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1. Muscle fibres developing during limb regeneration were examined for responsiveness to acetylcholine (ACh) applied iontophoretically along the membrane. 2. Fibres which were uninnervated as well as those with non-transmitting synapses had high over-all sensitivities, with only minor variations along their length. 3. Functionally innervated fibres in which depolarization did not yet elicit action potentials had high over-all sensitivities, even when the synaptic potentials ahd amplitudes of 40-50 mV. In these the membrane in the vicinity of synapses tended to have sensitivities above the background level. 4. Upon the appearance of action potentials, several weeks after fibre innervation, the responsiveness to ACh began to decline in synapse free regions of the membrane. In mature muscle the sensitivity to ACh is restricted to sites of synaptic contact.