Calmodulin, phospholipase, and exocytosis. p-Bromophenacyl bromide inhibits but mepacrine stimulates secretion in rat mast cells.

Abstract

p-Bromophenacyl bromide (10-50 microM), a reagent that reacts with and inhibits phospholipase A2, completely suppressed exocytotic responses in rat serosal mast cells elicited by A23187. In contrast, mepacrine (0.01-0.1 mM), an antimalarial drug which has likewise been reported to inhibit phospholipase and additionally calmodulin, did not inhibit exocytosis elicited by concanavalin A (con A) or ionophore A23187. Rather, mepacrine (0.1-0.5 mM) alone increased histamine release up to 70%. Light microscopic observations following ruthenium red staining demonstrated the mepacrine-induced response to be exocytosis. Like the response to compound 48/80, that elicited by mepacrine was maximal within 40 s and was inhibited by phosphatidylserine. In the absence of extracellular calcium, secretion elicited by mepacrine and by con A was inhibited by 87 and 90%, respectively, whereas that elicited by 48/80 was unaffected. Incubation of mast cells in the presence of 2 mM EDTA for 2 h inhibited responses to 48/809 by 76% and nearly abolished those to mepacrine and to con A. 5,8,11,14-Eicosatetraynoic acid (50 and 100 microM) inhibited secretion elicited by con A but not that evoked by mepacrine or polymyxin B. The phenothiazines thioridazine (10-50 microM) and chlorpromazine (25-125 microM) inhibited secretory responses to mepacrine, to 48/80 and to con A. Both phenothiazines inhibited secretion elicited by con A more effectively than that elicited by mepacrine or 48/80. The results indicate that mepacrine, like con A, elicits exocytosis in mast cells by drawing on extracellular sources of calcium, but the early events of secretion initiated by these two secretagogues are pharmacologically distinct.(ABSTRACT TRUNCATED AT 250 WORDS)