Acceleration of desensitization by agonist pre-treatment in the snake.

1. The kinetics of carbachol-induced desensitization have been studied in snake twitch-muscle fibres maintained in an isotonic potassium propionate solution and voltage clamped to +50 mV. 2. Microperfusion of carbachol (162-756 microM) induces a transient outward current which peaks within a few seconds and then slowly decays towards the base line. The time course of current decay estimates the time course of desensitization onset. 3. Brief exposure (30 s) to a 'conditioning' concentration of agonist (10.8 microM) accelerates the desensitization onset produced by exposure to higher 'test' concentrations of agonist (162-756 microM). 4. The acceleration of desensitization by pre-treatment with 10.8 microM-carbachol was independent of the duration of exposure between 15 and 60 s. This observation indicated that the mechanism responsible for the alteration in desensitization kinetics by treatment with 10.8 microM-carbachol differed from that responsible for the time-dependent development of desensitization produced in the presence of higher carbachol concentrations. 5. Pre-treatment with the muscarinic agonists, methylcholine and bethanechol, did not accelerate 216 microM-carbachol-induced desensitization, suggesting that the alteration of desensitization kinetics by pre-treatment was specific for nicotinic agonists. 6. The conditioning concentrations of carbachol (5.4-10.8 microM) produced no measurable outward current in muscle fibres voltage clamped to +50 mV. Further, in patch-clamp recordings it was observed that, with these concentrations of carbachol, there was no channel activity in many successful patches voltage clamped to +50 mV and, when present, the frequency of channel activity was very low. These results demonstrated that the alteration in desensitization was not the consequence of significant amounts of either receptor activation or desensitization produced by the conditioning concentration. 7. Exposure to 10.8 microM-carbachol for periods of up to 150 s did not change the amplitude of miniature end-plate currents recorded at end-plates voltage clamped to +50 mV. These results also demonstrated that the acceleration of desensitization by pre-treatment with conditioning concentrations of agonist was not due to partial desensitization occurring during the pre-treatment period. 8. Our results are consistent with the view that there are distinct populations of agonist binding sites on the acetylcholine receptor which separately regulate desensitization and channel opening.(ABSTRACT TRUNCATED AT 400 WORDS)