Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1993 Apr 1;291(Pt 1):263–267. doi: 10.1042/bj2910263

Cyclic AMP enhances agonist-induced Ca2+ entry into endothelial cells by activation of potassium channels and membrane hyperpolarization.

W F Graier 1, W R Kukovetz 1, K Groschner 1
PMCID: PMC1132511  PMID: 8385935

Abstract

The mechanism underlying cyclic AMP (cAMP)-mediated amplification of agonist-induced Ca2+ responses in endothelial cells was investigated in pig endothelial cells. Forskolin, adenosine and isoprenaline, as well as the membrane-permeant cAMP analogue dibutyryl cAMP, enhanced bradykinin-induced rises in intracellular free Ca2+ as well as bradykinin-induced Mn2+ entry. These agents were also found to hyperpolarize endothelial cells without increasing intracellular Ca2+ by itself, i.e. in the absence of bradykinin. Both amplification of bradykinin effects and the hyperpolarizing action was blocked by the protein kinase inhibitor H-8. The involvement of K+ channels in the hyperpolarizing effects of forskolin was consequently studied in perforated outside-out vesicles. Two different types of K+ channels were recorded, one of which had a large conductance (170 pS) and was activated by forskolin. We suggest that stimulation of endothelial adenylate cyclase results in activation of large-conductance K+ channels and consequently in membrane hyperpolarization, which in turn enhances bradykinin-induced entry of Ca2+ by increasing its electrochemical gradient.

