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Thromb Haemost 1992; 67(03): 331-334
DOI: 10.1055/s-0038-1648442
Original Articles
Schattauer GmbH Stuttgart

Cell Shape Change and Cytosolic Ca2+ in Human Umbilical-Vein Endothelial Cells Stimulated with Thrombin

Haruhiko Sago
The Clinical Dysmorphology Laboratory, National Children’s Medical Research Center, Tokyo, Japan
,
Kazuso linuma
The Clinical Dysmorphology Laboratory, National Children’s Medical Research Center, Tokyo, Japan
› Author Affiliations
Further Information

Publication History

Received 01 May 1991

Accepted after revision 12 September 1991

Publication Date:
03 July 2018 (online)

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Summary

We quantified thrombin-induced endothelial cells shape change and investigated the role of Ca2+ in such shape change. We used the fluorescent Ca2+ indicator, fura2, to measure both shape change as cell size and intracellular free Ca2+ ([Ca2+]i), in cultured human umbilical-vein endothelial cells (HUVEC). Thrombin induced concentration-dependent decreases in cell size (percentage of cell size at 6 min after stimulation with 0.01 U/ml, 0.1 U/ml, or 1 U/ml thrombin) was 90.1 ± 1.5%, 78.1 ± 2.4%, and 40.9 ± 2.4%, respectively. Thrombin also increased [Ca2+]i in a concentration-dependent manner. Both depletion of extracellular Ca2+, and also the addition of W5, a calmodulin antagonist, inhibited thrombin-induced size reduction. These results indicate an association between shape change and [Ca2+]i mobilization in human endothelial cells stimulated by thrombin.

 

  • References

  • 1 Majno G, Shea SM, Leventhal M. Endothelial contraction induced by histamine-type mediators. J Cell Biol 1969; 42: 647-672
    CrossrefPubMedGoogle Scholar
  • 2 Miller FN, Sims DE. Contractile elements in the regulation of macromolecular permeability. Fed Proc 1986; 45: 84-88
    PubMedGoogle Scholar
  • 3 Lerner RG, Cheong LC, Nelson JC. Thrombin-induced endothelial cell retraction. Thromb Haemostas 1979; 42: 244
    PubMedGoogle Scholar
  • 4 Laposota M, Dovnarsky DK, Shin HS. Thrombin-induced gap formation in confluent endothelial cell monolayers in vitro. Blood 1983; 62: 549-556
    PubMedGoogle Scholar
  • 5 McDonald RI, Shepro D, Rosenthal M, Booyse FM. Properties of cultured endothelial cells. Ser Haematol 1973; 4: 469-478
    PubMedGoogle Scholar
  • 6 Gaidai KS, Evensen SA, Brosstad F. Effects of thrombin on the integrity of monolayers of cultured human endothelial cells. Thromb Res 1982; 27: 575-584
    CrossrefPubMedGoogle Scholar
  • 7 Hamilton KK, Sims PJ. Changes in cytosolic Ca2+ associated with von Willebrand factor release in human endothelial cells exposed to histamine. J Clin Invest 1987; 79: 600-608
    CrossrefPubMedGoogle Scholar
  • 8 Jaffe EA, Grulich J, Weksler BB, Hampel G, Watanabe K. Correlation between thrombin-induced prostacyclin production and inositol triphosphate and cytosolic free calcium levels in cultured human endothelial cells. J Biol Chem 1987; 262: 8557-8565
    PubMedGoogle Scholar
  • 9 Pollock WK, Wreggett KA, Irvine RF. Inositol phosphate production and Ca2+ mobilization in human umbilical-vein endothelial cells stimulated by thrombin and histamine. Biochem J 1988; 256: 371-376
    CrossrefPubMedGoogle Scholar
  • 10 Northover AM. The effects of TMB-8 on the shape changes of vascular endothelial cells resulting from exposure to various inflammatory agents. Agents Actions 1989; 26: 367-371
    CrossrefPubMedGoogle Scholar
  • 11 Jaffe EA, Nachman RL, Becker CG, Minick CR. Culture of human endothelial cells derived from umbilical veins. J Clin Invest 1973; 52: 2745-2756
    CrossrefPubMedGoogle Scholar
  • 12 Tsien RY, Poenie M. Fluorescence ratio imaging: a new window into intracellular ionic signaling. Trends Biochem Sci 1986; 11: 450-455
    CrossrefPubMedGoogle Scholar
  • 13 Luckhoff A, Busse R. Increased free calcium in endothelial cells under stimulation with adenine nucleotides. J Cell Physiol 1986; 126: 414-420
    CrossrefPubMedGoogle Scholar
  • 14 Lum H, Del Vecchio PJ, Schneider AS, Goligorsky MS, Malik AB. Calcium dependence of the thrombin-induced increase in endothelial albumin permeability. J Appl Physiol 1989; 66: 1471-1476
    PubMedGoogle Scholar
  • 15 Garcia JGN, Siflinger-Birnboim A, Bizios R, Del Vecchio PJ, Fenton II JW, Malik AB. Thrombin-induced increase in albumin permeability across the endothelium. J Cell Physiol 1986; 128: 96-104
    CrossrefPubMedGoogle Scholar
  • 16 Del Vecchio PJ, Siflinger-Birnboim A, Shepard JM, Bizios R, Cooper JA, Malik AB. Endothelial monolayer permeability to macromolecules. Fed Proc 1987; 46: 2511-2515
    PubMedGoogle Scholar
  • 17 Ross R, Glomset J. The pathogenesis of atherosclerosis. N Engl J Med 1976; 295: 420-425
    CrossrefPubMedGoogle Scholar
  • 18 Schwartz SM. Role of endothelial integrity in atherosclerosis. Artery 1980; 8: 305-314
    PubMedGoogle Scholar
  • 19 Northover AM, Northover BJ. Changes of vascular endothelial cell shape and of membrane potential in response to the ionophore A23187. Int J Microcirc Clin Exp 1987; 6: 137-148
    PubMedGoogle Scholar
  • 20 Gaidai KS, Evensen SA, Nilsen E. Thrombin-induced shape changes of cultured endothelial cells: Metabolic and functional observations. Thromb Res 1983; 32: 57-66
    CrossrefPubMedGoogle Scholar
  • 21 Kakiuchi S, Sobue K. Control of the cytoskeleton by calmodulin and calmodulin-binding proteins. Trends Biochem Sci 1983; 8: 59-62
    CrossrefPubMedGoogle Scholar
  • 22 Maruyama K, Morimoto K, Kijima Y, Sobue K, Kakiuchi S. Calcium-dependent interaction of actin filaments with actin binding protein in the presence of calmodulin and caldesmon. J Biochem 1985; 97: 1517-1520
    CrossrefPubMedGoogle Scholar