Pflügers Archiv - European Journal of Physiology

, Volume 470, Issue 11, pp 1633–1645 | Cite as

Training state and fasting-induced PDH regulation in human skeletal muscle

  • Anders Gudiksen
  • Lærke Bertholdt
  • Tomasz Stankiewicz
  • Ida Villesen
  • Jens Bangsbo
  • Peter Plomgaard
  • Henriette PilegaardEmail author
Muscle physiology
Part of the following topical collections:
  1. Muscle physiology


The aim of the present study was to examine the influence of training state on fasting-induced skeletal muscle pyruvate dehydrogenase (PDH) regulation, including PDH phosphorylation. Trained and untrained subjects, matched for skeletal muscle CS activity and OXPHOS protein, fasted for 36 h after receiving a standardized meal. Respiratory exchange ratio (RER) was measured and blood as well as vastus lateralis muscle biopsies were obtained 2, 12, 24, and 36 h after the meal. RER decreased with fasting only in untrained individuals, while PDHa activity decreased from 12 h after the meal in untrained, but only tended to decrease at 36 h in trained. PDH-E1α, PDP1 protein, PDH phosphorylation, and PDH acetylation in skeletal muscle was higher in trained than untrained subjects, but did not change with fasting, while PDK4 protein was higher at 36 h than at 2 h after the meal in both groups. In conclusion, the present results suggest that endurance exercise training modifies the fasting-induced regulation of PDHa activity in skeletal muscle and the substrate switch towards fat oxidation. PDH phosphorylation could not explain the fasting-induced regulation of PDHa activity suggesting other post translational modifications.


Pyruvate dehydrogenase Skeletal muscle Acetylation Phosphorylation Fasting Exercise training 



We would like to thank the involved subjects for participation in the study.

Funding information

This study was funded by the Danish Ministry of Culture for Sports Research (1095421001), the Danish Council for Independent Research (36723-104353), and the Danish Diabetes Academy (1105701001). The Centre for Physical Activity Research is supported by a grant from TrygFonden, and the Centre of Inflammation and Metabolism was supported by a grant from the Danish National Research Foundation (DNRF55).


