Expression of two zona pellucida genes is regulated by 17α-ethinylestradiol in adult rare minnow Gobiocypris rarus

Zona pellucida (ZP) proteins are glycoproteins synthesized in liver, ovary or in both tissues in fish. In the present study, we aimed to determine the responsiveness of ZP2 and ZP3 to 17α-ethinylestradiol (EE2) in adult rare minnow Gobiocypris rarus. The full length of ZP3 cDNA was firstly characterized and its tissue distribution revealed that ZP3 mRNA was predominantly expressed in ovary of G. rarus. The gene expression profiles of ZP2, ZP3 and vitellogenin (VTG) were analyzed in gonad and liver of adult G. rarus exposed to EE2 at 1, 5, 25, and 125 ng/L for 3 and 6 days. The results show that ZP2 is more sensitive than ZP3 in gonads of both genders, and VTG in liver is extremely sensitive to EE2 in male fish. However, at lower concentrations (1 and 5 ng/L), the ZP2 in testes shows higher responsiveness to EE2 compared with VTG in rare minnow. The 5′ flanking regions of ZP2 and ZP3 were isolated and the comparison of transcription factors in the regions of ZP2 and ZP3 suggested that the disparity for the responsiveness of ZP2 and ZP3 to EE2 could partly be a result of differential cis-elements such as oocyte-specific protein (Osp1) binding sites or/and sex-determining region Y (SRY) binding site.