Skip to main content
SpringerLink
Log in

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

  • Skeletal Muscle
  • Published:
Pflügers Archiv Aims and scope Submit manuscript

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Bornman L, Polla BS, Gericke GS (1996) Heat-shock protein 90 and ubiquitin: developmental regulation during myogenesis. Muscle Nerve 19:574–580

    Article  PubMed  Google Scholar 

  2. Carlson BM, Faulkner JA (1983) The regeneration of skeletal muscle fibers following injury: a review. Med Sci Sports Exerc 15:187–198

    CAS  PubMed  Google Scholar 

  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–C1448

    PubMed  Google Scholar 

  4. Chang J, Wasser JS, Cornelussen RN, Knowlton AA (2001) Activation of heat-shock factor by stretch-activated channels in rat hearts. Circulation 104:209–214

    PubMed  Google Scholar 

  5. Clarke MS, Feeback DL (1996) Mechanical load induces sarcoplasmic wounding and FGF release in differentiated human skeletal muscle cultures. FASEB J 10:502–509

    CAS  PubMed  Google Scholar 

  6. Clarkson PM, Sayers SP (1999) Etiology of exercise-induced muscle damage. Can J Appl Physiol 24:234–248

    CAS  PubMed  Google Scholar 

  7. Cotto JJ, Morimoto RI (1999) Stress-induced activation of heat-shock response: cell and molecular biology of heat-shock factors. Biochem Soc Symp 64:105–118

    CAS  PubMed  Google Scholar 

  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 82:460–468

    CAS  PubMed  Google Scholar 

  9. Ecochard L, Lhenry F, Sempore B, Favier R (2000) Skeletal muscle HSP72 level during endurance training: influence of peripheral arterial insufficiency. Pflugers Arch 440:918–924

    Article  CAS  PubMed  Google Scholar 

  10. Einat MF, Haberfeld A, Shamay A, Horev G, Hurwitz S, Yahav S (1996) A novel 29-kDa chicken heat shock protein. Poult Sci 75:1528–1530

    CAS  PubMed  Google Scholar 

  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 194:323–334

    Article  CAS  PubMed  Google Scholar 

  12. Gonzalez B, Hernando R, Manso R (2000) Stress proteins of 70 kDa in chronically exercised skeletal muscle. Pflugers Arch 440:42–49

    Article  CAS  PubMed  Google Scholar 

  13. Grounds MD (1998) Age-associated changes in the responses of skeletal muscle cells to exercise and regeneration. Ann NY Acad Sci 854:78–91

    CAS  PubMed  Google Scholar 

  14. Guharay F, Sachs F (1984) Stretch-activated single ion channel currents in tissue-cultured embryonic chick skeletal muscle. J Physiol (Lond) 352:685–701

    Google Scholar 

  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–R309

    CAS  Google Scholar 

  16. Harris MB, Starnes JW (2001) Effects of body temperature during exercise training on myocardial adaptations. Am J Physiol 280:H2271–H2280

    CAS  Google Scholar 

  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 243:460–467

    CAS  PubMed  Google Scholar 

  18. Jansen W, Haveman J (1990) Histopathological changes in the skin and subcutaneous of mouse legs after treatment with hyperthermia. Pathol Res Pract 186:247–253

    CAS  PubMed  Google Scholar 

  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-307

    Google Scholar 

  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 90:1031–1035

    Google Scholar 

  21. Kilgore JL, Musch TI, Ross CR (1998) Physical activity, muscle, and the HSP70 response. Can J Appl Physiol 23:245–260

    CAS  PubMed  Google Scholar 

  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 12:246–251

    CAS  PubMed  Google Scholar 

  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–C1374

    CAS  PubMed  Google Scholar 

  24. Latchman DS (2001) Heat shock proteins and cardiac protection. Cardiovasc Res 51:637–646

    Article  CAS  PubMed  Google Scholar 

  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 21:351–355

    Article  CAS  PubMed  Google Scholar 

  26. Locke M, Noble ETG (1995) Stress proteins: the exercise response. Can J Appl Physiol 20:155–167

    PubMed  Google Scholar 

  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–C1394

    CAS  PubMed  Google Scholar 

  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–R1200

    CAS  Google Scholar 

  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 88:359–363

    Google Scholar 

  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 172:123–130

    Article  CAS  PubMed  Google Scholar 

  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 92:1097–1103

    CAS  PubMed  Google Scholar 

  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 103:877–881

    CAS  PubMed  Google Scholar 

  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 270:2099–2106

    Article  CAS  PubMed  Google Scholar 

  34. Puntschart A, Vogt M, Widmer HR, Hoppeler H, Billeter R (1996) Hsp70 expression in human skeletal muscle after exercise. Acta Physiol Scand 157:411–417

    CAS  PubMed  Google Scholar 

  35. Sadoshima J, Izumo S (1997) The cellular and molecular response of cardiac myocytes to mechanical stress. Annu Rev Physiol 59:551–571

    CAS  PubMed  Google Scholar 

  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 267:10551–10560

    CAS  PubMed  Google Scholar 

  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 11:239–246

    Article  CAS  PubMed  Google Scholar 

  38. Samelman TR (2000) Heat shock protein expression is increased in cardiac and skeletal muscle of Fisher 344 rats after endurance training. Exp Physiol 85:82–102

    Article  Google Scholar 

  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 54:195–204

    CAS  PubMed  Google Scholar 

  40. Segnitz B, Gehring U (1997) The function of steroid hormone receptors is inhibited by the hsp90-specific compound geldanamycin. J Biol Chem 272:18694–18701

    Article  CAS  PubMed  Google Scholar 

  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–R97

    CAS  PubMed  Google Scholar 

  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 88:61–65

    CAS  PubMed  Google Scholar 

  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 88:2097–2106

    Google Scholar 

  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 171:187–193

    Article  CAS  PubMed  Google Scholar 

  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 190:613–622

    Google Scholar 

  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 163:285–294

    CAS  PubMed  Google Scholar 

  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 159:252–256

    CAS  PubMed  Google Scholar 

  48. Wong HR (1998) Potential protective role of the heat shock response in sepsis. New Horiz 6:194–200

    CAS  PubMed  Google Scholar 

Download references

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.

Author information

Authors and Affiliations

  1. School of Health and Sport Sciences, Osaka University, 560-0043, Toyonaka City, Osaka, Japan

    Y. Ohira

  2. Department of Physiology, St. Marianna University School of Medicine, 216-8511, Kawasaki, Japan

    K. Goto, R. Okuyama, H. Sugiyama, M. Honda, T. Kobayashi, K. Uehara, T. Akema & T. Yoshioka

  3. Faculty of Education, Yamaguchi University, 753-8513, Yamaguchi, Japan

    T. Sugiura

  4. Department of Life Science, Graduate School of Arts and Science, University of Tokyo, 153-8902, Tokyo, Japan

    S. Yamada

  5. Aomori University of Health and Welfare, 030-8505, Aomori, Japan

    T. Yoshioka

Authors
  1. K. Goto
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. R. Okuyama
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. H. Sugiyama
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. M. Honda
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. T. Kobayashi
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. K. Uehara
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. T. Akema
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. T. Sugiura
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. S. Yamada
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Y. Ohira
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. T. Yoshioka
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Y. Ohira.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Goto, K., Okuyama, R., Sugiyama, H. et al. Effects of heat stress and mechanical stretch on protein expression in cultured skeletal muscle cells. Pflugers Arch - Eur J Physiol 447, 247–253 (2003). https://doi.org/10.1007/s00424-003-1177-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00424-003-1177-x

Keywords

Search

Navigation