  1. 1.
    Ahn BH, Kim HS, Song S, Lee IH, Liu J, Vassilopoulos A, Deng CX, Finkel T (2008) A role for the mitochondrial deacetylase Sirt3 in regulating energy homeostasis. Proc Natl Acad Sci U S A 105:14447–14452CrossRefPubMedCentralGoogle Scholar
  2. 2.
    Bergmeyer HU, Moellering H (1965) Acylphosphate: D-glucose-6-phosphotransferase. Biochem Z 343:97–102PubMedPubMedCentralGoogle Scholar
  3. 3.
    Bergstrom J (1975) Percutaneous needle biopsy of skeletal muscle in physiological and clinical research. Scand J Clin Lab Invest 35:609–616CrossRefPubMedCentralGoogle Scholar
  4. 4.
    Bertholdt L, Gudiksen A, Stankiewicz T, Villesen I, Tybirk J, van HG, Bangsbo J, Plomgaard P, Pilegaard H (2017) Impact of training state on fasting-induced regulation of adipose tissue metabolism in humans. CrossRefPubMedCentralGoogle Scholar
  5. 5.
    Bienso RS, Knudsen JG, Brandt N, Pedersen PA, Pilegaard H (2014) Effects of IL-6 on pyruvate dehydrogenase regulation in mouse skeletal muscle. Pflugers Arch 466:1647–1657CrossRefPubMedCentralGoogle Scholar
  6. 6.
    Bienso RS, Olesen J, Gliemann L, Schmidt JF, Matzen MS, Wojtaszewski JF, Hellsten Y, Pilegaard H (2015) Effects of exercise training on regulation of skeletal muscle glucose metabolism in elderly men. J Gerontol A Biol Sci Med Sci 70:866–872CrossRefPubMedCentralGoogle Scholar
  7. 7.
    Bowker-Kinley MM, Davis WI, Wu P, Harris RA, Popov KM (1998) Evidence for existence of tissue-specific regulation of the mammalian pyruvate dehydrogenase complex. Biochem J 329(Pt 1):191–196CrossRefPubMedCentralGoogle Scholar
  8. 8.
    Canto C, Jiang LQ, Deshmukh AS, Mataki C, Coste A, Lagouge M, Zierath JR, Auwerx J (2010) Interdependence of AMPK and SIRT1 for metabolic adaptation to fasting and exercise in skeletal muscle. Cell Metab 11:213–219CrossRefPubMedCentralGoogle Scholar
  9. 9.
    Cederblad G, Carlin JI, Constantin-Teodosiu D, Harper P, Hultman E (1990) Radioisotopic assays of CoASH and carnitine and their acetylated forms in human skeletal muscle. Anal Biochem 185:274–278CrossRefPubMedCentralGoogle Scholar
  10. 10.
    Chomczynski P, Sacchi N (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162:156–159CrossRefGoogle Scholar
  11. 11.
    Consitt LA, Saxena G, Saneda A, Houmard JA (2016) Age-related impairments in skeletal muscle PDH phosphorylation and plasma lactate are indicative of metabolic inflexibility and the effects of exercise training. Am J Physiol Endocrinol Metab 311:E145–E156CrossRefPubMedCentralGoogle Scholar
  12. 12.
    Constantin-Teodosiu D, Carlin JI, Cederblad G, Harris RC, Hultman E (1991a) Acetyl group accumulation and pyruvate dehydrogenase activity in human muscle during incremental exercise. Acta Physiol Scand 143:367–372CrossRefPubMedCentralGoogle Scholar
  13. 13.
    Constantin-Teodosiu D, Cederblad G, Hultman E (1991b) A sensitive radioisotopic assay of pyruvate dehydrogenase complex in human muscle tissue. Anal Biochem 198:347–351CrossRefPubMedCentralGoogle Scholar
  14. 14.
    Constantin-Teodosiu D, Peirce NS, Fox J, Greenhaff PL (2004) Muscle pyruvate availability can limit the flux, but not activation, of the pyruvate dehydrogenase complex during submaximal exercise in humans. J Physiol 561:647–655CrossRefPubMedCentralGoogle Scholar
  15. 15.
    Edgett BA, Hughes MC, Matusiak JB, Perry CG, Simpson CA, Gurd BJ (2016) SIRT3 gene expression but not SIRT3 subcellular localization is altered in response to fasting and exercise in human skeletal muscle. Exp Physiol 101:1101–1113CrossRefPubMedCentralGoogle Scholar
  16. 16.
    Evans WJ, Phinney SD, Young VR (1982) Suction applied to a muscle biopsy maximizes sample size. Med Sci Sports Exerc 14:101–102PubMedPubMedCentralGoogle Scholar
  17. 17.
