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Blood, 1 January 2002, Vol. 99, No. 1, pp. 159-167
HEMOSTASIS, THROMBOSIS, AND VASCULAR BIOLOGY
Shear-dependent tether formation during platelet translocation on
von Willebrand factor
Sacha M. Dopheide,
Mhairi
J. Maxwell, and
Shaun P. Jackson
From the Australian Centre for Blood Diseases,
Department of Medicine, Monash Medical School, Australia.
The adhesion and aggregation of platelets at sites of vascular
injury is dependent on the initial binding of the GP Ib/V/IX receptor
complex to immobilized von Willebrand factor (VWF). Under flow
conditions, this interaction supports platelet translocation that is
characteristically stop-start in nature. High resolution imaging of
platelets during surface translocation on immobilized VWF revealed that
thin membrane tethers (length: 0.91 µm-47.90 µm) were pulled from
the surface of these cells. Membrane tethers were dynamic structures
that extended from small, localized adhesion contacts under the
influence of flow. Perfusion of platelets in the presence of blocking
antibodies against integrin  IIb 3, or over
isolated A1 domains, demonstrated that the VWF-GP Ib interaction was
sufficient to induce membrane tether formation. The rate and extent of
tether elongation was shear-dependent (shear range: 150 s 1-10 000 s1), with mean tether length
ranging from 3.23 µm to 16.55 µm, tether frequency from 2.67% to
97.33%, and tether growth rate from 0.04 µm/sec to 8.39 µm/sec.
Tether formation and retraction did not require platelet activation;
however, the growth rate, lifetime, and dimensions were
significantly affected by the actin polymerization inhibitor,
cytochalasin D, and by chelating intracellular calcium. Single-cell
analysis revealed that formation of membrane tethers regulates the
stop-start phases of platelet translocation on VWF, suggesting a
potentially important role for this phenomenon in regulating the
dynamics of the platelet-VWF interaction under flow.
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