Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1993 Feb 2;120(4):1003–1010. doi: 10.1083/jcb.120.4.1003

Spreading of human endothelial cells on fibronectin or vitronectin triggers elevation of intracellular free calcium

PMCID: PMC2200079  PMID: 7679387

Abstract

Intracellular calcium ([Ca2+]i) was measured in FURA 2-loaded endothelial cells plated on fibronectin or vitronectin. Average values for [Ca2+]i increased to approximately twofold above basal levels by approximately 1 h after plating, and then declined. The increase in [Ca2+]i required extracellular calcium. Substituting potassium for sodium in the medium reduced the elevation of [Ca2+]i, a result that rules out the involvement of Na-Ca exchangers or voltage-dependent calcium channels, but that is consistent with the involvement of voltage-independent calcium channels. Plating cells on an anti-integrin beta 1 subunit antibody gave a similar [Ca2+]i response, but clustering beta 1 integrins with the same antibody, or occupying integrins with RGD (arg-gly-asp) peptides had no effect. Time course measurements on single cells revealed that in each cell [Ca2+]i rose abruptly at some point during spreading, from the basal level to a higher steady-state level that was maintained for some time. The elevated [Ca2+]i was unrelated to previously observed changes in intracellular pH, because chelating the Ca2+ in the medium failed to inhibit the elevation of pHi that occurred during cell spreading. In conclusion, these results show that integrin-mediated cell spreading can regulate [Ca2+]i, and the pathways involved are distinct from those that regulate intracellular pH.

Full Text

The Full Text of this article is available as a PDF (1.3 MB).

