Publication Date:
2022-05-26
Description:
© 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.
Description:
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.
Description:
This work was supported by Swedish Research Council Formas grants 2011-963 (EW) and 2008-1249 (MJ), by a European Commission Horizon 2020 grant, Project ID 634880 (MJ), by a National Institute of Environmental Health Sciences (NIEHS) grant P42ES007381 (JJS and MEH), R01ES006272 (MEH) and F32ES017585 (ART-L), by Japan Society for the Promotion of Science Postdoctoral Fellowship for Research Abroad194313 (AK), by Grant-in-Aids for Research Activity Start-up26881001 (AK) and for Young Scientists (A)15H05334 (AK).
Keywords:
Aryl hydrocarbon receptor
;
Cytochrome P4501
;
6-Formylindolo[3,2-b]carbazole
;
Enzyme inhibition
;
Zebrafish embryo toxicity
;
Synergistic receptor activation
Repository Name:
Woods Hole Open Access Server
Type:
Article
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