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The Journal of General Physiology, Vol 60, 248-262, Copyright © 1972 by The Rockefeller University Press


ARTICLE

Acetylcholine Receptors

Distribution and extrajunctional density in rat diaphragm after denervation correlated with acetylcholine sensitivity



H. Criss Hartzell 1 and Douglas M. Fambrough 1

1 From the Department of Biology, The Johns Hopkins University, Baltimore, Maryland 21218 and the Department of Embryology, Carnegie Institution of Washington, Baltimore, Maryland 21210

Using 125iodine-labeled alpha-bungarotoxin (alpha-BGT-125I) and quantitative radioautography, we have studied the time-course of the change in acetylcholine (ACh) receptor distribution and density occurring in rat diaphragm after denervation. In innervated fibers, ACh receptors are localized at the neuromuscular junction and the extrajunctional receptor density is less than five receptors per square micrometer. The extrajunctional receptor density begins to increase between 2 and 3 days after denervation and increases approximately linearly to 1695 receptors/µm2 at 14 days, subsequently decreasing to 529 receptors/µm2 at 45 days. We have isolated plasma membranes from rat leg muscles at various times after denervation and find that the change in concentration of ACh receptors in the membranes measured by alpha-BGT-125I binding and scintillation counting follows a time-course similar to the change in ACh receptor density measured radioautographically. Furthermore, we have correlated extrajunctional ACh receptor density measured by radioautography with extrajunctional ACh sensitivity measured by iontophoretic application of ACh and intracellular recording and find that the log of ACh receptor density is related to 0.53 times the log of ACh sensitivity. These results are discussed in terms of the electrophysiological experiments on the ACh receptor and the recent, more biochemical approaches to the study of ACh receptor control and function.

Submitted on March 10, 1972


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