Expression of two cytochrome P450 aromatase genes is regulated by endocrine disrupting chemicals in rare minnow Gobiocypris rarus juveniles

Abstract

To elucidate the effects of endocrine disrupting chemicals (EDCs) on aromatase, the rare minnow ovarian and brain P450 aromatase ( cyp19a1a and cyp19a1b) cDNA and their 5′-flanking regions were isolated and characterized. RT-PCR analysis revealed that the rare minnow cyp19a1a mRNA was predominantly expressed in ovary while cyp19a1b was predominantly expressed in brain. Sequences for binding sites of steroidogenic factor-1, peroxisome proliferators-activated receptor, aryl hydrocarbon receptor, CCAAT/enhancer binding protein, estrogen responsive element, glucocorticoid responsive element, and retinoic acid receptor were identified on promoter regions of cyp19a1 genes . The influence of several EDCs on the transcript abundance of cyp19a1a and cyp19a1b was investigated in rare minnow juveniles. Clofibrate did not influence the expression of either cyp19a1 genes. Exposure to 1 nM ethinylestradiol (EE2) for 3 days significantly downregulated the expression of cyp19a1a gene, however 0.1 and 1 nM EE2 significantly increased the gene expression of cyp19a1b. Exposure to 100 and 1000 nM 4-nonylphenol (NP) significantly suppressed the cyp19a1a expression, but it had no effect on the expression of cyp19a1b gene. Bisphenol A (BPA) strongly suppressed the cyp19a1b gene expression from 0.1 to 10 nM and significantly suppressed the gene expression of cyp19a1a only at 10 nM. These results indicate that EDCs may influence the expression of cyp19a1 genes through differential transcriptional modulation in rare minnow juveniles.