Histochem Cell Biol (2002) 117:113–122

DOI 10.1007/s00418-001-0368-9

REVIEW

Jan A. van Mourik · Thalia Romani de Wit
Jan Voorberg

Biogenesis and exocytosis of Weibel-Palade bodies

Accepted: 16 November 2001 / Published online: 19 January 2002
© Springer-Verlag 2002

Abstract Vascular endothelial cells contain typical, elongated vesicles, the so-called Weibel-Palade bodies, which
serve as a storage compartment for von Willebrand factor (VWF), a plasma protein that plays an essential role
in controlling the adhesion and aggregation of platelets
at sites of vascular injury. Upon activation of endothelial
cells by agonists such as thrombin, epinephrine or histamine, the Weibel-Palade bodies fuse with the plasma
membrane and release their contents into the blood circulation. This process provides an adequate means by
which endothelial cells can actively participate in controlling the arrest of bleeding upon vascular damage. Besides VWF, Weibel-Palade bodies contain a subset of
other proteins, including interleukin-8 (IL-8), P-selectin
and endothelin. Similar to VWF, these proteins are transported to the outside of the cell upon stimulation and
may control local or systemic biological effects, including inflammatory and vasoactive responses. Apparently,
endothelial cells are able to create a storage pool for a
variety of bioactive molecules which can be mobilised
upon demand. Endothelial cells that are deficient of
VWF synthesis are not only unable to form WeibelPalade bodies, but also lack the ability to store IL-8 or
P-selectin or release these proteins in a regulated manner. It thus appears that VWF not only plays a prominent
role in controlling primary haemostasis, but also may
modulate inflammatory processes through its ability to
target inflammatory mediators to the regulated secretion
pathway of the endothelium.

J.A. van Mourik (✉) · T. Romani de Wit · J. Voorberg
Department of Blood Coagulation, CLB, Plesmanlaan 125,
1066 CX Amsterdam, The Netherlands
e-mail: J_van_Mourik@clb.nl
Tel.: +31-20-5123120, Fax: +31-20-5123680
J.A. van Mourik · T. Romani de Wit · J. Voorberg
Department of Plasma Proteins, CLB, Amsterdam
J.A. van Mourik
Department of Vascular Medicine, Academic Medical Center,
University of Amsterdam, Amsterdam, The Netherlands

Keywords von Willebrand factor · Weibel-Palade
bodies · Endothelial cells

Introduction
Vascular endothelial cells are equipped with a machinery
that, upon perturbation, allows prompt delivery of a
number of bioactive substances, including hormones, receptors and adhesive molecules, to the surface of the
cell. In this way endothelial cells are able to selectively
alter the microenvironment of perturbed regions of the
vascular bed and, as such, may modulate and control a
variety of physiological processes such as haemostasis
and inflammatory responses. A distinct subset of proteins bound to be released upon stimulation of the endothelium, stems from Weibel-Palade bodies, typical and
morphologically highly organised storage vesicles that
release their contents by regulated exocytosis. In the
early 1960s, Weibel and Palade observed that endothelial
cells of virtually all blood vessel types contain elongated
organelles not found in other cell types (Weibel and
Palade 1964). More than 20 years later, immunohistochemical and cell fractionation studies revealed that
these organelles, so-called Weibel-Palade bodies, serve
as a storage vehicle for von Willebrand factor (VWF), a
plasma protein that plays an essential role in events that
lead to normal arrest of bleeding. It mediates the adhesion and aggregation of platelets at sites of vascular
injury and also functions as a stabilising chaperone protein of factor VIII, an essential cofactor of the coagulation system. The physiological significance of VWF is
underscored by the well-known observations that qualitative or quantitative defects, or von Willebrand’s disease, may predispose to a severe bleeding diathesis, because of both defective platelet plug formation and abnormal blood clotting at sites of vascular injury.
Accumulated evidence indicates that VWF is not only
stored in Weibel-Palade bodies, but also plays a critical
role in directing the formation of these granules. In addition, recent studies suggest that VWF also serves a role

