Novel staining pattern of skeletal muscle M-lines upon incubation with antibodies against MM-creatine kinase

T. Wallimann, Thomas C Doetschman, H. M. Eppenberger

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Abstract

Incubation of chicken skeletal muscle fibers with an excess of anti-M-creatine kinase (CK) immunoglobulin G and an excess of anti-M-CK Fab fragments leads to heavy decoration of the M-line (Wallimann, T., D.C. Turner, and H.M. Eppenberger, 1977, J. Cell Biol. 75:297-317) and to removal of the electron-dense M-line structure (Walliman, T., G.W. Pelloni, D.C. Turner, and H.M. Eppenberger, 1978, Proc. Natl. Acad. Sci. USA., 75:4296-4300), respectively. On the other hand, incubation with concentrations of monovalent anti-M-CK Fab did not extract but rather decorated the M-line, giving rise to a distinct two-line staining pattern. A similar double-line staining pattern, although less pronounced, was also observed within the M-line of paraformaldehyde-prefixed myogenic cells, which after permeabilization were incubated with low concentrations of divalent anti-M-CK antibody. In both cases, the two decorated lines appearing in the middle of the A-band were spaced axially 42-44 nm apart and correspond most likely to the two M4 and M4' m-bridge rows described by Sjostrom and Squire (1977, J. Mol. Biol., 109:49-68; 1977, J. Microscopy., 111:329-278). It is concluded that the muscle-specific form of creatine kinase, MM-CK, contributes mainly to the electron density of these M4 and M4' m-bridges within the M-line structure. This specific labeling pattern is a further demonstration that CK is an integral part of the M-line.

Original languageEnglish (US)
Pages (from-to)1772-1779
Number of pages8
JournalJournal of Cell Biology
Volume96
Issue number6
StatePublished - 1983
Externally publishedYes

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MM Form Creatine Kinase
Skeletal Muscle
Staining and Labeling
Antibodies
Electrons
Immunoglobulin Fab Fragments
Skeletal Muscle Fibers
Creatine Kinase
Microscopy
Chickens
Immunoglobulin G
Muscles

ASJC Scopus subject areas

  • Cell Biology

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Novel staining pattern of skeletal muscle M-lines upon incubation with antibodies against MM-creatine kinase. / Wallimann, T.; Doetschman, Thomas C; Eppenberger, H. M.

In: Journal of Cell Biology, Vol. 96, No. 6, 1983, p. 1772-1779.

Research output: Contribution to journalArticle

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abstract = "Incubation of chicken skeletal muscle fibers with an excess of anti-M-creatine kinase (CK) immunoglobulin G and an excess of anti-M-CK Fab fragments leads to heavy decoration of the M-line (Wallimann, T., D.C. Turner, and H.M. Eppenberger, 1977, J. Cell Biol. 75:297-317) and to removal of the electron-dense M-line structure (Walliman, T., G.W. Pelloni, D.C. Turner, and H.M. Eppenberger, 1978, Proc. Natl. Acad. Sci. USA., 75:4296-4300), respectively. On the other hand, incubation with concentrations of monovalent anti-M-CK Fab did not extract but rather decorated the M-line, giving rise to a distinct two-line staining pattern. A similar double-line staining pattern, although less pronounced, was also observed within the M-line of paraformaldehyde-prefixed myogenic cells, which after permeabilization were incubated with low concentrations of divalent anti-M-CK antibody. In both cases, the two decorated lines appearing in the middle of the A-band were spaced axially 42-44 nm apart and correspond most likely to the two M4 and M4' m-bridge rows described by Sjostrom and Squire (1977, J. Mol. Biol., 109:49-68; 1977, J. Microscopy., 111:329-278). It is concluded that the muscle-specific form of creatine kinase, MM-CK, contributes mainly to the electron density of these M4 and M4' m-bridges within the M-line structure. This specific labeling pattern is a further demonstration that CK is an integral part of the M-line.",
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