Gene Cloning and Expression Analysis of Growth Hormone Gene from Blotched Snakehead (Channa maculata)
GAO Dan-Dan1,2, OU Mi1, WU Yan-Duo1,2, CHEN Kun-Ci1, LIU Hai-Yang1, LUO Qing1, ZHAO Jian1,2,*
1 Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences/Key Laboratory of Tropical and Subtropical Fishery Resources Application and Cultivation, Ministry of Agriculture and Rural Affairs, Guangzhou 510380, China; 2 College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China
Abstract：Growth hormone (GH) is a polypeptide secreted by pituitary, which plays an important role in the development of teleost and the growth of somatic cells. In order to explore the role of GH in the growth dimorphism between male and female blotched snakehead (Channa maculata) and the mechanism of its interaction with sex hormones, in this study, GH gene (GenBank No. MW574005) of blotched snakehead was cloned and its expression pattern and response to exogenous steroid hormones were analyzed. The results showed that, the cDNA sequence of blotched snakehead GH gene was 882 bp in length, with an open reading frame (ORF) of 639 bp, encoding 212 amino acids. The length of GH sequence of blotched snakehead was 1 604 bp and contained 5 exons and 4 introns. A 1 199 bp flanking sequence was identified at the upstream of 5'-terminal, which contains pre-B-cell leukemia transcription factor 3 (PBX3), signal transducers and activators of transcription 1 (STA1), forkhead box O3 (FOXO3) and other transcription factors binding sites. Tissues expression analysis showed that GH was highly expressed in the pituitary of blotched snakehead, and the expression in male blotched snakehead was extremely significant higher than that in female (P<0.01); Expression analysis in different developmental stages showed that GH gene expression in the pituitary of male blotched snakehead was the highest at 365 d post hatching, which was extremely significant higher than that in female (P<0.01). The results of ELISA showed that the serum GH content in male blotched snakehead was higher than that in female in each developmental period. Hormone induction experiment showed that 17α-ethynylestradiol (EE2) inhibited GH gene expression in female and male blotched snakehead; 17α-methyltestosterone (MT) promoted GH gene expression in male pituitary, but decreased GH gene expression in female pituitary. This study provides the reference for further exploring the growth dimorphism mechanism of male and female blotched snakehead and the molecular mechanism of growth regulation.
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