Skip to main content
SpringerLink
Log in

Polymorphisms and DNA methylation level in the CpG site of the GHR1 gene associated with mRNA expression, growth traits and hormone level of half-smooth tongue sole (Cynoglossus semilaevis)

  • Published:
Fish Physiology and Biochemistry Aims and scope Submit manuscript

Abstract

The objectives of the present study were to estimate the GHR1 gene mutations and methylation status of CpGs, and whether those mutations and methylation were involved in the regulation of GHR1 gene expression, hormone level and growth traits in half-smooth tongue sole (Cynoglossus semilaevis). Identification of single-nucleotide polymorphisms was performed on 43 male fish. Through polymerase chain reaction-single-strand conformation polymorphism and sequencing, two SNPs were found. SNP1 [c.G1357A (p.Val376Ile)] creating one CpG site located in exon 8 was named L1 locus, and SNP2 (c.G1479A) located in exon 9 was named L2 locus. Individuals were divided into three genotypes, AA, AG and GG according to L1 locus (GG genotype had one more CpG site because of the mutation), and into two genotypes, AA- and GG-based on L2 locus. The results showed that only L1 locus was significantly associated with body weight (P < 0.01), gonad weight (P ≤ 0.05), triiodothyronine (T3) level (P ≤ 0.05) and mRNA expression (P < 0.01). At L1 locus, newly created CpG site in GG genotype was highly methylated (93.3 %), while there was no difference of methylation level in the other two CpG sites among three genotypes. AA genotype and AG genotype having higher T3 level were significantly different (P ≤ 0.05) from GG genotype. There were significant differences among body weights of AA, AG and GG genotypes (P < 0.01). Gonad weights of AA genotype and AG genotype were significantly lower than GG genotype. The GHR1 mRNA expression of GG genotype was significantly lower than AA and AG genotypes (P < 0.01). These implied that mutations and methylation status of GHR1 gene might influence the hormone level, growth traits and gene expression in male half-smooth tongue sole and the L1 locus could be regarded as a potential candidate genetic and epigenetic marker in half-smooth tongue sole selection.

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
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  • Bahrami A, Behzadi S, Miraei-Ashtiani SR, Roh S-G, Katoh K (2013) Genetic polymorphisms and protein structures in growth hormone, growth hormone receptor, ghrelin, insulin-like growth factor 1 and leptin in Mehraban sheep. Gene 527(1):397–404

    Article  CAS  PubMed  Google Scholar 

  • Burton JL, McBride BW, Block E, Glimm DR, Kenelly J (1994) A review of bovine growth hormone. Can J Anim Sci 74:167–201

    Article  CAS  Google Scholar 

  • Calduch-Giner JA, Mingarro M, Vega-Rubin de Celis S, Boujard D, Perez-Sanchez J (2003) Molecular cloning and characterization of gilthead sea bream (Sparus aurata) growth hormone receptor (GHR). Assessment of alternative splicing. Comp Biochem Physiol B: Biochem Mol Biol 136:1–13

    Article  Google Scholar 

  • Chen SL, Zhang GJ, Shao CW, Huang QF, Liu G, Zhang P, Song WT, An N, Chalopin D, Volff J-N, Hong YH, Li QY, Sha ZX, Zhou HL, Xie MS, Yu QL, Liu Y, Xiang H, Wang N, Wu K, Yang CG, Zhou Q, Liao XL, Yang LF, Hu QM, Zhang JL, Meng L, Jin LJ, Tian YS, Lian JM, Yang JF, Miao GD, Liu SS, Liang Z, Yan F, Li YZ, Sun B, Zhang H, Zhang J, Zhu Y, Du M, Zhao YW, Schart M, Tang QS, Wang J (2014) Whole-genome sequence of a flatfish provides insights into ZW sex chromosome evolution and adaptation to a benthic lifestyle. Nat Genet 46(3):53–260

    Article  Google Scholar 

  • Devlin AM, Singh R, Wade RE, Innis SM, Bottiglieri T, Lentz SR (2005) Hypermethylation of Fads2 and altered hepatic fatty acid and phospholipid metabolism in mice with hyperhomocysteinemia. J Biol Chem 282:37082–37090

    Article  Google Scholar 

  • Ding YX, He F, Wen HS, Li JF, Qian K, Chi ML, Ni M (2012) Polymorphism in exons CpG rich regions of the cyp17-II gene affecting its mRNA expression and reproductive endocrine levels in female Japanese flounder (Paralichthys olivaceus). Gen Comp Endocrinol 179:107–114

