title: Diphenyleneiodonium chloride, an inhibitor of reduced nicotinamide adenine dinucleotide phosphate oxidase, suppresses light-dependent induction of clock and DNA repair genes in zebrafish.
abstract: In most species, solar light is both a DNA-damaging agent and the key entraining stimulus for the endogenous circadian clock. The zebrafish is an attractive vertebrate system in which to study the influence of light on gene expression because the DNA repair proteins and circadian oscillators in this species are light-responsive. At the molecular level, light treatment of zebrafish cells induces the production of reactive oxygen species (ROS). ROS both alters the reduction-oxidation (redox) state of these cells and stimulates intracellular extracellular signal-regulated kinase (ERK)/mitogen activated protein kinase (MAPK) cascades that transduce photic signals activating the transcription of particular light-responsive genes, including some clock genes and some DNA repair genes involved in photoreactivation. To date, however, the phototransducing molecules responsible for light-dependent ROS production have not been identified. Flavin-containing oxidases, such as reduced nicotinamide adenine dinucleotide phosphate (NADPH) oxidase, are versatile flavoenzymes that catalyze molecular oxidation in numerous metabolic pathways. Importantly, light induces the photoreduction of the flavin adenine dinucleotide (FAD) moiety in these oxidases, leading to ROS production. Here, we show in cultured zebrafish cells that diphenyleneiodonium chloride (DPI), an inhibitor of NADPH oxidase, both suppresses ERK/MAPK activation and efficiently reduces light-dependent expression of clock and photoreactivation genes. Our results suggest that flavin-containing oxidases may be responsible for light-dependent ROS production and thus light-dependent gene expression in zebrafish. Our findings also support the existence of a regulatory link between photoreactivation and the circadian clock in this species.
authors.affiliation: Department of Developmental and Regenerative Biology, Medical Research Institute, Tokyo Medical and Dental University, Japan.
authors.forename: Tomomi
authors.initials: T
authors.lastname: Osaki
authors.forename: Yoshimi
authors.initials: Y
authors.lastname: Uchida
authors.forename: Jun
authors.initials: J
authors.lastname: Hirayama
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Nishina
mesh_terms: D000818:Animals; D056926:CLOCK Proteins; D002478:Cells, Cultured; D002712:Chlorides / Q000494:pharmacology; D057906:Circadian Clocks / Q000235:genetics*; D004260:DNA Repair / Q000235:genetics*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D048049:Extracellular Signal-Regulated MAP Kinases / Q000037:antagonists & inhibitors; D005182:Flavin-Adenine Dinucleotide / Q000378:metabolism; D005415:Flavins / Q000378:metabolism*; D005786:Gene Expression Regulation; D008027:Light; D020928:Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors; D019255:NADPH Oxidases / Q000037:antagonists & inhibitors*; D009861:Onium Compounds / Q000494:pharmacology*; D010084:Oxidation-Reduction; D017382:Reactive Oxygen Species / Q000378:metabolism; D015398:Signal Transduction; D015027:Zebrafish
chemical_list: D002712:Chlorides; D004791:Enzyme Inhibitors; D005415:Flavins; D009861:Onium Compounds; D017382:Reactive Oxygen Species; D005182:Flavin-Adenine Dinucleotide; C007517:diphenyleneiodonium; D019255:NADPH Oxidases; D056926:CLOCK Proteins; D048049:Extracellular Signal-Regulated MAP Kinases; D020928:Mitogen-Activated Protein Kinases