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J Biol Chem, Vol. 273, Issue 8, 4338-4344, February 20, 1998

Platelet Adhesion to Native Type I Collagen Fibrils
ROLE OF GPVI IN DIVALENT CATION-DEPENDENT AND -INDEPENDENT ADHESION AND THROMBOXANE A₂ GENERATION

Takashi Nakamura, G. A. Jamieson^§, Minoru Okuma^¶, Jun-ichi Kambayashi, and Narendra N. Tandon

From the  Otsuka America Pharmaceutical Inc., Rockville, Maryland 20850, the ^§ American Red Cross, Rockville, Maryland 20855, and ^¶ Takashima General Hospital, Shiga, 520-11 Japan

Three glycoproteins (GPs), namely GPIa-IIa, GPVI, and GPIV, have been recently implicated in platelet-collagen adhesive interactions. We have employed antibodies to these GPs to investigate further their role in platelet adhesion to immobilized monomeric and polymeric fibrillar collagen under static conditions in the presence and the absence of Mg²⁺. In the presence of Mg²⁺, each antibody inhibited platelet adhesion to fibrillar collagen from 70 to 85%, especially during the early phase (<15 min), but the inhibitory effects diminished dramatically to 25% or less by 60 min. Combination of anti-GPVI with anti-GPIa-IIa antibodies completely inhibited platelet adhesion at 60 min. Anti-GPIV and anti-GPIa-IIa or anti-GPVI antibodies in combinations were more effective in inhibiting adhesion than was anti-GPIa-IIa or anti-GPVI alone. In the absence of Mg²⁺, anti-GPVI completely inhibited adhesion at 60 min, while anti-GPIV antibody inhibited adhesion by about 50% and minimal effects were seen with anti-GPIa-IIa, suggesting that GPIa-IIa does not play a significant role in the divalent cation-independent platelet adhesion to immobilized fibrillar collagen. Under either divalent cation-dependent or -independent conditions, platelets adhered to fibrillar collagen were able to secrete contents of both -granules and dense granules and generate thromboxane A₂ (TXA₂), but platelets adhering to acid soluble monomeric collagen neither secreted their granular contents nor generated TXA₂. Although anti-GPVI antibodies were not able to inhibit Mg²⁺-dependent adhesion, they completely inhibited TXA₂ generation under both divalent cation-dependent and -independent conditions. With the other antibodies, TXA₂ generation corresponded with the amount of adhesion observed. These results suggest that GPVI is directly associated with the TXA₂ generating system during platelet-collagen interaction.

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Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.

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