Advertisement

Fish Physiology and Biochemistry

, Volume 42, Issue 2, pp 407–421 | Cite as

Genetic polymorphisms and DNA methylation in exon 1 CpG-rich regions of PACAP gene and its effect on mRNA expression and growth traits in half smooth tongue sole (Cynoglossus semilaevis)

  • Yufeng Si
  • Feng He
  • Haishen Wen
  • Jifang Li
  • Junli Zhao
  • Yuanyuan Ren
  • Meilin Zhao
  • Liqin Ji
  • Zhengju Huang
  • Mo Zhang
  • Songlin Chen
Article

Abstract

The pituitary adenylate cyclase activating polypeptide (PACAP) is a new type of hypophysiotropic hormone and plays an important role in regulating the synthesis and secretion of growth hormone and gonadotropin. The research on the relationship between PACAP and different growth traits would contribute to explain its function during the process of growth. Moreover, epigenetic modifications, especially DNA methylation at the CpG sites of the SNPs, play important roles in regulating gene expression. The results suggest that a SNP mutation (c.C151G) in the PACAP gene of male half smooth tongue sole (Cynoglossus semilaevis) is significantly associated with growth traits and serum physiological and biochemical parameters such as inorganic phosphorus (P < 0.05). The SNP is located in a CpG-rich region of exon 1. Intriguingly, the transition (C→G) added a new methylation site of PACAP gene. This SNP was also significantly related to the expression and methylation level of PACAP (P < 0.05). Individuals with GG genotype had faster growth rates than those of CG and CC genotypes. Moreover, GG genotype had significantly higher PACAP expression level and lower methylation level than CG and CC genotypes. In the serum indexes, only inorganic phosphorus content within GG genotypes was significantly higher than CC genotypes. This implied that the mutation and methylation status of PACAP gene could influence growth traits and this locus could be considered as a candidate genetic or epigenetic marker for Cynoglossus semilaevis molecular breeding.

Keywords

PACAP SNP DNA methylation Growth Molecular markers 

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).

