title: In Vivo Detection of Mitochondrial Dysfunction Induced by Clinical Drugs and Disease-Associated Genes Using a Novel Dye ZMJ214 in Zebrafish.
abstract: Mitochondrial dysfunction has been implicated in various drug-induced toxicities and genetic disorders. Recently, the zebrafish has emerged as a versatile animal model for both chemical and genetic screenings. Taking advantage of its transparency, various in vivo fluorescent imaging methods have been developed to identify novel functions of chemicals and genes in zebrafish. However, there have not been fluorescent probes that can detect mitochondrial membrane potential in living zebrafish. In this study, we identified a novel cyanine dye called ZMJ214 that detects mitochondrial membrane potential in living zebrafish from 4 to 8 days post fertilization and is administered by simple immersion. The fluorescence intensity of ZMJ214 in zebrafish was increased and decreased by oligomycin and FCCP, respectively, suggesting a positive correlation between ZMJ214 fluorescence and mitochondrial membrane potential. In vivo imaging of zebrafish stained with ZMJ214 allowed for the detection of altered mitochondrial membrane potential induced by the antidiabetic drug troglitazone and the antiepileptic drug tolcapone, both of which have been withdrawn from the market due to mitochondrial toxicity. In contrast, pioglitazone and entacapone, which are similar to troglitazone and tolcapone, respectively, and have been used commercially, did not cause a change in mitochondrial membrane potential in zebrafish stained with ZMJ214. Live imaging of zebrafish stained with ZMJ214 also revealed that knock-down of slc25a12, a mitochondrial carrier protein associated with autism, dysregulated the mitochondrial membrane potential. These results suggest that ZMJ214 can be a useful tool to identify chemicals and genes that cause mitochondrial dysfunction in vivo.
authors.affiliation: Department of Molecular and Cellular Pharmacology, Pharmacogenomics and Pharamacoinformatics, Mie University Graduate School of Medicine , Tsu, Mie 514-8507, Japan.
authors.forename: Shota
authors.initials: S
authors.lastname: Sasagawa
authors.affiliation: Department of Molecular and Cellular Pharmacology, Pharmacogenomics and Pharamacoinformatics, Mie University Graduate School of Medicine , Tsu, Mie 514-8507, Japan.|Mie University Medical Zebrafish Research Center , Tsu, Mie 514-8507, Japan.|Depertment of Systems Pharmacology, Mie University Graduate School of Medicine , Tsu, Mie 514-8507, Japan.|Department of Omics Medicine, Mie University Industrial Technology Innovation Institute , Tsu, Mie 514-8507, Japan.|Department of Bioinformatics, Mie University Life Science Research Center , Tsu, Mie 514-8507, Japan.
authors.forename: Yuhei
authors.initials: Y
authors.lastname: Nishimura
authors.affiliation: Department of Molecular and Cellular Pharmacology, Pharmacogenomics and Pharamacoinformatics, Mie University Graduate School of Medicine , Tsu, Mie 514-8507, Japan.
authors.forename: Junko
authors.initials: J
authors.lastname: Koiwa
authors.affiliation: Corporate R&D Headquarters, Canon Inc. , Ohta-ku, Tokyo 146-8501, Japan.
authors.forename: Tsuyoshi
authors.initials: T
authors.lastname: Nomoto
authors.affiliation: Corporate R&D Headquarters, Canon Inc. , Ohta-ku, Tokyo 146-8501, Japan.
authors.forename: Taichi
authors.initials: T
authors.lastname: Shintou
authors.affiliation: Department of Molecular and Cellular Pharmacology, Pharmacogenomics and Pharamacoinformatics, Mie University Graduate School of Medicine , Tsu, Mie 514-8507, Japan.
authors.forename: Soichiro
authors.initials: S
authors.lastname: Murakami
authors.affiliation: Department of Molecular and Cellular Pharmacology, Pharmacogenomics and Pharamacoinformatics, Mie University Graduate School of Medicine , Tsu, Mie 514-8507, Japan.
authors.forename: Mizuki
authors.initials: M
authors.lastname: Yuge
authors.affiliation: Department of Molecular and Cellular Pharmacology, Pharmacogenomics and Pharamacoinformatics, Mie University Graduate School of Medicine , Tsu, Mie 514-8507, Japan.
authors.forename: Koki
authors.initials: K
authors.lastname: Kawaguchi
authors.affiliation: Department of Molecular and Cellular Pharmacology, Pharmacogenomics and Pharamacoinformatics, Mie University Graduate School of Medicine , Tsu, Mie 514-8507, Japan.
authors.forename: Reiko
authors.initials: R
authors.lastname: Kawase
authors.affiliation: Corporate R&D Headquarters, Canon Inc. , Ohta-ku, Tokyo 146-8501, Japan.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Miyazaki
authors.affiliation: Department of Molecular and Cellular Pharmacology, Pharmacogenomics and Pharamacoinformatics, Mie University Graduate School of Medicine , Tsu, Mie 514-8507, Japan.|Mie University Medical Zebrafish Research Center , Tsu, Mie 514-8507, Japan.|Depertment of Systems Pharmacology, Mie University Graduate School of Medicine , Tsu, Mie 514-8507, Japan.|Department of Omics Medicine, Mie University Industrial Technology Innovation Institute , Tsu, Mie 514-8507, Japan.|Department of Bioinformatics, Mie University Life Science Research Center , Tsu, Mie 514-8507, Japan.
authors.forename: Toshio
authors.initials: T
authors.lastname: Tanaka
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000633:toxicity; D000927:Anticonvulsants / Q000633:toxicity; D001577:Benzophenones / Q000633:toxicity; D002232:Carbocyanines / Q000737:chemistry*; D002839:Chromans / Q000633:toxicity; D004195:Disease Models, Animal; D005456:Fluorescent Dyes / Q000737:chemistry*; D007004:Hypoglycemic Agents / Q000633:toxicity; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects*; D008928:Mitochondria / Q000187:drug effects* / Q000473:pathology*; D009596:Nitrophenols / Q000633:toxicity; D009840:Oligomycins / Q000633:toxicity; D061848:Optical Imaging / Q000379:methods; D000077205:Pioglitazone; D045162:Thiazolidinediones / Q000633:toxicity; D000077867:Tolcapone; D018675:Toxicity Tests / Q000379:methods; D000077288:Troglitazone; D015027:Zebrafish / Q000235:genetics; D029961:Zebrafish Proteins / Q000235:genetics
chemical_list: D000900:Anti-Bacterial Agents; D000927:Anticonvulsants; D001577:Benzophenones; D002232:Carbocyanines; D002839:Chromans; D005456:Fluorescent Dyes; D007004:Hypoglycemic Agents; D009596:Nitrophenols; D009840:Oligomycins; D045162:Thiazolidinediones; D029961:Zebrafish Proteins; D000077867:Tolcapone; D000077288:Troglitazone; D000077205:Pioglitazone