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

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 beta'-component (beta'-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 17 alpha-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.