Fiber typing human skeletal muscle with fluorescent immunohistochemistry

Kevin A. Murach, Cory M. Dungan, Kate Kosmac, Thomas B. Voigt, Timothy W. Tourville, Mark S. Miller, Marcas M. Bamman, Charlotte A. Peterson, Michael J. Toth

Research output: Contribution to journalReview articlepeer-review

40 Scopus citations

Abstract

Skeletal muscle myosin heavy chain (MyHC) fiber type composition is a critical determinant of overall muscle function and health. Various approaches interrogate fiber type at the single cell, but the two most commonly utilized are single-muscle fiber sodium dodecyl sulfate-polyacrylamide gel electrophoresis (smfSDS-PAGE) and fluorescent immunohistochemistry (IHC). Although smfSDS-PAGE is generally considered the "gold standard," IHC is more commonly used because of its time-effectiveness and relative ease. Unfortunately, there is lingering inconsistency on how best to accurately and quickly determine fiber type via IHC and an overall misunderstanding regarding pure fiber type proportions, specifically the abundance of fibers exclusively expressing highly glycolytic MyHC IIX in humans. We therefore 1) present information and data showing the low abundance of pure MyHC IIX muscle fibers in healthy human skeletal muscle and 2) leverage this information to provide straightforward protocols that are informed by human biology and employ inexpensive, easily attainable antibodies for the accurate determination of fiber type.

Original languageEnglish
Pages (from-to)1632-1639
Number of pages8
JournalJournal of Applied Physiology
Volume127
Issue number6
DOIs
StatePublished - 2019

Bibliographical note

Funding Information:
This work was supported by grants from the National Institutes of Health to K. A. Murach (AR-071753), C. A. Peterson (AR-060701 and AG-046920), M. S. Miller (AG-031303), T. W. Tourville (AR-066729), and M. J. Toth (AG-033547, HL-077418, AR-069199).

Publisher Copyright:
© 2019 the American Physiological Society.

Keywords

  • Immunohistochemistry
  • MyHC
  • SDS-PAGE

ASJC Scopus subject areas

  • Medicine (all)

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