Acetylcholine receptor: modification of synaptic gating mechanism after treatment with a disulfide bond reducing agent

Abstract

Reduction of a 'reactive' disulfide bond in the postsyraptic membrane of the frog neuromuscular junction by dithiothreitol (DTT) decreases both the sensitivity of the membrane to applied acetylcholine (ACh) and the amplitude of the single 'shot effect'. Analysis of ACh induced conductance fluctuations in voltage clamped frog endplates indicates that DTT reduces both the amplitude gamma and duration tau of the elementary conductance events. The mean control value of gamma was 18.5-10(-12) omega-1 with no significant dependence on temperature. The mean control values pi were 2.3 msec at 7-9 degrees C and 0.94 msec at 20-22 degrees C. At 7-9 degrees C 1m7 DTT (20-50 min after application) reduced gamma to 61% of the control value and at 20-22 degrees C to 39%, while tau was reduced to 70% at both temperature ranges. The dose-response curve for iontophoretically applied ACh indicates that neither the total number of ionic channels nor the cooperativity within the receptors are changed. However, the affinity of ACh for the receptor sites was reduced. All effects of DTT were fully reversed by the oxidizing agent 5,5'-dithie-bis-(2-nitro-benzoic acid) (DTNB).