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Published November 1, 1979 | Published
Journal Article Open

Demonstration of the transmembrane nature of the acetylcholine receptor by labeling with anti-receptor antibodies


Antibodies raised in rabbits to Triton-solubilized, purified acetylcholine receptor from Torpedo californica were used to immunospecifically label intact T. californica electroplaque membrane vesicles attached to cover slips and oriented with the extracellular face of the synaptic membrane facing outward. Hemocyanin conjugated to Protein A was then used as a marker, making the antibody binding visible at the electron microscope level. Parallel labeling experiments were performed on vesicles attached to cover slips and sheared by sonication, leaving their cytoplasmic faces fully exposed to the labeling solution. While differences in antibody populations among different rabbits were observed, antigenic determinants of the receptor were present on both faces of the membrane, with those on the extracellular side more numerous than those on the cytoplasmic side, demonstrating the transmembrane nature of the receptor molecule.

Additional Information

© 1979 Rockefeller University Press. Received for publication 22 March 1979, and in revised form 29 June 1979. We are grateful to Dr. Marcia Miller for many helpful discussions. Anti-AcChR antibodies were the generous gifts of Dr. Toni Claudio, Cara Hsieh, and Dr. Richard Vandlen. We wish to thank Pat Koen and John Racs for expert technical assistance and Carolyn Sprague and Valerie Purvis for typing the manuscript. This research was supported by U. S. Public Health Service grants NS 10294 and GM 06965, by a grant from the Muscular Dystrophy Association of America, and by a National Institutes of Health predoctoral Training Grant.

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