Structural Basis for Endothelial Cell Function: Role of Calcium, Polyphosphoinositide Turnover and G-Proteins

  • Una S. Ryan


In 1980 it was shown that the vasodilator effect of acetylcholine on the aorta was dependent on an intact endothelium (Furchgott and Zawadski, 1980), later the requirement for intact endothelium was demonstrated for other vasodilator agents, such as bradykinin, ATP, thrombin, and histamine (Furchgott, 1984). In response to these agonists, endothelial cells secrete prostacyclin (PGI2) and endothelium-derived relaxing factor (EDRF) (Furchgott, 1984; Ryan et al., 1988b; Johns et al., 1987; Moncada and Vane, 1978; McIntyre et al., 1985). The latter is closely related to nitric oxide, which has been reported to account for many of its effects (Palmer et al., 1987). Both PGI2, and EDRF cause relaxation of vascular smooth muscle cells and have anti-aggregatory effects on platelets (Furchgott, 1984; Moncada and Vane, 1978; Murray et al., 1986). Some of the same agonists also elicit release of platelet activating factor (PAF) from endothelial cells (Mclntyre et al., 1985), a substance with pro-aggregatory and constrictor actions.


Endothelial Cell Cholera Toxin Guanine Nucleotide Pertussis Toxin Inositol Phosphate 
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  1. Adamson, A.W. Physical chemistry of surfaces. New York, Interscience Publ., p 629, 1960.Google Scholar
  2. Altura, B.U. and Chand, N. Bradykinin-induced relaxation of renal and pulmonary arteries is dependent upon intact endothelial cells. Br. J. Pharmacol. 74:10–11, 1981.PubMedCrossRefGoogle Scholar
  3. Aub, D.L., Frey, E.A., Sekura, R.D. and Cote, T.E. Coupling of the thyrotropin- phospholi-pase C by GTP-binding protein distinct from the inhibitory or stimulatory GTP-binding protein. J. Biol Chem. 261:9333–9340, 1986.PubMedGoogle Scholar
  4. Avdonin, P.V., Cheglakov, I.B., Boogry, E.M., Svitina-Ulitina I.V., Mazaev A.V. and Tkachuk VA. Evidence for the receptor-operated calcium channels in human platelet plasma membrane. Thromb. Res. 46:29–37, 1987.PubMedCrossRefGoogle Scholar
  5. Avdonin, P.V., Hayes, B.A., Pozin, E.Y., Popov, E.G., Gavrilov, I.Y., Tkachuk, V.A. and Ryan U.S. Dual-phase response of bovine pulmonary artery endothelial cells to the agonists increasing free cytoplasmic calcium concentration. Tissue & Cell (in press), 1989.Google Scholar
  6. Avdonin, P.V., Menshikov, M.Y., Orlov, S.N., Pokudin, N.I. and Tkachuk, V.A. Mechanisms of elevation of Ca2+ in platelet cytoplasm induced by aggregation factors. Biokhimia (Russian) 50:1241–1248, 1985.Google Scholar
  7. Beny, J.L., Brunet, P. and Huggel, H. Interaction of bradykinin and des-Arg9-bradykinin with isolated pig coronary arteries: mechanical and electrophysiological events. Regulatory Peptides 17:181–190, 1987.PubMedCrossRefGoogle Scholar
  8. Berridge, M.J. Inositoltrisphosphate and diacylglycerol as second messengers. Biochem. J. 220:345–360, 1984.PubMedGoogle Scholar
  9. Berridge, M.J. and Irvine, R.F. Inositol trisphosphate, a novel second messenger in cellular signal transduction. Nature 312:315–321, 1984.PubMedCrossRefGoogle Scholar
  10. Berridge, M.J., Dawson, R.U.C., Downes, C.P., Heslop, G.P. and Irvine, R. Changes in the level of inositol phosphates after agonist-dependent hydrolysis of membrane phospho-inositides. Biochem. J. 212:473–482, 1983.PubMedGoogle Scholar
