Advertisement

Pflügers Archiv

, Volume 447, Issue 2, pp 247–253 | Cite as

Effects of heat stress and mechanical stretch on protein expression in cultured skeletal muscle cells

  • K. Goto
  • R. Okuyama
  • H. Sugiyama
  • M. Honda
  • T. Kobayashi
  • K. Uehara
  • T. Akema
  • T. Sugiura
  • S. Yamada
  • Y. Ohira
  • T. Yoshioka
Skeletal Muscle

Abstract

Effects of heat stress, mechanical stretching or a combination of both on the expression of heat shock proteins (HSPs) and total protein level were studied in a culture system. Rat skeletal muscle cells (L6) were cultured on flexible-bottomed culture plates. They were subjected to one of the four following conditions: (1) 97 h incubation at 37 °C, (2) 1 h incubation at 41 °C followed by 96 h incubation at 37 °C, (3) 1 h incubation at 37 °C followed by 96 h cyclic stretching (18% of initial length, 2-s stretch and 4-s release) at 37 °C or (4) 1 h incubation at 41 °C followed by 96 h cyclic stretching at 37 °C. The expression of HSP72 and HSP90 and total protein was determined in the crude homogenates, supernatant and pellets. Cellular protein concentrations in the homogenates and pellets were increased by heat stress and/or mechanical stress (stretch). A cumulative effect of the combination of heating and stretch on the protein concentration in the homogenates and in the pellets was noted. The expressions of HSP72 and HSP90 in the pellets were also increased by heat stress and/or stretch. However, HSP90 in the supernatant did not change following heat stress and/or stretch. The regulation of HSP72 and HSP90 expression in skeletal muscle cells may be closely related to total protein, the abundance of which is also stimulated by mechanical and heat stresses. These observations suggest strongly that heating and passive stretch of muscle may be useful as a means of increasing muscle mass, not only in athletes but also in patients during rehabilitation.

Keywords

Skeletal muscle cells Proteins Culture Heat Stretch 

Notes

Acknowledgements

This study was supported, in part, by a Grant-in-Aid for Encouragement of Young Scientists (B, 14780025) from the Ministry of Education, Culture, Sports, Science and Technology (MEXT) to KG, and the Grant-in-Aid for Scientific Research (A, 15200049) from Japan Society for the Promotion of Science to Y.O.

