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Molecular and Cellular Biochemistry

, Volume 160, Issue 1, pp 231–239 | Cite as

HSP25 in isolated perfused rat hearts: Localization and response to hyperthermia

  • Brigitte Hoch
  • Gudrun Lutsch
  • Wolfgang-Peter Schlegel
  • Joachim Stahl
  • Gerd Wallukat
  • Sabine Bartel
  • Ernst-Georg Krause
  • Rainer Benndorf
  • Peter Karczewski
Article

Abstract

Recent investigations concentrate on the correlation between the myocardial expression of the inducible 70-kDa heat shock protein (HSP70i) by different stress conditions and its possible protective effects. Only few studies have focused on the involvement of small heat shock proteins in this process. We analyzed the location of the small heat shock protein HSP25 in isolated cardiomyocytes as well as its location and induction in isolated perfused hearts of rats. By immunofluorescence microscopy HSP25 was found to colocalize with actin in the I-band of myofibrils in cardiomyocytes of isolated perfused hearts as well as in isolated neonatal and adult cardiomyocytes. Hyperthermic perfusion of isolated hearts for 45 min resulted in modulation of different parameters of heart function and in induction of HSP25 and HSP70i. Temperatures higher than 43°C (44–46°C) were lethal with respect to the contractile function of the hearts. Compared to control hearts perfused at 37°C, significant increases during hyperthermic perfusion at 42°C and 43°C were obtained for heart rate, contraction velocity and relaxation velocity. In response to hyperthermia at 43°C and after subsequent normothermic perfusion for 135 min at 37°C, left ventricular pressure, contraction velocity and relaxation velocity remained significantly elevated. However, heart rate returned to control values immediately after the period of heat treatment. HSP25 is constitutively expressed even in normothermic perfused hearts as shown by Western blotting. Hyperthermia increased the content of HSP25 only in the left ventricular tissue. In contrast, HSP70i was strongly induced in all analyzed parts of the myocardium (left ventricle, right ventricle, septum). Our findings suggest a differential regulation of HSP25 and HSP70i expression in response to hyperthermia in isolated perfused hearts. The constitutively expressed HSP25 seems to be located adjacent to the myofibrils which implies a specific role of this protein even under unstressed conditions for the contractile function of the myocardium.

Key words

stress protein induction HSP25 intracellular location isolated perfused heart hyperthermia contractile function 

