The effects of alterations of transmembrane Na + and K + gradients by ionophores (nigericin, monensin) on serotonin transport in human blood platelets

Abstract

Ionophores (monensin, nigericin) capable of transporting both Na + and K + across cell membranes down their concentration gradients reduce the rate and total magnitude of serotonin uptake by platelets. The effect of the ionophores was time dependent, so that inhibition increased progressively until eventually uptake ceased entirely. Nigericin and monensin produced loss of platelet K + and an equivalent molar uptake of Na + thereby abolishing the normal transmembrane Na + and K + gradients. The time course of these ionophore-induced cation shifts at 37° C corresponded to the rate at which inhibition of serotonin transport developed. The ionophores did not affect total ATP concentration of platelets nor the metabolic pool of ATP formed from [ 14C] adenine. Nigericin and monensin released about 80% of platelet 14C and endogenous serotonin over a 30 min period, without release of platelet adenine nucleotides, calcium or β-glucuronidase. The ionophores did not elicit platelet aggregation nor did they prevent maximal aggregation brought about by ADP, collagen or A23187. Replacement of Na + in the medium by K + abolished serotonin uptake but only 10–20% of endogenous serotonin was released. In KCl medium the Na + gradient was initially reversed, but quickly dissipated as Na + reequilibrated with the extracellular fluid. At 37° C the ionophores did not affect either the rate of Na + reequilibration or the efflux of [ 14C] serotonin. Na + reequilibration was slower at 20° C and the ionophores significantly increased platelet Na + loss and strongly inhibited the efflux of [ 14C] serotonin. The data support a mechanism of serotonin transport due to a Na +-dependent carrier-mediated process which need not be directly dependent on metabolic energy, but which does require metabolic energy to maintain normal Na + + K + gradients.