Abstract:The purpose of this study was to investigate the role of Wnt4(wingless-type MMTV integration site family,member 4)gene in sex development of northern pike Esox lucius. The Wnt4 gene fragment was cloned by RT-PCR, the homology of amino acids and the evolutionary relationship of species were analyzed by Clustal X and MEGA 6.1 software. The length of Wnt4 gene fragment was 1505 bp, with the open reading frame of 1059 bp, encoding 352 amino acids. Amino acid homology analysis showed that northern pike had high homology with rainbow trout Oncorhynchus mykiss and medaka Oryzias latipes, but low similarity with Xenopus laevis and Rattus norvegicus. Phylogenetic analysis showed that the phylogenetic relationship between northern pike and Myripristis murdjan was the hithermost, and the phylogenetic relationship between northern pike and Paracentrotus lividus, Azumapecten farreri and Mytilus coruscus was the farthest. Real time fluorescent quantitative PCR analysis showed that Wnt4 gene was expressed in ovary, testis, intestine, muscle, gill and brain. The expression of Wnt4 gene in ovary was the highest, and the expression of Wnt4 gene in ovary was significantly higher than that in testis. With the development of ovary, the expression of Wnt4 gene was decreased. Wnt4 gene was involved in the formation and maintenance of some organs in the early stage of ovarian development. The findings provide reference for the sex development mechanism of northern pike.
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