    Fan J, Shan C, Kang HB, Elf S, Xie J, Tucker M, Gu TL, Aguiar M, Lonning S, Chen H, Mohammadi M, Britton LM, Garcia BA, Aleckovic M, Kang Y, Kaluz S, Devi N, Van Meir EG, Hitosugi T, Seo JH, Lonial S, Gaddh M, Arellano M, Khoury HJ, Khuri FR, Boggon TJ, Kang S, Chen J (2014) Tyr phosphorylation of PDP1 toggles recruitment between ACAT1 and SIRT3 to regulate the pyruvate dehydrogenase complex. Mol Cell 53:534–548CrossRefPubMedCentralGoogle Scholar
  18. 18.
    Frayn KN (1983) Calculation of substrate oxidation rates in vivo from gaseous exchange. Physiol Rep 55:628–634Google Scholar
  19. 19.
    Gudiksen A, Pilegaard H (2017) PGC-1alpha and fasting-induced PDH regulation in mouse skeletal muscle. Physiol Rep 5:e13222CrossRefPubMedCentralGoogle Scholar
  20. 20.
    Gudiksen A, Bertholdt L, Stankiewicz T, Tybirk J, Plomgaard P, Bangsbo J, Pilegaard H (2017a) Effects of training status on PDH regulation in human skeletal muscle during exercise. Pflugers Arch 469:1615–1630. CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Gudiksen A, Bertholdt L, Vingborg MB, Hansen HW, Ringholm S, Pilegaard H (2017b) Muscle interleukin-6 and fasting-induced PDH regulation in mouse skeletal muscle. Am J Physiol Endocrinol Metab 312:E204–E214CrossRefPubMedCentralGoogle Scholar
  22. 22.
    Hirschey MD, Shimazu T, Goetzman E, Jing E, Schwer B, Lombard DB, Grueter CA, Harris C, Biddinger S, Ilkayeva OR, Stevens RD, Li Y, Saha AK, Ruderman NB, Bain JR, Newgard CB, Farese RV Jr, Alt FW, Kahn CR, Verdin E (2010) SIRT3 regulates mitochondrial fatty-acid oxidation by reversible enzyme deacetylation. Nature 464:121–125CrossRefPubMedCentralGoogle Scholar
  23. 23.
    Jeoung NH, Wu P, Joshi MA, Jaskiewicz J, Bock CB, Depaoli-Roach AA, Harris RA (2006) Role of pyruvate dehydrogenase kinase isoenzyme 4 (PDHK4) in glucose homoeostasis during starvation. Biochem J 397:417–425CrossRefPubMedCentralGoogle Scholar
  24. 24.
    Jing E, O'Neill BT, Rardin MJ, Kleinridders A, Ilkeyeva OR, Ussar S, Bain JR, Lee KY, Verdin EM, Newgard CB, Gibson BW, Kahn CR (2013) Sirt3 regulates metabolic flexibility of skeletal muscle through reversible enzymatic deacetylation. Diabetes 62:3404–3417CrossRefPubMedCentralGoogle Scholar
  25. 25.
    Kiilerich K, Adser H, Jakobsen AH, Pedersen PA, Hardie DG, Wojtaszewski JF, Pilegaard H (2010) PGC-1alpha increases PDH content but does not change acute PDH regulation in mouse skeletal muscle. Am J Phys Regul Integr Comp Phys 299:R1350–R1359Google Scholar
  26. 26.
    Korotchkina LG, Patel MS (2001) Site specificity of four pyruvate dehydrogenase kinase isoenzymes toward the three phosphorylation sites of human pyruvate dehydrogenase. J Biol Chem 276:37223–37229CrossRefPubMedCentralGoogle Scholar
  27. 27.
    LeBlanc PJ, Peters SJ, Tunstall RJ, Cameron-Smith D, Heigenhauser GJ (2004) Effects of aerobic training on pyruvate dehydrogenase and pyruvate dehydrogenase kinase in human skeletal muscle. J Physiol 557:559–570CrossRefPubMedCentralGoogle Scholar
  28. 28.
    LeBlanc PJ, Harris RA, Peters SJ (2007) Skeletal muscle fiber type comparison of pyruvate dehydrogenase phosphatase activity and isoform expression in fed and food-deprived rats. Am J Physiol Endocrinol Metab 292:E571–E576CrossRefPubMedCentralGoogle Scholar
  29. 29.
    Lombard DB, Alt FW, Cheng HL, Bunkenborg J, Streeper RS, Mostoslavsky R, Kim J, Yancopoulos G, Valenzuela D, Murphy A, Yang Y, Chen Y, Hirschey MD, Bronson RT, Haigis M, Guarente LP, Farese RV Jr, Weissman S, Verdin E, Schwer B (2007) Mammalian Sir2 homolog SIRT3 regulates global mitochondrial lysine acetylation. Mol Cell Biol 27:8807–8814CrossRefPubMedCentralGoogle Scholar
  30. 30.
    Lowry OH, Passonneau JV (1972) A flexible system of enzymatic analysis. Academic Press, New YorkGoogle Scholar
  31. 31.
    Passonneau JV, Lauderdale VR (1974) A comparison of three methods of glycogen measurement in tissues. Anal Biochem 60:405–412CrossRefPubMedCentralGoogle Scholar
