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Fish Physiology and Biochemistry

, Volume 41, Issue 4, pp 853–865 | Cite as

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)

  • J. L. Zhao
  • Y. F. Si
  • F. He
  • H. S. Wen
  • J. F. Li
  • Y. Y. Ren
  • M. L. Zhao
  • Z. J. Huang
  • S. L. Chen
Article

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.

Keywords

GHR gene SNPs DNA methylation Gene expression Growth traits Half-smooth tongue sole (Cynoglossus semilaevis

Notes

Acknowledgments

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

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Copyright information

© Springer Science+Business Media Dordrecht 2015

Authors and Affiliations

  • J. L. Zhao
    • 1
    • 2
  • Y. F. Si
    • 1
    • 2
  • F. He
    • 1
    • 2
  • H. S. Wen
    • 1
    • 2
  • J. F. Li
    • 1
    • 2
  • Y. Y. Ren
    • 1
    • 2
  • M. L. Zhao
    • 1
    • 2
  • Z. J. Huang
    • 1
    • 2
  • S. L. Chen
    • 3
  1. 1.The Key Laboratory of Mariculture, Ministry of EducationOcean University of ChinaQingdaoChina
  2. 2.Fisheries CollegeOcean University of ChinaQingdaoChina
  3. 3.Yellow Sea Fisheries Research InstituteChinese Academy of Fishery SciencesQingdaoChina

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