Skip to main content
PMC home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1994 Dec;94(6):2451–2456. doi: 10.1172/JCI117613

Evidence for deactivation of both ectosolic and cytosolic 5'-nucleotidase by adenosine A1 receptor activation in the rat cardiomyocytes.

M Kitakaze 1, M Hori 1, T Minamino 1, S Takashima 1, K Komamura 1, K Node 1, T Kurihara 1, T Morioka 1, H Sato 1, M Inoue 1, et al.
PMCID: PMC330077  PMID: 7989602

Abstract

Adenosine, an important regulator of many cardiac functions, is produced by ectosolic and cytosolic 5'-nucleotidase. The activity of these enzymes is influenced by several ischemia-sensitive metabolic factors, e.g., ATP, ADP, H+, and inorganic phosphate. However, there is no clear evidence that adenosine itself affects 5'-nucleotidase activity. This study tested whether adenosine decreases the activity of ectosolic and cytosolic 5'-nucleotidase. Cardiomyocytes were isolated from adult male Wistar rats and suspended in the modified Hepes-Tyrode buffer solution. After stabilization, isolated cardiomyocytes were incubated with and without adenosine (10(-9) - 10(-4) M). Ectosolic and cytosolic 5'-nucleotidase activity was decreased by exogenous adenosine (ectosolic 5'-nucleotidase activity, 20.6 +/- 2.3 vs. 8.6 +/- 1.6 mumol/min per 10(6) cells [P < 0.05]; cytosolic 5'-nucleotidase activity, 2.47 +/- 0.58 vs. 1.61 +/- 0.54 mumol/min per 10(6) cells [P < 0.05] at 10(-6) M adenosine) after 30 min. The decrease in ectosolic and cytosolic 5'-nucleotidase activity was inhibited by 8-phenyltheophylline and pertussis toxin, and was mimicked by N6-cyclohexyladenosine, an adenosine A1 receptor agonist. Neither CGS21680C, and A2 receptor agonist, nor cycloheximide deactivated ectosolic and cytosolic 5'-nucleotidase. Thus, we conclude that activation of adenosine A1 receptors is coupled to Gi proteins and attenuates ectosolic and cytosolic 5'-nucleotidase activity in rat cardiomyocytes.

