FUNCTIONAL REGULATION OF OPIOID SIGNALLING

持续的吗啡治疗可增加兴奋性Gs蛋白偶联的浓度。 神经调节剂(如PGE2和强啡肽)和增加脊髓中疼痛神经递质的释放 电源线。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 前列腺素E_2和α-前列腺素E_2对培养的新生大鼠初级感觉神经元神经递质的释放 脊髓强啡肽的非阿片类片段，dyn2-13。此外，自早些时候以来，我们已经表明了持续的 吗啡处理导致Raf-1介导的腺酰环化酶敏化(S)(AC超激活) 对于重组细胞中的兴奋性物质，在项目C中，我们还将研究 RAF-1介导的AC超激活在基础和/或辣椒素诱导的降钙素基因相关肽释放中的敏化 持续吗啡治疗后的感觉神经元。我们假设Raf-1介导的AC 过度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。要评估这一点 假设是这样的。研究Raf-1在培养的新生儿cAMP形成敏化中的作用。 大鼠背根神经节神经元对Gs蛋白偶联的兴奋性神经调节剂PGE2和dyn2-13的作用；ii.测试 CAMP、cAMP依赖的蛋白激酶(PKA)和Raf-1在基础和/或辣椒碱调节中的作用 前列腺素E_2和Dyn2-13对培养的新生大鼠背根神经节神经元释放CGRP的影响 持续的吗啡治疗；以及III.研究选定的新化合物的作用-在 人工合成核心和A项目-cAMP浓度以及基础和辣椒素诱导的CGRP释放 培养新生大鼠背根神经节神经元，持续阿片类激动剂治疗前后。

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.