Differential expression of inward and outward potassium currents in the macrophage-like cell line J774.1.

AUTOR(ES)

Gallin, E K

RESUMO

J774.1 cells, a mouse-derived macrophage-like tumour cell line, were voltage clamped using whole-cell patch-clamp techniques. Cells were maintained in suspension cultures and plated at varying times before recording. The average zero-current potential of long-term adherent (greater than 24 h) cells was -77.6 mV. A tenfold increase in [K]o produced a 49 mV shift in zero-current potential. Freshly plated cells (less than 24 h) expressed two voltage-dependent currents: an outward current expressed transiently from 1 to 12 h post-plating and an inward current expressed 2-4 h post-plating which persisted in 100% of long-term adherent cells. Inward current was dependent upon voltage, time and [K]o 1/2, similar to the anomalous rectifier of other tissues. The conductance activated at potentials negative to -50 mV and plateaued at potentials negative to -110 mV. Inactivation was evident at potentials negative to -100 mV. Both the rate and extent of inactivation increased with hyperpolarization. Inward rectification was blocked by external BaCl2 or CsCl. The outward current was time- and voltage-dependent. The instantaneous I/V curves derived from tail experiments reversed at the potassium equilibrium potential (EK). A tenfold change of [K]o shifted the reversal potential 52 mV, indicating that the current was carried by potassium. This conductance activated at potentials positive to -50 mV, plateaued at potentials positive to -10 mV and inactivated completely with an exponential time course at all potentials. At voltages positive to -25 mV the rate of inactivation was independent of voltage. The outward current was blocked by 4-aminopyridine or D600. During the first 10 min after attaining a whole-cell recording, the conductance/voltage relation of the outward current shifted to more negative voltages and peak conductance showed a slight increase; recordings then stabilized. The voltage dependence of the inward current did not shift with time but wash-out of inward current was observed in some cells. The J774.1 cell line can serve as a model for the study of the role of voltage-dependent ionic conductances in macrophages.

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