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1. Micro-electrodes were used to record membrane potential and associated noise at the end-plate region of cat tenuissimus muscle (37 degrees C), during applications of acetylcholine (ACh) in continuously flowing Krebs solution containing eserine and tetrodotoxin. 2. Densensitization was assessed from the frequency of channel opening calculated from the noise variance. 3. At higher concentrations of ACh (10-50 microM), desensitization occurred with an exponential fall to a plateau. 4. At low concentrations of ACh (1-2 microM) only slight desensitization occurred and at a much lower rate. Frequency of channel opening decreased at the rate of 0.045 +/- 0.024 min-1. Maximum frequency was (33 +/- 9) X 10(7)/sec while maximum depolarization was 20.5 +/- 1.4 mV (n = 11 cats). Depolarization was well maintained. 5. This slow rate of desensitization at low concentrations of ACh was confirmed in experiments where voltage clamped current, its associated noise, and miniature end-plate current amplitude were measured. 6. At low concentrations of ACh (1-2 microM) in the presence of eserine there was sustained block in neuromuscular transmission when twitch tension was measured. 8. It is concluded that the mechanism of neuromuscular block by ACh at around 1 microM concentration is by depolarization itself, not desensitization.