Skip to main content
Biochemical Journal logoLink to Biochemical Journal
. 1990 Nov 15;272(1):217–221. doi: 10.1042/bj2720217

Homologous desensitization of ATP-mediated elevations in cytoplasmic calcium and prostacyclin release in human endothelial cells does not involve protein kinase C.

T D Carter 1, J S Newton 1, R Jacob 1, J D Pearson 1
PMCID: PMC1149679  PMID: 2264825

Abstract

Single human umbilical-vein endothelial cells in culture loaded with the Ca2(+)-sensitive dye fura-2 exhibited characteristic increases in cytosolic Ca2+ concentrations [( Ca2+]i) in response to extracellular ATP. The rapid decline of [Ca2+]i to prestimulated levels in the continued presence of ATP, with in most cells no sustained or oscillatory increase in [Ca2+]i, indicated desensitization. This was agonist-specific, and contrasted with the [Ca2+]i response to histamine, though each agonist mobilized Ca2+ from the same internal store. In populations of cells, when desensitization was variably induced by a second challenge with ATP after different times, desensitization of the initial peak [Ca2+]i was directly related to desensitization of prostacyclin release. This was not affected by treatment with the protein kinase C inhibitor staurosporine, under conditions where a similar degree of desensitization of peak [Ca2+]i induced by phorbol 12-myristate 13-acetate was blocked. Sequential addition of ATP to cell populations cumulatively desensitized the peak elevation of [Ca2+]i, but did not block the second, sustained, phase of the response. We conclude that desensitization of prostacyclin synthesis by ATP is likely to be due to uncoupling of the P2Y purinoceptor from phosphoinositidase C, but does not involve protein kinase C activation.

