Perturbation of cultured human vascular endothelial cells by phorbol ester or thrombin alters the cellular von Willebrand factor distribution

Reinders, Jan Hendrik; Vervoorn, Richard C.; Verweij, Cornelis L.; Mourik, Jan A. van; Groot, Philip G. de

doi:10.1002/jcp.1041330110

Cited by 35 publications

(18 citation statements)

References 47 publications

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“…Similarly, replenishment of stored vWF was not evident until 48 hours after PMA stimulation of endothelial cells in culture. 29 To resolve these differences, it will be necessary to correlate the morphological changes noted here with more direct and less subjective evidence of de novo synthesis of vWF. The model described lends itself to approaches such as mRNA quantification and in situ hybridization with speciesspecific vWF probes.…”

mentioning

confidence: 99%

Morphological alterations in endothelial cells associated with the release of von Willebrand factor after thrombin generation in vivo.

Richardson

Tinlin

et al. 1994

Arterioscler Thromb

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von Willebrand factor (vWF) is synthesized by endothelial cells and stored in endothelium-specific granules, the Weibel-Palade (WP) bodies. The release of vWF from endothelial cells in vitro in response to secretagogues such as thrombin is considered to result in the loss of WP bodies through the fusion of the WP bodies with the plasma membrane. Biochemical and morphological techniques, including transmission (TEM) and scanning (SEM) electron microscopy, were used to examine the plasma profile of vWF in parallel with morphological alterations in endothelial cells associated with the generation of thrombin in vivo. There was a rapid loss of high-molecular-weight multimers of the circulating vWF, with full recovery within 1 hour. Simultaneously, TEM demonstrated that the endothelial cells lost WP bodies and became V on Willebrand factor (vWF) is a large multimeric plasma protein that plays a central role in hemostasis. 12 It is synthesized in megakaryocytes 34 and endothelial cells 56 and stored in the a-granules of platelets 7 and in the endothelial cell storage organelles, the Weibel-Palade (WP) bodies. 8 P-selectin is a component of both the platelet a-granule and WP body membrane, 9 but other components of the a-granules have not been identified in WP bodies. 10 vWF is released from endothelial cells both constitutively and via the regulated pathway from WP bodies after endothelial cell stimulation." Release from WP bodies makes available the highly platelet-interactive, high-to abnormally high-molecular-weight multimeric forms of vWF. 2 vWF is released from endothelial cells in culture in response to stimulation by a variety of secretagogues, including thrombin.1112 It is generally considered that the mechanism of release involves the fusion of the limiting membrane of the WP bodies with either the luminal or abluminal membranes of the endothelial cell. This process has been inferred from observation in two settings. In vitro studies, using immunofluorescence 8 " 13 or electron mi- 10 demonstrated that the release of vWF was associated with a reduction in the numbers of WP bodies and the appearance of vWF in the supernatant. In studies in vivo, WP bodies appeared to release their contents into the circulation after fusing with the luminal membrane of endothelial cells.14 Although this observation was reported after the recognition that WP bodies stored vWF, 8 this was not appreciated by the authors, who considered the observation to indicate that WP bodies were secretory in nature but of unknown function.14 To our knowledge, there has been no in vivo study in which plasma vWF levels have been quantified in parallel with alterations in the WP bodies and vWF distribution within endothelial cells.We have developed a model for the induction of thrombin generation in vivo in various animal species. This is achieved through the infusion of two defined components of the prothrombinase complex 15 : factor Xa (activated factor X [FXa]) and a coagulant active phospholipid surface provided by phosphatidylchol...

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confidence: 99%

Morphological alterations in endothelial cells associated with the release of von Willebrand factor after thrombin generation in vivo.

Richardson

Tinlin

et al. 1994

Arterioscler Thromb

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“…vWF antigen was determined by ELISA (21). The amount of vWF and fibronectin in the extracellular matrix was determined as described (20).…”

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confidence: 99%

“…Cell lysate and medium fractions were prepared and, after preclearing of the fractions with gelatin coupled to Sepharose to remove fibronectin, vWF was immunoprecipitated with an anti-vWF monoclonal antibody coupled to Sepharose (21 (22).…”

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von Willebrand factor synthesized by endothelial cells from a patient with type IIB von Willebrand disease supports platelet adhesion normally but has an increased affinity for platelets.

