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J Appl Physiol (1985). 2019 Nov 7. doi: 10.1152/japplphysiol.00624.2019. [Epub ahead of print]

Cores of Reproducibility in Physiology (CORP): Fiber Typing Human Skeletal Muscle with Fluorescent Immunohistochemistry.

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Department of Physical Therapy, University of Kentucky, United States.
Department of Rehabilitation Sciences, University of Kentucky, United States.
Center for Muscle Biology, University of Kentucky, United States.
Department of Medicine and Molecular Physiology and Biophysics, University Of Vermont, United States.
Department of Orthopaedics and Rehabilitation, University of Vermont, College of Medicine, United States.
Department of Kinesiology, University of Massachusetts Amherst, Amherst, United States.
Department of Cell, Developmental, and Integrative Biology, University of Alabama at Birmingham, United States.
Department of Rehabilitation Sciences, the Center for Muscle Biology, University of Kentucky, United States.
Departments of Medicine, Molecular Physiology and Biophysics, and Orthopedics and Rehabilitation, College of Medicine, University of Vermont, United States.


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 due to 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.



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