Biological effects of 6-formylindolo[3,2-b]carbazole (FICZ) in vivo are enhanced by loss of CYP1A function in an Ahr2-dependent manner
Timme-Laragy, Alicia R.
Jonsson, Maria E.
Hahn, Mark E.
Stegeman, John J.
MetadataShow full item record
KeywordAryl hydrocarbon receptor; Cytochrome P4501; 6-Formylindolo[3,2-b]carbazole; Enzyme inhibition; Zebrafish embryo toxicity; Synergistic receptor activation
6-Formylindolo[3,2-b]carbazole (FICZ) is a potent aryl hydrocarbon receptor (AHR) agonist that is efficiently metabolized by AHR-regulated cytochrome P4501 enzymes. FICZ is a proposed physiological AHR ligand that induces its own degradation as part of a regulatory negative feedback loop. In vitro studies in cells show that CYP1 inhibition in the presence of FICZ results in enhanced AHR activation, suggesting that FICZ accumulates in the cell when its metabolism is blocked. We used zebrafish (Danio rerio) embryos to investigate the in vivo effects of FICZ when CYP1A is knocked down or inhibited. Embryos were injected with morpholino antisense oligonucleotides targeting CYP1A (CYP1A-MO), Ahr2, or a combination of both. FICZ exposure of non-injected embryos or embryos injected with control morpholino had little effect. In CYP1A-MO-injected embryos, however, FICZ dramatically increased mortality, incidence and severity of pericardial edema and circulation failure, reduced hatching frequency, blocked swim bladder inflation, and strongly potentiated expression of Ahr2-regulated genes. These effects were substantially reduced in embryos with a combined knockdown of Ahr2 and CYP1A, indicating that the toxicity was mediated at least partly by Ahr2. Co-exposure to the CYP1 inhibitor alpha-naphthoflavone (αNF) and FICZ had similar effects as the combination of CYP1A-MO and FICZ. HPLC analysis of FICZ-exposed embryos showed increased levels of FICZ after concomitant CYP1A-MO injection or αNF co-exposure. Together, these results show that a functioning CYP1/AHR feedback loop is crucial for regulation of AHR signaling by a potential physiological ligand in vivo and further highlights the role of CYP1 enzymes in regulating biological effects of FICZ.
© The Author(s), 2016. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Biochemical Pharmacology 110-111 (2016): 117-129, doi:10.1016/j.bcp.2016.04.012.
The following license files are associated with this item:
Except where otherwise noted, this item's license is described as Attribution-NonCommercial-NoDerivatives 4.0 International
Showing items related by title, author, creator and subject.
Involvement of Cytochrome P450 1A in the toxicity of aryl hydrocarbon receptor agonists : alteration arachidonic acid metabolism and production of reactive oxygen species Schlezinger, Jennifer J. (Massachusetts Institute of Technology and Woods Hole Oceanographic Institution, 1998-08)Two cytochrome P4501A-dependent mechanisms of aryl hydrocarbon receptor (AhR) agonist toxicity were examined in the marine teleost scup (Stenotomus chrysops), alteration of arachidonic acid (AA) metabolism and production ...
Hestermann, Eli V. (Massachusetts Institute of Technology and Woods Hole Oceanographic Institution, 1999-12)Aryl hydrocarbon receptor (AHR) expression and activity was characterized in the teleost hepatoma cell line, PLHC-1. This work was carried out in order to gain insights into mechanisms of halogenated aromatic hydrocarbon ...
Lapseritis, Joy M. (Massachusetts Institute of Technology and Woods Hole Oceanographic Institution, 2007-02)Marine mammals possess high body burdens of persistent organic pollutants, including PCBs and dioxin-like compounds (DLC). Chronic environmental or dietary exposure to these chemicals can disrupt the function of reproductive ...