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J Biol Chem, Vol. 274, Issue 51, 36241-36251, December 17, 1999
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From the Platelet adhesion to sites of vascular injury is
initiated by the binding of the platelet glycoprotein (GP) Ib-V-IX
complex to matrix-bound von Willebrand factor (vWf). This
receptor-ligand interaction is characterized by a rapid on-off rate
that enables efficient platelet tethering and rolling under conditions
of rapid blood flow. We demonstrate here that platelets adhering to
immobilized vWf under flow conditions undergo rapid morphological
conversion from flat discs to spiny spheres during surface
translocation. Studies of Glanzmann thrombasthenic platelets (lacking
integrin Australian Centre for Blood Diseases,
Department of Medicine, Monash Medical School, the ¶ Department of
Pathology, Box Hill Hospital, Victoria, Australia, and the
§ Etablissement de Transfusion Sanguine, INSERM,
Unite 311, 67065 Strasbourg Cedex, France
IIb
3) and Chinese hamster
ovary (CHO) cells transfected with GPIb/IX (CHO-Ib/IX) confirmed that
vWf binding to GPIb/IX was sufficient to induce actin polymerization
and cytoskeletal reorganization independent of integrin
IIb3. vWf-induced cytoskeletal
reorganization occurred independently of several well characterized
signaling processes linked to platelet activation, including calcium
influx, prostaglandin metabolism, protein tyrosine phosphorylation,
activation of protein kinase C or phosphatidylinositol 3-kinase but was
critically dependent on the mobilization of intracellular calcium.
Studies of Oregon Green 488 1,2-bis(o-amino-5-fluorophenoxy)ethane-N,N,N',N-tetraacetic acid tetraacetoxymethyl ester-loaded platelets and CHO-Ib/IX cells demonstrated that these cells mobilize intracellular calcium in a
shear-dependent manner during surface translocation on vWf. Taken together, these studies suggest that the vWf-GPIb interaction stimulates actin polymerization and cytoskeletal reorganization in
rolling platelets via a shear-sensitive signaling pathway linked to
intracellular calcium mobilization.
To whom correspondence should be addressed: Australian Centre
for Blood Diseases, Dept. of Medicine, Monash Medical School, Box Hill
Hospital, Arnold St., Box Hill, Victoria 3128, Australia. Tel.: 61 3 9895-0311; Fax: 61 3 9895-0332; E-mail:
Shaun.Jackson@med.monash.edu.au.
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