Electronic Measurement of the Intracellular Concentration and Net Flux of Sodium in the Squid Axon

Abstract

A unique, rapid, and non-destructive determination of the intracellular sodium concentration of a squid axon may be provided by the "voltage clamp" technique, in which the potential across the axon membrane is under electronic control. The potential at which the early component of ionic current reverses following a membrane potential step was used as an index of the intracellular sodium concentration. Several types of experiments were used to test the applicability of this method for measurement of intracellular sodium and its net flux. The concentration was found to increase from 38 mM for a fresh axon to 50 mM in about an hour. From this change, the net flux for a fresh resting axon was estimated to be 40 pmoles/cm² sec. Rapid stimulation of an unclamped axon produced a marked increase in the rate of sodium accumulation. Rapid pulsing of the membrane in a voltage clamp to potentials more positive than the sodium potential moved sodium out fast enough to produce a definite decrease in internal concentration. The agreement between the results with this method and those with more direct methods is quite satisfactory. An attractive feature of this method of intracellular sodium determination is that the physiological function of the axon is maintained and other measurements may be made concurrently.