Vanadate and fluoride effects on Na+-K+-Cl- cotransport in squid giant axon

The effects of vanadate and fluoride on the Na+-K+-Cl- cotransporter of the squid giant axon were assessed. In axons not treated with these agents, intracellular dialysis with ATP-depleting fluids caused bumetanide-inhibitable 36Cl influx to fall with a half time of ∼16 min. In the presence of either 40 μM vanadate or 5 mM fluoride, the decay of bumetanide-inhibitable 36Cl influx was significantly slowed; half time for vanadate-treated axons is 45 min and four fluoride-treated axons is 37 min. These agents are not exerting their effects on Na+-K+Cl- cotransport by influencing the rate of ATP depletion of the axon, since they had no effect on the ATP hydrolysis rate of an optic ganglia homogenate. We therefore suggest that these data support the hypothesis that Na+-K+-Cl- cotransport in squid axons is regulated by a phosphorylation-dephosphorylation mechanism and that vanadate and fluoride reduce the rate of dephosphorylation by inhibiting a protein phosphatase