References

  1. 1.
    Bornman L, Polla BS, Gericke GS (1996) Heat-shock protein 90 and ubiquitin: developmental regulation during myogenesis. Muscle Nerve 19:574–580CrossRefPubMedGoogle Scholar
  2. 2.
    Carlson BM, Faulkner JA (1983) The regeneration of skeletal muscle fibers following injury: a review. Med Sci Sports Exerc 15:187–198PubMedGoogle Scholar
  3. 3.
    Carson JA, Booth FW (1998) Effect of serum and mechanical stretch on skeletal α-actin gene regulation in cultured primary muscle cells. Am J Physiol 275:C1438–C1448PubMedGoogle Scholar
  4. 4.
    Chang J, Wasser JS, Cornelussen RN, Knowlton AA (2001) Activation of heat-shock factor by stretch-activated channels in rat hearts. Circulation 104:209–214PubMedGoogle Scholar
  5. 5.
    Clarke MS, Feeback DL (1996) Mechanical load induces sarcoplasmic wounding and FGF release in differentiated human skeletal muscle cultures. FASEB J 10:502–509PubMedGoogle Scholar
  6. 6.
    Clarkson PM, Sayers SP (1999) Etiology of exercise-induced muscle damage. Can J Appl Physiol 24:234–248PubMedGoogle Scholar
  7. 7.
    Cotto JJ, Morimoto RI (1999) Stress-induced activation of heat-shock response: cell and molecular biology of heat-shock factors. Biochem Soc Symp 64:105–118PubMedGoogle Scholar
  8. 8.
    Delcayre C, Samuel JL, Marotte F, Best-Belpomme M, Mercadier JJ, Rappaport L (1988) Synthesis of stress proteins in rat cardiac myocytes 2–4 days after imposition of hemodynamic overload. J Clin Invest 82:460–468PubMedGoogle Scholar
  9. 9.
    Ecochard L, Lhenry F, Sempore B, Favier R (2000) Skeletal muscle HSP72 level during endurance training: influence of peripheral arterial insufficiency. Pflugers Arch 440:918–924CrossRefPubMedGoogle Scholar
  10. 10.
    Einat MF, Haberfeld A, Shamay A, Horev G, Hurwitz S, Yahav S (1996) A novel 29-kDa chicken heat shock protein. Poult Sci 75:1528–1530PubMedGoogle Scholar
  11. 11.
    Goldspink G (1999) Changes in muscle mass and phenotype and the expression of autocrine and systemic growth factors by muscle in response to stretch and overload. J Anat 194:323–334CrossRefPubMedGoogle Scholar
  12. 12.
    Gonzalez B, Hernando R, Manso R (2000) Stress proteins of 70 kDa in chronically exercised skeletal muscle. Pflugers Arch 440:42–49CrossRefPubMedGoogle Scholar
  13. 13.
    Grounds MD (1998) Age-associated changes in the responses of skeletal muscle cells to exercise and regeneration. Ann NY Acad Sci 854:78–91PubMedGoogle Scholar
  14. 14.
    Guharay F, Sachs F (1984) Stretch-activated single ion channel currents in tissue-cultured embryonic chick skeletal muscle. J Physiol (Lond) 352:685–701Google Scholar
  15. 15.
    Halevy O, Krispin A, Leshem Y, McMurtry JP, Yahav S (2001) Early-age heat exposure affects skeletal muscle satellite cell proliferation and differentiation in chicks. Am J Physiol 281:R302–R309Google Scholar
  16. 16.
    Harris MB, Starnes JW (2001) Effects of body temperature during exercise training on myocardial adaptations. Am J Physiol 280:H2271–H2280Google Scholar
  17. 17.
    Hernando R, Manso R (1997) Muscle fibre stress in response to exercise: synthesis, accumulation and isoform transitions of 70-kDa heat-shock proteins. Eur J Biochem 243:460–467PubMedGoogle Scholar
  18. 18.
    Jansen W, Haveman J (1990) Histopathological changes in the skin and subcutaneous of mouse legs after treatment with hyperthermia. Pathol Res Pract 186:247–253PubMedGoogle Scholar
  19. 19.
    Jayakumar J, Suzuki K, Sammut IA, Smolenski RT, Khan M, Latif N, Abunasra H, Murtuza B, Amrani M, Yacoub MH (2001) Heat shock protein 70 gene transfection protects mitochondrial and ventricular function against ischemia-reperfusion injury. Circulation 104:I-303–I-307Google Scholar
  20. 20.
    Khassaf M, Child RB, McArdle A, Brodie DA, Esanu C, Jackson MJ (2001) Time course of responses of human skeletal muscle to oxidative stress induced by nondamaging exercise. J Appl Physiol 90:1031–1035Google Scholar
  21. 21.
    Kilgore JL, Musch TI, Ross CR (1998) Physical activity, muscle, and the HSP70 response. Can J Appl Physiol 23:245–260PubMedGoogle Scholar
  22. 22.
    Kippenberger S, Bernd A, Loitsch S, Muller J, Guschel M, Kaufmann R (1999) Cyclic stretch up-regulates proliferation and heat shock protein 90 expression in human melanocytes. Pigment Cell Res 12:246–251PubMedGoogle Scholar
  23. 23.
    Ku Z, Yang J, Menon V, Thomason DB (1995) Decreased polysomal HSP-70 may slow polypeptide elongation during skeletal muscle atrophy. Am J Physiol 268:C1369–C1374PubMedGoogle Scholar
  24. 24.
    Latchman DS (2001) Heat shock proteins and cardiac protection. Cardiovasc Res 51:637–646CrossRefPubMedGoogle Scholar
  25. 25.
    Liu Y, Lormes W, Baur C, Opitz-Gress A, Altenburg D, Lehmann M, Steinacker JM (2000) Human skeletal muscle HSP70 response to physical training depends on exercise intensity. Int J Sports Med 21:351–355CrossRefPubMedGoogle Scholar
  26. 26.
    Locke M, Noble ETG (1995) Stress proteins: the exercise response. Can J Appl Physiol 20:155–167PubMedGoogle Scholar
  27. 27.
    Locke M, Noble EG, Tanguay RM, Field MR, Ianuzzo SE, Ianuzzo CD (1995) Activation of heat-shock transcription factor in rat heart after heat shock and exercise. Am J Physiol 268:C1387–C1394PubMedGoogle Scholar
  28. 28.
