Sodium and potassium fluxes and membrane potential of human neutrophils: evidence for an electrogenic sodium pump.

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Abstract
Na and K ion contents and fluxes of isolated resting human peripheral polymorphonuclear leukocytes were measured. In cells kept at 37.degree. C, [Na]i was 25 mM and [K]i was 120 mM; both ions were completely exchangeable with extracellular isotopes. One-way Na and K fluxes, measured with 22Na and 42K, were all .apprx. 0.9 meq/l cell water .cntdot. min. Ouabain had no effect on Na influx or K efflux, but inhibited 95 .+-. 7% of Na efflux and 63% of K influx. Cells kept at 0.degree. C gained Na in exchange or K ([Na]i nearly tripled in 3 h); upon rewarming, ouabain-sensitive K influx into such cells was strongly enhanced. External K stimulated Na efflux (Km .apprx. 1.5 mM in 140-mM Na medium). The PNa/PK permeability rtio, estimated from ouabain-insensitive fluxes, was 0.10. Valinomycin (1 .mu.M) approximately doubled PK. Membrane potential (Vm) was estimated using the potentiometric indicator diS-C3(5) [3,3''-dipropylthiadicarbocyanine iodide]; calibration was based on the assumption of constant-field behavior. External K, but not Cl, affected Vm. Ouabain caused a depolarization whose magnitude depended on [Na]i. Na-depleted cells became hyperpolarized when exposed to the neutral exchange carrier monensin; this hyperpolarization was abolished by ouabain. The Na pump of human peripheral neutrophils is electrogenic. The size of the pump-induced hyperpolarization is consistent with the membrane conductance (3.7-4.0 .mu.S[microsiemens]/cm2) computed from the individual K and Na conductances.