title: In vivo micronucleus screening in zebrafish by flow cytometry.
abstract: Micronucleus (MN) assay is extensively used to biomonitor aneuploidy and clastogenicity of genotoxic compounds. However, the suitability of this assay for early life stages of model fish species is still poorly documented. In this study, the determination of MN using flow cytometry was successfully applied for the first time to zebrafish (Danio rerio) larvae. Mitomycin C (MMC), etoposide (ETO), cyclophosphamide, demecolcine (COL), benzo[a]pyrene (BP) and dibenzo[def,p]chrysene (DBC) were selected as model genotoxicants. The method was first confirmed in human HepG2 liver cells and then applied in vivo on isolated cells from exposed 4 days post fertilisation zebrafish larvae. All tested compounds induced MN formation. The flow cytometry results were validated by a strong correlation with results from a standard MN microscopy analysis (P = 0.002). Moreover, flow cytometry analysis enabled the detection of an up to 3.7-fold increase of hypodiploidy in zebrafish exposed to MMC, COL, BP or DBC. MMC, COL and DBC induced more than a 2-fold MN increase by flow cytometry and were therefore considered as the most suitable positive controls for in vivo zebrafish MN determination. These findings make important contribution by proposing a new reliable and sensitive method for using zebrafish as a model for genotoxicity monitoring.
authors.affiliation: Institute of Environmental Medicine, Karolinska Institutet, Nobels väg 13, 171 77 Stockholm, Sweden lebihanic@gmail.com.
authors.forename: Florane
authors.initials: F
authors.lastname: Le Bihanic
authors.affiliation: Institute of Environmental Medicine, Karolinska Institutet, Nobels väg 13, 171 77 Stockholm, Sweden.
authors.forename: Sebastiano
authors.initials: S
authors.lastname: Di Bucchianico
authors.affiliation: Institute of Environmental Medicine, Karolinska Institutet, Nobels väg 13, 171 77 Stockholm, Sweden.
authors.forename: Hanna L
authors.initials: HL
authors.lastname: Karlsson
authors.affiliation: Institute of Environmental Medicine, Karolinska Institutet, Nobels väg 13, 171 77 Stockholm, Sweden.
authors.forename: Kristian
authors.initials: K
authors.lastname: Dreij
mesh_terms: D000818:Animals; D004784:Environmental Monitoring / Q000379:methods*; D005434:Flow Cytometry; D056945:Hep G2 Cells; D006801:Humans; D007814:Larva / Q000187:drug effects* / Q000235:genetics; D048629:Micronuclei, Chromosome-Defective / Q000139:chemically induced*; D015162:Micronucleus Tests; D009153:Mutagens / Q000633:toxicity*; D015027:Zebrafish / Q000235:genetics*
chemical_list: D009153:Mutagens