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Role of cyclic nucleotides in rapid platelet adhesion to collagen
R Polanowska-Grabowska and AR Gear
Department of Biochemistry, University of Virginia, School of Medicine,
Charlottesville 22908.
Adhesion of human platelets to type I collagen under arterial flow
conditions is extremely fast, being mediated primarily by the alpha 2 beta
1 integrin (glycoprotein Ia/IIa). We have investigated the involvement of
cyclic nucleotides in platelet adhesion to soluble native collagen
immobilized on Sepharose beads using a new microadhesion assay under
arterial flow conditions. To prevent platelet stimulation by thromboxanes
and adenosine diphosphate (ADP), experiments were performed with
aspirin-treated platelets in the presence of ADP-removing enzyme systems
such as creatine phosphate/creatine phosphokinase or apyrase. Rapid
reciprocal changes in platelet adenosine 3'5'-cyclic monophosphate (cAMP)
and guanosine 3'5'-cyclic monophosphate (cGMP) occurred during adhesion.
cAMP levels in adherent platelets were 2.4-fold lower than in effluent
platelets or in static controls, whereas cGMP levels were increased
2.4-fold. These results suggest that contact between platelets and collagen
stimulates guanylate cyclase and inhibits adenylate cyclase. This occurs in
the absence of the platelet release reaction. We also studied short-term
effects of agents that regulate cyclic nucleotide synthesis, prostaglandin
E1 (PGE1) and sodium nitroprusside (SNP). After only 3.8 seconds at 10 to
30 dyne/cm2, PGE1 (10 mumol/L) increased cAMP 16.4- fold, whereas SNP (50
mumol/L) increased cGMP ninefold and caused a 3.2- fold increase in cAMP.
Both PGE1 and SNP rapidly (< 5 seconds) inhibited platelet adhesion in a
dose-dependent manner that was correlated with the increase in cyclic
nucleotides. Our data suggest that cAMP and cGMP play a regulatory role in
the initial phases of platelet adhesion to collagen mediated by the alpha 2
beta 1 integrin receptor.
Volume 83,
Issue 9,
pp. 2508-2515,
05/01/1994
Copyright © 1994 by The American Society of Hematology
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