Role of Internalization in Platelet Activation Induced by Collagen Fibers – Differential Effects of Aspirin, Cytochalasin D, and Prostaglandin E₁

Summary

Washed human platelets were stimulated with fibrillar collagen and platelet aggregation was prevented by non-stirring conditions. In these samples, electron microscopy revealed three fractions of platelets: 1) a majority without contacts to the collagen fibers, 2) with focal contacts to collagen, and 3) a small fraction of platelets with internalized collagen. All platelets had undergone shape change, and exhibited an internal contraction and granule release. However, only those with internalized collagen were completely degranulated. The internalized collagen was found to be in close contact to the contractile sphere in the platelet center, as it was demonstrable with computer assisted 3-D reconstruction from serial sections. Aspirin inhibited neither the adhesion to collagen nor its internalization by internal contraction. Also it did not impair the shape change and degranulation in the platelet fractions that internalized collagen. However, aspirin blocked the shape change and internal contraction of the other platelets and inhibited the [³H]serotonin release. Cytochalasin D 0.1 uM also suppressed the internalization of collagen, the shape change, the formation of a contractile gel, the degranulation, and the [³H]serotonin release in all platelets, whereas the number of platelets that adhered to collagen remained unchanged. The same effects were produced by prostaglandin E₁. If the platelets were stimulated with the TXA₂ mimetic, U46619, cytochalasin D at 0.1 εM had no effect; but at 20 εM it strongly enhanced the degranulation and the [³H]serotonin release, although the platelets remained discoid. It is concluded that collagen triggers a focal activation of an adherent platelet at the site of its initial contact to collagen. This subthreshold activation proceeds to degranulation and TXA₂ formation via a feedback cascade which involves internal contraction, internalization of collagen and multiplication of the collagen membrane contacts.

• References

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