Attali B, Saya D, Nah S Y, Vogel Z
Department of Neurobiology, Weizmann Institute of Science, Rehovot, Israel.
J Biol Chem. 1989 Jan 5;264(1):347-53.
The aim of the present study has been to characterize the regulation by opiates of 45Ca2+ influx in rat spinal cord-dorsal root ganglion cocultures. We have demonstrated that K+-induced depolarization, in the presence of the Ca2+ channel agonist Bay K8644, stimulated Ca2+ influx (3-4-fold) via the dihydropyridine class of voltage-dependent Ca2+ channels. While mu and delta opiates had no effect, kappa opiate agonists (e.g. U50488, dynorphin) profoundly depressed the stimulated Ca2+ influx (86% inhibition at 100 microM U50488). The kappa agonist action was stereospecific and could be reversed by the opiate antagonist naloxone. The inhibition produced by kappa agonists was greatly diminished following pertussis toxin treatment, and this effect was accompanied by toxin-induced ADP-ribosylation of a 40-41-kDa protein. This suggests that kappa opiate receptors are negatively coupled to voltage-dependent Ca2+ channels, via a pertussis toxin-sensitive GTP-binding protein. Basal 45Ca2+ uptake, stimulated by adenylate cyclase activators (forskolin and cholera toxin), was potently inhibited by kappa opiates suggesting that, under conditions of neurohormonal stimulation of adenylate cyclase, kappa receptors are coupled to Ca2+ channels indirectly via the adenylate cyclase complex. In addition, cAMP-independent coupling pathways may also be involved.
本研究的目的是描述阿片类药物对大鼠脊髓-背根神经节共培养物中45Ca2+内流的调节作用。我们已经证明,在Ca2+通道激动剂Bay K8644存在的情况下,K+诱导的去极化通过二氢吡啶类电压依赖性Ca2+通道刺激Ca2+内流(增加3 - 4倍)。虽然μ和δ阿片类药物没有作用,但κ阿片类激动剂(如U50488、强啡肽)能显著抑制刺激后的Ca2+内流(100μM U50488时抑制率达86%)。κ激动剂的作用具有立体特异性,且可被阿片类拮抗剂纳洛酮逆转。用百日咳毒素处理后,κ激动剂产生的抑制作用大大减弱,且这种作用伴随着毒素诱导的一种40 - 41 kDa蛋白质的ADP核糖基化。这表明κ阿片受体通过一种百日咳毒素敏感的GTP结合蛋白与电压依赖性Ca2+通道负性偶联。由腺苷酸环化酶激活剂(福斯高林和霍乱毒素)刺激的基础45Ca2+摄取被κ阿片类药物强烈抑制,这表明在神经激素刺激腺苷酸环化酶的条件下,κ受体通过腺苷酸环化酶复合物间接与Ca2+通道偶联。此外,可能还涉及不依赖cAMP的偶联途径。