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Prepublished online as a Blood First Edition Paper on May 8, 2003; DOI 10.1182/blood-2003-01-0331.
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Blood, 1 September 2003, Vol. 102, No. 5, pp. 1716-1722
HEMOSTASIS, THROMBOSIS, AND VASCULAR BIOLOGY
Granule stores from cellubrevin/VAMP-3 null mouse platelets exhibit normal stimulus-induced release
Todd D. Schraw,
Tara W. Rutledge,
Garland L. Crawford,
Audrey M. Bernstein,
Amanda L. Kalen,
Jeffery E. Pessin, and
Sidney W. Whiteheart
From the Department of Molecular and Cellular Biochemistry, University of
Kentucky College of Medicine, Lexington; and Department of Physiology and
Biophysics, University of Iowa, Iowa City.
It is widely accepted that the platelet release reaction is mediated by
heterotrimeric complexes of integral membrane proteins known as SNAREs (SNAP
receptors). In an effort to define the precise molecular machinery required
for platelet exocytosis, we have analyzed platelets from cellubrevin/VAMP-3
knockout mice. Cellubrevin/VAMP-3 has been proposed to be a critical v-SNARE
for human platelet exocytosis; however, data reported here suggest that it is
not required for platelet function. Upon stimulation with increasing
concentrations of thrombin, collagen, or with thrombin for increasing time
there were no differences in secretion of [3H]-5HT (dense core
granules), platelet factor IV (alpha granules), or hexosaminidase (lysosomes)
between null and wild-type platelets. There were no gross differences in
bleeding times nor in agonist-induced aggregation measured in platelet-rich
plasma or with washed platelets. Western blotting of wild-type, heterozygous,
and null platelets confirmed the lack of cellubrevin/VAMP-3 in nulls and
showed that most elements of the secretion machinery are expressed at similar
levels. While the secretory machinery in mice was similar to humans, mice did
express apparently higher levels of synaptobrevin/VAMP-2. These data show that
the v-SNARE, cellubrevin/VAMP-3 is not a requirement for the platelet release
reaction in mice.
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