    Article  CAS  PubMed  Google Scholar 

  • Fox BK, Breves JP, Davis LK, Pierce AL, Hirano T, Grau EG (2010) Tissue-specific regulation of the growth hormone/insulin-like growth factor axis during fasting and re-feeding: importance of muscle expression of IGF-I and IGF-II mRNA in the tilapia. Gen Comp Endocrinol 166(3):573–580

    Article  CAS  PubMed  Google Scholar 

  • Fuentes EN, Valdés JA, Molina A, Björnsson BT (2013) Regulation of skeletal muscle growth in fish by the growth hormone–insulin-like growth factor system. Gen Comp Endocrinol 192:36–148

    Article  Google Scholar 

  • Fukada H, Ozaki Y, Pierce AL, Adachi S, Yamauchi Y, Hara A, Swanson P, Dickhoff WW (2004) Salmon growth hormone receptor: molecular cloning, ligand specificity, and response to fasting. Gen Comp Endocrinol 139:61–71

    Article  CAS  PubMed  Google Scholar 

  • Glenn KL, Grapes L, Suwanasopee T, Harris DL, Li Y, Wilson K, Rothschild MF (2005) SNP analysis of AMY2 and CTSL genes in Litopenaeus vannamei and Penaeus monodon shrimp. Anim Genet 36:235–236

    Article  CAS  PubMed  Google Scholar 

  • Godowski PJ, Leung DW, Meacham LR, Galgani JP, Hellmiss R, Keret R, Rotwein PS, Parks JS, Laron Z, Wood WI (1989) Characterization of the human growth hormone receptor gene and demonstration of a partial gene deletion in two patients with Laron-type dwarfism. Proc Natl Acad Sci USA 86:8083–8087

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Jiao B, Huang X, Chan CB, Zhang L, Wang D, Cheng CH (2006) The co-existence of two growth hormone receptors in teleost fish and their differential signal transduction, tissue distribution and hormonal regulation of expression in sea bream. J Mol Endocrinol 36:23–40

    Article  CAS  PubMed  Google Scholar 

  • Kopchick JJ, Andry JM (2000) Growth hormone (GH), GH receptor, and signal transduction. Mol Genet Metab 71(1):293–314

    Article  CAS  PubMed  Google Scholar 

  • Korytko AI, Cuttler L (1997) Thyroid hormone and glucocorticoid regulation of pituitary growth hormone-releasing hormone receptor gene expression. J Endocrinol 152:R13–R17

    Article  CAS  PubMed  Google Scholar 

  • Li Y, Liu XC, Zhang Y, Zhu P, Lin HR (2007) Molecular cloning, characterization and distribution of two types of growth hormone receptor in orange-spotted grouper (Epinephelus coioides). Gen Comp Endocrinol 152(1):111–122

    Article  CAS  PubMed  Google Scholar 

  • Liu Y, Pan D, Xu J (2012) Tissue expression, polymorphism identification of IL10 gene and their relationship with serum IL10 level in pigs. Livest Sci 144:247–252

    Article  Google Scholar 

  • Ma XL, Liu XC, Zhang Y (2007) Two growth hormone receptors in Nile tilapia (Oreochromis niloticus): molecular characterization, tissue distribution and expression profiles during different gonad developing stages. Comp Biochem Physiol B: Biochem Mol Biol 147:325–339

    Article  Google Scholar 

  • Ma Q, Liu SF, Zhuang ZM, Sun ZZ, Liu CL, Tang QS (2012a) The co-existence of two growth hormone receptors and their differential expression profiles between female and male tongue sole (Cynoglossus semilaevis). Gene 511(2):341–352

    Article  CAS  PubMed  Google Scholar 

  • Ma RQ, He F, Wen HS, Li JF, Mu WJ, Liu M, Zhang YQ, Hu J, Liu Q (2012b) Polymorphysims of CYP17-Gene in the exons were associated with the reproductive endocrine of Japanese founder (Paralichthys olivaceus). Asian Aust J Anim Sci 25(6):794–799

    Article  CAS  Google Scholar 

  • Maj A, Strzałkowska N, Słoniewski K, Krzyzewski J, Oprzadek J, Zwierzchowski L (2004) Single nucleotide polymorphism (SNP) in the 5′-noncoding region of the bovine growth hormone receptor gene and its association with dairy production traits in Polish Black-and-White cattle, Czech. J Anim Sci 49:419–429