into the regulated secretion pathway of endothelial cells. thereby. 1979). Hollestelle et al. (For comprehensive reviews on the cell biology and biochemical and functional properties of VWF. A Cross-section of an artery of the kidney immunostained for VWF. Sadler 1998. see Wagner 1990. Similarly. It is possible that VWF serves a role in controlling Fig. an event that most likely precedes the genesis of a WeibelPalade body. 2001). 1973. M mitochondrion. Large differences in the level of VWF gene expression were observed in different tissues (Yamamoto et al. Most of our insights into the biosynthesis of VWF has been derived from studies with cultured umbilical vein endothelial cells.114 in targeting a selected subset of proteins. strongly labelled with antibody. In the early 1970s immunohistochemical studies revealed that VWF was associated with the endothelium of a variety of tissues (Fig. Folkman et al. 1991). G Golgi apparatus. Ruggeri 1999. Weibel-Palade bodies. into Weibel-Palade bodies and. 1998. the Weibel-Palade bodies. including the leukocyte-adhesion receptor P-selectin and the chemokine interleukin-8 (IL-8). Hoyer et al. 1. B Intracellular localisation of VWF in cultured human umbilical vein endothelial cells (HUVECs). the storage vehicle of VWF. This picture shows budding of a VWF-containing vesicle. Bars represent a micron or fractions thereof. Bloom et al. (1993). 1A–D Cellular and subcellular localisation of von Willebrand factor (VWF). Biosynthesis of VWF VWF is synthesised exclusively by endothelial cells and megakaryocytes. endothelial cells-specific. (1988) and panel D from Voorberg et al. green). storage and secretion of VWF by endothelial cells and the potential role and significance of VWF in controlling the formation of Weibel-Palade bodies. More recent studies have demonstrated that VWF is less uniformly expressed than previously thought. Note the typical rod-shaped secretory vesicles. Specific staining of the endothelial lining is indicated (arrow. The significance of these observations is not clear. elongated. D Ultrathin frozen section of a HUVEC immunostained with anti-VWF. It seems likely that the biosynthesis in megakaryocytes is similar to that in endothelial cells. The presence of typical. Panel C of this figure was from Reinders et al. Hop and Pannekoek 1996. WP Weibel-Palade body. electrondense vesicles is indicated. Romani de Wit and van Mourik 2001). C Electron micrograph of a HUVEC. from the trans-Golgi network. we will briefly discuss the major molecular events associated with the biosynthesis. with permission . are heterogeneously distributed along the vascular tree (Gebrane-Younès et al. In this review. 1973. 1973) and that endothelial cells derived from various blood vessel types synthesise and secrete VWF (Jaffe et al.

