Effects of endocrine disruptors on Bombina orientalis P450 aromatase activity.
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.
Lee
Kyung-min
KM
Department of Life Science, Hanyang University, Seoul, Korea.
Yang
Wonseok
W
Rhee
Jae-Sung
JS
Hwang
Dae-Sik
DS
Park
Chan Jin
CJ
Gye
Myung Chan
MC
Lee
Jae-Seong
JS
Shin
Incheol
I
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
D001141:Aromatase; D001559:Benzhydryl Compounds; D052244:Endocrine Disruptors; D010636:Phenols; C025256:nonylphenol; C006780:bisphenol A; D000098826:Bisphenol A Compounds