Summary. Background: Recent studies have shown that platelet adhesion and subsequent aggregation can occur in vivo in the absence of the two principal platelets adhesive ligands, von Willebrand factor and fibrinogen. These results highlight a possible role for fibronectin in supporting thrombus formation. Objective and methods: To evaluate the platelet integrins and subsequent activation pathways associated with fibronectin-dependent platelet adhesion utilizing both human and murine platelets. Results: Platelets can adhere to fibronectin via the integrin αIIbβ3, leading to formation of lamellipodia. This is mediated through an interaction with the tenth type III domain in fibronectin. Spreading on fibronectin promotes αIIbβ3-mediated Ca2+ mobilization and tyrosine phosphorylation of focal adhesion kinase and phospholipase C γ2. In contrast, studies with blocking antibodies and mice demonstrate that α5β1 and αvβ3 support adhesion and promote formation of filopodia but not lamellipodia or tyrosine phosphorylation of these proteins. Further, neither α5β1 nor αvβ3 is able to induce formation of lamellipodia in the presence of platelets agonists, such as collagen-related-peptide (CRP). Conclusions: These observations demonstrate that integrins α5β1 and αvβ3 support platelet adhesion and the generation of filopodia but that, in contrast to the integrin αIIbβ3, are unable to promote formation of lamellipodia.