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Low-protein diet fed to crossbred sows during pregnancy and lactation enhances myostatin gene expression through epigenetic regulation in skeletal muscle of weaning piglets

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Abstract

Purpose

This study was aimed to investigate the effects of a maternal low-protein diet on transcriptional regulation of the myostatin (MSTN) gene in skeletal muscle of weaning piglets.

Methods

Sows were fed either a standard-protein (SP, 15 and 18 % crude protein) or a low-protein (LP, 50 % protein level of SP) diet throughout pregnancy and lactation. Longissimus dorsi muscle was sampled from male piglets at 28 days of age. The mRNA was determined by RT-PCR, and protein was measured by Western blot. Chromatin immunoprecipitation assay was used to determine the binding of transcription factors and histone H3 modifications on the MSTN gene promoter.

Results

The maternal LP diet significantly decreased body weight and average daily gain (P < 0.05), which was associated with significantly lower plasma concentration of urea nitrogen and total protein (P < 0.05), as well as decreased muscle RNA content (P < 0.05). MSTN mRNA (P < 0.05) was significantly increased, together with enhanced (P < 0.05) mRNA and protein expression of forkhead box class O family member protein 3 (FoxO3), and a tendency of an increase (P = 0.10) in glucocorticoid receptor (GR) mRNA in the muscle of LP piglets. Furthermore, the binding of both FoxO3 and GR to the MSTN gene promoter was significantly higher (P < 0.05) in muscle of LP piglets, together with significantly enriched (P < 0.05) gene activation markers, H3K9Ac and H3K4me3.

Conclusion

These results indicate that MSTN mediates maternal LP diet-induced growth retardation, through epigenetic regulation involving FoxO3 and GR binding to its promoter.

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Abbreviations

ChIP:

Chromatin immunoprecipitation

FoxO3:

Forkhead box class O family member protein 3

GR:

Glucocorticoid receptor

LM:

Longissimus dorsi muscle

LP:

Low protein

MSTN:

Myostatin

SP:

Standard protein

References

  1. Joulia D, Bernardi H, Garandel V, Rabenoelina F, Vernus B, Cabello G (2003) Mechanisms involved in the inhibition of myoblast proliferation and differentiation by myostatin. Exp Cell Res 286:263–275

    Article  CAS  Google Scholar 

  2. Welle S, Burgess K, Thornton CA, Tawil R (2009) Relation between extent of myostatin depletion and muscle growth in mature mice. Am J Physiol Endocrinol Metab 297:E935–E940

    Article  CAS  Google Scholar 

  3. Wang Q, McPherron AC (2012) Myostatin inhibition induces muscle fibre hypertrophy prior to satellite cell activation. J Physiol 590:2151–2165

    Article  CAS  Google Scholar 

  4. Hennebry A, Berry C, Siriett V, O’Callaghan P, Chau L, Watson T, Sharma M, Kambadur R (2009) Myostatin regulates fiber-type composition of skeletal muscle by regulating MEF2 and MyoD gene expression. Am J Physiol Cell Physiol 296:C525–C534

    Article  CAS  Google Scholar 

  5. Reisz-Porszasz S, Bhasin S, Artaza JN, Shen R, Sinha-Hikim I, Hogue A, Fielder TJ, Gonzalez-Cadavid NF (2003) Lower skeletal muscle mass in male transgenic mice with muscle-specific overexpression of myostatin. Am J Physiol Endocrinol Metab 285:E876–E888

    Article  CAS  Google Scholar 

  6. Zhu X, Topouzis S, Liang LF, Stotish RL (2004) Myostatin signaling through Smad2, Smad3 and Smad4 is regulated by the inhibitory Smad7 by a negative feedback mechanism. Cytokine 26:262–272

    Article  CAS  Google Scholar 

  7. Yang W, Chen Y, Zhang Y, Wang X, Yang N, Zhu D (2006) Extracellular signal-regulated kinase 1/2 mitogen-activated protein kinase pathway is involved in myostatin-regulated differentiation repression. Cancer Res 66:1320–1326

    Article  CAS  Google Scholar 

  8. Sandri M, Sandri C, Gilbert A, Skurk C, Calabria E, Picard A, Walsh K, Schiaffino S, Lecker SH, Goldberg AL (2004) Foxo transcription factors induce the atrophy-related ubiquitin ligase atrogin-1 and cause skeletal muscle atrophy. Cell 117:399–412