Selected References

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

  1. Curry F. E. Modulation of venular microvessel permeability by calcium influx into endothelial cells. FASEB J. 1992 Apr;6(7):2456–2466. doi: 10.1096/fasebj.6.7.1563597. [DOI] [PubMed] [Google Scholar]
  2. Donjacour A. A., Cunha G. R. Stromal regulation of epithelial function. Cancer Treat Res. 1991;53:335–364. doi: 10.1007/978-1-4615-3940-7_16. [DOI] [PubMed] [Google Scholar]
  3. Dustin M. L., Springer T. A. Lymphocyte function-associated antigen-1 (LFA-1) interaction with intercellular adhesion molecule-1 (ICAM-1) is one of at least three mechanisms for lymphocyte adhesion to cultured endothelial cells. J Cell Biol. 1988 Jul;107(1):321–331. doi: 10.1083/jcb.107.1.321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Godinich M., LaPointe M. S., Batlle D. C. Free cytosolic calcium regulation via Na(+)-Ca2+ exchange in cultured vascular smooth muscle cells. Ann N Y Acad Sci. 1991;639:561–565. doi: 10.1111/j.1749-6632.1991.tb17351.x. [DOI] [PubMed] [Google Scholar]
  5. Grzesiak J. J., Davis G. E., Kirchhofer D., Pierschbacher M. D. Regulation of alpha 2 beta 1-mediated fibroblast migration on type I collagen by shifts in the concentrations of extracellular Mg2+ and Ca2+. J Cell Biol. 1992 Jun;117(5):1109–1117. doi: 10.1083/jcb.117.5.1109. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hynes R. O. Integrins: versatility, modulation, and signaling in cell adhesion. Cell. 1992 Apr 3;69(1):11–25. doi: 10.1016/0092-8674(92)90115-s. [DOI] [PubMed] [Google Scholar]
  7. Ingber D. E., Folkman J. How does extracellular matrix control capillary morphogenesis? Cell. 1989 Sep 8;58(5):803–805. doi: 10.1016/0092-8674(89)90928-8. [DOI] [PubMed] [Google Scholar]
  8. Ingber D. E., Prusty D., Frangioni J. V., Cragoe E. J., Jr, Lechene C., Schwartz M. A. Control of intracellular pH and growth by fibronectin in capillary endothelial cells. J Cell Biol. 1990 May;110(5):1803–1811. doi: 10.1083/jcb.110.5.1803. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kimura M., Gardner J. P., Aviv A. Agonist-evoked alkaline shift in the cytosolic pH set point for activation of Na+/H+ antiport in human platelets. The role of cytosolic Ca2+ and protein kinase C. J Biol Chem. 1990 Dec 5;265(34):21068–21074. [PubMed] [Google Scholar]
  10. Kornberg L. J., Earp H. S., Turner C. E., Prockop C., Juliano R. L. Signal transduction by integrins: increased protein tyrosine phosphorylation caused by clustering of beta 1 integrins. Proc Natl Acad Sci U S A. 1991 Oct 1;88(19):8392–8396. doi: 10.1073/pnas.88.19.8392. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Lakos Z., Somogyi B., Balázs M., Matkó J., Damjanovich S. The effect of transmembrane potential on the dynamic behavior of cell membranes. Biochim Biophys Acta. 1990 Mar 30;1023(1):41–46. doi: 10.1016/0005-2736(90)90007-b. [DOI] [PubMed] [Google Scholar]
  12. Levin E. G., Santell L. Association of a plasminogen activator inhibitor (PAI-1) with the growth substratum and membrane of human endothelial cells. J Cell Biol. 1987 Dec;105(6 Pt 1):2543–2549. doi: 10.1083/jcb.105.6.2543. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Manganel M., Turner R. J. Agonist-induced activation of Na+/H+ exchange in rat parotid acinar cells is dependent on calcium but not on protein kinase C. J Biol Chem. 1990 Mar 15;265(8):4284–4289. [PubMed] [Google Scholar]
  14. Miekka S. I., Ingham K. C., Menache D. Rapid methods for isolation of human plasma fibronectin. Thromb Res. 1982 Jul 1;27(1):1–14. doi: 10.1016/0049-3848(82)90272-9. [DOI] [PubMed] [Google Scholar]
  15. Ng-Sikorski J., Andersson R., Patarroyo M., Andersson T. Calcium signaling capacity of the CD11b/CD18 integrin on human neutrophils. Exp Cell Res. 1991 Aug;195(2):504–508. doi: 10.1016/0014-4827(91)90402-g. [DOI] [PubMed] [Google Scholar]
  16. Ober S. S., Pardee A. B. Both protein kinase C and calcium mediate activation of the Na+/H+ antiporter in Chinese hamster embryo fibroblasts. J Cell Physiol. 1987 Aug;132(2):311–317. doi: 10.1002/jcp.1041320216. [DOI] [PubMed] [Google Scholar]
  17. Pardi R., Bender J. R., Dettori C., Giannazza E., Engleman E. G. Heterogeneous distribution and transmembrane signaling properties of lymphocyte function-associated antigen (LFA-1) in human lymphocyte subsets. J Immunol. 1989 Nov 15;143(10):3157–3166. [PubMed] [Google Scholar]
  18. Schwartz M. A., Both G., Lechene C. Effect of cell spreading on cytoplasmic pH in normal and transformed fibroblasts. Proc Natl Acad Sci U S A. 1989 Jun;86(12):4525–4529. doi: 10.1073/pnas.86.12.4525. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Schwartz M. A., Ingber D. E., Lawrence M., Springer T. A., Lechene C. Multiple integrins share the ability to induce elevation of intracellular pH. Exp Cell Res. 1991 Aug;195(2):533–535. doi: 10.1016/0014-4827(91)90407-l. [DOI] [PubMed] [Google Scholar]
  20. Schwartz M. A., Lechene C. Adhesion is required for protein kinase C-dependent activation of the Na+/H+ antiporter by platelet-derived growth factor. Proc Natl Acad Sci U S A. 1992 Jul 1;89(13):6138–6141. doi: 10.1073/pnas.89.13.6138. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Schwartz M. A., Lechene C., Ingber D. E. Insoluble fibronectin activates the Na/H antiporter by clustering and immobilizing integrin alpha 5 beta 1, independent of cell shape. Proc Natl Acad Sci U S A. 1991 Sep 1;88(17):7849–7853. doi: 10.1073/pnas.88.17.7849. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Shimizu Y., Shaw S. Lymphocyte interactions with extracellular matrix. FASEB J. 1991 Jun;5(9):2292–2299. doi: 10.1096/fasebj.5.9.1860621. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES