Fiber Typing Human Skeletal Muscle with Fluorescent Immunohistochemistry

Overview

Journal J Appl Physiol (1985)

Publisher American Physiological Society

Date 2019 Nov 8

PMID 31697594

Citations 40

Authors

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

Affiliations

Soon will be listed here.

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 ) present information and data showing the low abundance of pure MyHC IIX muscle fibers in healthy human skeletal muscle and ) 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.

Citing Articles

Human skeletal muscle fiber heterogeneity beyond myosin heavy chains.

Moreno-Justicia R, Van der Stede T, Stocks B, Laitila J, Seaborne R, Van de Loock A Nat Commun. 2025; 16(1):1764.

PMID: 39971958 PMC: 11839989. DOI: 10.1038/s41467-025-56896-6.

Impact of physical activity on physical function, mitochondrial energetics, ROS production, and Ca handling across the adult lifespan in men.

Cefis M, Marcangeli V, Hammad R, Granet J, Leduc-Gaudet J, Gaudreau P Cell Rep Med. 2025; 6(2):101968.

PMID: 39933528 PMC: 11866497. DOI: 10.1016/j.xcrm.2025.101968.

Abnormal expression of myosin heavy chains in early postnatal stages of spinal muscular atrophy type I at single fibre level.

Hedberg-Oldfors C, Jennions E, Visuttijai K, Oldfors A Acta Myol. 2024; 43(3):89-94.

PMID: 39468964 PMC: 11537715. DOI: 10.36185/2532-1900-558.

Effects of age on human skeletal muscle: a systematic review and meta-analysis of myosin heavy chain isoform protein expression, fiber size, and distribution.

Lee C, Woods P, Paluch A, Miller M Am J Physiol Cell Physiol. 2024; 327(6):C1400-C1415.

PMID: 39374077 PMC: 11684863. DOI: 10.1152/ajpcell.00347.2024.

Fast and slow myofiber nuclei, satellite cells, and size distribution with lifelong endurance exercise in men and women.

Montenegro C, Skiles C, Kuszmaul D, Gouw A, Minchev K, Chambers T Physiol Rep. 2024; 12(13):e16052.

PMID: 38987200 PMC: 11236482. DOI: 10.14814/phy2.16052.

References

Larsson L, Moss R . Maximum velocity of shortening in relation to myosin isoform composition in single fibres from human skeletal muscles. J Physiol. 1993; 472:595-614. PMC: 1160504. DOI: 10.1113/jphysiol.1993.sp019964. View

Larsson L, Li X, Frontera W . Effects of aging on shortening velocity and myosin isoform composition in single human skeletal muscle cells. Am J Physiol. 1997; 272(2 Pt 1):C638-49. DOI: 10.1152/ajpcell.1997.272.2.C638. View

Serrano N, Colenso-Semple L, Lazauskus K, Siu J, Bagley J, Lockie R . Extraordinary fast-twitch fiber abundance in elite weightlifters. PLoS One. 2019; 14(3):e0207975. PMC: 6436686. DOI: 10.1371/journal.pone.0207975. View

Bloemberg D, Quadrilatero J . Rapid determination of myosin heavy chain expression in rat, mouse, and human skeletal muscle using multicolor immunofluorescence analysis. PLoS One. 2012; 7(4):e35273. PMC: 3329435. DOI: 10.1371/journal.pone.0035273. View

Luden N, Hayes E, Galpin A, Minchev K, Jemiolo B, Raue U . Myocellular basis for tapering in competitive distance runners. J Appl Physiol (1985). 2010; 108(6):1501-9. DOI: 10.1152/japplphysiol.00045.2010. View

Klitgaard H, Zhou M, Schiaffino S, Betto R, Salviati G, Saltin B . Ageing alters the myosin heavy chain composition of single fibres from human skeletal muscle. Acta Physiol Scand. 1990; 140(1):55-62. DOI: 10.1111/j.1748-1716.1990.tb08975.x. View

Trappe S, Harber M, Creer A, Gallagher P, Slivka D, Minchev K . Single muscle fiber adaptations with marathon training. J Appl Physiol (1985). 2006; 101(3):721-7. DOI: 10.1152/japplphysiol.01595.2005. View