  32. 32.
    Patel MS, Korotchkina LG (2001) Regulation of mammalian pyruvate dehydrogenase complex by phosphorylation: complexity of multiple phosphorylation sites and kinases. Exp Mol Med 33:191–197CrossRefPubMedCentralGoogle Scholar
  33. 33.
    Pilegaard H, Ordway GA, Saltin B, Neufer PD (2000) Transcriptional regulation of gene expression in human skeletal muscle during recovery from exercise. Am J Physiol Endocrinol Metab 279:E806–E814CrossRefPubMedCentralGoogle Scholar
  34. 34.
    Pilegaard H, Saltin B, Neufer PD (2003) Effect of short-term fasting and refeeding on transcriptional regulation of metabolic genes in human skeletal muscle. Diabetes 52:657–662CrossRefPubMedCentralGoogle Scholar
  35. 35.
    Pilegaard H, Birk JB, Sacchetti M, Mourtzakis M, Hardie DG, Stewart G, Neufer PD, Saltin B, van HG, Wojtaszewski JF (2006) PDH-E1alpha dephosphorylation and activation in human skeletal muscle during exercise: effect of intralipid infusion. Diabetes 55:3020–3027CrossRefPubMedCentralGoogle Scholar
  36. 36.
    Rodgers JT, Lerin C, Haas W, Gygi SP, Spiegelman BM, Puigserver P (2005) Nutrient control of glucose homeostasis through a complex of PGC-1alpha and SIRT1. Nature 434:113–118CrossRefPubMedCentralGoogle Scholar
  37. 37.
    Schwer B, North BJ, Frye RA, Ott M, Verdin E (2002) The human silent information regulator (Sir)2 homologue hSIRT3 is a mitochondrial nicotinamide adenine dinucleotide-dependent deacetylase. J Cell Biol 158:647–657CrossRefPubMedCentralGoogle Scholar
  38. 38.
    Spriet LL, Tunstall RJ, Watt MJ, Mehan KA, Hargreaves M, Cameron-Smith D (2004) Pyruvate dehydrogenase activation and kinase expression in human skeletal muscle during fasting. J Appl Physiol (1985 ) 96:2082–2087CrossRefGoogle Scholar
  39. 39.
    St Amand TA, Spriet LL, Jones NL, Heigenhauser GJ (2000) Pyruvate overrides inhibition of PDH during exercise after a low-carbohydrate diet. Am J Physiol Endocrinol Metab 279:E275–E283CrossRefPubMedCentralGoogle Scholar
  40. 40.
    Sugden MC, Holness MJ (2006) Mechanisms underlying regulation of the expression and activities of the mammalian pyruvate dehydrogenase kinases. Arch Physiol Biochem 112:139–149CrossRefPubMedCentralGoogle Scholar
  41. 41.
    Sugden MC, Kraus A, Harris RA, Holness MJ (2000) Fibre-type specific modification of the activity and regulation of skeletal muscle pyruvate dehydrogenase kinase (PDK) by prolonged starvation and refeeding is associated with targeted regulation of PDK isoenzyme 4 expression. Biochem J 346 Pt 3:651–657CrossRefPubMedCentralGoogle Scholar
  42. 42.
    Verdin E, Hirschey MD, Finley LW, Haigis MC (2010) Sirtuin regulation of mitochondria: energy production, apoptosis, and signaling. Trends Biochem Sci 35:669–675CrossRefPubMedCentralGoogle Scholar
  43. 43.
    Wu P, Inskeep K, Bowker-Kinley MM, Popov KM, Harris RA (1999) Mechanism responsible for inactivation of skeletal muscle pyruvate dehydrogenase complex in starvation and diabetes. Diabetes 48:1593–1599CrossRefPubMedCentralGoogle Scholar
  44. 44.
    Wu P, Blair PV, Sato J, Jaskiewicz J, Popov KM, Harris RA (2000) Starvation increases the amount of pyruvate dehydrogenase kinase in several mammalian tissues. Arch Biochem Biophys 381:1–7CrossRefPubMedCentralGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Section for Cell Biology and Physiology, Department of BiologyUniversity of CopenhagenCopenhagenDenmark
  2. 2.Section of Integrative Physiology, Department of Nutrition, Exercise and SportsUniversity of CopenhagenCopenhagenDenmark
  3. 3.Department of Clinical Biochemistry, Rigshospitalet, Centre of Inflammation and Metabolism and Centre for Physical Activity ResearchUniversity of CopenhagenCopenhagenDenmark

Personalised recommendations