    Luo G, Sun X, Hungness E, Hasselgren P-O (2001) Heat shock protects L6 myotubes from catabolic effects of dexamethasone and prevents downregulation of NF-κB. Am J Physiol 281:R1193–R1200Google Scholar
  29. 29.
    Naito H, Powers SK, Demirel HA, Sugiura T, Dodd SL, Aoki J (2000) Heat stress attenuates skeletal muscle atrophy in hindlimb-unweighted rats. J Appl Physiol 88:359–363Google Scholar
  30. 30.
    Oishi Y, Ishihara A, Talmadge RJ, Ohira Y, Taniguchi K, Matsumoto H, Roy RR, Edgerton VR (2001) Expression of HSP72 in atrophied rat skeletal muscles. Acta Physiol Scand 172:123–130CrossRefPubMedGoogle Scholar
  31. 31.
    Oishi Y, Taniguchi K, Matsumoto H, Ishihara A, Ohira Y, Roy RR (2002) Muscle type-specific response of HSP60, HSP72 and HSC73 during recovery after the elevation of muscle temperature. J Appl Physiol 92:1097–1103PubMedGoogle Scholar
  32. 32.
    Okubo S, Wildner O, Shah MR, Chelliah JC, Hess ML, Kukreja RC (2001) Gene transfer of heat-shock protein 70 reduces infarct size in vivo after ischemia/reperfusion in the rabbit heart. Circulation 103:877–881PubMedGoogle Scholar
  33. 33.
    Perrone CE, Fenwick-Smith D, Vandenburgh HH (1995) Collagen and stretch modulate autocrine secretion of insulin-like growth facot-1 and insulin-like growth factor binding proteins from differentiated skeletal muscle cells. J Biol Chem 270:2099–2106CrossRefPubMedGoogle Scholar
  34. 34.
    Puntschart A, Vogt M, Widmer HR, Hoppeler H, Billeter R (1996) Hsp70 expression in human skeletal muscle after exercise. Acta Physiol Scand 157:411–417PubMedGoogle Scholar
  35. 35.
    Sadoshima J, Izumo S (1997) The cellular and molecular response of cardiac myocytes to mechanical stress. Annu Rev Physiol 59:551–571PubMedGoogle Scholar
  36. 36.
    Sadoshima J, Jahn L, Takahashi T, Kulik TJ, Izumo S (1992) Molecular characterization of the stretch-induced adaptation of cultured cardiac cells. An in vitro model of load-induced cardiac hypertrophy. J Biol Chem 267:10551–10560PubMedGoogle Scholar
  37. 37.
    Salo DC, Donovan CM, Davies KJ (1991) HSP70 and other possible heat shock or oxidative stress proteins are induced in skeletal muscle, heart, and liver during exercise. Free Radic Biol Med 11:239–246CrossRefPubMedGoogle Scholar
  38. 38.
    Samelman TR (2000) Heat shock protein expression is increased in cardiac and skeletal muscle of Fisher 344 rats after endurance training. Exp Physiol 85:82–102CrossRefGoogle Scholar
  39. 39.
    Sass JB, Weinberg ES, Krone PH (1996) Specific localization of zebrafish hsp90 alpha mRNA to myoD-expressing cells suggests a role for hsp90 alpha during normal muscle development. Mech Dev 54:195–204PubMedGoogle Scholar
  40. 40.
    Segnitz B, Gehring U (1997) The function of steroid hormone receptors is inhibited by the hsp90-specific compound geldanamycin. J Biol Chem 272:18694–18701CrossRefPubMedGoogle Scholar
  41. 41.
    Skidmore R, Gutierrez JA, Guerriero V Jr, Kregel KC (1995) HSP70 induction during exercise and heat stress in rats: role of internal temperature. Am J Physiol 268:R92–R97PubMedGoogle Scholar
  42. 42.
    Takarada Y, Nakamura Y, Aruga S, Onda T, Miyazaki S, Ishii N (2000) Rapid increase in plasma growth hormone after low-intensity resistance exercise with vascular occlusion. J Appl Physiol 88:61–65PubMedGoogle Scholar
  43. 43.
    Takarada Y, Takazawa H, Sato Y, Takebayashi S, Tanaka Y, Ishii N (2000) Effects of resistance exercise combined with moderate vascular occlusion on muscular function in humans. J Appl Physiol 88:2097–2106Google Scholar
  44. 44.
    Thompson HS, Scordilis SP, Clarkson PM, Lohrer WA (2001) A single bout of exercise increases HSP27 and HSC/HSP70 in human skeletal muscle. Acta Physiol Scand 171:187–193CrossRefPubMedGoogle Scholar
  45. 45.
    Yang H, Alnaqeeb M, Simpson H, Goldspink G (1997) Changes in muscle fibre type, muscle mass and IGF-I gene expression in rabbit skeletal muscle subjected to stretch. J Anat 190:613–622Google Scholar
  46. 46.
    Vandenburgh HH, Shansky J, Solerssi R, Chromiak J (1995) Mechanical stimulation of skeletal muscle increases prostaglandin F2 alpha production, cyclooxygenase activity, and cell growth by a pertussis toxin sensitive mechanism. J Cell Physiol 163:285–294PubMedGoogle Scholar
  47. 47.
    Weitzel G, Pilatus U, Rensing L (1985) Similar dose response of heat shock protein synthesis and intracellular pH change in yeast. Exp Cell Res 159:252–256PubMedGoogle Scholar
  48. 48.
    Wong HR (1998) Potential protective role of the heat shock response in sepsis. New Horiz 6:194–200PubMedGoogle Scholar

Copyright information

© Springer-Verlag  2003

Authors and Affiliations

  • K. Goto
    • 2
  • R. Okuyama
    • 2
  • H. Sugiyama
    • 2
  • M. Honda
    • 2
  • T. Kobayashi
    • 2
  • K. Uehara
    • 2
  • T. Akema
    • 2
  • T. Sugiura
    • 3
  • S. Yamada
    • 4
  • Y. Ohira
    • 1
  • T. Yoshioka
    • 2
    • 5
  1. 1.School of Health and Sport SciencesOsaka UniversityToyonaka City, OsakaJapan
  2. 2.Department of PhysiologySt. Marianna University School of MedicineKawasakiJapan
  3. 3.Faculty of EducationYamaguchi UniversityYamaguchiJapan
  4. 4.Department of Life Science, Graduate School of Arts and ScienceUniversity of TokyoTokyoJapan
  5. 5.Aomori University of Health and WelfareAomoriJapan

Personalised recommendations