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JOURNAL ARTICLE| 01 November 1974

Acetylcholine Receptors of Cultured Muscle Cells Demonstrated with Ferritin-α-Bungarotoxin Conjugates

B. T. HOURANI,

B. T. HOURANI

Laboratory of Experimental Pathology, National Institute of Arthritis, Metabolism, and Digestive Diseases, and Behavioural Biology Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20014, U.S.A.; Present address: University Hospital, San Diego, California, U.S.A.

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B. F. TORAIN,

B. F. TORAIN

Laboratory of Experimental Pathology, National Institute of Arthritis, Metabolism, and Digestive Diseases, and Behavioural Biology Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20014, U.S.A.

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M. P. HENKART,

M. P. HENKART

Laboratory of Experimental Pathology, National Institute of Arthritis, Metabolism, and Digestive Diseases, and Behavioural Biology Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20014, U.S.A.

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R. L. CARTER,

R. L. CARTER

Laboratory of Experimental Pathology, National Institute of Arthritis, Metabolism, and Digestive Diseases, and Behavioural Biology Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20014, U.S.A.

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V. T. MARCHESI,

V. T. MARCHESI

Laboratory of Experimental Pathology, National Institute of Arthritis, Metabolism, and Digestive Diseases, and Behavioural Biology Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20014, U.S.A.; Present address: Yale University School of Medicine, New Haven, Connecticut, U.S.A.

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G. D. FISCHBACH

G. D. FISCHBACH

Laboratory of Experimental Pathology, National Institute of Arthritis, Metabolism, and Digestive Diseases, and Behavioural Biology Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20014, U.S.A.; Present address: Harvard Medical School, Boston, Massachusetts, U.S.A.

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Author and article information

B. T. HOURANI

Laboratory of Experimental Pathology, National Institute of Arthritis, Metabolism, and Digestive Diseases, and Behavioural Biology Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20014, U.S.A.; Present address: University Hospital, San Diego, California, U.S.A.

B. F. TORAIN

Laboratory of Experimental Pathology, National Institute of Arthritis, Metabolism, and Digestive Diseases, and Behavioural Biology Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20014, U.S.A.

M. P. HENKART

Laboratory of Experimental Pathology, National Institute of Arthritis, Metabolism, and Digestive Diseases, and Behavioural Biology Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20014, U.S.A.

R. L. CARTER

Laboratory of Experimental Pathology, National Institute of Arthritis, Metabolism, and Digestive Diseases, and Behavioural Biology Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20014, U.S.A.

V. T. MARCHESI

Laboratory of Experimental Pathology, National Institute of Arthritis, Metabolism, and Digestive Diseases, and Behavioural Biology Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20014, U.S.A.; Present address: Yale University School of Medicine, New Haven, Connecticut, U.S.A.

G. D. FISCHBACH

Laboratory of Experimental Pathology, National Institute of Arthritis, Metabolism, and Digestive Diseases, and Behavioural Biology Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20014, U.S.A.; Present address: Harvard Medical School, Boston, Massachusetts, U.S.A.

Received: 29 Mar 1974

Online ISSN: 1477-9137

Print ISSN: 0021-9533

Copyright © 1974 The Company of Biologists Ltd.

1974

J Cell Sci (1974) 16 (2): 473–479.

https://doi.org/10.1242/jcs.16.2.473

α-Bungarotoxin-ferritin conjugates were used to visualize by freeze-fracture and thin-section electron microscopy toxin-binding sites, presumably acetylcholine (ACh) receptors, in membranes of muscle cells grown in tissue culture. Toxin conjugated to ferritin by a glutaraldehyde reaction and purified by column chromatography in a buffer of high ionic strength remains active in blocking the effect of iontophoretically applied ACh. The potency of the conjugates was decreased 5-10 times compared to native α-bungarotoxin. Toxin-ferritin conjugates were identified in small clusters which were not uniformly distributed over the surface membrane. Binding was inhibited by preincubation in D-tubocurare or unconjugated toxin. The relation of the clusters to the non-uniform distribution of ACh sensitivity and α-bungarotoxin binding on cultured muscle fibres is discussed.

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Copyright © 1974 The Company of Biologists Ltd.

1974

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