  11. Bregestovski, P.D. and Ryan, U.S. Voltage-gated and receptor-mediated ionic currents in the membrane of endothelial cells. J. Molec. Cell Cardiol., (in press), 1988.Google Scholar
  12. Burch, R.U. and Axelrod, J. Dissociation of bradykinin-induced prostaglandin formation from phosphatidylinositol turnover in Swiss 3T3 fibroblasts: Evidence for G protein regulation of phospholipase A2 Proc. Natl Acad. Sci. U.S.A. 84:6374–6378, 1987.PubMedCrossRefGoogle Scholar
  13. Chand, N. and Altura, B.U. Acetylcholine and bradykinin relax intrapulmonary arteries by acting on endothelial cells: Role in lung vascular diseases. Science 213:1376–1379, 1981.PubMedCrossRefGoogle Scholar
  14. Cherry, P.D., Furchgott, R.F., Sadawski, Y.V. and Yothianandan, D. The role of endothelial cells in the relaxation of isolated arteries by bradykinin. Proc. Natl Acad. Sci. USA. 79:2106–2110, 1982.PubMedCrossRefGoogle Scholar
  15. Cockroft, S. and Gamperts, B.D. Role of guanine nucleotide binding protein in the activation of polyphosphoinositide phosphodiesterase. Nature (Lond). 314:534–536, 1985.CrossRefGoogle Scholar
  16. Colden-Stanfield M., Schilling, W.P., Ritchie, A.K., Eskin, S.G., Navarro, L.T. and Kunze, D.N. Bradykinin-induced increases in cytosolic calcium and ionic currents in cultured bovine aortic endothelial cells. Circulation Res. 61:632–640, 1987.PubMedCrossRefGoogle Scholar
  17. Crutchley, D.J., Ryan, J.W., Ryan, U.S. and Fisher, G.H. Bradykinin-induced release of prostacyclin and thromboxanes from bovine pulmonary artery endothelial cells. Studies with lower homologs and calcium antagonists. Biochem. Biophys. Acta. 751:99–107, 1983.PubMedCrossRefGoogle Scholar
  18. Furchgott, R.F. 1984. The role of endothelium in the responses of vascular smooth muscle to drugs. Ann. Rev. Pharmacol. Toxicol. 24:175–197, 1984.Google Scholar
  19. Furchgott, R. and Zawadski, J. The obligatory role of endothelial cells in the relaxation of arterial smooth muscle by acetylcholine. Nature. 288:373–376, 1980.PubMedCrossRefGoogle Scholar
  20. Gamperts B.D. Involvement of guanine-nucleotide-binding protein in the gating of Ca2+ by receptors. Nature (Lond) 306:64–66, 1983.CrossRefGoogle Scholar
  21. Griffith, T.M., Edward, D.H., Newby, A.C., Lewis, M.J. and Henderson, A.H. Production of endothelium derived relaxant factor is dependent on oxidative phosphorylation and extracellular calcium. Cardiovasc. Res. 20:7–12, 1986.PubMedCrossRefGoogle Scholar
  22. Gilman, A.G .G proteins: transducers of receptor-generated signals. Annu. Rev. Biochem. 56:615–649, 1987.PubMedCrossRefGoogle Scholar
  23. Grigorian, G.Y. and Ryan, U.S. Platelet activating factor effects on bovine pulmonary artery endothelial cells. Cira Res. 61:389–395, 1987.CrossRefGoogle Scholar
  24. Hallam, T.J. and Rink, T.J. Agonists stimulate divalent cation channels in the plasma membrane of human platelets. FEBS Lett. 186:175–179, 1985.PubMedCrossRefGoogle Scholar
  25. Hallam, T.J. and Pearson, J.D. Exogeneous ATP raises cytoplasmic free calcium in FURA-2 loaded piglet aortic endothelial cells. FEBS Lett. 207:95–99, 1986.PubMedCrossRefGoogle Scholar
  26. Hamilton, K.K. and Sims, P.J. Changes in cytosolic Ca2+ associated with von Willebrand factor release in human endothelial cells exposed to histamine. Study on microcarrier cell monolayers using the fluorescent probe INDO-1. J. Clin. Invest. 79:600–609, 1987.PubMedCrossRefGoogle Scholar
  27. Haslam R.J. and Davidson, M.M.L. Guanine nucleitudes decrease the free Ca2+ required for secretion of serotonin from permeabilized platelets. Evidence of a role for a GTP-binding protein in platelet activation. FEBS Lett. 174:90–95, 1984.CrossRefGoogle Scholar