Full text

PDF
217

Selected References

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

  1. Brock T. A., Capasso E. A. Thrombin and histamine activate phospholipase C in human endothelial cells via a phorbol ester-sensitive pathway. J Cell Physiol. 1988 Jul;136(1):54–62. doi: 10.1002/jcp.1041360107. [DOI] [PubMed] [Google Scholar]
  2. Carter T. D., Hallam T. J., Cusack N. J., Pearson J. D. Regulation of P2y-purinoceptor-mediated prostacyclin release from human endothelial cells by cytoplasmic calcium concentration. Br J Pharmacol. 1988 Dec;95(4):1181–1190. doi: 10.1111/j.1476-5381.1988.tb11754.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Carter T. D., Hallam T. J., Pearson J. D. Protein kinase C activation alters the sensitivity of agonist-stimulated endothelial-cell prostacyclin production to intracellular Ca2+. Biochem J. 1989 Sep 1;262(2):431–437. doi: 10.1042/bj2620431. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Crouch M. F., Lapetina E. G. Dual mechanisms of platelet hormone receptor desensitization. Differential importance between agonists of protein kinase C-dependent and -independent pathways. J Biol Chem. 1989 Jan 5;264(1):584–588. [PubMed] [Google Scholar]
  5. Demolle D., Lecomte M., Boeynaems J. M. Pattern of protein phosphorylation in aortic endothelial cells. Modulation by adenine nucleotides and bradykinin. J Biol Chem. 1988 Dec 5;263(34):18459–18465. [PubMed] [Google Scholar]
  6. Gonzalez F. A., Rozengurt E., Heppel L. A. Extracellular ATP induces the release of calcium from intracellular stores without the activation of protein kinase C in Swiss 3T6 mouse fibroblasts. Proc Natl Acad Sci U S A. 1989 Jun;86(12):4530–4534. doi: 10.1073/pnas.86.12.4530. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hallam T. J., Jacob R., Merritt J. E. Evidence that agonists stimulate bivalent-cation influx into human endothelial cells. Biochem J. 1988 Oct 1;255(1):179–184. doi: 10.1042/bj2550179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hallam T. J., Pearson J. D. Exogenous ATP raises cytoplasmic free calcium in fura-2 loaded piglet aortic endothelial cells. FEBS Lett. 1986 Oct 20;207(1):95–99. doi: 10.1016/0014-5793(86)80019-9. [DOI] [PubMed] [Google Scholar]
  9. Hallam T. J., Pearson J. D., Needham L. A. Thrombin-stimulated elevation of human endothelial-cell cytoplasmic free calcium concentration causes prostacyclin production. Biochem J. 1988 Apr 1;251(1):243–249. doi: 10.1042/bj2510243. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Halldórsson H., Kjeld M., Thorgeirsson G. Role of phosphoinositides in the regulation of endothelial prostacyclin production. Arteriosclerosis. 1988 Mar-Apr;8(2):147–154. doi: 10.1161/01.atv.8.2.147. [DOI] [PubMed] [Google Scholar]
  11. Halldórsson H., Thorgeirsson G. Desensitization of inositol phosphate production after agonist stimulation of endothelial cells is not mediated by protein kinase C. Biochem Biophys Res Commun. 1989 Jun 30;161(3):1064–1069. doi: 10.1016/0006-291x(89)91351-x. [DOI] [PubMed] [Google Scholar]
  12. Jacob R. Agonist-stimulated divalent cation entry into single cultured human umbilical vein endothelial cells. J Physiol. 1990 Feb;421:55–77. doi: 10.1113/jphysiol.1990.sp017933. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Jacob R. Calcium oscillations in electrically non-excitable cells. Biochim Biophys Acta. 1990 May 22;1052(3):427–438. doi: 10.1016/0167-4889(90)90152-4. [DOI] [PubMed] [Google Scholar]
  14. Jacob R., Merritt J. E., Hallam T. J., Rink T. J. Repetitive spikes in cytoplasmic calcium evoked by histamine in human endothelial cells. Nature. 1988 Sep 1;335(6185):40–45. doi: 10.1038/335040a0. [DOI] [PubMed] [Google Scholar]
  15. Kelm M., Feelisch M., Spahr R., Piper H. M., Noack E., Schrader J. Quantitative and kinetic characterization of nitric oxide and EDRF released from cultured endothelial cells. Biochem Biophys Res Commun. 1988 Jul 15;154(1):236–244. doi: 10.1016/0006-291x(88)90675-4. [DOI] [PubMed] [Google Scholar]
  16. Lewis M. J., Henderson A. H. A phorbol ester inhibits the release of endothelium-derived relaxing factor. Eur J Pharmacol. 1987 Jun 4;137(2-3):167–171. doi: 10.1016/0014-2999(87)90218-4. [DOI] [PubMed] [Google Scholar]
  17. Long C. J., Stone T. W. The release of endothelium-derived relaxant factor is calcium dependent. Blood Vessels. 1985;22(4):205–208. doi: 10.1159/000158602. [DOI] [PubMed] [Google Scholar]
  18. Lückhoff A., Zeh R., Busse R. Desensitization of the bradykinin-induced rise in intracellular free calcium in cultured endothelial cells. Pflugers Arch. 1988 Oct;412(6):654–658. doi: 10.1007/BF00583768. [DOI] [PubMed] [Google Scholar]
  19. Martin W., Cusack N. J., Carleton J. S., Gordon J. L. Specificity of P2-purinoceptor that mediates endothelium-dependent relaxation of the pig aorta. Eur J Pharmacol. 1985 Feb 5;108(3):295–299. doi: 10.1016/0014-2999(85)90452-2. [DOI] [PubMed] [Google Scholar]
  20. Pearson J. D., Gordon J. L. P2 purinoceptors in the blood vessel wall. Biochem Pharmacol. 1989 Dec 1;38(23):4157–4163. doi: 10.1016/0006-2952(89)90509-1. [DOI] [PubMed] [Google Scholar]
  21. Pollock W. K., Wreggett K. A., Irvine R. F. Inositol phosphate production and Ca2+ mobilization in human umbilical-vein endothelial cells stimulated by thrombin and histamine. Biochem J. 1988 Dec 1;256(2):371–376. doi: 10.1042/bj2560371. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Sugiya H., Putney J. W., Jr Protein kinase C-dependent and -independent mechanisms regulating the parotid substance P receptor as revealed by differential effects of protein kinase C inhibitors. Biochem J. 1988 Dec 1;256(2):677–680. doi: 10.1042/bj2560677. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Tamaoki T., Nomoto H., Takahashi I., Kato Y., Morimoto M., Tomita F. Staurosporine, a potent inhibitor of phospholipid/Ca++dependent protein kinase. Biochem Biophys Res Commun. 1986 Mar 13;135(2):397–402. doi: 10.1016/0006-291x(86)90008-2. [DOI] [PubMed] [Google Scholar]
  24. Toothill V. J., Needham L., Gordon J. L., Pearson J. D. Desensitization of agonist-stimulated prostacyclin release in human umbilical vein endothelial cells. Eur J Pharmacol. 1988 Nov 22;157(2-3):189–196. doi: 10.1016/0014-2999(88)90382-2. [DOI] [PubMed] [Google Scholar]
  25. Voyno-Yasenetskaya T. A., Tkachuk V. A., Cheknyova E. G., Panchenko M. P., Grigorian G. Y., Vavrek R. J., Stewart J. M., Ryan U. S. Guanine nucleotide-dependent, pertussis toxin-insensitive regulation of phosphoinositide turnover by bradykinin in bovine pulmonary artery endothelial cells. FASEB J. 1989 Jan;3(1):44–51. doi: 10.1096/fasebj.3.1.2535990. [DOI] [PubMed] [Google Scholar]

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

RESOURCES