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Myogenesis pp 283-300 | Cite as

Measuring Both Glucose Uptake and Myosin Heavy Chain Isoform Expression in Single Rat Skeletal Muscle Fibers

  • Mark W. Pataky
  • Edward B. Arias
  • Gregory D. CarteeEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1889)

Abstract

Glucose uptake by skeletal muscle is important for metabolic health. Because skeletal muscle is composed of multiple fiber types that have differing metabolic and contractile properties, studying glucose uptake in whole muscle tissue does not elucidate differences at the cellular level. Here, we describe a procedure that enables the measurement of both glucose uptake and fiber type (by myosin heavy chain isoform expression) in individual rat epitrochlearis muscle fibers.

Key words

Muscle fiber type Myosin heavy chain Glucose transport Glucose uptake SDS-PAGE 

Notes

Acknowledgments

This work was supported by grants from the National Institutes of Health (R01 DK71771 and R01 AG10026). We thank Jim MacKrell and Haiyan Wang for providing valuable suggestions about the manuscript.

References

  1. 1.
    Pette D, Staron RS (1990) Cellular and molecular diversities of mammalian skeletal muscle fibers. Rev Physiol Biochem Pharmacol 116:1–76PubMedGoogle Scholar
  2. 2.
    Pandorf CE, Garland T, Aoi W, Handschin C, Gorza L, Garcia-Roves PM, Copp SW, Tipton CM, Caiozzo VJ, Haddad F (2010) A rationale for SDS-PAGE of MHC isoforms as a gold standard for determining contractile phenotype. J Appl Physiol 108(1):222–225CrossRefGoogle Scholar
  3. 3.
    Fitzsimons DP, Diffee GM, Herrick RE, Baldwin KM (1990) Effects of endurance exercise on isomyosin patterns in fast-and slow-twitch skeletal muscles. J Appl Physiol 68(5):1950–1955CrossRefGoogle Scholar
  4. 4.
    Röckl KS, Hirshman MF, Brandauer J, Fujii N, Witters LA, Goodyear LJ (2007) Skeletal muscle adaptation to exercise training: AMP-activated protein kinase mediates muscle fiber type shift. Diabetes 56(8):2062–2069CrossRefGoogle Scholar
  5. 5.
    Korhonen MT, Cristea A, Alén M, Häkkinen K, Sipilä S, Mero A, Viitasalo JT, Larsson L, Suominen H (2006) Aging, muscle fiber type, and contractile function in sprint-trained athletes. J Appl Physiol 101(3):906–917CrossRefGoogle Scholar
  6. 6.
    Nilwik R, Snijders T, Leenders M, Groen BB, van Kranenburg J, Verdijk LB, van Loon LJ (2013) The decline in skeletal muscle mass with aging is mainly attributed to a reduction in type II muscle fiber size. Exp Gerontol 48(5):492–498CrossRefGoogle Scholar
  7. 7.
    Hansen PA, Nolte LA, Chen MM, Holloszy JO (1998) Increased GLUT-4 translocation mediates enhanced insulin sensitivity of muscle glucose transport after exercise. J Appl Physiol 85(4):1218–1222CrossRefGoogle Scholar
  8. 8.
    Lee AD, Hansen PA, Holloszy JO (1995) Wortmannin inhibits insulin-stimulated but not contraction-stimulated glucose transport activity in skeletal muscle. FEBS Lett 361(1):51–54CrossRefGoogle Scholar
  9. 9.
    Wallberg-Henriksson H, Constable S, Young D, Holloszy J (1988) Glucose transport into rat skeletal muscle: interaction between exercise and insulin. J Appl Physiol 65(2):909–913CrossRefGoogle Scholar
  10. 10.
    Wojtaszewski JF, Higaki Y, Hirshman MF, Michael MD, Dufresne SD, Kahn CR, Goodyear LJ (1999) Exercise modulates postreceptor insulin signaling and glucose transport in muscle-specific insulin receptor knockout mice. J Clin Invest 104(9):1257–1264CrossRefGoogle Scholar
  11. 11.
