Kinetic analysis of acetylcholine-induced chloride current in isolated Aplysia neurones.

Abstract

(1) Kinetics of activation and desensitization phases of the ACh-induced chloride current (ICl) were studied in isolated single neurones of Aplysia kurodai, using the 'concentration clamp' technique which combines internal perfusion and rapid exchange of the external solution within a few milliseconds (2) The dose-response curve for the peak ICl gave a dissociation constant of 6.7 X 10(-6) M and a Hill coefficient of 1.7. (3) The current-voltage relationship was linear in the voltage range examined (-70 to +30 mV). The reversal potential (EACh) was -7.1 +/- 1.8 mV (n = 14). The value was close to the calculated equilibrium potential for chloride ions (ECl). (4) The activation phase of the ICl was single exponential and the desensitization proceeded double exponentially to a steady state level. The time constants of both phases decreased with increasing concentrations of ACh but showed no potential dependency. The desensitizing component of the ICl was generated by activation of a single population of the receptor-channel complex. (5) The recovery from desensitization of the ICl induced by 6 X 10(-6) M ACh proceeded double exponentially, with time constants of 6.5 and 43 s at a holding potential of -30 mV. (6) Noise analysis performed on the steady state of ICl induced by low concentrations of ACh (3 X 10(-7) M to 3 X 10(-6) M) showed that the steady ICl was due to activation of a single population of the receptor-channel complex with a single channel conductance of 23.3 +/- 4.3 pS (n = 9).(ABSTRACT TRUNCATED AT 250 WORDS)