2 Assembly. an event generally associated with secretion of proteins in eukaryotes (Fig. is occasionally found in close apposition to the TGN (Fig. the pro-VWF dimers polymerise during their travel through the Golgi compartments through amino-terminal “head-to-head” disulphide bonds. is only secreted upon proper stimulation of the cell (see below). 1988) but. Vischer et al. including binding to platelet surface receptors. VWF (subunit) is composed of a number of highly conserved domains that exert autonomous functions.1 µm thick (Weibel and Palade 1964. pro-VWF (Fig. Borchiellini et al. 1997). Polymerisation of VWF is one of the most characteristic post-translational One of the most remarkable features of endothelial cells is their ability to store de novo synthesised VWF. primarily consisting of the functionally most potent (high molecular weight) multimers. Accumulating evidence suggests that VWF by itself is the driving force behind the formation of Weibel-Palade . where further processing takes place. 1987. A portion of mature VWF. “tail-to-tail” dimerisation at the carboxy-terminal region. frozen sections of cultured endothelial cells showed that. The left panel shows the polymeric structure of VWF as visualised by polyacrylamide gel electrophoresis. The propeptide plays an essential role in the assembly of multimers in that it promotes interchain disulphide bond formation and polymerisation. After endoproteolytic cleavage of pro-VWF in the TGN (Vischer and Wagner 1994). The top panel depicts a schematic representation of the primary structure of pro-VWF. This figure is adapted from van Mourik and Romani de Wit (2001). 1). extracellular matrix components and factor VIII. Wagner et al. In cultured endothelial cells part of the VWF and propeptide generated upon proteolytic maturation is secreted in a constitutive manner. where further multimerisation and endoproteolytic cleavage occur. together with some unprocessed proVWF (Sporn et al. Storage of VWF Fig. These observations support the view that VWF-containing storage vesicles emerge through budding from the Golgi complex. In addition. Our current insights into the mechanisms and signals involved in the biogenesis of Weibel-Palade bodies and in directing VWF (and other soluble secretory proteins. most likely representing VWF packed in a virtually crystalloid state. Pro-VWF monomers dimerise through cysteine residues located at the carboxy-terminus. may also catalyse disulphide-linked crosslinking (Mayadas and Wagner 1992). Electron microscopic examination suggests that in Weibel-Palade bodies VWF is densely packed in a highly organised manner (Fig. 1. Wise et al. yielding mature VWF multimers and propeptide dimers. Wagner et al. Electron microscopic studies also revealed that Weibel-Palade bodies probably originate from the TGN (Sengel and Stoebner 1970). 2). pro-VWF undergoes N-linked glycosylation and disulphide-linked. which appear as elongated. particularly at high hydrodynamic shear forces (Ruggeri 1999). surrounded by a limited membrane.115 events that occurs during its voyage to the outside of the cell. The propeptide is essential for proper multimerisation in that it not only aligns the amino-termini of two pro-VWF dimers for interdimer crosslinking (Verweij et al. Inside the Weibel-Palade bodies are several longitudinally arranged tubules. This multifunctional nature of VWF is markedly amplified by polymerisation. Simultaneously with these post-translational processing steps. Voorberg et al. Immunostaining of ultrathin. Arvan and Castle 1998). The location of conserved structural domains and the propeptide and mature VWF moiety are indicated. After cleavage of the signal peptide during translocation in the endoplasmic reticulum. together with its propeptide. The pro-VWF dimers are subsequently transported to the Golgi apparatus and postGolgi compartments. 1987. it has been proposed that the propeptide serves a role in targeting VWF (and other proteins) into the Weibel-Palade bodies (see below). The molecular events associated with the biosynthesis of VWF occur in a highly ordered manner. with permission haemostasis in a vascular bed-specific manner (Rosenberg and Aird 1999). In these granules VWF and propeptide are found in equimolar amounts (Ewenstein et al. The subtilisin-like endoprotease furin most likely catalyses the cleavage of the propeptide portion of the molecule. mature VWF and its propeptide partition between two different secretion pathways. 1996. and processing of VWF by endothelial cells. see below) into these storage vesicles is rather limited. Pro-VWF polymers are subsequently transported to the trans-Golgi network (TGN). probably by virtue of its intrinsic protein disulphide isomerase activity. is stored in unique secretory vesicles. 1986). 1987. VWF originates from a 360-kDa precursor. 1. 1993). Griffiths and Simons 1986. 1982). Before VWF leaves the cell it undergoes a number of posttranslational modifications which are of importance for the function of VWF. Multimers are formed via cysteines in the D′ and D3 domain. densely packed VWF. the constitutive and regulated pathway. the Weibel-Palade bodies. rod-shaped vesicles which can be up to 4 µm long and 0. including sulphation and O-linked glycosylation. The latter pool. a process that may enhance the typical multi-interactive nature of VWF. together with apparently mature Weibel-Palade bodies. It is likely that these structures determine the dimension of Weibel-Palade bodies. The lower panel schematically depicts the multimeric structure of VWF.