References

  1. Amills M, Jimenez N, Villalba D, Tor M, Molina E, Cubilo D, Marcos C, Francesch A, Sanchez A, Estany J (2003) Identification of three single nucleotide polymorphisms in the chicken insulin-like growth factor 1 and 2 genes and their associations with growth and feeding traits. Poult Sci 82:1485–1493CrossRefPubMedGoogle Scholar
  2. Aubert N, Falluel-Morel A, Vaudry D, Xifro X, Rodriguez-Alvarez J, Fisch C, De Jouffrey S, Lebigot J, Fournier A, Vaudry H, Gonzalez BJ (1996) PACAP and C2-ceramide generate different AP-1 complexes through a MAP-kinase-dependent pathway: involvement of c-Fos in PACAP-induced Bcl-2 expression. J Neurochem 99:1237–1250CrossRefGoogle Scholar
  3. Bahrami A, Behzadi S, Miraei-Ashtiani SR, Roh SG, 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 527:397–404CrossRefPubMedGoogle Scholar
  4. Chen SL, Li J, Deng SP, Tian YS, Wang QY, Zhuang ZM, Sha ZX, Xu JY (2007) Isolation of female-specific AFLP markers and molecular identification of genetic sex in half-smooth tongue sole (Cynoglossus semilaevis). Mar Biotechnol 9:273–280CrossRefPubMedGoogle Scholar
  5. Chen CF, Wen HS, Wang ZP, He F, Zhang JR, Chen XY, Jin GX, Shi B, Shi D, Yang YP, Li JF, Qi BX, Li N (2010) Cloning and expression of P450c17-I (17α-hydroxylase/17,20-lyase) in brain and ovary during gonad development in Cynoglossus semilaevis. Fish Physiol Biochem 36:1001–1012CrossRefPubMedGoogle Scholar
  6. Cummings KJ, Gray SL, Simmonsa CJT, Kozak CA, Sherwood NM (2002) Mouse pituitary adenylate cyclase-activating polypeptide (PACAP): gene, expression and novel splicing. Mol Cell Endocrinol 192:133–145CrossRefPubMedGoogle Scholar
  7. Dai XY, Zhang W, Zhuo ZJ, He JY, Yin Z (2015) Neuroendocrine regulation of somatic growth in fishes. Sci China Life Sci 58:137–147CrossRefPubMedGoogle Scholar
  8. Ding YX, He F, Wen HS, Li JF, Qian K, Chi ML, Ni M, Yin XH, Bu Y, Zhao YJ, Zhang DQ (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 179:107–114CrossRefPubMedGoogle Scholar
  9. Gardiner-Garden M, Frommer M (1987) CpG islands in vertebrate genomes. J Mol Biol 196(2):261–282CrossRefPubMedGoogle Scholar
  10. Ge W, Davis ME, Hines HC, Irvin KM, Simmen RCM (2003) Association of single nucleotide polymorphisms in the growth hormone and growth hormone receptor genes with blood serum insulin-like growth factor I concentration and growth traits in Angus cattle. J Anim Sci 81:641–648PubMedGoogle Scholar
  11. Gui JF, Zhu ZY (2012) Molecular basis and genetic improvement of economically important traits in aquaculture animals. Chin Sci Bull 57:1751–1760CrossRefGoogle Scholar
  12. He F, Wen HS, Dong SL, Shi B, Chen CF, Wang LS, Yao J, Mu XJ, Zhou YG (2008a) Identification of single nucleotide polymorphism cytochrome P450-c19a and its relation to reproductive traits in Japanese flounder (Paralichthys olivaceus). Aquaculture 279:177–181CrossRefGoogle Scholar
  13. He F, Wen HS, Dong SL, Wang LS, Chen CF, Shi B, Mu XJ, Yao J, Zhou YG (2008b) Identification of estrogen receptor α gene polymorphisms by SSCP and its effect on reproductive traits in Japanese flounder (Paralichthys olivaceus). Comp Biochem Physiol B Biochem Mol Biol 150:278–283CrossRefPubMedGoogle Scholar
  14. He F, Wen HS, Li JF, Yu DH, Ma RQ, Shi D, Mu WJ, Zhang YQ, Hu JA, Liu MA, Han WG, Zhang JN, Wang QQ, Yuan YR, Liu Q (2011) Single Nucleotide Polymorphisms of the GnRHR gene associated with reproductive traits of Japanese flounder (Paralichthys olivaceus). Asian Australas J Anim 24(4):463–470CrossRefGoogle Scholar
  15. Herman JG, Latif F, Weng Y, Lerman MI, Zbar B, Liu S, Samid D, Duan DS, Gnarra JR, Linehan WM (1994) Silencing of the VHL tumor-suppressor gene by DNA methylation in renal carcinoma. PNAS 91:9700–9704CrossRefPubMedPubMedCentralGoogle Scholar
  16. Huang H, Wei Y, Meng Z, Zhang Y, Liu X, Guo L, Jian L, Chen G, Lin H (2014) Polymorphisms of Leptin-b gene associated with growth traits in Orange-spotted grouper (Epinephelus coioides). Int J Mol Sci 15:11996–12006CrossRefPubMedPubMedCentralGoogle Scholar
