Role of internalization in platelet activation induced by collagen fibers--differential effects of aspirin, cytochalasin D, and prostaglandin E1.
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 microM 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 microM had no effect; but at 20 microM 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.(ABSTRACT TRUNCATED AT 250 WORDS)
Ruf
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Institute for Clinical Chemistry, Klinikum Mannheim University of Heidelberg, FRG.
Patscheke
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Morgenstern
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D000527:Alprostadil / Q000494:pharmacology*; D001241:Aspirin / Q000494:pharmacology*; D001699:Biometry; D003094:Collagen / Q000037:antagonists & inhibitors*; D015638:Cytochalasin D / Q000494:pharmacology*; D006801:Humans; D007091:Image Processing, Computer-Assisted; D008854:Microscopy, Electron; D015539:Platelet Activation / Q000187:drug effects*; D011450:Prostaglandin Endoperoxides, Synthetic / Q000494:pharmacology; D012701:Serotonin / Q000097:blood; D013928:Thromboxane A2 / Q000502:physiology
D011450:Prostaglandin Endoperoxides, Synthetic; D015638:Cytochalasin D; D012701:Serotonin; D013928:Thromboxane A2; D003094:Collagen; D000527:Alprostadil; D001241:Aspirin