Changes in cytosolic Ca2+ associated with von Willebrand factor release in human endothelial cells exposed to histamine. Study of microcarrier cell monolayers using the fluorescent probe indo-1
- PMID: 3492515
- PMCID: PMC424139
- DOI: 10.1172/JCI112853
Changes in cytosolic Ca2+ associated with von Willebrand factor release in human endothelial cells exposed to histamine. Study of microcarrier cell monolayers using the fluorescent probe indo-1
Abstract
A method for measuring fluorescence in anchored monolayers of human endothelial cells is described and used to demonstrate changes in the cytosolic free-calcium concentration ([Ca2+]c) in these cells exposed to histamine and thrombin; some endothelial responses to both agonists (e.g., mitogenesis) have been suggested to be Ca2+-mediated. Umbilical vein endothelial cells were cultured on microcarriers and loaded with the Ca2+ indicator, indo-1. Enzymatic cell detachment was avoided by monitoring the indo-1 fluorescence ratio (400/480 nm) of a stirred suspension of cell-covered microcarriers. Basal [Ca2+]c was estimated to be 70-80 nM. Thrombin induced a transient dose-dependent increase in [Ca2+]c, which was active-site dependent. Histamine stimulated a dose-dependent increase in [Ca2+]c, which was reversed by removal of histamine and inhibited competitively by the H1-receptor antagonist pyrilamine, but not by the H2-receptor antagonist cimetidine. Furthermore, histamine induced a dose-dependent secretion of von Willebrand factor, which paralleled the rise in [Ca2+]c and was similarly blocked by the H1-receptor antagonist, and which may contribute to platelet deposition at sites of inflammation.
Similar articles
-
Histamine type I receptor occupancy increases endothelial cytosolic calcium, reduces F-actin, and promotes albumin diffusion across cultured endothelial monolayers.J Cell Biol. 1986 Dec;103(6 Pt 1):2379-87. doi: 10.1083/jcb.103.6.2379. J Cell Biol. 1986. PMID: 3782301 Free PMC article.
-
Thrombin-induced release of von Willebrand factor from endothelial cells is mediated by phospholipid methylation. Prostacyclin synthesis is independent of phospholipid methylation.J Biol Chem. 1984 Nov 10;259(21):13329-33. J Biol Chem. 1984. PMID: 6436246
-
Cell shape change and cytosolic Ca2+ in human umbilical-vein endothelial cells stimulated with thrombin.Thromb Haemost. 1992 Mar 2;67(3):331-4. Thromb Haemost. 1992. PMID: 1641824
-
Prolonged peak elevations in cytoplasmic free calcium ions, derived from intracellular stores, correlate with the extent of thrombin-stimulated exocytosis in single human umbilical vein endothelial cells.J Cell Physiol. 1994 Sep;160(3):545-54. doi: 10.1002/jcp.1041600318. J Cell Physiol. 1994. PMID: 7521337
-
Measurement of [Ca2+]i in cell suspensions using indo-1.Methods Mol Biol. 1999;114:41-7. doi: 10.1385/1-59259-250-3:41. Methods Mol Biol. 1999. PMID: 10081009 Review. No abstract available.
Cited by
-
Possible involvement of adenosine 3':5'-cyclic monophosphate and extracellular calcium ions in histamine stimulation of interleukin-1 release from macrophage-like P388D1 cells.Immunology. 1990 Jun;70(2):186-90. Immunology. 1990. PMID: 2165034 Free PMC article.
-
Hypoxia and inflammatory synovitis: observations and speculation.Ann Rheum Dis. 1991 Feb;50(2):124-32. doi: 10.1136/ard.50.2.124. Ann Rheum Dis. 1991. PMID: 1705416 Free PMC article. Review. No abstract available.
-
Effect of vasoactive amines on Weibel-Palade bodies in capillary endothelial cells.Experientia. 1991 Aug 15;47(8):830-2. doi: 10.1007/BF01922465. Experientia. 1991. PMID: 1915765
-
GMP-140, a platelet alpha-granule membrane protein, is also synthesized by vascular endothelial cells and is localized in Weibel-Palade bodies.J Clin Invest. 1989 Jul;84(1):92-9. doi: 10.1172/JCI114175. J Clin Invest. 1989. PMID: 2472431 Free PMC article.
-
Defect in regulated secretion of P-selectin affects leukocyte recruitment in von Willebrand factor-deficient mice.Proc Natl Acad Sci U S A. 2001 Mar 27;98(7):4072-7. doi: 10.1073/pnas.061307098. Epub 2001 Mar 6. Proc Natl Acad Sci U S A. 2001. PMID: 11274431 Free PMC article.
References
Publication types
MeSH terms
Substances
Grants and funding
LinkOut - more resources
Full Text Sources
Other Literature Sources
Research Materials
Miscellaneous