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Reduced expression of MyHC slow isoform in rat soleus during unloading is accompanied by alterations of endogenous inhibitors of calcineurin/NFAT signaling pathway

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An Erratum to this article was published on 07 December 2015

Abstract

Under muscle disuse conditions decrease of expression of MyHC of slow type, and sometimes of type IIa, as well as upregulation of expression of IIb and IId/x isoforms were observed. Through dephosphorylation and entry of NFAT molecules to the nucleus calcineurin/NFATc1 signaling pathway promotes upregulation of the slow MyHC expression. We supposed that downregulation of calcineurin pathway took place during unloading. The study was aimed to analyze the states of the myonuclear NFAT inhibitors calsarcin I (CSI) and calsarcin II (CSII) (also referred to as myozenin II and I) and GSK3β in rat soleus during hindlimb suspension (HS). Male Wistar rats were subjected to 3, 7 and 14 day of HS. We found that after 3 days of HS the content of CSII mRNA twofold increased in soleus as compared to the controls. This level was increased by more than fivefold (as compared to control) after 2 weeks of HS. The increase of CSII mRNA expression may be explained as the mechanism of stabilization of fast phenotype. We found that from the 3 day till 14 day of HS the content of MuRF-1 and MuRF-2 in the nuclear fraction fourfold to fivefold increased in HS soleus. We supposed that nuclear import of the MuRFs allows to promote CSII expression during unloading. We also observed the decline of the phosphorylated GSK3β content in the nuclear extract of the soleus tissue. Thus decline of slow MyHC expression characteristic for the unloading conditions is accompanied with the increased expression and activation of the factors known to prevent NFAT accumulation in the myonuclei.

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References

  • Blaauw B, Schiaffino S, Reggiani C (2013) Mechanisms modulating skeletal muscle phenotype. Compr Physiol 3:1645–1687

    Article  PubMed  Google Scholar 

  • Drenning JA, Lira VA, Simmons CG, Soltow QA, Sellman JE, Criswell DS (2008) Nitric oxide facilitates NFAT-dependent transcription in mouse myotubes. Am J Physiol Cell Physiol 294:1088–1095

    Article  Google Scholar 

  • Dupont-Versteegden EE, Knox M, Gurley CM, Houle JD, Peterson CA (2002) Maintenance of muscle mass is not dependent on the calcineurin-NFAT pathway. Am J Physiol Cell Physiol 282(6):C1387–C1395

    Article  CAS  PubMed  Google Scholar 

  • Frey N, Richardson JA, Olson EN (2000) Calsarcins, a novel family of sarcomeric calcineurin-binding proteins. Proc Natl Acad Sci USA 97:14632–14637

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Frey N, Frank D, Lippl S, Kuhn C, Kögler H, Barrientos T, Rohr C, Will R, Müller OJ, Weiler H, Bassel-Duby R, Katus HA, Olson EN (2008) Calsarcin-2 deficiency increases exercise capacity in mice through calcineurin/NFAT activation. J Clin Investig 118:3598–3608

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Gallagher P, Trappe S, Harber M, Creer A, Mazzetti S, Trappe T, Alkner B, Tesch P (2005) Effects of 84-days of bedrest and resistance training on single muscle fibre myosin heavy chain distribution in human vastus lateralis and soleus muscles. Acta Physiol Scand 185:61–69

    Article  CAS  PubMed  Google Scholar 

  • Giger JM, Bodell PW, Zeng M, Baldwin KM, Haddad F (2009) Rapid muscle atrophy response to unloading: pretranslational processes involving MHC and actin. J Appl Physiol 107:1204–1212

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hirner S, Krohne C, Schuster A, Hoffmann S, Witt S, Erber R, Sticht C, Gasch A, Labeit S, Labeit D (2008) MuRF1-dependent regulation of systemic carbohydrate metabolism as revealed from transgenic mouse studies. J Mol Biol 379:666–677

    Article  CAS  PubMed  Google Scholar 

  • Hortobagyi T, Dempsey L, Fraser D, Zheng D, Hamilton G, Lambert J, Dohm L (2000) Changes in muscle strength, muscle fibre size and myofibrillar gene expression after immobilization and retraining in humans. J Physiol 524(1):293–304

