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Molecular response and association analysis of Megalobrama amblycephala fih-1 with hypoxia

Abstract

Hypoxia is one of the most important environmental factors which affect fish growth, development and survival, but regulation mechanisms of hypoxia in fish remain unclear. Therefore, to further understand molecular functions of factor inhibiting HIF-1 (Fih-1), an essential hypoxia sensor, the full-length cDNA of fih-1 was cloned from Megalobrama amblycephala, a hypoxia-sensitive cyprinid fish. The deduced amino acid sequence showed high homology with that of other vertebrates, and all structural and functional domains were highly conserved. The mRNA level in different tissues and developmental stages indicated that M. amblycephala fih-1 expression was higher in liver and muscle, followed by gill, intestine and spleen. During embryogenesis, the fih-1 mRNA was highly expressed in the early embryonic development, then decreased to a very low level, and maintained a relative high level of expression after hatching. In most tissues, the fih-1 mRNA was down-regulated at 2 h but up-regulated at 4 h after hypoxia treatment. In addition, the promoter sequence of M. amblycephala fih-1 was obtained using thermal asymmetric interlaced PCR. Three single nucleotide polymorphism (SNP) sites were found in the cDNA and promoter sequences, and identified significant association with hypoxia trait by correlation analysis in hypoxia-sensitive group and hypoxia-tolerant group. These results demonstrated that M. amblycephala fih-1 plays important roles in embryo development and hypoxia response, which will contribute to systematic understanding of the molecular mechanisms of fish in response to hypoxia, and provide help for fish genetic breeding with hypoxia-tolerant strains or breeds.

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Acknowledgments

This research was supported by the Fundamental Research Funds for the Central Universities (2013PY067 and 2662015PY205), Key Technologies R&D Program of China (no. 2012BAD26B00), and Modern Agroindustry Technology Research System entitled “Staple Freshwater Fishery Industry Technology System” (no. CARS-46-05).

Author information

Correspondence to Huanling Wang.

Ethics declarations

All the experimental procedures were approved by the institution of animal care and use committee of Huazhong Agricultural University.

Conflict of interest

The authors declare no conflicts of interest.

Additional information

Communicated by J. Cerdá.

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Zhang, B., Chen, N., Huang, C. et al. Molecular response and association analysis of Megalobrama amblycephala fih-1 with hypoxia. Mol Genet Genomics 291, 1615–1624 (2016). https://doi.org/10.1007/s00438-016-1208-x

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Keywords

  • Megalobrama amblycephala
  • Fih-1
  • SNPs
  • Hypoxia
  • Expression