title: Evaluation of relative potencies for in vitro transactivation of the baikal seal aryl hydrocarbon receptor by dioxin-like compounds.
abstract: To evaluate the sensitivity and responses to dioxins and related compounds (DRCs) via aryl hydrocarbon receptor (AHR) in Baikal seals (Pusa sibirica), we constructed an in vitro reporter gene assay system. Baikal seal AHR (BS AHR) expression plasmid and a reporter plasmid containing CYP1A1 promoter were transfected in COS-7 cells. The cells were treated with six representative congeners, and dose-dependent responses were obtained for all the congeners. EC50 values of 2,3,7,8-TCDD, 1,2,3,7,8-PeCDD, 2,3,7,8-TCDF, 2,3,4,7,8-PeCDF, and PCB126 were found to be 0.021, 1.8, 0.16, 2.4, and 2.5 nM, respectively. As the response did not reach the maximal plateau, EC50 value for PCB118 could not be obtained. The TCDD-EC50 for BS AHR was as high as that for dioxin sensitive C57BL/6 mouse AHR. The in vitro dose responses were further analyzed following an established systematic framework and multiple (20, 50, and 80%) relative potencies (REPs) to the maximum TCDD response. The estimates revealed lower REP ranges (20-80%) of PeCDD and PeCDF for BS AHR than for mouse AHR. Average of the 20, 50, and 80% REPs was designated as Baikal seal specific TCDD induction equivalency factor (BS IEF). The BS IEFs of PeCDD, TCDF, PeCDF, PCB126, and PCB118 were estimated as 0.010, 0.018, 0.0078, 0.0059, and 0.00010, respectively. Total TCDD induction equivalents (IEQs) that were calculated using BS IEFs and hepatic concentrations in wild Baikal seals corresponded to only 12-31% of 2005 WHO TEF-derived TEQs. Nevertheless, about 50% of Baikal seals accumulated IEQs over the TCDD-EC50 obtained in this study. This assessment was supported by the enhanced CYP1A1 mRNA expression found in 50% of the specimens contaminated over the TCDD-EC50. These findings suggest that the IEFs proposed from this in vitro assay could be used to predict AHR-mediated responses in wild seals.
authors.affiliation: Department of Life and Nanopharmaceutical Science and Department of Biology, Kyung Hee University, Hoegi-Dong, Dongdaemun-Gu, Seoul 130-701, Korea.
authors.forename: Eun-Young
authors.initials: EY
authors.lastname: Kim
authors.forename: Tomoko
authors.initials: T
authors.lastname: Suda
authors.forename: Shinsuke
authors.initials: S
authors.lastname: Tanabe
authors.forename: Valeriy B
authors.initials: VB
authors.lastname: Batoev
authors.forename: Evgeny A
authors.initials: EA
authors.lastname: Petrov
authors.forename: Hisato
authors.initials: H
authors.lastname: Iwata
mesh_terms: D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000187:drug effects; D001572:Benzofurans; D019556:COS Cells; D002522:Chlorocebus aethiops; D019363:Cytochrome P-450 CYP1A1 / Q000235:genetics / Q000378:metabolism; D004147:Dioxins / Q000378:metabolism / Q000494:pharmacology* / Q000633:toxicity; D004785:Environmental Pollutants / Q000378:metabolism / Q000493:pharmacokinetics* / Q000633:toxicity; D005544:Forecasting; D008099:Liver / Q000378:metabolism; D051379:Mice; D018336:Receptors, Aryl Hydrocarbon / Q000187:drug effects* / Q000378:metabolism; D046048:Seals, Earless / Q000378:metabolism*
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D001572:Benzofurans; D019363:Cytochrome P-450 CYP1A1; D004147:Dioxins; D004785:Environmental Pollutants; D018336:Receptors, Aryl Hydrocarbon; C484722:Ahr protein, mouse; C014211:2,3,7,8-tetrachlorodibenzofuran