Damaged muscle fibers might masquerade as hybrid fibers – a cautionary note on immunophenotyping mouse muscle with mouse monoclonal antibodies

Submitted: 5 February 2018
Accepted: 6 July 2018
Published: 24 July 2018
Abstract Views: 924
PDF: 468
Supplementary: 232
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Authors

We report that, labeling mouse muscle tissue, with mouse monoclonal antibodies specific to slow or fast myosin heavy chain (sMyHC and fMyHC, respectively), can lead to artefactual labeling of damaged muscle fibers, as hybrid fibers (sMyHC+ and fMyHC+).  We demonstrate that such erroneous immunophenotyping of muscle may be avoided, by performing colabeling or serial-section-labeling, to identify damaged fibers. The quadriceps femoris muscle group (QF) in 7-month-old, male, C57BL/6J mice had: 1.21 ± 0.21%, 98.34 ± 1.06%, 0.07 ± 0.01%, and 0.53 ± 0.85% fibers, that were, sMyHC+, fMyHC+, hybrid, and damaged, respectively.  All fibers in the tibialis anterior muscle (TA) of 3-month-old, male, C57BL/6J mice were fMyHC+; and at 3 days after injurious eccentric contractions, there was no fiber-type shift, but ~ 18% fibers were damaged.

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Supporting Agencies

Alliance for Regenerative Rehabilitation Research and Training (AR3T), NIH 1R03HD091648-01 from NICHD, Jain Foundation Inc., Faculty Research Awards Program (FRAP) grant from EACPHS Wayne State University, faculty startup package from Wayne State University, to JAR, AR3T is supported by the Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD), National Institute of Neurological Disorders and Stroke (NINDS), and National Institute of Biomedical Imaging and Bioengineering (NIBIB) of the National Institutes of Health under Award Number P2CHD086843
Morium Begam, Wayne State University, Department of Health Care Sciences

Department of Health Care Sciences

Physical Therapy Program

Research Assistant

Joseph A. Roche, Wayne State University, Department of Health Care Sciences

Department of Health Care Sciences

Physical Therapy Program

Assistant Professor

How to Cite

Begam, M., & Roche, J. A. (2018). Damaged muscle fibers might masquerade as hybrid fibers – a cautionary note on immunophenotyping mouse muscle with mouse monoclonal antibodies. European Journal of Histochemistry, 62(3). https://doi.org/10.4081/ejh.2018.2896