The cytotoxicity and genotoxicity of hexavalent chromium in medaka (Oryzias latipes) cells.
Chromium is an increasing health concern for aquatic environments, however, the mechanism of chromium toxicity in aquatic species is yet unknown. We used a medaka (Oryzias latipes) fin cell line to investigate the cytotoxicity and genotoxicity of sodium chromate, a soluble form of hexavalent chromium. We used a clonogenic cytotoxicity assay to measure sodium chromate cytotoxicity, gamma-H2A.X immunofluoresence to measure DNA double-strand breaks, and chromosome damage to measure clastogenicity. We found that sodium chromate is cytotoxic to medaka fin cells, with toxicity increasing in a concentration-dependent manner. Treatments of 0.5, 1, 5, 10, 25, 50 and 100 microM sodium chromate caused 100, 103.5, 87.8, 77.5, 40.9, 15 and 2.7% survival, respectively, relative to the control. We visualized DNA double-strand breaks in medaka cells through the formation of gamma-H2A.X foci. Breaks could be detected at concentrations as low as 1 microM. We also found that sodium chromate induces chromosomal aberrations, causing chromatid lesions and exchanges that increase with concentration. Treatments of 0, 1, 5, 10 and 25 microM sodium chromate damaged 10.3, 17, 32.3, 43 and 51.6% of metaphases and induced 13, 23, 44, 69 and 118 total aberrations in 100 metaphases, respectively. These data show that hexavalent chromium is both cytotoxic and genotoxic to fish cells. Our results set the context for future work in the medaka cell culture model and provide important tools for investigating mechanisms of toxicity in aquatic organisms.
Goodale
Britton C
BC
Wise Laboratory of Environmental and Genetic Toxicology, University of Southern Maine, 96 Falmouth Street, P.O. Box 9300, Portland, ME 04104, United States.
Walter
Ron
R
Pelsue
Stephen R
SR
Thompson
W Douglas
WD
Wise
Sandra S
SS
Winn
Richard N
RN
Mitani
Hiroshi
H
Wise
John Pierce
JP
D000818:Animals; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D002840:Chromates / Q000633:toxicity*; D004305:Dose-Response Relationship, Drug; D009990:Oryzias / Q000502:physiology*; D017670:Sodium Compounds / Q000633:toxicity*; D014410:Tumor Stem Cell Assay; D014874:Water Pollutants, Chemical / Q000633:toxicity*
D002840:Chromates; D017670:Sodium Compounds; D014874:Water Pollutants, Chemical; C028982:sodium chromate(VI)