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Ionic strength and pH both influence the binding of neutrotransmitters to the platelet membrane. The present study was designed to examine the effect of these factors on the stimulation of phospholipase C activity (PLC) by 12-O-tetradecanoylphorbol 13-acetate (TPA) and arachidonic acid (AA) in human platelets. TPA stimulated PLC activity in a dose-dependent manner. Both ionic strength and pH influenced the extent of stimulation. Below 150 mM ionic strength, AA stimulated PLC in this system. However, at this ionic strength AA failed to stimulate PLC above the level observed with TPA. The presence of 1 mM Na+ completely blocked the stimulation of PLC by TPA, but did not prevent the potentiation of TPA-induced PLC by AA. The inhibition of TPA stimulation of PLC by Na+ is probably due to the interference of Na+ with the membrane receptor complex, since TPA did not stimulate AA metabolism under these conditions. Above 150 mM ionic strength AA stimulated PLC (and, as a result, the AA response) in a dose-dependent manner. The presence of 25 mM Na+ did not inhibit stimulation of PLC by TPA, but it did inhibit the potentiation of PLC by AA. In addition, AA inhibited PLC induced by 1 microM platelet activating factor (PAF). The effects of ionic strength on PLC and AA responses were not due to the mere addition of Na+ to

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