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Blood, 1 July 2001, Vol. 98, No. 1, pp. 117-124
HEMOSTASIS, THROMBOSIS, AND VASCULAR BIOLOGY
Assembly of a fibronectin matrix by adherent platelets
stimulated by lysophosphatidic acid and other agonists
Olufunke E. Olorundare,
Olivier Peyruchaud,
Ralph M. Albrecht, and
Deane F. Mosher
From the Department of Medicine and the Department of
Animal Health and Biomedical Sciences, University of Wisconsin,
Madison; Department of Pharmacology and Therapeutics, University of
Ilorin, Nigeria; and INSERM U403, Hopital Edouard Herriot, Lyon,
France.
Lysophosphatidic acid (LPA) and sphingosine-1-phosphate (S1P) are
agonists of the endothelial differentiation gene (Edg) family of G-protein-coupled receptors. LPA and S1P are generated by platelet activation during blood coagulation. Both lipids induce assembly of
exogenous fibronectin (FN) by fibroblasts. This study examined whether
LPA and S1P stimulate binding and assembly of fluoresceinated FN
(FITC-FN) by adherent platelets. LPA enhanced deposition of FITC-FN
into linear arrays overlying platelet surfaces and on edges of
platelets adherent to FN or vitronectin (VN). Deposition was greater
when platelets were adherent to FN than to VN and was elicited by
platelet agonists with the following order of potency:
thrombin > LPA = ADP (adenosine diphosphate) > S1P. The linear pattern of FITC-FN deposition was different from the
more diffuse pattern of Alexa-fibrinogen (Alexa-FGN) binding to
adherent platelets. FITC-FN was deposited by adherent platelets that
had dense arrays of cytoskeletal actin when stained with
rhodamine-phalloidin. The 70-kd N-terminal fragment of FN or L8
monoclonal antibody to a self-association domain of FN abolished
deposition of FITC-FN but had no effect on binding of Alexa-FGN.
Conversely, integrilin did not attenuate deposition of FITC-FN but
abolished binding of Alexa-FGN. RGDS (Arg-Gly-Asp-Ser) or antibodies to
 5 1 or IIb3 integrins caused a partial decrease in
LPA-induced deposition of FITC-FN. Correlative electron microscopy with
anti-FITC coupled to gold beads revealed linear arrays on platelet
surfaces associated with less than 20-nm-diameter filaments. These
observations demonstrate that LPA, thrombin, ADP, and S1P induce
adherent platelets to bind and assemble FN and suggest that platelets
may contribute to early deposition of FN matrix after vascular injury.
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