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References

  1. 1.
    Yellon DM, Latchman DS: Stress proteins and myocardial protection. J Mol Cell Cardiol 24: 113–124, 1992Google Scholar
  2. 2.
    Morimoto RI, Tissières A, Georgopoulos C: The Biology of Heat Shock Proteins and Molecular Chaperones. Cold Spring Harbor Laboratory Press, New York, 1994Google Scholar
  3. 3.
    Hahn GM, Li GC: Thermotolerance, thermoresistance and thermosensitization. In: RI Morimoto, A Tissières, C Georgopoulos (eds). Stress Proteins in Biology and Medicine. Cold Spring Harbor Laboratory Press, New York, 1990, pp 79–100Google Scholar
  4. 4.
    Polla BS, Mili N, Kantengwa S: Heat shock and oxidative injury in human cells. In: B Maresca, S Lindquist (eds). Heat Shock. Berlin, Springer-Verlag, 1991, pp 279–290Google Scholar
  5. 5.
    Iwaki K, Chi S-H, Dillmann WH, Mestril R: Induction of HSP70 in cultured rat neonatal cardiomyocytes by hypoxia and metabolic stress. Circulation 87: 2023–2032, 1993Google Scholar
  6. 6.
    Mestril R, Chi S-H, Sayen MR, O'Reilly K, Dillmann WH: Expression of inducible stress protein 70 in rat heart myogenic cells confers protection against simulated ischemia-induced injury. J Clin Invest 93:759–767,1994Google Scholar
  7. 7.
    Heads RJ, Latchman DS, Yellon DM: Stable high level expression of a transfected human HSP70 gene protects a heart-derived muscle cell line against thermal stress. J Mol Cell Cardiol 26: 695–699, 1994Google Scholar
  8. 8.
    Heads RJ, Yellon DM, Latchman DS: Differential cytoprotection against heat stress or hypoxia following expression of specific stress protein genes in myogenic cells. J Mol Cell Cardiol 27: 1669–1678, 1995Google Scholar
  9. 9.
    Currie RW, Karmazyn M, Kloc M, Mailer K: Heat shock response is associated with enhanced postischemic ventricular recovery. Circ Res 63:543–549,1988Google Scholar
  10. 10.
    Donnelly TJ, Sievers RE, Vissern FLJ, Welch WJ, Wolfe CL: Heat shock protein induction in rat hearts. Circulation 85: 769–778, 1992Google Scholar
  11. 11.
    Currie RW, Tanguay RM, Kingma Jr JG: Heat shock response and limitation of tissue necrosis during occlusion/reperfusion in rabbit hearts. Circulation 87: 963–971, 1993Google Scholar
  12. 12.
    Marber MS, Walker JM, Latchman DS, Yellon DM: Myocardial protection after whole body heat stress in the rabbit is dependent on metabolic substrate and is related to the amount of the inducible 70-kD heat stress protein. J Clin Invest 93: 1087–1094, 1994Google Scholar
  13. 13.
    Hutter MM, Sievers RE, Barbosa V, Wolfe CL: Heat-shock protein induction in rat hearts. Circulation 89: 355–360, 1994Google Scholar
  14. 14.
    Currie RW: Effects of ischemia and perfusion temperature on the synthesis of stress-induced (heat shock) proteins in isolated and perfused rat hearts. J Mol Cell Cardiol 19: 795–808, 1987Google Scholar
  15. 15.
    Plumier J-CL, Ross BM, Currie RW, Angelidis CE, Kazlaris H, Kollias G, Pagoulatos GN: Transgenic mice expressing the human heat shock protein 70 have improved post-ischemic myocardial recovery. J Clin Invest 95: 1854–1860, 1995Google Scholar
  16. 16.
    Marber MS, Mestril R, Chi S-H, Sayen MR, Yellon DM, Dillmann W H: Overexpression of the rat inducible 70-kD heat stress protein in a transgenic mouse increases the resistance of the heart to ischemic injury. J Clin Invest 95: 1446–1456, 1995Google Scholar
  17. 17.
    Currie RW, Ross BM, Davis TA: Induction of the heat shock response in rats modulates heart rate, creatine kinase and protein synthesis after a subsequent hyperthermic treatment. Cardiovasc Res 24: 87–93, 1990Google Scholar
  18. 18.
    Amrani M, Allen NJ, O'Shea J, Corbett J, Dunn MJ, Tadjkarimi S, Theodoropoulos S, Pepper J, Yacoub MH: Role of catalase and heat shock protein on recovery of endothelial and mechanical function. Cardioscience 4: 193–198, 1993Google Scholar
  19. 19.
    Karmazyn M, Mailer K, Currie RW: Acquisition and decay of heatshock-enhanced postischemic ventricular recovery. Am J Physiol 259: H424-H431,1990Google Scholar
  20. 20.
