title: Effects of endocrine disruptors on Bombina orientalis P450 aromatase activity.
abstract: To assess the effects of endocrine-disrupting chemicals on the expression and activity of aromatase in the gonads of Bombina orientalis, a common amphibian, we intraperitoneally injected nonylphenol or bisphenol-A and then examined aromatase mRNA levels by RT-PCR as well as aromatase enzymatic activity by tritiated water release assays. To design primers for the RT-PCR, we cloned the B. orientalis aromatase gene using RT-PCR and degenerate primers. The full-length cDNA was obtained by 5'- and 3'-RACE PCR. The complete sequence of the B. orientalis aromatase gene revealed an open reading frame of 1500 bp encoding a deduced protein of 500 amino acids. Semi-quantitative RT-PCR indicated that nonylphenol or bisphenol-A injection did not significantly affect the expression of B. orientalis aromatase mRNA. However, a 48-hr treatment with nonyphenol or bisphenol A reduced aromatase activity to 47% and 32% of the control, respectively. These results suggest that endocrine disrupters can effectively modulate the activity of B. orientalis aromatase without affecting its mRNA levels.
authors.affiliation: Department of Life Science, Hanyang University, Seoul, Korea.
authors.forename: Kyung-min
authors.initials: KM
authors.lastname: Lee
authors.forename: Wonseok
authors.initials: W
authors.lastname: Yang
authors.forename: Jae-Sung
authors.initials: JS
authors.lastname: Rhee
authors.forename: Dae-Sik
authors.initials: DS
authors.lastname: Hwang
authors.forename: Chan Jin
authors.initials: CJ
authors.lastname: Park
authors.forename: Myung Chan
authors.initials: MC
authors.lastname: Gye
authors.forename: Jae-Seong
authors.initials: JS
authors.lastname: Lee
authors.forename: Incheol
authors.initials: I
authors.lastname: Shin
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001001:Anura / Q000235:genetics / Q000378:metabolism*; D001141:Aromatase / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D001559:Benzhydryl Compounds; D052244:Endocrine Disruptors / Q000633:toxicity; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects*; D008969:Molecular Sequence Data; D010636:Phenols / Q000633:toxicity*; D010802:Phylogeny; D000098826:Bisphenol A Compounds
chemical_list: D001141:Aromatase; D001559:Benzhydryl Compounds; D052244:Endocrine Disruptors; D010636:Phenols; C025256:nonylphenol; C006780:bisphenol A; D000098826:Bisphenol A Compounds