Acetylcholine receptor site density affects the rising phase of miniature endplate currents.

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The relationship between acetylcholine receptor (AcChoR) site density (sigma) and the rising phase of the miniature endplate current was determined in esterase-inactivated lizard intercostal neuromuscular junctions. The currents were recorded by using a voltage clamp. The receptor site density was determined by electron microscope autoradiography after labeling with 125I-labeled alpha-bungarotoxin in normal endplates and in those partially inactivated with nonradioactive alpha-bungarotoxin. We found that as sigma is decreased the rise time in increased and the amplitude is decreased. These results are compatible with a previously stated "saturating disk" model, which suggests that a quantum of acetylcholine (AcCho) acts on a small postsynaptic area at saturating concentration. We conclude that in the normal neuromuscular junction the most likely number of AcCho molecules needed to open an ion channel is 2, and that the 20--80% rise time of < 100 musec is influenced both by the sigma-dependent factors such as diffusion and binding of AcCho to AcChoR and by the sigma-independent time delays such as the conformation change time to open the ion channels. From our data we calculate the lower limits to the forward rate constant of AcCho binding to AcChoR greater than or equal to 3 X 10(7) M-1 sec-1 and the diffusion constant for AcCho in the cleft greater than or equal to 4 X 10(-6) cm2 sec-1.

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