    CAS  Google Scholar 

  • Malveiro E, Pereira M, Marques PX, Santos IC, Belo C, Renavile R, Cravador A (2011) Polymorphisms at the five exons of the growth hormone gene in the algarvia goat: the possible association with milk traits. Small Rumin Res 41:163–170

    Article  Google Scholar 

  • Ni J, You F, Yu SH, Zhang PJ, Xu DD, Wu ZH, Wen AY, Xu YL (2008) Microsatellite polymorphism analysis of GHR gene promoter region in Paralichthys olivaceus hatchery stocks and its association with growth traits. Periodic Ocean Univ China 38(5):719–725

    CAS  Google Scholar 

  • Ozaki Y, Fukada H, Kazeto Y, Adachi S, Hara A, Yamauchi K (2006) Molecular cloning and characterization of growth hormone receptor and its homologue in the Japanese eel (Auguilla japonica). Comp Biochem Physiol B: Biochem Mol Biol 143:422–431

    Article  Google Scholar 

  • Porterfield SP, Henderson CE (1993) The role of thyroid hormones in prenatal and neonatal neurological development current perspectives. Endocr Rev 14:94–106

    CAS  PubMed  Google Scholar 

  • Saera-Vila A, Calduch-Giner JA, Perez-Sanchez J (2005) Duplication of growth hormone receptor (GHR) in fish genome: gene organization and transcriptional regulation of GHR type I and II in gilthead sea bream (Sparus aurata). Gen Comp Endocrinol 142:193–203

    Article  CAS  PubMed  Google Scholar 

  • Saera-Vila A, Calduch-Giner JA, Pérez-Sánchez J (2007) Co-expression of IGFs and GH receptors (GHRs) in gilthead sea bream (Sparus aurata L.): sequence analysis of the GHR-flanking region. J Endocrinol 194(2):361–372

    Article  CAS  PubMed  Google Scholar 

  • Saera-Vila A, Calduch-Giner JA, Prunet P, Pérez-Sánchez J (2009) Dynamics of liver GH/IGF axis and selected stress markers in juvenile gilthead sea bream (Sparus aurata) exposed to acute confinement: differential stress response of growth hormone receptors. Comp Biochem Physiol A: Mol Integr Physiol 154(2):197–203

    Article  Google Scholar 

  • Schmid AC, Lutz L, Kloas W, Reinecke M (2003) Thyroid hormone stimulates hepatic IGF-I mRNA expression in a bony fish, the tilapia Oreochromis mossambicus, in vitro and in vivo. Gen Comp Endocrinol 130:129–134

    Article  CAS  PubMed  Google Scholar 

  • Sharma A, Dutt G, Sivakingam J, Singh MK, Pathodiya OP, Khadda BS, Dixit SP (2013) Novel SNPs in IGF1, GHR and IGFBP-3 genes reveal significant association with growth traits in India goat breeds. Small Rumin Res 115:7–14

    Article  Google Scholar 

  • Tanamati F, da Silva SCC, Rodriguez MDPR, Schuroff GP, do Nascimento CS CS, Del Vesco Ap, Gasparino E (2015) GHR and IGF-I gene expression and production characteristics associated with GH gene polymorphism in Nile tilapia. Aquaculture 435:195–199

    Article  CAS  Google Scholar 

  • Tao WJ, Ma LJ, Ruan RX, Yu JH (2011) SNP loci associated with weight gain on growth hormone receptor genes in Cyprinus carpio var. Jian. Acta Hydrobiol Sin 35(4):622–629

    CAS  Google Scholar 

  • Tollet P, Enberg B, Mode A (1990) Growth hormone (GH) regulation of cytochrome P-450IIC1, insulin-like growth factor-I (IGF-I), and GH receptor messenger RNA expression in primary rat hepatocytes: a hormonal interplay with insulin, IGF-I, and thyroid hormone. Mol Endocrinol 4:1934–1942

    Article  CAS  PubMed  Google Scholar 

  • Vanselow J, Fürbass R (2011) The bovine genome contains three differentially methylated paralogous copies of the P450c17 encoding gene (CYP17A1). Gen Comp Endocrinol 170:475–479

    Article  CAS  PubMed  Google Scholar 

  • Wang LF, Li J, Ruan YY (2013) Sequencing ASMT identifies rare mutations in Chinese Han patients with autism. PLoS ONE 8:e53727

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Xu YX, Zhu ZY, Lo LC, Wang CM, Lin G, Feng F, Yue GH (2006) Characterization of two parvalbumin genes and their association with growth traits in Asian seabass (Lates calcarifer). Anim Genet 37:266–268

