Von Willebrand factor (vWF), fibronectin (FN) and 13-hydroxy-octadecadienoic acid (13-HODE) are known to influence the regulation of the adhesive properties of vascular surfaces. In the present study vWF, FN and 13-HODE were comparatively localized in endothelial cells (EC) and in the extracellular matrix (ECM) produced by EC. An indirect immunofluorescent technique was applied to coverslips containing human EC cultures previously fixed and permeabilized following different procedures: A. Alcohol/acetone; B. Paraformaldehyde alone and C. Paraformaldehyde followed by Triton X-100. vWF was observed inside EC (A), on the ECM produced by EC (B) or in EC and ECM (C) depending on the fixation procedures used. FN was mainly localized in the ECM despite the fixation procedures employed. FN was only seen in relation to cell bodies after strong permeabilization (A). Under our experimental conditions 13-HODE was never found in ECM. This latter antigen was observed randomly dispersed in those preparations fixed with alcohol/acetone, indicating that it is probably extracted by this fixative. 13-HODE was detected in granular shaped structures in EC after permeabilization with detergent (C). These results suggest that the cellular localization of vWF and FN is compatible with an adhesive role related to the abluminal side of ECs. 13-HODE was readily observed after mild permeabilization. This finding would be morphologically consistent with its contribution to the regulation of the vessel wall thromboresistance.
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Bastida E, Almirall L, Ordinas A (1989) Platelet and shear rate promote tumor cell adhesion to human endothelial extracellular matrix. Abscence of a role for platelet cyclooxygenase. Thromb Haemost 61:485–489
Beckstead JH, Stenberg PE, McEver RP, Schuman MA, Bainton DF (1986) Immunohistochemical localization of membrane and alpha-granule proteins in human megakaryocytes: application to plastic-embedded bone marrow specimens. Blood 67:285–293
Buchanan MR, Bastida E (1987) The role of 13-HODE and HETEs in vessel wall/circulating blood cells interactions. Agents Actions 22:1–3
Buchanan MR, Haas TA, Lagarde M, Guichardant M (1985) 13-hydroxy-octadecadienoic acid is the vessel wall chemorepellant, LOX. J Biol Chem 260:16056–16059
Buchanan MR, Bastida E, Escolar G, Haas TA, Weber E (1989) Vessel wall reactivity and 13-hydroxy-octadecadienoic acid synthesis. In: Samuelsson B, Wong PYK, Sun FF (eds) Advances in prostaglandin, thromboxane and leukotriene research, vol 119. Raven Press, New York, pp 293–298
Castagnino CG, Berria MI (1986) Influence of cell fixatives and incubation schedule on PAP labelling of fibronectin in cultivated fibroblast. J Histochem Cytochem 34:1331–1333
De Groot PG, Sixma JJ (1987) Role of von Willebrand Factor in the vessel wall. Semin Thromb Hemost 13:416–424
Holund B, Clauren PP, Clemmensen I (1981) The influence of fixation and tissue preparation on the immunohistochemical demonstration of fibronectin in human tissue. Histochemistry 72:291–299
Hormia M, Lehto VP, Virtanen I (1983) Factor VIII-related antigen. A pericellular matrix component of cultured endothelial cells. Exp Cell Res 149:483–497
Houdijk WPM, Sixma JJ (1985) Fibronectin in artery subendothelium is important for platelet adhesion. Blood 65:598–604
Jaffe EA, Nachman RL, Becker CG, Minick CR (1973) Culture of human endothelial cells derived from umbilical veins. Identification by morphologic and immunologic criteria. J Clin Invest 52:2745–2756
Laurie GW, Leblond CP, Martin GR (1982) Localization of type IV collagen, laminin, heparan sulfate protcoglycan, and fibronectin to the basal lamina of basement membranes. J Cell Biol 95:340–344
Moake JL, Turner NA, Stathopoulos NA, Nolasco LH, Hellums JD (1986) Involvement of large plasma von Willebrand Factor (vWF) multimers and unusualy large vWF forms derived from endothelial cells in shear stress-induced platelet aggregation. J Clin Invest 78:1456–1461
Mueller SC, Hasenaga T, Yamada SS, Yamada KM, Chen W (1988) Transmembrane orientation of the fibronectin receptor complex (integrin) demonstrated directly by a combination of immunocytochemical approaches. J Histochem Cytochem 36:297–306
Rand JH, Sussman II, Gordon RE, Shu SU, Solomon V (1980) Localization of factor VIII-related antigen in human vascular subendothelium. Blood 55:752–756
Reinders JH, De Groot PG, Gonsalves MD, Zanderberger J, Loesberg J, Van Mourik JA (1982) Arachidonate acid metabolism in cultured vascular endothelial cells. Biochim Biophys Acta 713:581–588
Reinders JH, De Groot PG, Gonsalves MD, Zanderbergen J, Loesberg C, Van Mourik JA (1984) Isolation of a storage and secretory organelle containing von Willebrand protein from cultured human endothelial cells. Biochim Biophys Acta 804:361–369
Reinders JH, de Groot PG, Dawes J, Hunter NR, van Heugten HAA, Zandbergen J, Gonsalves MD, van Mourik JA (1985) Comparison of secretion and subcellular localization of von Willebrand protein with that of thrombospondin and fibronectin in cultured human vascular endothelial cells. Biochim Biophys Acta 844:306–313
Ruggeri ZM, Zimmerman TS (1987) Von Willebrand factor and von Willebrand disease. Blood 70:895–904
Sporn LA, Marder VJ, Wagner DD (1986) Inducible secretion of large biologically potent von Willebrand multimers. Cell 46:185–190
Van Wachem PB, Reinders JH, Van Buul-Wortelbuer MF, De Groot PG, Van Aken WG, Van Mourik JA (1986) Von Willebrand Factor in cultured human vascular endothelial cells from adult and umbilical cord arteries and veins. Thromb Haemost 56:189–192
Wagner DD, Olmsted JB, Marder VJ (1982) Immunolocalization of von Willebrand protein in Weibel-Palade bodies of human endothelial cells. J Cell Biol 95:355–360
Wagner DD, Urban-Pickering M, Marder V (1984) Von Willebrand protein binds to extracellular matrices independently of collagen. Proc Natl Acad Sci USA 81:471–475
Yamada KM, Olden K (1978) Fibronectin-adhesive glycoproteins of cell surface and blood. Nature 275:179–184
Yamada KM, Akiyama SK, Humphries MJ, Yamada S, Santoro S (1986) Platelet and fibroblast interactions with fibronectin and other adhesive proteins. In: Jolles G, Legrand YJ, Nurden A (eds) Biology and pathology of platelet-vessel wall interactions. Academic Press, pp 53–74
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Aznar-Salatti, J., Bastida, E., Buchanan, M.R. et al. Differential localization of von Willebrand factor, fibronectin and 13-HODE in human endothelial cell cultures. Histochemistry 93, 507–511 (1990). https://doi.org/10.1007/BF00266409
- Human Endothelial Cell
- Fixation Procedure
- Human Vascular Endothelial Cell
- Indirect Immunofluorescent Technique