title: In vitro transactivation potencies of black-footed albatross (Phoebastria nigripes) AHR1 and AHR2 by dioxins to predict CYP1A expression in the wild population.
abstract: Our previous studies have detected high levels of dioxins and related compounds (DRCs) including polychlorinated dibenzo-p-dioxins (PCDDs), furans (PCDFs), and coplanar PCBs (Co-PCBs) in the black-footed albatross (BFA), Phoebastria nigripes, from the North Pacific region. We have also cloned two aryl hydrocarbon receptors, AHR1 and AHR2, of the BFA. To evaluate the sensitivity to DRCs in the BFA and to assess the status of cytochrome P450 1A (CYP1A) induction in the wild population, this study investigated the mRNA expression levels of BFA AHR1 and AHR2 and also the transactivation potencies of each AHR by 15 selected DRC congeners. Quantitative real-time PCR of BFA AHR mRNAs showed that hepatic AHR1 is more highly expressed than AHR2. Transactivation by graded concentrations of individual DRCs was measured in COS-7 cells, where BFA AHR1 or AHR2 was transiently transfected. For congeners that exhibited AHR-mediated dose-dependent activities, 50% effective concentration (EC(50)) and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) relative potencies (REPs) were estimated. Based on the estimates of the REPs, TCDD induction equivalency factors (IEFs) were determined. For BFA AHR1, PeCDF was equipotent to TCDD, but other congeners exhibited lower IEFs. For BFA AHR2, PCDD/F congeners except OCDD/F showed IEFs ≥ 1.0. Using BFA AHR1- or AHR2-IEFs and hepatic concentrations of DRCs in North Pacific BFAs, TCDD induction equivalents (IEQs) were calculated. We further constructed nonlinear regression models on the relationships between BFA AHR1- or AHR2-IEF derived total IEQ or WHO-TEF derived total TEQ and ethoxyresorufin-O-deethylase activity (EROD) in the liver of wild BFAs. The results indicated that the relationships of BFA AHR1- and AHR2-based IEQs and EROD were predictable from BFA AHR1- and AHR2-mediated transactivation by TCDD, respectively. Collectively, these results suggest that the in vitro assay incorporating the AHR of species of concern would be a useful tool to predict the sensitivity to DRCs in the species and CYP1A induction in the wild population.
authors.affiliation: Center for Marine Environmental Studies, Ehime University, Bunkyo-cho 2-5, Matsuyama 790-8577, Japan.
authors.forename: Thuruthippallil Leena
authors.initials: TL
authors.lastname: Mol
authors.forename: Eun-Young
authors.initials: EY
authors.lastname: Kim
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Ishibashi
authors.forename: Hisato
authors.initials: H
authors.lastname: Iwata
mesh_terms: D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D000835:Animals, Wild / Q000235:genetics*; D001717:Birds / Q000235:genetics*; D019363:Cytochrome P-450 CYP1A1 / Q000378:metabolism*; D004147:Dioxins / Q000633:toxicity*; D005786:Gene Expression Regulation / Q000187:drug effects; D008099:Liver / Q000187:drug effects / Q000201:enzymology; D017711:Nonlinear Dynamics; D018336:Receptors, Aryl Hydrocarbon / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D012044:Regression Analysis; D013329:Structure-Activity Relationship; D015533:Transcriptional Activation / Q000187:drug effects* / Q000235:genetics; D014944:World Health Organization
chemical_list: D000596:Amino Acids; D004147:Dioxins; D018336:Receptors, Aryl Hydrocarbon; D019363:Cytochrome P-450 CYP1A1