Cloning and expression patterns of espl1 gene in loach (Misgurnus anguillicaudatus) during gonad development

Abstract

espl1 (extra spindle pole bodies like 1), a cysteine endopeptidase, is a mitotic key player in chromosomal segregation and centriole duplication during mitosis and meiosis. Considering the espl1 gene has not been reported in aquatic organisms, we reported the isolation and expression of the espl1 gene from loach. In this study, the full-length cDNA of espl1 was cloned for the first time in loach. In loach, the full-length cDNA of espl1 consisted of 6948 bp, the open reading frame (ORF) is 6240 bp, and the espl1 gene encodes 2139 amino acids. Moreover, the deduced amino acid sequences of espl1 in loach shared the highest identity with those of Cyprinus carpio (78.39%) and Sinocyclocheilus anshuiensis (78.27%), and the sequence homology among the various separases is confined to the C-terminal region. Furthermore, tissue-specific checking results indicated that the espl1 gene of the loach gene was highly expressed in the ovary and testis, especially in stage Ⅳ oocytes and stage Ⅳ testis by Real-time quantitative PCR (qPCR). Then, whole-mount in situ hybridization analyses revealed the expression of espl1 in the early development of loach. We found that the positive signal of espl1 was observed in the notochord during the early embryo development of loach. Last but not least, when treated with luteinizing hormone-releasing hormone analog (LHRH-A2), the mRNA expression of espl1 was significantly increased in the testes and ovaries. These observations suggest that the espl1 gene had a distinct and important role in the gonads of Loach. This study will be of value for further studies into the function of the espl1 gene in fish.