  17. Ji XS, Chen SL, Jiang YL, Xu TJ, Yang JF, Tian YS (2011) Growth differences and differential expression analysis of pituitary adenylate cyclase activating polypeptide (PACAP) and growth hormone-releasing hormone (GHRH) between the sexes in half-smooth tongue sole Cynoglossus semilaevis. Gen Comp Endocrinol 170(1):99–109CrossRefPubMedGoogle Scholar
  18. Karin M, Liu Z, Zandi E (1997) AP-1 function and regulation. Curr Opin Cell Biol 9:240–246CrossRefPubMedGoogle Scholar
  19. Laird PW (2010) Principles and challenges of genome-wide DNA methylation analysis. Nat Rev Genet 11:191–203CrossRefPubMedGoogle Scholar
  20. Liu ZJ, Cordes JF (2004) DNA marker technologies and their applications in aquaculture genetics. Aquaculture 238:1–37CrossRefGoogle Scholar
  21. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25:402–408CrossRefPubMedGoogle Scholar
  22. Luo C, Zhou A, Han Y (2004) Effects of supplementary inorganic phosphorus on performance and nutrient digestibility of growing pigs fed wheat bran diets. Acta Zoonutrimenta Sin 3:50–55Google Scholar
  23. Ma Q, Liu SF, Zhuang ZM, Lin L, Sun ZZ, Liu CL, Su YQ, Tang QS (2011) Genomic structure, polymorphism and expression analysis of growth hormone-releasing hormone and pituitary adenylate cyclase activating polypeptide genes in the half-smooth tongue sole (Cynoglossus semilaevis). Genet Mol Res 10(4):3828–3846CrossRefPubMedGoogle Scholar
  24. Ma RQ, He F, Wen HS, Li JF, Mu WJ, Liu M, Zhang YQ, Hu J, Liu Q (2012) Polymorphysims of CYP17-I gene in the exons were associated with the reproductive endocrine of Japanese Flounder (Paralichthys olivaceus). Asian Australas J Anim 25(6):794–799CrossRefGoogle Scholar
  25. Miyata A, Arimura A, Dahl RR, Minamino N, Uehara A, Jiang L, Culler MD, Coy DH (1989) Isolation of a novel 38 residue-hypothalamic polypeptide which stimulates adenylate cyclase in pituitary cells. Biochem Biophys Res Commun 164:567–574CrossRefPubMedGoogle Scholar
  26. Miyata A, Jiang L, Dahl RD, Kitada C, Kubo K, Fujino M, Minamino N, Arimura A (1990) Isolation of a neuropeptide corresponding to the N-terminal 27 residues of the pituitary adenylate cyclase activating polypeptide with 38 residues (PACAP38). Biochem Biophys Res Commun 170:643–648CrossRefPubMedGoogle Scholar
  27. Montero M, Yon L, Rousseau K, Arimura A, Fournier A, Dufour S, Vaudry H (1998) Distribution, characterization, and growth hormone-releasing activity of pituitary adenylate cyclase-activating polypeptide in the European Eel, Anguilla anguilla. Endocrinology 139:4300–4310PubMedGoogle Scholar
  28. Parker DB, Power ME, Swanson P, Rivier J, Sherwood NM (1997) Exon skipping in the gene encoding pituitary adenylate cyclase-activating polypeptide in salmon alters the expression of two hormones that stimulate growth hormone release. Endocrinology 138:414–423PubMedGoogle Scholar
  29. Pfeifer GP (2000) p53 mutational spectra and the role of methylated CpG sequences. Mutat Res 450:155–166CrossRefPubMedGoogle Scholar
  30. Rousseau K, Le Belle N, Pichavant K, Marchelidon J, Chow BKC, Boeuf G, Dufour S (2001) Pituitary growth hormone secretion in the turbot, a phylogenetically recent teleost, is regulated by a species-specific pattern of neuropeptides. Neuroendocrinology 74:375–385CrossRefPubMedGoogle Scholar
  31. Schäfer H, Zheng J, Gundlach F, Günther R, Schmidt WE (1996) PACAP stimulates transcription of c-Fos and c-Jun and activates the AP-1 transcription factor in rat pancreatic carcinoma cells. Biochem Biophys Res Commun 221:111–116CrossRefPubMedGoogle Scholar
  32. Shi B, Wen HS, He F, Dong SL, Ma S, Chen CF, Wang LS, Yao J, Mu XJ, Zhou YG (2009) Association of reproductive performance with SNPs of FOXL2 gene by SSCP in Japanese flounder (Paralichthys olivaceus). Comp Biochem Physiol B Biochem Mol Biol 153:1–7CrossRefPubMedGoogle Scholar
  33. Si YF, Ding YX, He F, Wen HS, Li JF, Zhao JL, Huang ZJ (2015) DNA methylation level of cyp19a1a and Foxl2 gene related to their expression patterns and reproduction traits during ovary development stages of Japanese flounder (Paralichthys olivaceus). Gene. doi: 10.1016/j.gene.2015.09.006 Google Scholar