Murgia M, Toniolo L, Nagaraj N, Ciciliot S, Vindigni V, Schiaffino S . Single Muscle Fiber Proteomics Reveals Fiber-Type-Specific Features of Human Muscle Aging. Cell Rep. 2017; 19(11):2396-2409. DOI: 10.1016/j.celrep.2017.05.054. View

Andersen J, Terzis G, Kryger A . Increase in the degree of coexpression of myosin heavy chain isoforms in skeletal muscle fibers of the very old. Muscle Nerve. 1999; 22(4):449-54. DOI: 10.1002/(sici)1097-4598(199904)22:4<449::aid-mus4>3.0.co;2-2. View

10.

Harber M, Gallagher P, Trautmann J, Trappe S . Myosin heavy chain composition of single muscle fibers in male distance runners. Int J Sports Med. 2002; 23(7):484-8. DOI: 10.1055/s-2002-35067. View

11.

Konopka A, Trappe T, Jemiolo B, Trappe S, Harber M . Myosin heavy chain plasticity in aging skeletal muscle with aerobic exercise training. J Gerontol A Biol Sci Med Sci. 2011; 66(8):835-41. PMC: 3202903. DOI: 10.1093/gerona/glr088. View

12.

Miller M, Bedrin N, Callahan D, Previs M, Jennings 2nd M, Ades P . Age-related slowing of myosin actin cross-bridge kinetics is sex specific and predicts decrements in whole skeletal muscle performance in humans. J Appl Physiol (1985). 2013; 115(7):1004-14. PMC: 3798822. DOI: 10.1152/japplphysiol.00563.2013. View

13.

Malisoux L, Jamart C, Delplace K, Nielens H, Francaux M, Theisen D . Effect of long-term muscle paralysis on human single fiber mechanics. J Appl Physiol (1985). 2006; 102(1):340-9. DOI: 10.1152/japplphysiol.00609.2006. View

14.

Carroll C, Gallagher P, Seidle M, Trappe S . Skeletal muscle characteristics of people with multiple sclerosis. Arch Phys Med Rehabil. 2005; 86(2):224-9. DOI: 10.1016/j.apmr.2004.03.035. View

15.

Klitgaard H, Zhou M, Richter E . Myosin heavy chain composition of single fibres from m. biceps brachii of male body builders. Acta Physiol Scand. 1990; 140(2):175-80. DOI: 10.1111/j.1748-1716.1990.tb08989.x. View

16.

Toth M, Callahan D, Miller M, Tourville T, Hackett S, Couch M . Skeletal muscle fiber size and fiber type distribution in human cancer: Effects of weight loss and relationship to physical function. Clin Nutr. 2016; 35(6):1359-1365. PMC: 6411286. DOI: 10.1016/j.clnu.2016.02.016. View

17.

Sawano S, Komiya Y, Ichitsubo R, Ohkawa Y, Nakamura M, Tatsumi R . A One-Step Immunostaining Method to Visualize Rodent Muscle Fiber Type within a Single Specimen. PLoS One. 2016; 11(11):e0166080. PMC: 5096669. DOI: 10.1371/journal.pone.0166080. View

18.

Galpin A, Raue U, Jemiolo B, Trappe T, Harber M, Minchev K . Human skeletal muscle fiber type specific protein content. Anal Biochem. 2012; 425(2):175-82. PMC: 3358799. DOI: 10.1016/j.ab.2012.03.018. View

19.

Parcell A, Sawyer R, Drummond M, ONeil B, Miller N, Woolstenhulme M . Single-fiber MHC polymorphic expression is unaffected by sprint cycle training. Med Sci Sports Exerc. 2005; 37(7):1133-7. DOI: 10.1249/01.mss.0000170123.27209.e1. View

20.

Lutz G, Bremner S, Bade M, Lieber R . Identification of myosin light chains in Rana pipiens skeletal muscle and their expression patterns along single fibres. J Exp Biol. 2002; 204(Pt 24):4237-48. DOI: 10.1242/jeb.204.24.4237. View