Molecular cloning of vitellogenin gene and mRNA expression by 17α-ethinylestradiol from slender bitterling

Abstract

Indigenous aquatic population such as fish could be used as a successful test species for evaluating the ecological effects in aquatic environment. In the present study, vitellogenin (Vtg) from slender bitterling ( Acheilognathus yamatsutae), an indigenous aquatic species in Korea, was cloned and sequenced to determine if the Vtg gene possesses an important characteristic so as to act as a sensitive biomarker for estrogenic endocrine disrupting chemicals (EEDCs). The sbVtg cDNA is 5010 bp in length, containing a 4653 bp open reading frame, which encodes 1550 amino acid residues. The sbVtg cDNA was divided into lipovitellin heavy chain (LvH), phosvitin (Pv), lipovitellin light chain (LvL) as well as a β′-component (β′-c) domain, and belongs to VtgAo2. SbVtg has conserved important sequences for Vtg functions such as signal peptide, VtgR-binding region, and disulfide bond formation, all of which are consistent with those of other teleosts. In addition, the male slender bitterling aqueous exposed to 17α-ethinylestradiol (EE2, 12.5, 25, and 50 ng/L) produced a statistically significant and concentration-dependent increase in hepatic Vtg mRNA expression, which showed a similar pattern to biliary estrogenic activity, measured by ERE-reporter gene assay. Thus, this study clearly indicates that the induction of Vtg in slender bitterling might be a suitable biomarker in toxicological research of EEDCs.