  28. Johns, A., Lategan, T.W., Lodge, N.J., Ryan, U.S., Van Breemen, C. and Adams, D.J. Calcium entry through receptor-operated channels in bovine pulmonary artery endothelial cells. Tissue & Cell, 19:1–13, 1987.CrossRefGoogle Scholar
  29. Lambert, T.L., Kent, R.S. and Whorton, A.R. Bradykinin stimulation of inositol polyphosphate production in porcine aortic endothelial cells. J. Biol. Chem. 261:15288–15293, 1986.PubMedGoogle Scholar
  30. Litosch, I. Guanine nucleotide and NaF stimulation of phospholipase C activity in rat cerbral-cortical membranes. Biochem. J. 244:35–40, 1987.PubMedGoogle Scholar
  31. Litosch, I., Wallis, C. and Fain, J.N. 5-hydroxytryptamine stimulates inositol phosphate production in a cell-free system from blowfly salivary glands. Evidence for role of GTP in coupling receptor activation to phosphoinositide breakdown. J. Biol. Chem. 260:5464–5471, 1985.PubMedGoogle Scholar
  32. Lodge, N.J., Adams, D.J., Jones, A., Ryan, U.S. and Van Breemen, C. Calcium activation of endothelial cells. In: Resistance Arteries, (ed. W. Halpern), pp. 152–161, 1988.Google Scholar
  33. Long, C.J. and Stone, T.W. The release of endothelium-derived relaxant factor is calcium dependent. Blood Vessels 22:205–208, 1985.PubMedGoogle Scholar
  34. Luckhoff, A. and Busse, R. 1986. Increased free calcium in endothelial cells under stimulation with adenine nucleotides. J. Cell. Physiol. 126:414–420, 1986.Google Scholar
  35. Martin, T.F.Y., Lucas, D.O., Bajjalien, S.U. and Kowalchyk, Y.A. Thyrotropin-releasing hormone activates a Ca2+-dependent polyphosphoinositide phosphodiesterase in permeable GM3 cells. GTP7S potentiation by a cholera and pertussis toxin-insensitive mechanism. J. Biol. Chem. 261:2918–2927, 1986.PubMedGoogle Scholar
  36. Martin, U.W., Evans, T. and Harden, T.K. Further evidence that muscarinic cholinergic receptors of 1321N1 astrocytoma cells, couple to a guanine nucleotide regulatory protein that is not Ni. Biochem. J. 229:539–544, 1985.PubMedGoogle Scholar
  37. Mcintyre, T.M., Zimmerman, G.A., Satoh, K. and Prescott, S.M. Cultured endothelial cells synthesize both platelet-activating factor and prostacyclin in response to histamine, bradykinin and adenosine triphosphate. J. Clin. Invest. 76:271–280, 1985.PubMedCrossRefGoogle Scholar
  38. Moncada, S. and Vane, J.R. Prostacyclin (PGI2), the vascular wall and vasodilation. In: Mechanisms of Vasodilation, ed. by P.A. Vanhoutte and I. Leusen, pp 107–121. Karger, Basel, 1978.Google Scholar
  39. Murray, J.J., Fridovich, I., Makhoul, R.G. and Hagen, P. Stabilization and partial charactarization of endothelium derived relaxing factor from cultured bovine aortic endothelium cells. Biochem. Biophys. Res. Commun. 141:689–696, 1986.PubMedCrossRefGoogle Scholar
  40. Neer, E.J. and Clapham, D.E. Roles of G protein subunits in transmembrane signalling. Nature 333:129–134, 1988.PubMedCrossRefGoogle Scholar
  41. Palmer, R.M.J., Ferrige, A.G. and Moncada, S. Nitric oxide release accounts for the biological activity of endothelium-derived relaxing factor. Nature 327:524–526, 1987.PubMedCrossRefGoogle Scholar
  42. Poll, C., Kyrle, P. and Westwick, J. Activation of protein kinase C inhibits sodium fluoride-induced elevation of human platelet cytosolic free calcium and thromboxane B2 generation. Biochem. Biophys. Res. Commun. 136:381–389, 1986.PubMedCrossRefGoogle Scholar
