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Stimulation of neutrophil leucocytes with chemotactic factors is known to result in membrane permeability changes, as evidenced by fluxes of Na+ and K+ across the cell membrane together with an increased uptake of Ca2+ from the medium. These fluxes have been implicated in the transduction mechanisms of various responses, including locomotion and subsequent chemotaxis. We have previously reported that exposure of unstimulated, round neutrophils held in suspension, to the chemotactic peptide fMet-Leu-Phe confers morphological polarity on the neutrophils by stimulating waves of contraction, which are also intimately connected with locomotion on an appropriate substratum. As the acquisition of polarity is the important first step in the chemotactic response we have investigated the effects of modifying the external ionic environment and of various ion channel blockers on the polarizing response of neutrophils held in suspension. Removal and chelation of both Ca2+ and Mg2+ from the external medium did not inhibit the acquisition of polarity and a variety of inorganic Ca2+ channel blockers together with the organic Ca2+ antagonists, verapamil and D600, were ineffective in inhibiting the response. Replacement of Na+ in the external medium with choline inhibited the polarizing response completely but tetrodotoxin, which blocks fast Na+ channels, and amiloride, which inhibits Na+/K+ exchange, had no effect. Inhibition of the Na+/K+-ATPase with ouabain and also tetraethylammonium ions, which block potassium channels, had no inhibiting effect on polarization. These results indicate that while Ca2+ and Mg2+ are not required in the external medium, Na+ is essential, and therefore Na+/K+ fluxes across the cell membrane play a role in initiating locomotion.

Signal transduction in human neutrophil leucocytes: Effects of external Na+ and Ca2+ on cell polarity