title: Embryonic cardiotoxicity of weak aryl hydrocarbon receptor agonists and CYP1A inhibitor fluoranthene in the Atlantic killifish (Fundulus heteroclitus).
abstract: High affinity aryl hydrocarbon receptor (AHR) ligands, such as certain polychlorinated biphenyls and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), cause severe cardiac teratogenesis in fish embryos. Moderately strong AHR agonists, for example benzo[a]pyrene and β-naphthoflavone, are capable of causing similar cardiotoxic effects, particularly when coupled with cytochrome P450 1A (CYP1A) inhibitors (e.g., fluoranthene (FL). Additionally, some weaker AHR agonists (carbaryl, 2-methylindole, 3-methylindole, and phenanthrene) are known to also cause cardiotoxicity in zebrafish (Danio rerio) embryos when coupled with FL; however, the cardiotoxic effects were not mediated specifically by AHR stimulation. This study was performed to determine if binary exposure to weak AHR agonists and FL were also capable of causing cardiotoxicity in Atlantic killifish Fundulus heteroclitus embryos. Binary exposures were performed in both naïve and PAH-adapted killifish embryos to examine resistance to weak agonists and FL binary exposures. Weak agonists used in this study included the following: carbaryl, phenanthrene, 2-methylindole, 3-methylindole, indigo, and indirubin. Carbaryl, indigo, and indirubin induced the highest CYP1 activity levels in naïve killifish embryos, but no significant CYP1 induction was observed in the PAH-adapted killifish. Embryos were coexposed to subteratogenic levels of each agonist and 500μg/L FL to assess if binary administration could cause cardiotoxicity. Indigo and indirubin coupled with FL caused cardiac teratogenesis in naïve killifish, but coexposures did not produce cardiac chamber abnormalities in the PAH-adapted population. Knockdown of AHR2 in naïve killifish embryos did not prevent cardiac teratogenesis. The data suggest a unique mechanism of cardiotoxicity that is not driven by AHR2 activation.
authors.affiliation: Nicholas School of the Environment, Duke University, Durham, NC 27514, USA. Electronic address: daniel.r.browndu@gmail.com.
authors.forename: D R
authors.initials: DR
authors.lastname: Brown
authors.affiliation: Nicholas School of the Environment, Duke University, Durham, NC 27514, USA. Electronic address: bwclark8@gmail.com.
authors.forename: B W
authors.initials: BW
authors.lastname: Clark
authors.affiliation: Nicholas School of the Environment, Duke University, Durham, NC 27514, USA. Electronic address: lvtgarner@gmail.com.
authors.forename: L V T
authors.initials: LV
authors.lastname: Garner
authors.affiliation: Nicholas School of the Environment, Duke University, Durham, NC 27514, USA. Electronic address: richd@duke.edu.
authors.forename: R T
authors.initials: RT
authors.lastname: Di Giulio
mesh_terms: D000818:Animals; D066126:Cardiotoxicity; D065607:Cytochrome P-450 Enzyme Inhibitors / Q000633:toxicity*; D000072461:Cytochrome P450 Family 1 / Q000037:antagonists & inhibitors* / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004625:Embryo, Nonmammalian / Q000187:drug effects / Q000201:enzymology; D029941:Fish Proteins / Q000819:agonists* / Q000037:antagonists & inhibitors* / Q000378:metabolism; D005449:Fluorenes / Q000633:toxicity*; D023781:Fundulidae / Q000196:embryology / Q000235:genetics / Q000378:metabolism*; D005786:Gene Expression Regulation; D055785:Gene Knockdown Techniques; D006321:Heart / Q000187:drug effects*; D008024:Ligands; D009206:Myocardium / Q000201:enzymology*; D011084:Polycyclic Aromatic Hydrocarbons / Q000633:toxicity*; D018336:Receptors, Aryl Hydrocarbon / Q000819:agonists* / Q000235:genetics / Q000378:metabolism; D018570:Risk Assessment; D014874:Water Pollutants, Chemical / Q000633:toxicity*
chemical_list: D065607:Cytochrome P-450 Enzyme Inhibitors; D029941:Fish Proteins; D005449:Fluorenes; D008024:Ligands; D011084:Polycyclic Aromatic Hydrocarbons; D018336:Receptors, Aryl Hydrocarbon; D014874:Water Pollutants, Chemical; C007738:fluoranthene; D000072461:Cytochrome P450 Family 1
keywords: Aryl hydrocarbon receptor agonists; Cardiotoxicity; Cytochrome P450; Development; Fundulus heteroclitus; Morpholino