    Funai K, Cartee GD (2009) Inhibition of contraction-stimulated AMP-activated protein kinase inhibits contraction-stimulated increases in PAS-TBC1D1 and glucose transport without altering PAS-AS160 in rat skeletal muscle. Diabetes 58(5):1096–1104.  https://doi.org/10.2337/db08-1477CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Wright DC, Hucker KA, Holloszy JO, Han DH (2004) Ca2+ and AMPK both mediate stimulation of glucose transport by muscle contractions. Diabetes 53(2):330–335CrossRefGoogle Scholar
  13. 13.
    Mackrell JG, Cartee GD (2012) A novel method to measure glucose uptake and myosin heavy chain isoform expression of single fibers from rat skeletal muscle. Diabetes 61(5):995–1003.  https://doi.org/10.2337/db11-1299CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Deves R, Krupka R (1978) Cytochalasin B and the kinetics of inhibition of biological transport. A case of asymmetric binding to the glucose carrier. Biochim Biophys Acta 510(2):339–348CrossRefGoogle Scholar
  15. 15.
    Cartee GD, Arias EB, Carmen SY, Pataky MW (2016) Novel single skeletal muscle fiber analysis reveals a fiber type-selective effect of acute exercise on glucose uptake. Am J Physiol Endocrinol Metab 311(5):E818–E824CrossRefGoogle Scholar
  16. 16.
    Wang H, Arias EB, Yu CS, Verkerke AR, Cartee GD (2017) Effects of calorie restriction and fiber type on glucose uptake and abundance of electron transport chain and oxidative phosphorylation proteins in single fibers from old rats. J Gerontol Ser A: Biomed Sci Med Sci 72(12):1638–1646CrossRefGoogle Scholar
  17. 17.
    Pataky MW, Wang H, Yu CS, Arias EB, Ploutz-Snyder RJ, Zheng X, Cartee GD (2017) High-fat diet-induced insulin resistance in single skeletal muscle fibers is Fiber type selective. Sci Rep 7(1):13642.  https://doi.org/10.1038/s41598-017-12682-zCrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Castorena CM, Arias EB, Sharma N, Bogan JS, Cartee GD (2015) Fiber type effects on contraction-stimulated glucose uptake and GLUT4 abundance in single fibers from rat skeletal muscle. Am J Physiol Endocrinol Metab 308(3):E223–E230CrossRefGoogle Scholar
  19. 19.
    Wallberg-Henriksson H (1987) Glucose transport into skeletal muscle. Influence of contractile activity, insulin, catecholamines and diabetes mellitus. Acta Physiol Scand Suppl 564:1–80PubMedGoogle Scholar
  20. 20.
    Delp MD, Duan C (1996) Composition and size of type I, IIA, IID/X, and IIB fibers and citrate synthase activity of rat muscle. J Appl Physiol 80(1):261–270CrossRefGoogle Scholar
  21. 21.
    Mackrell JG, Arias EB, Cartee GD (2012) Fiber type-specific differences in glucose uptake by single fibers from skeletal muscles of 9- and 25-month-old rats. J Gerontol A Biol Sci Med Sci 67(12):1286–1294.  https://doi.org/10.1093/gerona/gls194CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Castorena CM, Mackrell JG, Bogan JS, Kanzaki M, Cartee GD (2011) Clustering of GLUT4, TUG, and RUVBL2 protein levels correlate with myosin heavy chain isoform pattern in skeletal muscles, but AS160 and TBC1D1 levels do not. J Appl Physiol (1985) 111(4):1106–1117.  https://doi.org/10.1152/japplphysiol.00631.2011CrossRefPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Mark W. Pataky
    • 1
  • Edward B. Arias
    • 1
  • Gregory D. Cartee
    • 1
    • 2
    • 3
    Email author
  1. 1.Muscle Biology Laboratory, School of KinesiologyUniversity of MichiganAnn ArborUSA
  2. 2.Department of Molecular and Integrative PhysiologyUniversity of MichiganAnn ArborUSA
  3. 3.Institute of GerontologyUniversity of MichiganAnn ArborUSA

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