Opening rate of acetylcholine receptor channels

The nicotinic acetylcholine (ACh) receptor is responsible for rapid conversion of chemical signals to electrical signals at the neuromuscular junction. Because the receptor and its ion channel are components of a single transmembrane protein, the time between ACh binding and channel opening can be minimized. To determine just how quickly the channel opens, we made rapid (100-400 microseconds) applications of 0.1-10 mM ACh to outside-out, multichannel membrane patches from BC3H-1 cells, while measuring the onset of current flow through the channels at 11 degrees C. Onset time is steeply dependent upon ACh concentration when channel activation is limited by binding of ACh (0.1-1 mM). At +50 mV, the 20-80% onset time reaches a plateau near 110 microseconds above 5 mM ACh as channel opening becomes rate limiting. Thus, we calculate the opening rate, beta = 12/ms, without reference to specific channel activation schemes. At -50 mV, the combination of a rapid, voltage-dependent block of channels by ACh with a finite solution exchange time distorts onset. To determine opening rate at -50 mV, we determine the kinetic parameters of block from "steady-state" current and noise analyses, assume a sequential model of channel activation/block, and numerically simulate current responses to rapid perfusion of ACh. Using this approach, we find beta = 15/ms. In contrast to the channel closing rate, the opening rate is relatively insensitive to voltage.