  43. Popov, E.G., Gavrilov, I.Y., Pozin, E.Y. and Gabbasov, ZA. Multiwavelength method for measuring concentration of free cytosolic calcium using the fluorescent probe INDO-1. Arch. Biochem Biophys. 261:91–96, 1988.PubMedCrossRefGoogle Scholar
  44. Resink, T.J., Grigorian, G.Y., Moldabaeva, A.K., Danilov, S.M. and Buehler, F.R. Histamine--induced polyphosphoinositide metabolism in cultured human endothelial cells: association with thromboxane and prostacyclin release. Biochem. Biophys. Res. Commun. 144:438–446, 1987.PubMedCrossRefGoogle Scholar
  45. Roth, B.L. Modulation of phosphatidylinositol-4,5-bisphosphate hydrolysis in rat aorta by guanyl nucleotides, calcium and magnesium. Life Science 41:622–634, 1987.Google Scholar
  46. Rotrosen, D. and Gallin, J.I. Histamine type 1 receptor occupancy increases endothelial cytosolic calcium, reduces F-actin, and promotes albumin diffusion across cultured endothelial monolayers. J. Cell Biol. 103:2379 – 2387, 1986.PubMedCrossRefGoogle Scholar
  47. Ryan, U.S. Processing of angiotensin and other peptides by the lungs. In Handbook of Physiology — The Respiratory System 1 (eds. A.P. Fishman and A.B. Fisher), Amer. Physiol. Soc, Bethesda, MD, Chapter 10, pp. 351–364, 1985.Google Scholar
  48. Ryan, U.S. and Maxwell, G. Isolation, culture and subculture of bovine pulmonary artery endothelial cells: mechanical methods. J. Tissue Cult. Methods 10:3–5, 1986.CrossRefGoogle Scholar
  49. Ryan, U.S., Avdonin, P.V., Posin, E.Y., Popov, E.G., Davilov, S.M. and Tkachuk, V.A. Influence of vasoactive agents on cytoplasmic free calcium concentration in INDO-1 loaded endothelial cells. J. Applied Physiol. 65:2221–2227, 1988a.Google Scholar
  50. Ryan, U.S., Johns, A. and Van Breeman, C. Role of calcium in receptor-mediated endothelial cell responses. Chest 93:105S–109S, 1988b.PubMedCrossRefGoogle Scholar
  51. Ryan, U.S., Mortara, M. and Whitaker, C. Methods for microcarrier culture of bovine pulmonary artery endothelial cells avoiding the use of enzymes. Tissue & Cell, 12:619–635, 1980.CrossRefGoogle Scholar
  52. Stryer, L. and Bowine, H.R. G proteins: a family of signal transducers. Ann. Rev. Cell Biol. 2:391–419, 1986.PubMedCrossRefGoogle Scholar
  53. Uhing, R.Y., Prpic, V., Yiang, H. and Exton, Y.H. Hormone-stimulated polyphosphoinositide breakdown in rat liver plasma membranes. Roles of guanine nucleotides and calcium. J. Biol. Chem. 261:2140–2146, 1986.PubMedGoogle Scholar
  54. Voyno-Yasenetskaya, T.A., Tkachuk, V.A., Cheknyova, E.G., Panchenko, M.P., Grigorian, G.Y., Vavrek, R.J., Stewart, J.M. and Ryan U.S. Guanine nucleotide dependent, pertussis toxin insensitive regulation of phosphoinositide turnover by bradykinin in bovine pulmonary artery endothelial cells. FASEB J. 3:44–51, 1989.PubMedGoogle Scholar
  55. Weksler, B.B., Ley, C.W. and Jaffe, E.A. Stimulation of endothelial prostacyclin production by thrombin, trypsin, and ionophore A23187. J. Clin. Invest. 62:923–930, 1978.PubMedCrossRefGoogle Scholar
  56. Whorton, A.R., Young, S.L., Data, Y.L., Barchowsky, H. and Kent, R.S. Mechanism of bradykinin-stimulated prostacyclin synthesis in porcine aortic endothelial cells. Biochem. Biophys. Acta. 712:79–87, 1982.PubMedCrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1989

Authors and Affiliations

  • Una S. Ryan
    • 1
  1. 1.Department of MedicineUniversity of Miami School of MedicineMiamiUSA

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