DNA methylation in the 5′ flanking region of cytochrome P450 17 in adult rare minnow Gobiocypris rarus — Tissue difference and effects of 17α-ethinylestradiol and 17α-methyltestoterone exposures

Cytochrome P450 17 (CYP17) plays a vital role in hormone production in the body. In our previous study, mRNA expression of cyp17a1 was regulated by endocrine disrupting chemicals in rare minnow Gobiocypris rarus. However, the mechanism underlying the regulation is unclear. In the present study, we aim to explore whether the differential expression of cyp17a1 in distinct tissues and the modulation of its expression upon 17α-ethinylestradiol (EE2) and 17α-methyltestoterone (MT) are related to the DNA methylation status in G. rarus. The 732-bp fragment of 5′ flanking region of cyp17a1 gene was isolated in G. rarus. The bisulfite sequencing PCR result showed that DNA methylation levels in 5′ flanking of cyp17a1 in the gonads were significantly lower than those in the brains, which is negatively related to its mRNA expression in the 2 tissues in the previous study. The 7-day EE2 exposure of 25 ng/L caused a significant increase of methylation levels of cyp17a1 gene and a significant decrease of its transcript in testis. While 100 ng/L MT exposure for 7 days caused a significant decrease of methylation levels of cyp17a1 gene and a significant increase of its transcript in the ovary. The present findings indicate that the methylation status of cyp17a1 gene is negatively correlated with its mRNA expression in response to EE2 and MT in G. rarus. We hypothesize that the regulation of cyp17a1 expression by EE2 and MT might attribute to the change of its DNA methylation status in G. rarus.