    Article  CAS  Google Scholar 

  9. Peiris HN, Ponnampalam AP, Osepchook CC, Mitchell MD, Green MP (2010) Placental expression of myostatin and follistatin-like-3 protein in a model of developmental programming. Am J Physiol Endocrinol Metab 298:E854–E861

    Article  CAS  Google Scholar 

  10. Reed SA, Raja JS, Hoffman ML, Zinn SA, Govoni KE (2014) Poor maternal nutrition inhibits muscle development in ovine offspring. J Anim Sci Biotechnol 5(1):43

    Article  Google Scholar 

  11. Liu X, Wang J, Li R, Yang X, Sun Q, Albrecht E, Zhao R (2011) Maternal dietary protein affects transcriptional regulation of myostatin gene distinctively at weaning and finishing stages in skeletal muscle of Meishan pigs. Epigenetics 6:899–907

    Article  CAS  Google Scholar 

  12. McMillen IC, Robinson JS (2005) Developmental origins of the metabolic syndrome: prediction, plasticity, and programming. Physiol Rev 85:571–633

    Article  CAS  Google Scholar 

  13. Jones PA (2012) Functions of DNA methylation: islands, start sites, gene bodies and beyond. Nat Rev Genet 13:484–492

    Article  CAS  Google Scholar 

  14. Shahbazian MD, Grunstein M (2007) Functions of site-specific histone acetylation and deacetylation. Annu Rev Biochem 76:75–100

    Article  CAS  Google Scholar 

  15. Gupta S, Kim SY, Artis S, Molfese DL, Schumacher A, Sweatt JD, Paylor RE, Lubin FD (2010) Histone methylation regulates memory formation. J Neurosci 30:3589–3599

    Article  CAS  Google Scholar 

  16. Huang Y, Greene E, Murray Stewart T, Goodwin AC, Baylin SB, Woster PM, Casero RA Jr (2007) Inhibition of lysine-specific demethylase 1 by polyamine analogues results in reexpression of aberrantly silenced genes. Proc Natl Acad Sci USA 104:8023–8028

    Article  CAS  Google Scholar 

  17. Bettini ML, Pan F, Bettini M, Finkelstein D, Rehg JE, Floess S, Bell BD, Ziegler SF, Huehn J, Pardoll DM, Vignali DA (2012) Loss of epigenetic modification driven by the Foxp3 transcription factor leads to regulatory T cell insufficiency. Immunity 36:717–730

    Article  CAS  Google Scholar 

  18. Watson ML, Baehr LM, Reichardt HM, Tuckermann JP, Bodine SC, Furlow JD (2012) A cell-autonomous role for the glucocorticoid receptor in skeletal muscle atrophy induced by systemic glucocorticoid exposure. Am J Physiol Endocrinol Metab 302:E1210–E1220

    Article  CAS  Google Scholar 

  19. Allen DL, Unterman TG (2007) Regulation of myostatin expression and myoblast differentiation by FoxO and SMAD transcription factors. Am J Physiol Cell Physiol 292:C188–C199

    Article  CAS  Google Scholar 

  20. Johnson LR, Chandler AM (1973) RNA and DNA of gastric and duodenal mucosa in antrectomized and gastrin-treated rats. Am J Physio 224:937–940

    CAS  Google Scholar 

  21. Dwyer CM, Stickland NC, Fletcher JM (1994) The influence of maternal nutrition on muscle fiber number development in the porcine fetus and on subsequent postnatal growth. J Anim Sci 72:911–917

    CAS  Google Scholar 

  22. Zambrano E, Bautista CJ, Deas M, Martinez-Samayoa PM, Gonzalez-Zamorano M, Ledesma H, Morales J, Larrea F, Nathanielsz PW (2006) A low maternal protein diet during pregnancy and lactation has sex- and window of exposure-specific effects on offspring growth and food intake, glucose metabolism and serum leptin in the rat. J Physiol 571:221–230

    Article  CAS  Google Scholar 

  23. Addis T, Barrett E, Poo LJ, Yuen DW (1947) The relation between the serum urea concentration and the protein consumption of normal individuals. J Clin Invest 26:869–874

    Article  CAS  Google Scholar 

  24. Wu G, Pond WG, Ott T, Bazer FW (1998) Maternal dietary protein deficiency decreases amino acid concentrations in fetal plasma and allantoic fluid of pigs. J Nutr 128:894–902

