Effects of acute exercise and long-term exercise on total Na+,K+-ATPase content and Na+,K+-ATPase isoform expression profile in equine muscle

Maarten M. M. van den Burg Department of Pathobiology, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 1, NL-3584 CL Utrecht, The Netherlands.

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Karin Eizema Department of Pathobiology, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 1, NL-3584 CL Utrecht, The Netherlands.

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Ellen de Graaf-Roelfsema Department of Equine Sciences, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 1, NL-3584 CL Utrecht, The Netherlands.

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Eric van Breda Department of Movement Sciences, Maastricht University, NL-6200 MD Maastricht, The Netherlands.

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Inge D. Wijnberg Department of Equine Sciences, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 1, NL-3584 CL Utrecht, The Netherlands.

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Johannes H. van der Kolk Department of Equine Sciences, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 1, NL-3584 CL Utrecht, The Netherlands.

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Maria E. Everts Department of Pathobiology, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 1, NL-3584 CL Utrecht, The Netherlands.

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Abstract

Objective—To investigate the effects of acute exercise and long-term training on Na+,K+-ATPase content, mRNA isoforms, and protein concentration in equine muscle.

Animals—6 Standardbreds.

Procedures—Horses performed a bout of exercise on a treadmill before and after 18 weeks of combined interval and endurance training. Muscle biopsy specimens were obtained from vastus lateralis muscle (VLM) and pectoralis descendens muscle (PDM) before and after exercise. The Na+,K+-ATPase content, mRNA isoforms, and protein concentrations were determined by use of [3H]ouabain binding, real-time PCR assay, and western blotting, respectively.

Results—6 Na+,K+-ATPase mRNA isoforms were present in equine muscle, but only A2 and B1 proteins were detected. Exercise before training resulted in increases of mRNA isoforms A1, A2, A3, and B2 in VLM and A1 and B3 in PDM. Training increased resting values for mRNA isoforms A3 and B1 in VLM and B3 in PDM. The Na+,K+-ATPase, [3H]ouabain binding, and proteins of mRNA A2 and B1 increased in VLM, whereas in PDM, only A2 protein increased as a result of training. After training, effects of strenuous exercise on mRNA expression were no longer detectable.

Conclusions and Clinical Relevance—Equine muscle contained all Na+,K+-ATPase mRNA isoforms, but only A2 and B1 proteins could be detected. Expression of these isoforms changed as a result of strenuous exercise and long-term training, representing an adaptive response. Determination of Na+,K+-ATPase gene expression may be relevant for understanding alterations in excitability during neuromuscular diseases.

Abstract

Objective—To investigate the effects of acute exercise and long-term training on Na+,K+-ATPase content, mRNA isoforms, and protein concentration in equine muscle.

Animals—6 Standardbreds.

Procedures—Horses performed a bout of exercise on a treadmill before and after 18 weeks of combined interval and endurance training. Muscle biopsy specimens were obtained from vastus lateralis muscle (VLM) and pectoralis descendens muscle (PDM) before and after exercise. The Na+,K+-ATPase content, mRNA isoforms, and protein concentrations were determined by use of [3H]ouabain binding, real-time PCR assay, and western blotting, respectively.

Results—6 Na+,K+-ATPase mRNA isoforms were present in equine muscle, but only A2 and B1 proteins were detected. Exercise before training resulted in increases of mRNA isoforms A1, A2, A3, and B2 in VLM and A1 and B3 in PDM. Training increased resting values for mRNA isoforms A3 and B1 in VLM and B3 in PDM. The Na+,K+-ATPase, [3H]ouabain binding, and proteins of mRNA A2 and B1 increased in VLM, whereas in PDM, only A2 protein increased as a result of training. After training, effects of strenuous exercise on mRNA expression were no longer detectable.

Conclusions and Clinical Relevance—Equine muscle contained all Na+,K+-ATPase mRNA isoforms, but only A2 and B1 proteins could be detected. Expression of these isoforms changed as a result of strenuous exercise and long-term training, representing an adaptive response. Determination of Na+,K+-ATPase gene expression may be relevant for understanding alterations in excitability during neuromuscular diseases.

  • 1.

    Rivero JL. A scientific background for skeletal muscle conditioning in equine practice. J Vet Med A Physiol Pathol Clin Med 2007;54:321332.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 2.

    Clausen T. Na+-K+ pump regulation and skeletal muscle contractility. Physiol Rev 2003;83:12691324.

  • 3.

    Rietbroek NJ, Dingboom EG, Joosten BJ, et al. Effect of show jumping training on the development of locomotory muscle in young horses. Am J Vet Res 2007;68:12321238.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 4.

    Suwannachot P, Joosten BJ, Klarenbeek A, et al. Effects of training on potassium homeostasis during exercise and skeletal muscle Na+,K(+)-ATPase concentration in young adult and middle-aged Dutch Warmblood horses. Am J Vet Res 2005;66:12521258.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 5.

    Green HJ. Cation pumps in skeletal muscle: potential role in muscle fatigue. Acta Physiol Scand 1998;162:201213.

  • 6.

    Murphy KT, Petersen AC, Goodman C, et al. Prolonged sub-maximal exercise induces isoform-specific Na+-K+-ATPase mRNA and protein responses in human skeletal muscle. Am J Physiol Regul Integr Comp Physiol 2006;290:R414R424.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 7.

    Murphy KT, Snow RJ, Petersen AC, et al. Intense exercise up-regulates Na+,K+-ATPase isoform mRNA, but not protein expression in human skeletal muscle. J Physiol 2004;556:507519.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 8.

