Cheng L, Precht P, Frank D, Liang C T
Laboratory of Biological Chemistry, National Institute on Aging, National Institutes of Health, Baltimore, Maryland 21224.
Am J Physiol. 1990 Apr;258(4 Pt 2):F877-82. doi: 10.1152/ajprenal.1990.258.4.F877.
Dopamine receptors have been identified in many tissues including the kidney. To establish an in vitro system as a model for dopamine action, we studied the effect of dopamine (DA) receptor agonists and antagonists on adenosine 3',5'-cyclic monophosphate (cAMP) formation in opossum kidney (OK) cells. The stimulation of cAMP production in these cells by dopamine was dose dependent, and markedly higher levels were observed in the presence of dopamine plus a phosphodiesterase inhibitor, 3-isobutyl-1-methylxanthine. Half-maximal stimulation was found with 1.15 +/- 0.22 microM dopamine. A DA1-receptor agonist, SKF 82526J, stimulated cAMP production, whereas a DA2-receptor agonist, Ly 171555, did not. The stimulatory effects of dopamine and SKF 82526J were abolished by a specific DA1-receptor antagonist, Sch 23390 with half-maximal inhibition concentrations of 1.24 +/- 0.18 and 4.0 +/- 0.5 nM, respectively. In contrast, the DA2-receptor antagonist, spiperone, had no inhibitory effect on dopamine- and SKF 82526J-stimulated cAMP production. Beta-Adrenergic antagonists failed to attenuate the stimulatory effects of dopamine and SKF 82526J on cAMP production. In addition, the beta-adrenergic receptor agonist, isoproterenol, did not stimulate cAMP production. These results suggest that the action of dopamine was not mediated through beta-adrenergic receptors. Furthermore, our results clearly demonstrated the existence of DA1-receptors linked to adenylate cyclase in OK cells.
多巴胺受体已在包括肾脏在内的许多组织中被鉴定出来。为了建立一个体外系统作为多巴胺作用的模型,我们研究了多巴胺(DA)受体激动剂和拮抗剂对负鼠肾(OK)细胞中3',5'-环磷酸腺苷(cAMP)形成的影响。多巴胺对这些细胞中cAMP产生的刺激呈剂量依赖性,并且在多巴胺加磷酸二酯酶抑制剂3-异丁基-1-甲基黄嘌呤存在的情况下观察到明显更高的水平。发现1.15±0.22微摩尔/升的多巴胺可产生半数最大刺激。DA1受体激动剂SKF 82526J刺激了cAMP的产生,而DA2受体激动剂Ly 171555则没有。多巴胺和SKF 82526J的刺激作用被特异性DA1受体拮抗剂Sch 23390消除,其半数最大抑制浓度分别为1.24±0.18和4.0±0.5纳摩尔/升。相比之下,DA2受体拮抗剂螺哌隆对多巴胺和SKF 82526J刺激的cAMP产生没有抑制作用。β-肾上腺素能拮抗剂未能减弱多巴胺和SKF 82526J对cAMP产生的刺激作用。此外,β-肾上腺素能受体激动剂异丙肾上腺素也没有刺激cAMP的产生。这些结果表明多巴胺的作用不是通过β-肾上腺素能受体介导的。此外,我们的结果清楚地证明了OK细胞中存在与腺苷酸环化酶相连的DA1受体。