Full text

PDF
263

Selected References

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

  1. Brock T. A., Dennis P. A., Griendling K. K., Diehl T. S., Davies P. F. GTP gamma S loading of endothelial cells stimulates phospholipase C and uncouples ATP receptors. Am J Physiol. 1988 Nov;255(5 Pt 1):C667–C673. doi: 10.1152/ajpcell.1988.255.5.C667. [DOI] [PubMed] [Google Scholar]
  2. Buchan K. W., Martin W. Modulation of agonist-induced calcium mobilisation in bovine aortic endothelial cells by phorbol myristate acetate and cyclic AMP but not cyclic GMP. Br J Pharmacol. 1991 Oct;104(2):361–366. doi: 10.1111/j.1476-5381.1991.tb12436.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cohen P. The structure and regulation of protein phosphatases. Annu Rev Biochem. 1989;58:453–508. doi: 10.1146/annurev.bi.58.070189.002321. [DOI] [PubMed] [Google Scholar]
  4. Colden-Stanfield M., Schilling W. P., Possani L. D., Kunze D. L. Bradykinin-induced potassium current in cultured bovine aortic endothelial cells. J Membr Biol. 1990 Jul;116(3):227–238. doi: 10.1007/BF01868462. [DOI] [PubMed] [Google Scholar]
  5. Fichtner H., Fröbe U., Busse R., Kohlhardt M. Single nonselective cation channels and Ca2+-activated K+ channels in aortic endothelial cells. J Membr Biol. 1987;98(2):125–133. doi: 10.1007/BF01872125. [DOI] [PubMed] [Google Scholar]
  6. Graier W. F., Groschner K., Schmidt K., Kukovetz W. R. Increases in endothelial cyclic AMP levels amplify agonist-induced formation of endothelium-derived relaxing factor (EDRF). Biochem J. 1992 Dec 1;288(Pt 2):345–349. doi: 10.1042/bj2880345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Graier W. F., Schmidt K., Kukovetz W. R. Effect of sodium fluoride on cytosolic free Ca2(+)-concentrations and cGMP-levels in endothelial cells. Cell Signal. 1990;2(4):369–375. doi: 10.1016/0898-6568(90)90067-k. [DOI] [PubMed] [Google Scholar]
  8. Groschner K., Graier W. F., Kukovetz W. R. Activation of a small-conductance Ca(2+)-dependent K+ channel contributes to bradykinin-induced stimulation of nitric oxide synthesis in pig aortic endothelial cells. Biochim Biophys Acta. 1992 Oct 27;1137(2):162–170. doi: 10.1016/0167-4889(92)90198-k. [DOI] [PubMed] [Google Scholar]
  9. Grynkiewicz G., Poenie M., Tsien R. Y. A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem. 1985 Mar 25;260(6):3440–3450. [PubMed] [Google Scholar]
  10. Hamill O. P., Marty A., Neher E., Sakmann B., Sigworth F. J. Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pflugers Arch. 1981 Aug;391(2):85–100. doi: 10.1007/BF00656997. [DOI] [PubMed] [Google Scholar]
  11. Hidaka H., Inagaki M., Kawamoto S., Sasaki Y. Isoquinolinesulfonamides, novel and potent inhibitors of cyclic nucleotide dependent protein kinase and protein kinase C. Biochemistry. 1984 Oct 9;23(21):5036–5041. doi: 10.1021/bi00316a032. [DOI] [PubMed] [Google Scholar]
  12. Kume H., Takai A., Tokuno H., Tomita T. Regulation of Ca2+-dependent K+-channel activity in tracheal myocytes by phosphorylation. Nature. 1989 Sep 14;341(6238):152–154. doi: 10.1038/341152a0. [DOI] [PubMed] [Google Scholar]
  13. Laskey R. E., Adams D. J., Johns A., Rubanyi G. M., van Breemen C. Membrane potential and Na(+)-K+ pump activity modulate resting and bradykinin-stimulated changes in cytosolic free calcium in cultured endothelial cells from bovine atria. J Biol Chem. 1990 Feb 15;265(5):2613–2619. [PubMed] [Google Scholar]
  14. Levitan E. S., Kramer R. H. Neuropeptide modulation of single calcium and potassium channels detected with a new patch clamp configuration. Nature. 1990 Dec 6;348(6301):545–547. doi: 10.1038/348545a0. [DOI] [PubMed] [Google Scholar]
  15. Lückhoff A., Mülsch A., Busse R. cAMP attenuates autacoid release from endothelial cells: relation to internal calcium. Am J Physiol. 1990 Apr;258(4 Pt 2):H960–H966. doi: 10.1152/ajpheart.1990.258.4.H960. [DOI] [PubMed] [Google Scholar]
  16. Mehrke G., Daut J. The electrical response of cultured guinea-pig coronary endothelial cells to endothelium-dependent vasodilators. J Physiol. 1990 Nov;430:251–272. doi: 10.1113/jphysiol.1990.sp018290. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Mehrke G., Pohl U., Daut J. Effects of vasoactive agonists on the membrane potential of cultured bovine aortic and guinea-pig coronary endothelium. J Physiol. 1991 Aug;439:277–299. doi: 10.1113/jphysiol.1991.sp018667. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Reiser G. Nitric oxide formation caused by Ca2+ release from internal stores in neuronal cell line is enhanced by cyclic AMP. Eur J Pharmacol. 1992 Sep 1;227(1):89–93. doi: 10.1016/0922-4106(92)90147-n. [DOI] [PubMed] [Google Scholar]
  19. Rink T. J., Montecucco C., Hesketh T. R., Tsien R. Y. Lymphocyte membrane potential assessed with fluorescent probes. Biochim Biophys Acta. 1980;595(1):15–30. doi: 10.1016/0005-2736(80)90243-6. [DOI] [PubMed] [Google Scholar]
  20. Rubanyi G. M., Romero J. C., Vanhoutte P. M. Flow-induced release of endothelium-derived relaxing factor. Am J Physiol. 1986 Jun;250(6 Pt 2):H1145–H1149. doi: 10.1152/ajpheart.1986.250.6.H1145. [DOI] [PubMed] [Google Scholar]
  21. Rusko J., Tanzi F., van Breemen C., Adams D. J. Calcium-activated potassium channels in native endothelial cells from rabbit aorta: conductance, Ca2+ sensitivity and block. J Physiol. 1992 Sep;455:601–621. doi: 10.1113/jphysiol.1992.sp019318. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Sage S. O., Merritt J. E., Hallam T. J., Rink T. J. Receptor-mediated calcium entry in fura-2-loaded human platelets stimulated with ADP and thrombin. Dual-wavelengths studies with Mn2+. Biochem J. 1989 Mar 15;258(3):923–926. doi: 10.1042/bj2580923. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Sauvé R., Chahine M., Tremblay J., Hamet P. Single-channel analysis of the electrical response of bovine aortic endothelial cells to bradykinin stimulation: contribution of a Ca2(+)-dependent K+ channel. J Hypertens Suppl. 1990 Dec;8(7):S193–S201. [PubMed] [Google Scholar]
  24. Schilling W. P., Elliott S. J. Ca2+ signaling mechanisms of vascular endothelial cells and their role in oxidant-induced endothelial cell dysfunction. Am J Physiol. 1992 Jun;262(6 Pt 2):H1617–H1630. doi: 10.1152/ajpheart.1992.262.6.H1617. [DOI] [PubMed] [Google Scholar]
  25. Steinberg S. F., Jaffe E. A., Bilezikian J. P. Endothelial cells contain beta adrenoceptors. Naunyn Schmiedebergs Arch Pharmacol. 1984 Apr;325(4):310–313. doi: 10.1007/BF00504374. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES