FUNCTIONAL EXPRESSION OF ADRENERGIC AND OPIOID RECEPTORS IN XENOPUS-OOCYTES - INTERACTION BETWEEN ALPHA(2)-ADRENERGIC AND BETA(2)-ADRENERGIC RECEPTORS
FUNCTIONAL EXPRESSION OF ADRENERGIC AND OPIOID RECEPTORS IN XENOPUS-OOCYTES - INTERACTION BETWEEN ALPHA(2)-ADRENERGIC AND BETA(2)-ADRENERGIC RECEPTORS
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DOI：
10.1016/0169-328x(94)00185-h
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发表时间：
1995-01-01
期刊：
影响因子：
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通讯作者：
WILCOX, GL
中科院分区：
文献类型：
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作者：
BIRNBAUM, AK;WOTTA, DR;WILCOX, GL
我们在非洲爪蟾卵母细胞中功能性地表达α 2肾上腺素能、β 2肾上腺素能和δ阿片受体。我们检测了受体功能的变化，由腺苷3 '，5'-环磷酸（cAMP）调节的氯离子通道提供的囊性纤维化跨膜电导调节器（CFTR）和记录的双电极电压钳电流。去应用毛喉素和异丁基甲基黄嘌呤（IBMX）或IBMX单独产生的电流与反转电位指示氯离子仅在卵母细胞先前注射的mRNA编码CFTR。异丙肾上腺素产生浓度依赖性的反应，在卵母细胞注射的mRNA编码β（2）-肾上腺素能受体和CFTR，和共同管理的普萘洛尔拮抗这些反应。同样，α 2肾上腺素能激动剂UK 14304仅在注射编码α 2肾上腺素能受体和CFTR的mRNA的卵母细胞中增加IBMX诱导的电流，而咪唑克生拮抗这些增强。δ-阿片受体激动剂DADLE仅在注射编码δ-阿片受体和CFTR的mRNA的卵母细胞中产生浓度相关的纳洛酮可逆性IBMX和毛喉素诱导电流增加。在共注射α（2）、β（2）和CFTR mRNA的卵母细胞中，等辐射图分析显示α（2）和β（2）肾上腺素能受体之间存在加性相互作用。这些研究确立了卵母细胞作为研究cAMP调节G蛋白偶联受体之间相互作用的细胞系统，并提供了α 2肾上腺素能受体和δ阿片受体与G蛋白（可能是G（s）蛋白，而不是G（i）蛋白）的替代偶联的另一个例子。
We functionally expressed alpha(2)-adrenergic, beta(2)-adrenergic, and delta-opioid receptors in Xenopus laevis oocytes. We detected receptor function as changes in currents carried by adenosine 3',5'-cyclic monophosphate (cAMP)-regulated chloride channels provided by the cystic fibrosis transmembrane conductance regulator (CFTR) and recorded by two-electrode voltage clamp. Go-application of forskolin and isobutylmethylxanthine (IBMX) or IBMX alone produced currents with a reversal potential indicative of chloride ions only in oocytes previously injected with mRNA encoding CFTR. Isoproterenol produced concentration-dependent responses in oocytes injected with mRNA encoding beta(2)-adrenergic receptors and CFTR, and co-administration of propranolol antagonized these responses. Similarly, the alpha(2)-adrenergic agonist UK14304 increased IBMX-induced currents only in oocytes injected with mRNA encoding alpha(2)-adrenergic receptors and CFTR, and idazoxan antagonized these enhancements. The delta-opioid agonist DADLE produced concentration-related, naloxone-reversible increases in IBMX- and forskolin-induced currents only in oocytes injected with mRNA encoding delta-opioid receptors and CFTR. In oocytes co-injected with alpha(2), beta(2), and CFTR mRNAs, isobolographic analysis revealed an additive interaction between alpha(2)- and beta(2)-adrenergic receptors. These studies establish the oocyte as a cell system for studying the interactions among cAMP-modulating G protein-coupled receptors and provide another example of alternative coupling of alpha(2)-adrenergic and delta-opioid receptors to G proteins, possibly G(s) proteins, other than G(i) proteins.