these proteins are both structurally and functionally clearly distinct. Halban and Irminger 1994. Thus. when an epithelial cell line which synthesises P-selectin. 2000). rather than multimerisation as a component of sorting. Exit of VWF from the Golgi complex The structural features of VWF that trigger the formation of VWF-containing condensing vesicles which subsequently mature into Weibel-Palade bodies have not been clearly identified. exocytosis of WeibelPalade bodies is of broader biological significance than previously thought. results in the formation of WeibelPalade body-like vesicles (Voorberg et al. It thus appears that VWF plays a prominent role in the formation of Weibel-Palade bodies (or WeibelPalade body-like vesicles). The second hypothesis considers that the propeptide moiety of VWF contains a targeting signal that interacts with a carrier protein or receptor in the Golgi membrane to mediate targeting of VWF (and the propeptide itself) to storage (Haberichter et al. Pertinent to this point is the observation that the propeptide may bind to (mature) VWF in a milieu thought to prevail in the TGN (Vischer and Wagner 1994). Weibel-Palade bodies have been shown to contain a number of other proteins (Table 1). it has been shown that when expressed alone in AtT-20 cells. is poorly understood. 1993) but not in AtT-20 (mouse pituitary cell line. Weibel-Palade body-like organ- .e. Journet et al. to the plasma membrane of endothelial cells is dramatically reduced in VWF-deficient mice (Denis et al. 1991. but does not retain this receptor in storage vesicles. It is possible that the sorting mechanism is dependent on the cell type employed. despite considerable efforts to unravel the mechanism responsible for targeting of VWF to storage vesicles. 1991. These observations suggest that VWF carries within its structure one or more domains or structural features that allows its segregation in the TGN from the bulk flow of constitutively released protein. it has been shown that also dimeric. 1991. was transfected with VWF-cDNA. It is thought that only through association with the propeptide VWF is brought into storage (Haberichter et al. 2000). leukocyte recruitment at vascular inflammatory sites is impaired (Denis et al. indicating the absence of its storage vehicle. irrespective of the cell type by which VWF is expressed. 1993. Mayadas and Wagner 1992). How these discrepancies can be explained remains uncertain. together with VWF. This “multimerisation hypothesis” is primarily based on the observation that only in cells that synthesise polymerised VWF. 2001). Taken together. Polymerisation is apparently not the only requirement for storage. It is also not clear whether or not cleavage of the propeptide is required for sorting. Obviously. Hop et al. but may also have a helper function in the sorting of other proteins to these secretory vesicles. the process of VWF storage is still not fully understood. Pertinent to this point is the observation that in mice deficient of Weibel-Palade bodies. cells that produce VWF mutants that are unable to multimerise are devoid of VWF-containing storage vesicles. On the other hand. Denis et al. Most studies have in common that at least the propeptide of VWF is required for sorting. the WeibelPalade body. Hop et al. It has been postulated that the targeting of VWF into Weibel-Palade bodies occurs as a consequence of multimerisation of this protein in the TGN (Voorberg et al. 2000). For instance. these proteins are transported to the outside of the cell upon stimulation and may control local or systemic biological effects. Accumulated evidence suggests that VWF not only plays a role in directing the formation of Weibel-Palade bodies (vide supra). Despite their common subcellular localisation. Voorberg et al. Similarly. stimulus-induced translocation of P-selectin. including inflammatory and vasoactive responses. 2000). Arvan and Castle 1998). the propeptide harbours a signal required for targeting to storage vesicles. Possibly the propeptide contains an anchor point that allows retention in the TGN and serves as a condensation nucleus for VWF molecules and the propeptide itself. a cell of the regulated type) or RIN 5F cells (rat insulin-secreting β cell line. 2001). not multimerisation of VWF per se. It has been postulated that this signal is also required for targeting of VWF to storage granules. 2001). For instance. different experimental conditions (for example. another resident of Weibel-Palade bodies (see below). In this respect. Additional Weibel-Palade body constituents In addition to VWF and its propeptide. a process that underlies the sorting and targeting of many other proteins and hormones destined for regulated secretion (reviewed in Kelly 1985. 1993). transfection of pro-VWF cDNA into cells devoid of VWF synthesis. The mechanism responsible for sorting of these “foreign” proteins into Weibel-Palade bodies. multimerisation) and condensation. 2000). unpolymerised VWF can be targeted to storage vesicles (Wagner et al. 1993. resulted in the formation of VWF-containing vesicles that are morphologically very similar to Weibel-Palade bodies (Wagner et al. Electron microscopic and immunohistochemical studies showed that in pigs or mice deficient of VWF the endothelium is devoid of Weibel-Palade bodies (Gebrane-Younès et al. In monkey kidney CV-1 cells expression of a VWF mutant that lacks the propeptide cleavage site but is able to form covalently linked polymers. cell lines and VWF mutants used to study expression) may underlie these differences. this hypothesis advocates that storage of VWF is directed by selective aggregation (i. In addition. 1997. VWF-containing granules are formed. Two possible mechanisms have been proposed for targeting VWF into the regulated pathway of secretion. Similar to VWF and the propeptide. Apparently.116 bodies. the propeptide is sorted into granular vesicles (Haberichter et al.