    Walker DM, Pasini E, Kucukoglu S, Marber MS, Iliodromitis E, Ferrari R, Yellon DM: Heat stress limits infarct size in the isolated perfused rabbit heart. Cardiovasc Res 27: 962–967, 1993Google Scholar
  21. 21.
    McCully JD, Myrmel T, Lotz MM, Krukenkamp IB, Levitsky S: The rapid expression of myocardial HSP70 mRNA and the heat shock 70 kDa protein can be achieved after only a brief period of retrograde hyperthermic perfusion. J Mol Cell Cardiol 27: 873–882, 1995Google Scholar
  22. 22.
    Das DK, Maulik N, Moraru II: Gene expression in acute myocardial stress. Induction by hypoxia, ischemia, reperfusion, hyperthermia and oxidative stress. J Mol Cell Cardiol 27: 181–193, 1995Google Scholar
  23. 23.
    Uoshima K, Handelman B, Cooper LF: Isolation and characterization of a rat HSP27 gene. Biochem Biophys Res Commun 197: 1388–1395, 1993Google Scholar
  24. 24.
    Knowlton AA: The role of heat shock proteins in the heart. J Mol Cell Cardiol 27: 121–131, 1995Google Scholar
  25. 25.
    Mestril R, Dillmann WH: Heat shock proteins and protection against myocardial ischemia. J Mol Cell Cardiol 27: 45–52, 1995Google Scholar
  26. 26.
    Welch WJ: Phorbol ester, calcium ionophore, or serum added to quiescent rat fibroblast cells all result in the elevated phosphorylation of two 28000 dalton mammalian stress proteins. J Biol Chem 260: 3058–3062,1985Google Scholar
  27. 27.
    Arrigo A-P: Tumor necrosis factor induces the rapid phosphorylation of the mammalian heat shock protein hsp28. Mol Cell Biol 10: 1276–1280, 1980Google Scholar
  28. 28.
    Gaestel M, Schröder W, Benndorf R, Lippmann C, Buchner K, Hucho F, Erdmann VA, Bielka H: Identification of the phosphorylation sites of the murine small heat shock protein hsp25. J Biol Chem 266: 14721–14724, 1991Google Scholar
  29. 29.
    Saklatvala J, Kaur P, Guesdon F: Phosphorylation of the small heatshock protein is regulated by interleukin 1, tumour necrosis factor, growth factors, bradykinin and ATP. Biochem J 277: 635–642, 1991Google Scholar
  30. 30.
    Mehlen P, Kretz-Remy C, Briolay J, Fostan P, Mirault M-E, Arrigo AP: Intracellular reactive oxygen species as apparent modulators of heatshock protein 27 (hsp27) structural organization and phosphorylation in basal and tumour necrosis factor α-treated T47D human carcinoma cells. Biochem J 312: 367–375, 1995Google Scholar
  31. 31.
    Behlke J, Dube P, van Heel M, Wieske M, Hayeß K, Benndorf R, Lutsch G: Supramolecular structure of the small heat shock protein HSP25. Prog Colloid Polym Sci 99: 87–93, 1995Google Scholar
  32. 32.
    Miron T, Vancompernolle K, Vandekerckhove J, Wilchek M, Geiger B: A 25-kD inhibitor of actin polymerization is a low molecular mass heat shock protein. J Cell Biol 114: 255–261, 1991Google Scholar
  33. 33.
    Rahman DRJ, Bentley N, Tuite MF: The Saccharomyces cerevisiae small heat shockprotein HSP26 inhibits actin polymerization. Biochem Society Transactions 23: 77S, 1995Google Scholar
  34. 34.
    Lavoie JN, Hickey E, Weber LA, Landry J: Modulation of actin microfilament dynamics and fluid phase pinocytosis by phosphorylation of heat-shock protein 27. J Biol Chem 268: 24210–24214, 1993Google Scholar
  35. 35.
    Benndorf R, Hayeß K, Ryazantsev S, Wieske M, Behlke J, Lutsch G: Phosphorylation and supramolecular organization of murine small heat shock protein HSP25 abolish its actin polymerization-inhibiting activity. J Biol Chem 269: 20780–20784, 1994Google Scholar
  36. 36.
    Lavoie JN, Lambert H, Hickey E, Weber LA, Landry J: Modulation of cellular thermoresistance and actin filament stability accompanies phosphorylation-induced changes in the oligomeric structure of heat shock protein 27. Mol Cell Biol 15: 505–516, 1995Google Scholar
  37. 37.
    Kato K, Shinohara H, Goto S, Inaguma Y, Morishita R, Asano T: Copurification of small heat shock protein with αB-crystallin from human skeletal muscle. J Biol Chem 267: 7718–7725, 1992Google Scholar
  38. 38.
    Das DK, Engelman RM, Kimura Y: Molecular adaptation of cellular defences following preconditioning of the heart by repeated ischemia. Cardiovasc Res 27: 578–584, 1993Google Scholar