    CAS  Google Scholar 

  25. Millward DJ, Garlick PJ, James WP, Nnanyelugo DO, Ryatt JS (1973) Relationship between protein synthesis and RNA content in skeletal muscle. Nature 241:204–205

    Article  CAS  Google Scholar 

  26. Schakman O, Kalista S, Barbe C, Loumaye A, Thissen JP (2013) Glucocorticoid-induced skeletal muscle atrophy. Int J Biochem Cell Biol 45:2163–2172

    Article  CAS  Google Scholar 

  27. Ma K, Mallidis C, Artaza J, Taylor W, Gonzalez-Cadavid N, Bhasin S (2001) Characterization of 5′-regulatory region of human myostatin gene: regulation by dexamethasone in vitro. Am J Physiol Endocrinol Metab 281:E1128–E1136

    CAS  Google Scholar 

  28. Ma K, Mallidis C, Bhasin S, Mahabadi V, Artaza J, Gonzalez-Cadavid N, Arias J, Salehian B (2003) Glucocorticoid-induced skeletal muscle atrophy is associated with upregulation of myostatin gene expression. Am J Physiol Endocrinol Metab 285:E363–E371

    Article  CAS  Google Scholar 

  29. Du R, An XR, Chen YF, Qin J (2007) Some motifs were important for myostatin transcriptional regulation in sheep (Ovis aries). J Biochem Mol Biol 40:547–553

    Article  CAS  Google Scholar 

  30. Qin J, Du R, Yang YQ, Zhang HQ, Li Q, Liu L, Guan H, Hou J, An XR (2013) Dexamethasone-induced skeletal muscle atrophy was associated with upregulation of myostatin promoter activity. Res Vet Sci 94:84–89

    Article  CAS  Google Scholar 

  31. Mammucari C, Milan G, Romanello V, Masiero E, Rudolf R, Del Piccolo P, Burden SJ, Di Lisi R, Sandri C, Zhao J, Goldberg AL, Schiaffino S, Sandri M (2007) FoxO3 controls autophagy in skeletal muscle in vivo. Cell Metab 6:458–471

    Article  CAS  Google Scholar 

  32. Han DS, Huang HP, Wang TG, Hung MY, Ke JY, Chang KT, Chang HY, Ho YP, Hsieh WY, Yang WS (2010) Transcription activation of myostatin by trichostatin A in differentiated C2C12 myocytes via ASK1-MKK3/4/6-JNK and p38 mitogen-activated protein kinase pathways. J Cell Biochem 111:564–573

    Article  CAS  Google Scholar 

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Acknowledgments

We thank Shanghai Farm of Bright Food (Group) Co., Ltd., for providing the experimental site and Rongkui Zhang for care of animals. In addition, we thank Dr. Wanyne J. Kuenzel of University of Arkansas for his suggestions on paper writing, especially for language revision. This work was supported by the National Basic Research Program of China (2012CB124703), the Special Fund for Agro-scientific Research in the Public Interest (201003011), and the Priority Academic Program Development of Jiangsu Higher Education Institutions.

Conflict of interest

Yimin Jia, Guichao Gao, Haogang Song, Demin Cai, Xiaojing Yang, and Ruqian Zhao declare no conflicts of interest.

Ethical standards

Operating procedures for animal breeding and husbandry were in accordance with Technical Regulations for Commercial Pig Production for Intensive Pig Farms (GB/T 17824.2-2008). The experimental protocol was approved by the Animal Ethics Committee of Nanjing Agricultural University with the project number 2012CB124703. The slaughter and sampling procedures complied with the “Guidelines on Ethical Treatment of Experimental Animals” (2006) No. 398 set by the Ministry of Science and Technology, China.

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Authors and Affiliations

  1. Key Laboratory of Animal Physiology and Biochemistry, Nanjing Agricultural University, Nanjing, 210095, People’s Republic of China

    Yimin Jia, Guichao Gao, Haogang Song, Demin Cai, Xiaojing Yang & Ruqian Zhao

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  1. Yimin Jia
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  2. Guichao Gao
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  3. Haogang Song
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Correspondence to Ruqian Zhao.

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Jia, Y., Gao, G., Song, H. et al. Low-protein diet fed to crossbred sows during pregnancy and lactation enhances myostatin gene expression through epigenetic regulation in skeletal muscle of weaning piglets. Eur J Nutr 55, 1307–1314 (2016). https://doi.org/10.1007/s00394-015-0949-3

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