    Article  CAS  PubMed  Google Scholar 

  • Yamano K, Takano-Ohmuro H, Obinata T, Inui Y (1994) Effect of thyroid hormone on developmental transition of myosin light chains during flounder metamorphosis. Gen Comp Endocrinol 93:321–326

    Article  CAS  PubMed  Google Scholar 

  • Zeng C, Liu XL, Wang WM, Tong JG, Luo W, Zhang J, Gao ZX (2014) Characterization of GHRs, IGFs and MSTNs, and analysis of their expression relationships in blunt snout bream, Megalobrama amblycephala. Gene 535(2):239–249

    Article  CAS  PubMed  Google Scholar 

  • Zhang L, Huang XG, Jiao BW, Wu FR, Wang DS (2006) The structure analysis, tissue distribution and hormonal regulation of two distinct growth hormone receptors in southern catfish Silurus meridionalis. Acta Zool Sin 52(6):1096–1106

    CAS  Google Scholar 

  • Zhang Y, Zhu Z, Xu Q, Chen G (2014) Association of polymorphisms of exon 2 of the growth hormone gene with production performance in huoyan goose. Int J Mol Sci 15(1):670–683

    Article  PubMed Central  PubMed  Google Scholar 

  • Zhao R, Muehlbauer E, Decuypere E, Grossmann R (2004) Effect of genotype–nutrition interaction on growth and somatotropic gene expression in the chicken. Gen Comp Endocrinol 136(1):2–11

  • Zhong H, Xiao J, Chen W, Zhou Y, Tang Z, Guo Z, Luo Y, Lin Z, Gan X, Zhang M (2014) DNA methylation of pituitary growth hormone is involved in male growth superiority of Nile tilapia (Oreochromis niloticus). Comp Biochem Physiol B: Biochem Mol Biol 171:42–48

    Article  CAS  Google Scholar 

  • Zhou Y, Jiang H (2006) A milk trait-associated polymorphism in the bovine growth hormone receptor does not affect receptor signaling. J Dairy Sci 89:1761–1764

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by State 863 High-Technology R&D Project of China (2012AA10A403) and National Nature Science Foundation of China (31130057).

Author information

Author notes

    Authors and Affiliations

    1. The Key Laboratory of Mariculture, Ministry of Education, Ocean University of China, Qingdao, China

      J. L. Zhao, Y. F. Si, F. He, H. S. Wen, J. F. Li, Y. Y. Ren, M. L. Zhao & Z. J. Huang

    2. Fisheries College, Ocean University of China, Qingdao, China

      J. L. Zhao, Y. F. Si, F. He, H. S. Wen, J. F. Li, Y. Y. Ren, M. L. Zhao & Z. J. Huang

    3. Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao, China

      S. L. Chen

    Authors
    1. J. L. Zhao
      View author publications

      You can also search for this author in PubMed Google Scholar

    2. Y. F. Si
      View author publications

      You can also search for this author in PubMed Google Scholar

    3. F. He
      View author publications

      You can also search for this author in PubMed Google Scholar

    4. H. S. Wen
      View author publications

      You can also search for this author in PubMed Google Scholar

    5. J. F. Li
      View author publications

      You can also search for this author in PubMed Google Scholar

    6. Y. Y. Ren
      View author publications

      You can also search for this author in PubMed Google Scholar

    7. M. L. Zhao
      View author publications

      You can also search for this author in PubMed Google Scholar

    8. Z. J. Huang
      View author publications

      You can also search for this author in PubMed Google Scholar

    9. S. L. Chen
      View author publications

      You can also search for this author in PubMed Google Scholar

    Corresponding author

    Correspondence to F. He.

    Additional information

    J. L. Zhao and Y. F. Si have contributed equally to this work.

    Rights and permissions

    Reprints and permissions

    About this article

    Check for updates. Verify currency and authenticity via CrossMark

    Cite this article

    Zhao, J.L., Si, Y.F., He, F. et al. Polymorphisms and DNA methylation level in the CpG site of the GHR1 gene associated with mRNA expression, growth traits and hormone level of half-smooth tongue sole (Cynoglossus semilaevis). Fish Physiol Biochem 41, 853–865 (2015). https://doi.org/10.1007/s10695-015-0052-y

    Download citation

    • Received:

    • Accepted:

    • Published:

    • Issue Date:

    • DOI: https://doi.org/10.1007/s10695-015-0052-y

    Keywords

    Search

    Navigation