼痛神经递质的释放 绳索。在项目 C 中，我们将研究 cAMP 调节的信号通路在疼痛调节中的作用 PGE2 和 a 蛋白从培养的新生大鼠初级感觉 (DRG) 神经元中释放神经递质 (CGRP) 脊髓强啡肽的非阿片类片段，dyn2-13。此外，自早些时候以来，我们已经表明，持续 吗啡治疗导致 Raf-1 介导的腺苷酸环化酶致敏（AC 超激活） 对于重组细胞中的兴奋剂，在项目 C 中，我们还将研究 Raf-1 介导的 AC 超激活对基础和/或辣椒素诱发的 CGRP 释放的敏化 持续吗啡治疗后的感觉神经元。我们假设 Raf-1 介导的 AC 超激活使初级感觉神经元对 Gs 蛋白偶联神经调节剂敏感，从而导致 持续吗啡治疗后基础和/或诱发的 CGRP 释放增加。为了评价这一点 假设我们将 I. 研究 Raf-1 在培养新生儿 cAMP 形成致敏中的作用 大鼠 DRG 神经元朝向 Gs 蛋白偶联的兴奋性神经调节剂 PGE2 和 dyn2-13；二.测试 cAMP、cAMP 依赖性蛋白激酶 (PKA) 和 Raf-1 在基础和/或辣椒素诱发调节中的作用 培养的新生大鼠 DRG 神经元前后 PGE2 和 dyn2-13 释放的 CGRP 持续吗啡治疗；和三。研究所选新型化合物的效果 - 制备于 合成核心和项目 A - 关于 cAMP 浓度以及基础和辣椒素诱发的 CGRP 释放 在持续阿片类激动剂治疗前后培养新生大鼠 DRG 神经元。