Platelet-activating factor stimulates a rapid accumulation of inositol (1,4,5)trisphosphate in guinea pig eosinophils: relationship to calcium mobilization and degranulation.

The effect of platelet-activating factor (PAF) on inositol (1,4,5)trisphosphate (Ins[1,4,5]P3) mass, calcium mobilization, and the release of granule enzymes was studied on guinea pig peritoneal eosinophils (EOSs). PAF evoked a concentration-dependent accumulation of Ins(1,4,5)P3 with a drug concentration that elicits 50% of the maximum attainable response (EC50) of 10 nmol/L; the production of this second messenger was maximal at 1 mumol/L of PAF. Kinetic analysis of PAF (1 mumol/L)-induced Ins(1,4,5)P3 accumulation demonstrated it to be transient with a 3.8-fold increase over resting levels observed at 5 seconds. Thereafter, the level of Ins(1,4,5)P3 declined, returning to vehicle-treated levels 60 seconds after PAF challenge. Lyso-PAF, the inactive precursor and metabolite of PAF, was inactive at all concentrations examined. PAF also induced a rapid, concentration-dependent (EC50, 12 nmol/L) rise in the cytosolic-free calcium concentration ([Ca++]i) in fura 2-AM-loaded EOSs that was transient, peaking after the maximum increase in Ins(1,4,5)P3 mass was observed. A highly significant positive correlation was found between the peak increase in Ins(1,4,5)P3 and the peak rise in [Ca++]i. Functionally, PAF evoked a concentration-dependent release of granule constituents from both the small (arylsulfatase B; EC50, 3 nmol/L) and specific (EOS peroxidase; EC50, 2.7 nmol/L) granules that lagged, temporally, behind both Ins(1,4,5)P3 accumulation and the rise in [Ca++]i. Both the biochemical and functional effects of PAF examined in this study were antagonized by WEB 2086 (300 nmol/L), a selective PAF receptor-blocking drug. It is concluded that stimulus (PAF)-response coupling in guinea pig peritoneal EOSs may involve the receptor-mediated formation of Ins(1,4,5)P3 and subsequent release of intracellularly stored Ca++. This sequence of events may link PAF receptor activation to Ca(++)-dependent cellular responses, such as degranulation.