(1985) Ewenstein et al. a process that may facilitate their retention and condensation within the Weibel-Palade bodies (Arvan and Castle 1998). 3.and P-selectin RNA of T24 cells transfected with VWF cDNA or mock-transfected cells (control).3-Fucosyltransferase VI Tissue-type plasminogen activator Membrane surface glycosylation Fibrinolysis Wagner et al. These observations suggest that VWF may trigger sequestering of structurally unrelated proteins into Weibel-Palade bodies. It is possible that IL-8 and other Weibel-Palade body residents interact or coaggregate with VWF in TGN. with permission elles. Cells that synthesise IL-8 in substantial amounts. (VWF von Willebrand factor) Protein Function Referencesa Mature VWF Haemostasis VWF propeptide VWF polymerisation chaperone of VWF inflammation P-selectin Leukocyte adhesion Endothelin CD63/lamp3 Interleukin-8 Vasoconstriction Cell adhesion/migration Inflammatory reactions α1. The left panel shows double-labelling immunofluorescent staining of T24 cells devoid of VWF synthesis (control) and T24 cells transfected with VWF cDNA (VWF cDNA). (2000). (1999) Datta et al. (1998) Wolff et al. (1989) McEver et al. This figure is adapted from Hop et al. Only cells transfected with VWF cDNA display colocalisation of VWF and P-selectin.117 Table 1 Contents of Weibel-Palade bodies. (1999) a Refer to cellular localisation of Weibel-Palade body proteins Fig. 3 Co-targeting of VWF and P-selectin into Weibel-Palade bodies in T24 cells (human bladder carcinoma cell line). containing both VWF and P-selectin. (1982) Ewenstein et al. 2000). (1989) Hattori et al. Hop et al. A VWF mutant that is unable to polymerise (VWFdel cDNA) does not colocalise with P-selectin. (1998) Vischer and Wagner (1993) Utgaard et al. (1987) McCarroll et al. This mutant distributes primarily in the rough endoplasmic reticulum and the Golgi apparatus whereas only faint staining of P-selectin can be seen. with anti-VWF and anti-P-selectin antibodies. (1987) Bonfanti et al. Similarly VWF also targets IL-8 to Weibel-Palade bodies (Romani de Wit et al. (1989) Russell et al. (2000) Rosnoblet et al. 2001). do not store this cytokine in Weibel-Palade body-like vesicles. (1998) Schnyder-Candrian et al. The right panel shows northern blot analysis for VWF. were formed (Fig. As VWF is able to bind a variety of . unless VWF and IL-8 are coexpressed.

