title: Sensitivity of the vitellogenin assay to diagnose exposure of fathead minnows to 17α-ethynylestradiol.
abstract: Vitellogenin is frequently used as a biomarker of exposure to environmental estrogens due to its specificity and sensitivity. Appropriate incorporation of this biomarker into environmental monitoring and assessment necessitates evaluation of its critical performance parameters. In this study, we characterize the sensitivity of both vitellogenin gene (vtg) mRNA transcripts in liver and protein (VTG) in plasma over a range of concentrations and exposure durations. Male fathead minnows were exposed to 17α-ethynylestradiol (EE2) in a flow-through system for 2, 4 and 7 days at multiple EE2 concentrations in order to provide information regarding the sensitivity of each of these biomarkers to diagnose exposure to this representative estrogen. Measurements of the expression of the vitellogenin gene and protein both reliably detected exposures to EE2 at concentrations of 5ng/l and higher at all time points. Vtg mRNA and plasma VTG appear to have similar sensitivities, though the lower variability in VTG in control fish may make it more sensitive to small changes in expression compared to vtg. For lower concentrations, sensitivity may be improved by increasing exposure duration. A sample size of ∼12 fish was sufficient in many cases to produce a statistically significant increase in vitellogenin. Larger sample sizes may provide more sensitivity at low concentrations, but detecting exposure to estrogens in the lower range of environmentally relevant concentrations may need larger sample sizes. These data will assist in designing experiments that have sufficient statistical power necessary to determine if fish have been exposed to estrogens.
authors.affiliation: US Environmental Protection Agency, Office of Research and Development, National Exposure Research Laboratory, Ecological Exposure Research Division, 26 W. Martin Luther King Drive, Cincinnati, OH 45268, USA. Electronic address: flick.robert@epa.gov.
authors.forename: Robert W
authors.initials: RW
authors.lastname: Flick
authors.affiliation: US Environmental Protection Agency, Office of Research and Development, National Exposure Research Laboratory, Ecological Exposure Research Division, 26 W. Martin Luther King Drive, Cincinnati, OH 45268, USA. Electronic address: bencic.david@epa.gov.
authors.forename: David C
authors.initials: DC
authors.lastname: Bencic
authors.affiliation: US Environmental Protection Agency, Office of Research and Development, National Exposure Research Laboratory, Ecological Exposure Research Division, 26 W. Martin Luther King Drive, Cincinnati, OH 45268, USA. Electronic address: see.maryjean@epa.gov.
authors.forename: Mary J
authors.initials: MJ
authors.lastname: See
authors.affiliation: US Environmental Protection Agency, Office of Research and Development, National Exposure Research Laboratory, Ecological Exposure Research Division, 26 W. Martin Luther King Drive, Cincinnati, OH 45268, USA. Electronic address: biales.adam@epa.gov.
authors.forename: Adam D
authors.initials: AD
authors.lastname: Biales
mesh_terms: D000818:Animals; D001681:Biological Assay / Q000592:standards; D003530:Cyprinidae / Q000235:genetics / Q000502:physiology*; D004784:Environmental Monitoring / Q000379:methods* / Q000592:standards; D004997:Ethinyl Estradiol / Q000633:toxicity*; D005786:Gene Expression Regulation / Q000187:drug effects*; D008297:Male; D012680:Sensitivity and Specificity; D014819:Vitellogenins / Q000235:genetics*; D014874:Water Pollutants, Chemical / Q000633:toxicity*
chemical_list: D014819:Vitellogenins; D014874:Water Pollutants, Chemical; D004997:Ethinyl Estradiol
keywords: 17α-Ethynylestradiol (EE2); Biomarker; Estrogens; Fathead minnow; Vitellogenin