  39. 39.
    Currie RW, Tanguay RM: Analysis of RNA for transcripts for catalase and SP 71 in rat hearts after in vivo hyperthermia. Biochem Cell Biol 69:375–382,1991Google Scholar
  40. 40.
    Andres J, Sharma HS, Knöll R, Stahl J, Sassen LMA, Verdouw PD, Schaper W: Expression of heat shock proteins in the normal and stunned porcine myocardium. Cardiovasc Res 27: 1421–1429, 1993Google Scholar
  41. 41.
    Volz A, Piper HM, Siegmund B, Schwartz P: Longevity of adult ventricular rat heart muscle cells -in serum-free primary culture. J Mol Cell Cardiol 23: 161–173, 1991Google Scholar
  42. 42.
    Halle W, Wollenberger A: Differentiation and behaviour of isolated embryonic and neonatal heart cells in a chemically definite medium. Am J Cardiol 25: 292–299, 1970Google Scholar
  43. 43.
    Wallukat G, Morwinski R, Kuhn H: Modulation of the β adrenergic response of cardiomyocytes by specific lipoxygenase products involves their incorporation into phosphatidylinositol and activation of protein kinase C. J Biol Chem 269: 29055–29060, 1994Google Scholar
  44. 44.
    Schlegel W-P, Bartel S, Karczewski P, Krause E-G: Interaction of distinct signal transduction systems in the heart: Studies on contractility in isolated rat heart. In: Freiburg Focus on Biomeasurement Vol. 9: Pharmacological Evaluation of Cardioprotective Substances. Experimental Induction and Indicators of Myocardial Injury and Myocardial Protection. Publisher: Gesellschaft für Erfahrungstransfer in der Biomesstechnik e.V, Buchenbach, Germany; Biomesstechnik-Verlag, March, Germany, 1995, pp 71–77Google Scholar
  45. 45.
    Vielkind U, Swierenga SH: A simple fixation procedure for immunofluorescent detection of different cytoskeletal components within the same cell. Histochem 91: 81–88, 1989Google Scholar
  46. 46.
    Stahl J, Wobus AM, Ihrig S, Lutsch G, Bielka H: The small heat shock protein HSP25 is accumulated in P19 embryonal carcinoma cells and embryonic stem cells of line BLC6 during differentiation. Differentiation 51: 33–37, 1992Google Scholar
  47. 47.
    Tokuyasu KT: Application of cryoultramicrotomy to immunocytochemistry. J Microscopy 143: 139–149, 1986Google Scholar
  48. 48.
    Benndorf R, Nürnberg P, Bielka H: Growth phase-dependent proteins of the Ehrlich ascites tumor analyzed by one- and two-dimensional electrophoresis. Exp Cell Res 174: 130–138, 1988Google Scholar
  49. 49.
    Bitar KN, Kaminski MS, Hailat N, Cease KB, Strahler JR: HSP27 is a mediator of sustained smooth muscle contraction in response to bombesin. Biochem Biophys Res Commun 181: 1192–1200, 1991Google Scholar
  50. 50.
    Lutsch G, Vetter R, Stahl J, Benndorf R: Localization of HSP25 in the Iband of cardiac myofibrils of adult rat. J Mol Cell Cardiol 27: A111, 1995Google Scholar
  51. 51.
    Yamaguchi N, Champlain J, Nadeau R: Correlation between the response of the heart to sympathetic stimulation and the release of endogenous catecholamine into coronary sinus of the dog. Ore Res 36: 662–668, 1975Google Scholar
  52. 52.
    Wollenberger A, Krause E-G, Shahab L: Endogenous catecholamine mobilization and the shift to anaerobic energy production in the acutely ischemic myocardium. Int Symp Coronary Circulation and Energetics of the Myocardium, Karger, Basel/New York, 1966, pp 200–219Google Scholar
  53. 53.
    Krause E-G, Lehmann I, Mest H-J, Taube C, Förster W: Electrically induced tachyarrhythmia and the effect of propranolol on the release of cyclic AMP and prostaglandin E by the canine left ventricle. Adv Myocardiol 3: 185–191, 1982Google Scholar

Copyright information

© Kluwer Academic Publishers 1996

Authors and Affiliations

  • Brigitte Hoch
    • 1
  • Gudrun Lutsch
    • 1
  • Wolfgang-Peter Schlegel
    • 1
  • Joachim Stahl
    • 1
  • Gerd Wallukat
    • 1
  • Sabine Bartel
    • 1
  • Ernst-Georg Krause
    • 1
  • Rainer Benndorf
    • 1
  • Peter Karczewski
    • 1
  1. 1.Max Delbrück Centre for Molecular MedicineBerlin-BuchGermany

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