Voltage-clamp study of the conductance activated at fertilization in the starfish egg.

Abstract

Ionic currents underlying the fertilization potential of the egg of the starfish Mediaster aequalis were studied using a two-micro-electrode voltage clamp. Mature eggs were fertilized in vitro under voltage-clamp conditions. The fertilization current, here termed IF, was induced by adding sperm to sea water bathing the egg. At a holding potential of -70 mV, IF was inward. It reached a peak within 2-4 min and then decayed over the next approximately 20 min with a rate which depended on the holding membrane potential. Instantaneous current-voltage relations measured at different times during IF were approximately linear and reversed at a potential of +6.0 +/- 5.8 mV (mean +/- S.D., n = 11). Membrane chord conductance was highest at the peak of inward current and the declining phase of IF was due to a decrease in conductance towards the pre-fertilization level. When the membrane potential was rapidly stepped to levels more positive than about -45 mV, the conductance underlying IF decreased in a manner which depended on both membrane potential and time. The fertilization-specific conductance showed a sigmoidal activation curve between -50 and +10 mV with a half-activation level of -25 mV. Analysis of the steady-state voltage dependence indicated that at the peak of the fertilization potential (+10 to +15 mV) only 4-5% of the total available channels would be open. Current relaxations followed first-order kinetics and the relaxation time constant depended upon the membrane potential during the voltage pulse. The relation between the time constant and voltage was bell-shaped, decreasing at potentials more negative than -40 and more positive than 0 mV. Both the steady-state conductance-voltage relation and the kinetics of the current relaxations were consistent with a simple two-state gating model in which the probability of a channel being open is determined by a single gating particle with an effective valency of -1.7 moving through the entire membrane field. The shifts in reversal potential with changes in external Na (at 10 mM-external K) were analysed using the constant field expression, which gave a relative permeability of Na to K of approximately 0.6.(ABSTRACT TRUNCATED AT 400 WORDS)