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ingalls CP, Warren GL, Armstrong RB (1999) Intracellular Ca2+ transients in mouse soleus muscle after hindlimb unloading and reloading. J Appl Physiol 87(1):386–390

    CAS  PubMed  Google Scholar 

  • Ingalls CP, Wenke JC, Armstrong RB (2001) Time course changes in [Ca2+)i, force, and protein content in hindlimb-suspended mouse soleus muscles. Aviat Space Environ Med 72(5):471–476

    CAS  PubMed  Google Scholar 

  • Kachaeva EV, Shenkman BS (2012) Various jobs of proteolytic enzymes in skeletal muscle during unloading: facts and speculations. J Biomed Biotechnol 2012:493618. doi:10.1155/2012/493618

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lange S, Xiang F, Yakovenko A, Vihola A, Hackman P, Rostkova E, Kristensen J, Brandmeier B, Franzen G, Hedberg B, Gunnarsson LG, Hughes SM, Marchand S, Sejersen T, Richard I, Edström L, Ehler E, Udd B, Gautel M (2005) The kinase domain of titin controls muscle gene expression and protein turnover. Science 308:1599–1603

    Article  CAS  PubMed  Google Scholar 

  • Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(−∆∆C(T)) method. Methods 25:402–408

    Article  CAS  PubMed  Google Scholar 

  • Lomonosova YuN, Shenkman BS, Nemirovskaya TL (2009) Regulation of expression of myosin heavy chains with calcineurin in soleus during decrease of rats physical activity. I.M. Sechenov Physiol J 95(9):969–974

    Google Scholar 

  • Lomonosova YN, Kalamkarov GR, Bugrova AE, Shevchenko TF, Kartashkina NL, Lysenko EA, Shvets VI, Nemirovskaya TL (2011) Protective effect of l-arginine administration on proteins of unloaded m. soleus. Biochemistry (Mosc) 76(5):571–580

    Article  CAS  Google Scholar 

  • Martins KJ, St-Louis M, Murdoch GK, MacLean IM, McDonald P, Dixon WT, Putman CT, Michel RN (2012) Nitric oxide synthase inhibition prevents activity-induced calcineurin–NFATc1 signalling and fast-to-slow skeletal muscle fibre type conversions. J Physiol 590(6):1427–1442

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • McElhinny AS, Kakinuma K, Sorimachi H, Labeit S, Gregorio CC (2002) Muscle-specific RING finger-1 interacts with titin to regulate sarcomeric M-line and thick filament structure and may have nuclear functions via its interaction with glucocorticoid modulatory element binding protein-1. J Cell Biol 157(1):125–136

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Morey-Holton ER, Globus RK (2002) Hindlimb unloading rodent model: technical aspects. J Appl Physiol 92(4):1367–1377

    Article  PubMed  Google Scholar 

  • Moriscot A, Baptista IL, Bogomolovas J, Krohne C, Hirner S, Granzier H, Labeit S (2010) MuRF1 is a muscle fiber type II associated factor and together with MuRF2 regulates type II fiber trophicity and maintenance. J Struct Biol 170(2):344–353

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Novikov VE, Ilyin EA (1981) Age-related reactions of rat bones to their unloading. Aviat Space Environ Med 52(9):551–553

    CAS  PubMed  Google Scholar 

  • Ochala J, Gustafson A-M, Diez ML, Renaud G, Li M, Aare S, Qaisar R, Banduseela VC, Hedstrom Y, Tang X, Dworkin B, Ford GC, Sreekumaran Nair K, Perera S, Gautel M, Larsson L (2011) Preferential skeletal muscle myosin loss in response to mechanical silencing in a novel rat intensive care unit model: underlying mechanisms. J Physiol 589(8):2007–2026

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Pandorf CE, Haddad F, Roy RR, Qin AX, Edgerton VR, Baldwin KM (2006) Dynamics of myosin heavy chain gene regulation in slow skeletal muscle. Role of natural antisense RNA. J Biol Chem 281(50):38330–38342

    Article  CAS  PubMed  Google Scholar 

  • Pandorf CE, Haddad F, Wright C, Bodell PW, Baldwin KM (2009) Differential epigenetic modifications of histones at the myosin heavy chain genes in fast and slow skeletal muscle fibers and in response to muscle unloading. Am J Physiol Cell Physiol 297:6–16

