Cyclic nucleotides and intracellular-calcium homeostasis in human platelets.
The relationship between agonist-sensitive calcium compartments and those discharged by the Ca(2+)-ATPase inhibitor thapsigargin were studied in human platelets. In this context, calcium mobilization from intracellular pools and manganese influx was investigated in relation to the effect of altered cyclic-nucleotide levels. For maximal calcium release from intracellular stores, thapsigargin, compared to a receptor agonist like thrombin, requires the platelet's self-amplification mechanism, known to generate thromboxane A2. With this lipid mediator formed, thapsigargin released calcium and stimulated manganese influx in a manner similar to thrombin. Blocking the thromboxane receptor by addition of sulotroban (BM13.177) or, alternatively, increasing platelet cAMP or cGMP using prostacyclin or sodium nitroprusside, dramatically reduced the ability of thapsigargin to release calcium from intracellular compartments. The same experimental conditions significantly reduced the rate of manganese influx initiated by thapsigargin compared to thrombin. The experiments indicate that thapsigargin-sensitive compartments play only a minor role in inducing manganese influx compared to the receptor-sensitive compartment. Cyclic nucleotides accelerate the redistribution of an agonist-elevated platelet calcium into the thapsigargin-sensitive compartment, from which calcium can be released by inhibition of the Ca(2+)-ATPase. In human platelets, thapsigargin-induced calcium increase and influx were responsible for only part the calcium release resulting from inhibition of the corresponding ATPase; another part results from the indirect effect of thapsigargin acting via thromboxane-A2-receptor activation. Cyclic nucleotides are therefore an interesting regulatory device which can modify the thapsigargin response by not allowing the self-amplification mechanism of platelets to operate.
Brüne
B
B
Faculty of Biology, University of Konstanz, Federal Republic of Germany.
Ullrich
V
V
D001792:Blood Platelets / Q000378:metabolism*; D002118:Calcium / Q000097:blood*; D002451:Cell Compartmentation; D000242:Cyclic AMP / Q000097:blood*; D006152:Cyclic GMP / Q000097:blood*; D003600:Cytosol / Q000378:metabolism; D011464:Epoprostenol / Q000494:pharmacology; D006706:Homeostasis; D006801:Humans; D066298:In Vitro Techniques; D008345:Manganese / Q000494:pharmacology; D009599:Nitroprusside / Q000494:pharmacology; D013449:Sulfonamides / Q000494:pharmacology; D013729:Terpenes / Q000494:pharmacology; D019284:Thapsigargin; D013917:Thrombin / Q000494:pharmacology
D013449:Sulfonamides; D013729:Terpenes; D009599:Nitroprusside; D008345:Manganese; D019284:Thapsigargin; C040624:sulotroban; D011464:Epoprostenol; D000242:Cyclic AMP; D013917:Thrombin; D006152:Cyclic GMP; D002118:Calcium