  34. Tachibana M, Matsumura Y, Fukuda M, Kimura H, Shinkai Y (2008) G9a/GLP complexes independently mediate H3K9 and DNA methylation to silence transcription. EMBO J 27:2681–2690CrossRefPubMedPubMedCentralGoogle Scholar
  35. Tanamati F, Silva SCCDS, Rodriguez MDPR, Schuroff GP, Nascimento CSD, Vesco APD, Gasparino E (2015) GHR and IGF-I gene expression and production characteristics associated with GH gene polymorphism in Nile tilapia. Aquaculture 435:195–199CrossRefGoogle Scholar
  36. Tao W, Boulding E (2003) Associations between single nucleotide polymorphisms in candidate genes and growth rate in Arctic charr (Salvelinus alpinus L.). Heredity 91:60–69CrossRefPubMedGoogle Scholar
  37. Tian C, Yang M, Lv L, Yuan Y, Liang X, Guo W, Song Y, Zhao C (2014) Single nucleotide polymorphisms in growth hormone gene and their association with growth traits in Siniperca chuatsi (Basilewsky). Int J Mol Sci 15:7029–7036CrossRefPubMedPubMedCentralGoogle Scholar
  38. Tsai HY, Hamilton A, Guy DR, Houston RD (2014) Single nucleotide polymorphisms in the insulin-like growth factor 1 (IGF1) gene are associated with growth-related traits in farmed Atlantic salmon. Anim Genet 45:709–715CrossRefPubMedPubMedCentralGoogle Scholar
  39. Vaudry D, Falluel-Morel A, Bourgault S, Basille M, Burel D, Wurtz O, Fournier A, Chow BKC, Hashimoto H, Galas L, Vaudry H (2009) Pituitary adenylate cyclase-activating polypeptide and its receptors: 20 years after the discovery. Pharmacol Rev 61:283–357CrossRefPubMedGoogle Scholar
  40. Wang Y, Wang X, Meng X, Wang H, Jiang Z, Qiu X (2014) Identification of two SNPs in myostatin (MSTN) gene of Takifugu rubripesand their association with growth traits. Mol Cell Probe 28:200–203CrossRefGoogle Scholar
  41. Wen HS, Si YF, Zhang YQ, He F, Li JF (2014) Cloning and expression of follistatin gene in half-smooth tongue sole Cynoglossus semilaevis during the reproduction cycle. Chin J Oceanol Limnol 33(2):299–308CrossRefGoogle Scholar
  42. Wilkie GS, Dickson KS, Gray NK (2003) Regulation of mRNA translation by 5′- and 3′-UTR-binding factors. Trends Biochem Sci 28:182–188CrossRefPubMedGoogle Scholar
  43. Wong AOL, Leung MY, Shea WLC, Tse LY, Chang JP, Chow BKC (1998) Hypophysiotropic action of pituitary adenylate cyclase-activating polypeptide (PACAP) in the goldfish: immunohistochemical demonstration of PACAP in the pituitary, PACAP stimulation of growth hormone release from pituitary cells, and molecular cloning of pituitary Type I PACAP receptor. Endocrinology 139:3465–3479PubMedGoogle Scholar
  44. Wong NACS, Britton MP, Choi GS, Stanton TK, Bicknell DC, Wilding JL, Bodmer WF (2004) Loss of CDX1 expression in colorectal carcinoma: promoter methylation, mutation, and loss of heterozygosity analyses of 37 cell lines. PNAS 101:574–579CrossRefPubMedPubMedCentralGoogle Scholar
  45. Wong AOL, Li W, Leung CY, Huo L, Zhou H (2005) Pituitary adenylate cyclase-activating polypeptide (PACAP) as a growth hormone (GH)-releasing factor in grass carp. I. Functional coupling of cyclic adenosine 3′,5′-monophosphate and Ca2+/calmodulin-dependent signaling pathways in PACAP-induced GH secretion and GH gene expression in grass carp pituitary cells. Endocrinology 146:5407–5424CrossRefPubMedGoogle Scholar
  46. Zhang C, Wang Y, Chen H, Lan X, Lei C, Fang X (2009) Association between variants in the 5′-untranslated region of the bovine MC4R gene and two growth traits in Nanyang cattle. Mol Biol Rep 36:1839–1843CrossRefPubMedGoogle Scholar
  47. Zhao JL, Si YF, He F, Wen HS, Li JF, Ren YY, Zhao ML, Huang ZJ, Chen SL (2015) 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 41:853–865CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2015

Authors and Affiliations

  • Yufeng Si
    • 1
    • 2
  • Feng He
    • 1
    • 2
  • Haishen Wen
    • 1
    • 2
  • Jifang Li
    • 1
    • 2
  • Junli Zhao
    • 1
    • 2
  • Yuanyuan Ren
    • 1
    • 2
  • Meilin Zhao
    • 1
    • 2
  • Liqin Ji
    • 1
    • 2
  • Zhengju Huang
    • 1
    • 2
  • Mo Zhang
    • 1
    • 2
  • Songlin 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

Personalised recommendations