Distribution and kinetics of membrane dielectric polarization. II. Frequency domain studies of gating currents.

We have studied the admittance of the membrane of squid giant axon under voltage clamp in the absence of ionic conductances in the range of 0-12 kHz for membrane potentials (V) between --130 and 70 mV. The admittance was measured at various holding potentials (HP) or 155 ms after pulsing from a given holding potential. Standard P/4 procedure was used to study gating currents in the same axons. We found that the membrane capacity Cm (omega) is voltage as well as frequency dependent. For any given V, the voltage-dependent part of the membrane capacitance has a maximum as the frequency approaches zero and requires at least a two-time constant equivalent circuit to be described. When the holding potential is varied, the voltage-dependent capacitance follows a bell-shaped curve with a maximum change of 0.15 muF/cm2 at about --60 mV. With the pulse method, the maximum is at --40 mV for HP = --70 and it shifts to --70 mV for HP = 0. The shift in the maximum of the voltage-dependent capacitance is consistent with the shift in the charge (Q) vs. V curve observed in our experiments with regular P/4 procedure when the HP is varied. Our data can be explained qualitatively by a four-state model for the sodium channel gating, where a charged particle can move within the field and interact with another particle not affected by the field.