a GTPase-activating protein for Cdc42 and Rac. In both cell types. H. 1998). The molecular mechanism of cAMPmediated VWF secretion has not been identified. In addition. 1998. such as epinephrine or DDAVP (1-deamino-8-D-arginine vasopressin). independently of a rise of cytosolic Ca2+. [Ca2+]i-raising agents and agents that activate the cAMP pathway may act in a synergistic manner (Vischer and Wollheim 1997). 2000). it was shown that VWF-mediated targeting of factor VIII was due to binding of factor VIII to VWF in the TGN. and other constituents of the Weibel-Palade bodies. agents known for their ability to activate cAMP-dependent signalling independently of a rise in [Ca2+]i (Vischer and Wollheim 1997. Limited information is available concerning the mechanisms underlying these processes (for recent reviews on the regulated secretion in endothelial cells. agents known for their ability to activate G proteincoupled receptors and cAMP-dependent signalling. Regulated. 1997. resulting in a rise of cytoplasmic Ca2+. 1998). thrombin-induced cycling of Ral from its inactive GDP-bound to its . The hydrolysis of GTP on Cdc42 or Rac may result in cytoskeleton rearrangement. which may precede exocytosis of WeibelPalade bodies. Vischer et al. black circles) or exposed at the plasma membrane (for example P-selectin). This view is supported by coexpression studies of VWF and factor VIII (a coagulation factor that readily binds to VWF) in AtT-20 cells and vascular endothelial cells (Rosenberg et al. Ral in its GTPbound conformation interacts downstream with effector molecules such as RLIP76. Birch et al. van den Eijnden-Schrauwen et al.118 proteins and biological compounds (Sadler 1998). 4). Recently it has been shown that the small GTP-binding protein Ral is associated with Weibel-Palade bodies. V2R Vasopressin type 2 receptor. Calcium in complex with calmodulin (CaM) binds the small GTP-binding protein Ral in its GDP form. GDP on Ral is then exchanged for GTP by mediation of a Ral guanine nucleotide exchange factor (RalGEF). 1999. involves the translocation of the Weibel-Palade bodies from the cytoplasm towards the plasma membrane and the fusion of these vesicles with the plasma membrane (Fig. whereas no storage could be observed in the absence of VWF synthesis. 4 Machinery of stimulus-induced exocytosis of WeibelPalade bodies by endothelial cells. Datta and Ewenstein 2001). a small amount of which is associated with the Weibel-Palade bodies. 1992. it is tempting to speculate that Weibel-Palade body constituents may specifically interact with VWF preceding maturation of secretory vesicles. see Vischer and de Moerloose 1999. Thrombin signalling is mediated by a G protein-coupled proteaseactivated receptor (PAR). such as epinephrine. Upon stimulation of endothelial cells by agonists such as thrombin. de Leeuw. In addition. including thrombin and histamine. the phosphatidyl inositol pathway is activated. Regulated secretion of VWF can also be induced by secretagogues. The contents of Weibel-Palade bodies are released into the circulation (for example VWF. adapted version. Exocytosis of Weibel-Palade bodies The regulated secretion of VWF. Courtesy of Dr. adenosine or DDAVP (1-deamino-8-D-arginine vasopressin). de Leeuw et al. The cellular responses to the increased [Ca2+]i are most likely mediated through calmodulin and small GTPbinding proteins (Hamilton and Simps 1987. de Leeuw (2000) et al. colocalisation of factor VIII and VWF was observed in storage vesicles. receptor-mediated secretion of VWF can also be accomplished by agonists. β2-AR β2-adrenergic receptor. Increased levels of cytosolic free Ca2+ concentration ([Ca2+]i) have been implicated in the mechanism of exocytosis for a number of agonists. Hegeman Fig.

