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Substitution of heavy water (D2O) for H2O slows the Na+ and K+ currents in Myxicola giant axons by 40-50% at 5 degrees C. In contrast, the delays in Na+ K+ activation which can be produced by hyperpolarizing pre-pulses are not affected by solvent substitution. We suggest that in the cascade of transitions leading to the opening of Na+ and K+ channels, D2O affects only the final step that establishes the aqueous pore across the membrane.