Increasing Intracellular calcium of guinea pig ventricular myocytes induced by platelet activating factor through IP3 pathway.

We used Fluo-3/AM to examine the effect of platelet-activating factor on the intracellular Ca2+([Ca2+]i) levels in isolated myocytes of guinea pig ventricle. Myocytes were isolated with Langendorff perfusion technique and were challenged with platelet-activating factor. Addition of platelet-activating factor (1 pM to 10 nM) significantly increased the [Ca2+]i in the presence and absence of extracellular Ca2+. The notion that increases in intracellular Ca2+ induced by platelet-activating factor is the result of stimulation of intracellular Ca2+ pool rather than increasing Ca2+ influx was further supported by the whole cell patch-clamp experiments in which the platelet-activating factor did not alter the activity of L-type of Ca2+ channels (I(Ca-L)). Treatment of myocytes with ryanodine failed to abolish the stimulatory effect of platelet-activating factor on [Ca2+]i. In contrast, inhibition of IP3-sensitive Ca2+ release pool with 2-aminoethoxydiphenyl borate (2-APB) blocked the effect of platelet-activating factor. We conclude that the platelet-activating factor-induced increase in intracellular Ca2+ is mediated by stimulation of IP3 receptor but not by stimulation of I(Ca-L) and ryanodine-sensitive receptor.