    Article  Google Scholar 

  • Perera S, Mankoo B, Gautel M (2012) Developmental regulation of MURF E3 ubiquitin ligases in skeletal muscle. J Muscle Res Cell Motil 33:107–122

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Salanova M, Bortoloso E, Schiffl G, Gutsmann M, Belavý DL, Felsenberg D, Sandra Furlan S, Volpe P, Blottner D (2011) Expression and regulation of Homer in human skeletal muscle during neuromuscular junction adaptation to disuse and exercise. FASEB J 25:4312–4325

    Article  CAS  PubMed  Google Scholar 

  • Seward DJ, Haney JC, Rudnicki MA, Swoap SJ (2001) bHLH transcription factor MyoD affects myosin heavy chain expression pattern in a muscle-specific fashion. Am J Physiol Cell Physiol 85:408–413

    Google Scholar 

  • Shen T, Cseresnyes Z, Liu Y, Randall WR, Schneider MF (2007) Regulation of the nuclear export of the transcription factor NFATc1 by protein kinases after slow fibre type electrical stimulation of adult mouse skeletal muscle fibres. J Physiol 579(2):535–551

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Shenkman BS, Nemirovskaya TL (2008) Calcium-dependent signaling mechanisms and soleus fiber remodeling under gravitational unloading. J Muscle Res Cell Motil 29(6–8):221–230

    Article  CAS  PubMed  Google Scholar 

  • Shenkman BS, Podlubnaya ZA, Vikhlyantsev IM, Litvinova KS, Udaltsov SN, Nemirovskaya TL, Lemesheva YuS, Mukhina AM, Kozlovskaya IB (2004) Contractile characteristics and sarcomeric cytoskeletal proteins of human soleus fibers in muscle unloading: role of mechanical stimulation from the support surface. Biophysics 49(5):807–815

    Google Scholar 

  • Stevens L, Gohlsch B, Mounier Y, Pette D (1999a) Changes in myosin heavy chain mRNA and protein isoforms in single fibers of unloaded rat soleus muscle. FEBS Lett 463:15–18

    Article  CAS  PubMed  Google Scholar 

  • Stevens L, Sultan KR, Peuker H, Gohlsch B, Mounier Y, Pette D (1999b) Time-dependent changes in myosin heavy chain mRNA and protein isoforms in unloaded soleus muscle of rat. Am J Physiol Cell Physiol 46:1044–1049

    Google Scholar 

  • Talmadge RJ (2000) Myosin heavy chain isoform expression following reduced neuromuscular activity: potential regulatory mechanisms. Muscle Nerve 23:661–679

    Article  CAS  PubMed  Google Scholar 

  • Trappe S, Costill D, Gallagher P, Creer A, Peters JR, Evans H, Riley DA, Fitts RH (2009) Exercise in space: human skeletal muscle after 6 months aboard the International Space Station. J Appl Physiol 106:1159–1168

    Article  PubMed  Google Scholar 

  • Witt SH, Granzier H, Witt CC, Labeit S (2005) MURF-1 and MURF-2 target a specific subset of myofibrillar proteins redundantly: towards understanding MURF-dependent muscle ubiquitination. J Mol Biol 350(4):713–722

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

The authors would like to express their sincere gratitude to Dr. Siegfried Labeit (Mannheim, Germany) for generously sharing antibodies against MuRF-1 and MuRF-2. We would also like to extend our thanks to Anton Pokusaev and Alexandra Minailova who helped us with processing the biosamples. The work was supported by a Russian Science Foundation Grant No. 14-15-00358.

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  1. Institute of Biomedical Problems, Russian Academy of Sciences, 76a, Khoroshevskoye Shosse, Moscow, Russia, 123007

    Yulia N. Lomonosova, Olga V. Turtikova & Boris S. Shenkman

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  1. Yulia N. Lomonosova
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  2. Olga V. Turtikova
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  3. Boris S. Shenkman
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Correspondence to Boris S. Shenkman.

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Lomonosova, Y.N., Turtikova, O.V. & Shenkman, B.S. Reduced expression of MyHC slow isoform in rat soleus during unloading is accompanied by alterations of endogenous inhibitors of calcineurin/NFAT signaling pathway. J Muscle Res Cell Motil 37, 7–16 (2016). https://doi.org/10.1007/s10974-015-9428-y

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