Groot¹,

Federici²,

Boer³

et al. 1989

Proc. Natl. Acad. Sci. U.S.A.

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“…Recently, it was demonstrated that vWF re leased from the Weibel-Palade bodies is pre dominantly secreted to the luminal site of the endothelium [89]. Treatment of cultured endothelial cells with PMA and thrombin even blocked the deposition of vWF in the extracellular matrix [72], Thus, the WeibelPalade bodies contain a subclass of very large and biologically active vWF multimers that can be utilized in hemostasis upon phys iological stimulation. vWF released constitu tively is shown to be secreted both to the luminal and the basolateral sites of the endo thelium and may serve to maintain a con stant level of vWF in the blood under normal conditions.…”

Section: Biosynthesis Of Vwf By Human Endothelial Cellsmentioning

confidence: 99%

“…The majority of the newly synthesized protein is secreted constitutively which takes at least 2 h. About 5 % of the newly synthesized vWF follows the regu lated pathway [56], The rod-shaped organ elles, called Weibel-Palade bodies, are spe cific for endothelial cells [65][66][67] and most likely originate from the Golgi apparatus [68]. Release of vWF from these organelles can be induced by treatment of the cell with secretagogues such as thrombin, phorbol es ters such as phorbol myristate acetate (PMA), and calcium ionophore A23187 [69][70][71], The release of vWF induced by PMA was shown to parallel the capacity of PMA to activate proteinase kinase C [72], This obser vation may indicate that the activation of protein kinase C is involved in stimulusinduced release of vWF. After endothelial cells receive a stimulus, vWF is rapidly re leased (within 10 min), at a rate which is much higher than that required for the syn thesis and secretion of vWF by nonstimulated cells (about 2 h).…”

Section: Biosynthesis Of Vwf By Human Endothelial Cellsmentioning

confidence: 99%

Biosynthesis of Human von Willebrand Factor

Verweij¹

1988

Pathophysiol Haemos Thromb

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Endothelium forms the inner lining of all blood vessels and, as a consequence, is in direct contact with the blood. Because of this and the synthesis and secretion of hemostatic components, the endothelium is able to modulate coagulation and fibrinolysis. An important hemostatic factor synthesized by endothelial cells is the von Willebrand factor (vWF). vWF is a large plasma glycoprotein which promotes the adhesion of platelets to the vessel wall after a vascular injury. vWF is initially synthesized as a pre-pro-polypeptide. During its transport to the outside of the cell, the single-chain polypeptides are assembled into multimers. The pro-polypeptide can be cleaved and also be secreted. Free pro-polypeptide is identified as von Willebrand antigen II, a plasma glycoprotein of unknown function. Plasma vWF consists of a heterogenous series of multimers, composed of an apparently single-type glycoprotein subunit, linked together by disulfide bonds. The hemostatic potency of vWF was shown to increase with increasing multimer size. Therefore, the multimeric assembly of vWF is a crucial aspect in vWF biosynthesis. Furthermore, vWF synthesized by endothelial cells can either be secreted constitutively or stored and released upon stimulation of the endothelial cell. In this review, data are presented which contribute to the understanding of the biosynthetic pathway and complex processing which vWF has to undergo before it is secreted by the endothelial cell. These data have allowed a prediction of the sequential events underlying vWF biosynthesis, processing, multimer assembly, and secretion.

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Perturbation of cultured human vascular endothelial cells by phorbol ester or thrombin alters the cellular von Willebrand factor distribution

Cited by 35 publications

References 47 publications

Morphological alterations in endothelial cells associated with the release of von Willebrand factor after thrombin generation in vivo.

Morphological alterations in endothelial cells associated with the release of von Willebrand factor after thrombin generation in vivo.

von Willebrand factor synthesized by endothelial cells from a patient with type IIB von Willebrand disease supports platelet adhesion normally but has an increased affinity for platelets.

Biosynthesis of Human von Willebrand Factor

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