Full text

PDF
2451

Images in this article

2454Image
on p.2454

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Achterberg P. W., de Tombe P. P., Harmsen E., de Jong J. W. Myocardial S-adenosylhomocysteine hydrolase is important for adenosine production during normoxia. Biochim Biophys Acta. 1985 Jul 5;840(3):393–400. doi: 10.1016/0304-4165(85)90220-x. [DOI] [PubMed] [Google Scholar]
  2. Belardinelli L., Isenberg G. Actions of adenosine and isoproterenol on isolated mammalian ventricular myocytes. Circ Res. 1983 Sep;53(3):287–297. doi: 10.1161/01.res.53.3.287. [DOI] [PubMed] [Google Scholar]
  3. Berne R. M. The role of adenosine in the regulation of coronary blood flow. Circ Res. 1980 Dec;47(6):807–813. doi: 10.1161/01.res.47.6.807. [DOI] [PubMed] [Google Scholar]
  4. Cronstein B. N., Levin R. I., Belanoff J., Weissmann G., Hirschhorn R. Adenosine: an endogenous inhibitor of neutrophil-mediated injury to endothelial cells. J Clin Invest. 1986 Sep;78(3):760–770. doi: 10.1172/JCI112638. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. DeWitt D. F., Wangler R. D., Thompson C. I., Sparks H. V., Jr Phasic release of adenosine during steady state metabolic stimulation in the isolated guinea pig heart. Circ Res. 1983 Nov;53(5):636–643. doi: 10.1161/01.res.53.5.636. [DOI] [PubMed] [Google Scholar]
  6. Engler R. Consequences of activation and adenosine-mediated inhibition of granulocytes during myocardial ischemia. Fed Proc. 1987 May 15;46(7):2407–2412. [PubMed] [Google Scholar]
  7. Hedqvist P., Fredholm B. B. Inhibitory effect of adenosine on adrenergic neuroeffector transmission in the rabbit heart. Acta Physiol Scand. 1979 Jan;105(1):120–122. doi: 10.1111/j.1748-1716.1979.tb06321.x. [DOI] [PubMed] [Google Scholar]
  8. Honma M., Satoh T., Takezawa J., Ui M. An ultrasensitive method for the simultaneous determination of cyclic AMP and cyclic GMP in small-volume samples from blood and tissue. Biochem Med. 1977 Dec;18(3):257–273. doi: 10.1016/0006-2944(77)90060-6. [DOI] [PubMed] [Google Scholar]
  9. Hori M., Kitakaze M. Adenosine, the heart, and coronary circulation. Hypertension. 1991 Nov;18(5):565–574. doi: 10.1161/01.hyp.18.5.565. [DOI] [PubMed] [Google Scholar]
  10. Iwakura K., Hori M., Watanabe Y., Kitabatake A., Cragoe E. J., Jr, Yoshida H., Kamada T. Alpha 1-adrenoceptor stimulation increases intracellular pH and Ca2+ in cardiomyocytes through Na+/H+ and Na+/Ca2+ exchange. Eur J Pharmacol. 1990 Sep 4;186(1):29–40. doi: 10.1016/0014-2999(90)94057-5. [DOI] [PubMed] [Google Scholar]
  11. Katada T., Amano T., Ui M. Modulation by islet-activating protein of adenylate cyclase activity in C6 glioma cells. J Biol Chem. 1982 Apr 10;257(7):3739–3746. [PubMed] [Google Scholar]
  12. Kitakaze M., Hori M., Kamada T. Role of adenosine and its interaction with alpha adrenoceptor activity in ischaemic and reperfusion injury of the myocardium. Cardiovasc Res. 1993 Jan;27(1):18–27. doi: 10.1093/cvr/27.1.18. [DOI] [PubMed] [Google Scholar]
  13. Kitakaze M., Hori M., Sato H., Takashima S., Inoue M., Kitabatake A., Kamada T. Endogenous adenosine inhibits platelet aggregation during myocardial ischemia in dogs. Circ Res. 1991 Nov;69(5):1402–1408. doi: 10.1161/01.res.69.5.1402. [DOI] [PubMed] [Google Scholar]
  14. Kitakaze M., Hori M., Tamai J., Iwakura K., Koretsune Y., Kagiya T., Iwai K., Kitabatake A., Inoue M., Kamada T. Alpha 1-adrenoceptor activity regulates release of adenosine from the ischemic myocardium in dogs. Circ Res. 1987 May;60(5):631–639. doi: 10.1161/01.res.60.5.631. [DOI] [PubMed] [Google Scholar]
  15. Kohl C., Linck B., Schmitz W., Scholz H., Scholz J., Tóth M. Effects of carbachol and (-)-N6-phenylisopropyladenosine on myocardial inositol phosphate content and force of contraction. Br J Pharmacol. 1990 Dec;101(4):829–834. doi: 10.1111/j.1476-5381.1990.tb14165.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Newby A. C., Holmquist C. A., Illingworth J., Pearson J. D. The control of adenosine concentration in polymorphonuclear leucocytes, cultured heart cells and isolated perfused heart from the rat. Biochem J. 1983 Aug 15;214(2):317–323. doi: 10.1042/bj2140317. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Richardt G., Waas W., Kranzhöfer R., Mayer E., Schömig A. Adenosine inhibits exocytotic release of endogenous noradrenaline in rat heart: a protective mechanism in early myocardial ischemia. Circ Res. 1987 Jul;61(1):117–123. doi: 10.1161/01.res.61.1.117. [DOI] [PubMed] [Google Scholar]
  18. Schrader J., Baumann G., Gerlach E. Adenosine as inhibitor of myocardial effects of catecholamines. Pflugers Arch. 1977 Nov 25;372(1):29–35. doi: 10.1007/BF00582203. [DOI] [PubMed] [Google Scholar]
  19. Smith K., Varon H. H., Race G. J., Paulson D. L., Urschel H. C., Mallams J. T. Serum 5'-nucleotidase in patients with tumor in the liver. Cancer. 1966 Sep;19(9):1281–1285. doi: 10.1002/1097-0142(196609)19:9<1281::aid-cncr2820190914>3.0.co;2-0. [DOI] [PubMed] [Google Scholar]
  20. Sparks H. V., Jr, Bardenheuer H. Regulation of adenosine formation by the heart. Circ Res. 1986 Feb;58(2):193–201. doi: 10.1161/01.res.58.2.193. [DOI] [PubMed] [Google Scholar]
  21. Sullivan J. M., Alpers J. B. In vitro regulation of rat heart 5'-nucleotidase by adenine nucleotides and magnesium. J Biol Chem. 1971 May 10;246(9):3057–3063. [PubMed] [Google Scholar]
  22. Widnell C. C., Schneider Y. J., Pierre B., Baudhuin P., Trouet A. Evidence for a continual exchange of 5'-nucleotidase between the cell surface and cytoplasmic membranes in cultured rat fibroblasts. Cell. 1982 Jan;28(1):61–70. doi: 10.1016/0092-8674(82)90375-0. [DOI] [PubMed] [Google Scholar]
  23. Yamane R., Nakamura T., Matsuura E., Ishige H., Fujimoto M. A simple and sensitive radioimmunoassay for adenosine. J Immunoassay. 1991;12(4):501–519. doi: 10.1080/01971529108053277. [DOI] [PubMed] [Google Scholar]
  24. van den Berghe G., van Pottelsberghe C., Hers H. G. A kinetic study of the soluble 5'-nucleotidase of rat liver. Biochem J. 1977 Mar 15;162(3):611–616. doi: 10.1042/bj1620611. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES