Key result
Margatoxin blocks Kv1.3 currents and depolarizes alveolar macrophages to ~0 mV without affecting phagocytosis.
Why the study?
The role of Kv1.3 potassium channels in human alveolar macrophages and their effect on resting membrane potential and phagocytosis was unclear.
Kv1.3 channels are responsible for setting the resting membrane potential in human alveolar macrophages but are not essential for Fc receptor-mediated phagocytosis.
Kv1.3 sets macrophage resting potential without mediating phagocytosis; leaves open its immunomodulatory role in lung disease models.
Human alveolar macrophages were obtained from macroscopically normal lung tissue obtained at surgical resections, isolated by adherence, and identified by morphology. Whole cell recordings were made from cells 1-3 h in culture, using electrodes containing potassium chloride. From a holding potential of -100 mV, depolarizing pulses to -40 mV or greater activated an outward current. Tail current reversals showed that this current was potassium selective. Margatoxin completely blocked the current; the concentration giving half-maximal block was 160 pM. In current clamp recordings, the resting membrane potential was -34 mV; margatoxin depolarized cells to close to 0 mV. A pure macrophage population was isolated by fluorescence-activated cell sorting, using the phagocytosis of BODIPY-labeled zymosan particles. Reverse transcription-polymerase chain reaction showed that, of 13 voltage-gated K+ (Kv) potassium channels sought, only Kv1.3 mRNA was present. Margatoxin (1 nM) did not affect the percentage of cells showing phagocytosis sorted from the total population. Under these experimental conditions Kv1.3 sets the resting potential of the cells, but it is not required for Fc receptor-mediated phagocytosis.
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Mackenzie et al. (2003) studied Macroscopically normal lung tissue. Margatoxin was evaluated on Potassium current block, resting membrane potential, and phagocytosis. Margatoxin blocked the Kv1.3 potassium current and depolarized human alveolar macrophages to close to 0 mV, but did not affect Fc receptor-mediated phagocytosis.
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