    Keryanov S, Gardner KL. Physical mapping and characterization of the human Na,K-ATPase isoform, ATP1A4. Gene 2002;292:151166.

  • 9.

    Nordsborg N, Thomassen M, Lundby C, et al. Contraction-induced increases in Na+-K+-ATPase mRNA levels in human skeletal muscle are not amplified by activation of additional muscle mass. Am J Physiol Regul Integr Comp Physiol 2005;289:R84R91.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 10.

    Blanco G, Mercer RW. Isozymes of the Na-K-ATPase: heterogeneity in structure, diversity in function. Am J Physiol 1998;275:F633F650.

  • 11.

    Tsakiridis T, Wong PP, Liu Z, et al. Exercise increases the plasma membrane content of the Na+ -K+ pump and its mRNA in rat skeletal muscles. J Appl Physiol 1996;80:699705.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 12.

    Juel C, Nielsen JJ, Bangsbo J. Exercise-induced translocation of Na(+)-K(+) pump subunits to the plasma membrane in human skeletal muscle. Am J Physiol Regul Integr Comp Physiol 2000;278:R1107R1110.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 13.

    Green HJ, Duhamel TA, Holloway GP, et al. Muscle Na+-K+-ATPase response during 16 hours of heavy intermittent cycle exercise. Am J Physiol Endocrinol Metab 2007;293:E523E530.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 14.

    Murphy KT, Aughey RJ, Petersen AC, et al. Effects of endurance training status and sex differences on Na+,K+-pump mRNA expression, content and maximal activity in human skeletal muscle. Acta Physiol (Oxf) 2007;189:259269.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 15.

    Nordsborg N, Bangsbo J, Pilegaard H. Effect of high-intensity training on exercise-induced gene expression specific to ion homeostasis and metabolism. J Appl Physiol 2003;95:12011206.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 16.

    Aughey RJ, Murphy KT, Clark SA, et al. Muscle Na+,K+ATPase activity and isoform adaptations to intense interval exercise and training in well-trained athletes. J Appl Physiol 2007;103:3947.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 17.

    Dela F, Holten M, Juel C. Effect of resistance training on Na,K pump and Na+/H+ exchange protein densities in muscle from control and patients with type 2 diabetes. Pflugers Arch 2004;447:928933.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 18.

    Mohr M, Krustrup P, Nielsen JJ, et al. Effect of two different intense training regimens on skeletal muscle ion transport proteins and fatigue development. Am J Physiol Regul Integr Comp Physiol 2007;292:R1594R1602.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 19.

    Nielsen JJ, Mohr M, Klarskov C, et al. Effects of high-intensity intermittent training on potassium kinetics and performance in human skeletal muscle. J Physiol 2004;554:857870.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 20.

    McCutcheon LJ, Geor RJ, Shen H. Skeletal muscle Na(+)-K(+)-ATPase and K+ homeostasis during exercise: effects of short-term training. Equine Vet J Suppl 1999;30:303310.

    • Search Google Scholar
    • Export Citation
  • 21.

    Suwannachot P, Verkleij CB, Kocsis S, et al. Specificity and reversibility of the training effects on the concentration of Na+,K+-ATPase in foal skeletal muscle. Equine Vet J 2001;33:250255.

    • Search Google Scholar
    • Export Citation
  • 22.

    Suwannachot P, Verkleij CB, Weijs WA, et al. Effects of training on the concentration of Na+, K+-ATPase in foal muscle. Equine Vet J Suppl 1999;31:101105.

    • Search Google Scholar
    • Export Citation
  • 23.

    Bruin G, Kuipers H, Keizer HA, et al. Adaptation and overtraining in horses subjected to increasing training loads. J Appl Physiol 1994;76:19081913.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 24.

    Jemiolo B, Trappe S. Single muscle fiber gene expression in human skeletal muscle: validation of internal control with exercise. Biochem Biophys Res Commun 2004;320:10431050.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 25.

    Nørgaard A, Kjeldsen K, Clausen T. A method for the determination of the total number of 3H-ouabain binding sites in biopsies of human skeletal muscle. Scand J Clin Lab Invest 1984;44:509518.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 26.

    Nørgaard A, Kjeldsen K, Hansen O, et al. A simple and rapid method for the determination of the number of 3H-ouabain binding sites in biopsies of skeletal muscle. Biochem Biophys Res Commun 1983;111:319325.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 27.

    Orlowski J, Lingrel JB. Differential expression of the Na,K-ATPase alpha 1 and alpha 2 subunit genes in a murine myogenic cell line. Induction of the alpha 2 isozyme during myocyte differentiation. J Biol Chem 1988;263:1781717821.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 28.

    van Dam KG, van Breda E, Schaart G, et al. Investigation of the expression and localization of glucose transporter 4 and fatty acid translocase/CD36 in equine skeletal muscle. Am J Vet Res 2004;65:951956.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 29.

    Green HJ, Chin ER, Ball-Burnett M, et al. Increases in human skeletal muscle Na(+)-K(+)-ATPase concentration with short-term training. Am J Physiol 1993;264:C1538C1541.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 30.

    Nordsborg N, Goodmann C, McKenna MJ, et al. Dexamethasone up-regulates skeletal muscle maximal Na+,K+ pump activity by muscle group specific mechanisms in humans. J Physiol 2005;567:583589.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 31.

    Petersen AC, Murphy KT, Snow RJ, et al. Depressed Na+-K+-ATPase activity in skeletal muscle at fatigue is correlated with increased Na+-K+-ATPase mRNA expression following intense exercise. Am J Physiol Regul Integr Comp Physiol 2005;289:R266R274.

    • Crossref
    • Search Google Scholar
    • Export Citation

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