Conclusions and future perspectives This review has served to illustrate the role of VWF in controlling the genesis of Weibel-Palade bodies in endo- . it has been reported that VWF released from cultured cells is primarily derived from the storage pool (Tsai et al. 1989. It remains unclear.119 active GTP-bound state coincided with secretion of VWF (de Leeuw et al. 1992) and polar secretion predominantly to the basal side have been observed (Sporn et al. 1996). including activation of protein kinases that eventually lead to cellular secretion. unperturbed endothelial cells secrete VWF both to the apical and basolateral side of the cell (van Buul-Wortelboer et al. 2000). Most studies have shown that VWF is secreted by the endothelium in an organised fashion. 2001). Ral is most likely also involved in cytoskeleton dynamics and rearrangements (Bos 1998). tissue with a relatively high VWF expression level (Yamamoto et al. substantially polymerised and functionally mature VWF appeared to be stored in WeibelPalade bodies. alternatively. 2001). though the extent of secretion to either side seems to be dependent of culture conditions. Kaufmann et al. secretion and the ability of endothelial cells to store VWF. but also by microvascular cells of the lung. It seems clear. Ral interacts with calmodulin in a Ca2+-dependent manner. Narahara et al. This suggests that. On the other hand. The molecular mechanisms associated with [cAMP]idependent exocytosis of Weibel-Palade bodies remain to be identified. together with its propeptide. In vivo exocytosis of Weibel-Palade bodies and subsequent secretion of VWF seem to occur primarily towards the apical surface upon perturbation of the endothelium (Richardson et al. This vasopressin receptor subtype is primarily expressed in the kidney. it is possible that. 2000). Only fully processed. the observations are quite contradictory. Hakkert et al. Obviously a rather controversial issue. little is known about the molecular mechanisms distal to adenylate cyclase activation and cAMP generation. 1994). These observations indicate that Ral is a mediator of exocytosis of Weibel-Palade bodies. 1989). This conclusion is further supported by the observation that expression of a constitutively active (GTP-bound) Ral mutant in endothelial cells resulted in the disappearance of Weibel-Palade bodies (de Leeuw et al. under normal physiological conditions. converted into mature VWF and propeptide. Indeed. 2000). such as cellular support. coupled via G proteins to adenylate cyclase activation and cAMP generation. pro-VWF is rapidly cleared from the circulation or. VWF released through the constitutive pathway would be less efficient in this respect. 1986). pro-VWF. 2001). a mechanism that could be of significance in controlling plasma VWF levels under physiological conditions. after its release. Both secretion of VWF exclusively to the luminal side upon stimulation of the cell (van Buul-Wortelboer et al. respectively). In vitro studies have shown that a significant portion of de novo synthesised VWF is secreted through the constitutive pathway as partially processed. In any case. 1975. Besides the possibility that cAMP triggers a cascade of reactions. 1994). Only trace amounts of pro-VWF are detectable in plasma (Borchiellini et al. Similarly the β2 adrenergic receptors have been implicated in cAMP-mediated secretion of VWF (Vischer and Wollheim 1997). 1989. As to the polarity of regulated secretion. therefore. that activation of lung V2R by ligands such as vasopressin or DDAVP contributes to the acute increase of plasma VWF observed after administration of these drugs (Mannucci et al. It seems reasonable to assume that a subset of biologically potent VWF polymers become readily available in the vicinity of the injured vessel upon activation of endothelial cells and the subsequent exocytosis of Weibel-Palade bodies (for example. In particular. an event that also enhances binding of GTP to Ral several fold (Wang and Roufogalis 1999). incompletely polymerised and functionally incompetent protein (Sporn et al. 1989. therefore. whereas cAMPmediated secretion involves only vesicles located in the periphery of the cell (Vischer et al. To what extent the plasma VWF pool stems from the constitutive route in vivo is unknown. It has been proposed that in particular the latter cellular pool of VWF is of importance in maintaining plasma VWF levels (Vischer et al. at least VWF that is targeted into the regulated pathway. Sporn et al. 1991). that this small GTPase plays a key role in controlling regulated exocytosis of VWF by endothelial cells. animal experimental studies have shown that after infusion of pro-VWF the propeptide is rapidly cleaved from its precursor (Turecek et al. is the vasopressin V2 receptor (V2R. these VWF species play a prominent role in controlling platelet adhesion and aggregation. On the other hand. therefore. 1999). the constitutive pathway does not substantially contribute to the release of VWF. It is possible. Manucci 1997). is not (Borchiellini et al. their precursor. 1998. induced by thrombin formed locally as a result of vascular damage). Hollestelle et al. only mature VWF and its propeptide are detectable in plasma in significant amounts (about 50 and 5 nM. to what extent release through the constitutive pathway plays a role in controlling plasma VWF levels. Resting. Again different experimental conditions most likely underlie these striking discrepancies. Because of its limited degree of polymerisation. One of the candidate endothelial cell receptors. 1996). Physiological significance of VWF processing and secretion Little is known about the physiological significance of the diversity of processing. In one respect cAMP-mediated responses differ from regulated secretion elicited by rises in [Ca2+]i in that secretion induced by [Ca2+]i-raising agents involves the release of both peripheral and central granules.

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