Testing estrogenicity of known and novel (xeno-)estrogens in the MolDarT using developing zebrafish (Danio rerio).
The MolDarT is a novel short-term assay for testing mechanism-based molecular effects in developing zebrafish embryos. The objective of this study was to evaluate the inducibility of vitellogenin1 mRNA (Vtg1) by the estrogenically active compounds 17beta-Estradiol (E2), 17alpha-Ethinylestradiol (EE2), Nonylphenol (NP), Bisphenol A (BPA), Cyproconazol, and the suspected xeno-estrogen Atrazin in the MolDarT. Freshly fertilized zebrafish eggs were exposed semistatically for 120 h. Using reverse transcription real-time PCR, the relative abundance of Vtg1 was measured. For EE2 a dose-response relationship was established with EC50 = 60.7 ng/L (205 pM). Induction of Vtg1 was significant at concentrations of 84 pM EE2 (25 ng EE2/L) and above, 10 nM E2 (2.7 microg E2/L), 100 nM E2 (27 microg E2/L), 10 microM BPA (2280 microg BPA/L), and 15 microM BPA (3420 microg BPA/L). At NP concentrations of 0.75 microM (165 microg NP/L) and 1.5 microM (330 microg NP/L) Vtg1 was significantly down-regulated. Both atrazine and cyproconazol showed no effect on relative Vtg1 abundance. With this study we further characterize the MolDarT assay and show its applicability for effect screening of compounds.
Muncke
Jane
J
Eawag, Swiss Federal Institute of Aquatic Science and Technology, Environmental Toxicology, Ueberlandstrasse 133, 8600 Dübendorf, Switzerland.
Junghans
Marion
M
Eggen
Rik I L
RI
D000818:Animals; D001280:Atrazine / Q000633:toxicity; D001559:Benzhydryl Compounds; D001681:Biological Assay; D004967:Estrogens / Q000633:toxicity*; D004997:Ethinyl Estradiol / Q000633:toxicity; D005659:Fungicides, Industrial / Q000633:toxicity; D006540:Herbicides / Q000633:toxicity; D010636:Phenols / Q000633:toxicity; D011960:Receptors, Estrogen / Q000378:metabolism; D014230:Triazoles / Q000633:toxicity; D014819:Vitellogenins / Q000378:metabolism*; D015027:Zebrafish / Q000196:embryology / Q000378:metabolism*; D000098826:Bisphenol A Compounds
D001280:Atrazine; D001559:Benzhydryl Compounds; D004967:Estrogens; D004997:Ethinyl Estradiol; D005659:Fungicides, Industrial; D006540:Herbicides; D010636:Phenols; D011960:Receptors, Estrogen; D014230:Triazoles; D014819:Vitellogenins; C093628:cyproconazole; C025256:nonylphenol; C006780:bisphenol A; D000098826:Bisphenol A Compounds