Selective modification of sodium channel gating in lobster axons by 2, 4, 6-trinitrophenol: Evidence for two inactivation mechanisms.

Trinitrophernol (TNP) selectively alters the sodium conductance system of lobster giant axons as measured in current clamp and voltage clamp experiments using the double sucrose gap technique. TNP has no measurable effect on potassium currents but reversibly prolongs the time-course of sodium currents during maintained depolarizations over the full voltage range of observable currents. Action potential durations are increased also. Tm of the Hodgkin-Huxley model is not markedly altered during activation of the sodium conductance but is prolonged during removal of activation by repolarization, as observed in sodium tail experiments. The sodium inactivation versus voltage curve is shifted in the hyperpolarizing direction as is the inactivation time constant curve, measured with conditioning voltage steps. This shift speeds the kinetics of inactivation over part of the same voltage range in which sodium currents are prolonged, a contradiction incompatible with the Hodgkin-Huxley model. These results are interpreted as support for a hypothesis of two inactivation processes, one proceeding directly from the resting state and the other coupled to the active state of sodium conductance.

Studies on Calcium and Sodium in Uterine Smooth Muscle Excitation under Current-Clamp and Voltage-Clamp Conditions

The objective of these studies was to define the roles of calcium and sodium in uterine smooth muscle excitation. The double sucrose-gap technique was used for current-clamp and voltage-clamp experiments. It was shown that neither sodium nor calcium alone is capable of supporting excitation in estrogen-dominated uterine smooth muscle. Calcium dependence was explained in part by increased membrane "leakage" current in calcium-free solution and calcium control of the voltage dependence of the early transient conductance. High concentrations of TTX did not affect the magnitude of the peak transient current while La+++, Mn++, and Co++ greatly reduced or abolished it and decreased the steady-state current. From these and other data it was concluded that the regenerative mechanism in uterine smooth muscle has the functional characteristics of a single transient conductance channel whose activation requires the presence of both sodium and calcium. Insensitivity to TTX indicates that the molecular structure of the channel is unlike that in certain sodium-dependent systems, while the effects of La+++, Mn++, Co++, and Ca++ reveal a similar dependence of conductances on extracellular polyvalent cations.

Temperature Dependence of Accommodation and Excitation in Space-Clamped Axons

Accommodation and excitation in space-clamped squid axons were studied with the double sucrose gap technique, using linear current ramps, short (50 µsec) square wave pulses, and rheobasic square wave pulses as stimuli. The temperature was varied from 5° to 35°C. Experimental results showed a Q10 for accommodation which was 44% higher than that for excitation. Yet calculations on the basis of the Hodgkin-Huxley equations predict equal Q10's for excitation and accommodation. Although the Hodgkin-Huxley equations are spectacularly successful for so many nerve phenomena, the differences between calculations of accommodation and these experiments, which were designed to test the equations, show that the equations need modification in this area.