Effect of phorbol 12-myristate 13-acetate on collagen-induced signal transduction in rabbit platelet.
Investigations were made on the inhibitory effect of phorbol 12-myristate 13-acetate (PMA), a powerful activator on protein kinase C, on collagen-induced signal transduction in washed rabbit platelets. Upon activation of the platelets with a low-dose of collagen (5 micrograms/ml), which was suppressed by 10 microM indomethacin, pretreatment of the platelets with 2 nM PMA caused prolongation of lag phase (2 min) before the onsets of the aggregation and ATP secretion as compared with PMA-untreated platelets (30 sec). Under this condition, appearance of the cell responses including the phosphatidic acid formation, thromboxane (Tx) generation and Ca2+-influx was similarly retarded for 2-3 min, whereas arachidonic acid liberation from the membrane phospholipids was not significantly affected by the PMA pretreatment. After such lag phase, every response appeared rapidly and reached almost the control value (without PMA). Upon activation of the same platelets with a high-dose of collagen (50 micrograms/ml), which was only half suppressible by indomethacin, PMA in the presence of indomethacin almost completely suppressed the phosphatidic acid formation as well as the aggregation and ATP secretion. Thus, our results suggest that collagen-platelet interaction may elicit direct activation of phospholipase A2 and C, and that the latter enzyme activation may be regulated by a negative effect of protein kinase C. However, the phospholipase A2 activation may be regulated by a mechanism independent of such effect. In PMA-pretreated platelets in response to a low-dose of collagen, the prolonged lag phase for aggregation appears to be due to impaired conversion of liberated arachidonic acid to TxA2.
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D016718:Arachidonic Acid; D001095:Arachidonic Acids / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D002450:Cell Communication / Q000187:drug effects*; D003094:Collagen / Q000494:pharmacology*; D066298:In Vitro Techniques; D050356:Lipid Metabolism; D010712:Phosphatidic Acids / Q000096:biosynthesis; D010974:Platelet Aggregation / Q000187:drug effects*; D011817:Rabbits; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*; D013929:Thromboxane B2 / Q000096:biosynthesis
D001095:Arachidonic Acids; D010712:Phosphatidic Acids; D016718:Arachidonic Acid; D013929:Thromboxane B2; D000255:Adenosine Triphosphate; D003094:Collagen; D013755:Tetradecanoylphorbol Acetate; D002118:Calcium