Transgenic zebrafish as sentinels for aquatic pollution

Carvan III, M.J., Dalton, T.P., Stuart, G.W., and Nebert, D.W.
Annals of the New York Academy of Sciences   919: 133-147 (Journal)
Registered Authors
Carvan III, Michael J., Nebert, Daniel W., Stuart, Gary
MeSH Terms
  • Animals
  • Animals, Genetically Modified
  • Antioxidants/pharmacology
  • Cells, Cultured
  • Dioxins/analysis
  • Dioxins/pharmacology
  • Enhancer Elements, Genetic/genetics
  • Environmental Monitoring/methods*
  • Estrogens/pharmacology
  • Gene Expression Regulation*/drug effects
  • Genes, Reporter/genetics
  • Metals/pharmacology
  • Polychlorinated Biphenyls/analysis
  • Polychlorinated Biphenyls/pharmacology
  • Polycyclic Compounds/analysis
  • Polycyclic Compounds/pharmacology
  • Receptors, Aryl Hydrocarbon/metabolism
  • Response Elements/genetics
  • Transgenes/genetics
  • Tretinoin/pharmacology
  • Water Pollutants, Chemical/analysis
  • Water Pollutants, Chemical/pharmacology
  • Water Pollution/analysis*
  • Zebrafish/genetics*
11083105 Full text @ Ann N Y Acad Sci
Using the golden mutant zebrafish having a decrease in interfering pigmentation, we are developing transgenic lines in which DNA motifs that respond to selected environmental pollutants are capable of activating a reporter gene that can be easily assayed. We have begun with three response elements that recognize three important classes of foreign chemicals. Aromatic hydrocarbon response elements (AHREs) respond to numerous polycyclic hydrocarbons and halogenated coplanar molecules such as 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD; dioxin) and polychlorinated biphenyls. Electrophile response elements (EPREs) respond to quinones and numerous other potent electrophilic oxidants. Metal response elements (MREs) respond to heavy metal cations such as mercury, copper, nickel, cadmium, and zinc. Soon, we will include estrogen response elements (EREs) to detect the effects of environmental endocrine disruptors, and retinoic acid response elements (RARE, RXRE) to detect the effects of retinoids in the environment. Each of these substances is known to be bioconcentrated in fish to varying degrees; for example, 10(-17) M TCDD in a body of water becomes concentrated to approximately 10(-12) M TCDD in a fish, where it would act upon the AHRE motif and turn on the luciferase (LUC) reporter gene. The living fish as a sentinel will not only be assayed intact in the luminometer, but--upon several days or weeks of depuration--would be usable again. To date, we have established that zebrafish transcription factors are able to recognize both mammalian and trout AHRE, EPRE, and MRE sequences in a dose-dependent and chemical-class-specific manner, and that expression of both the LUC and jellyfish green fluorescent protein (GFP) reporter genes is easily detected in zebrafish cell cultures and in the intact live zebrafish. Variations in sensitivity of this model system can be achieved by increasing the copy number of response elements and perhaps by altering the sequence of each core consensus response element and flanking regions. This transgenic technology should allow for a simple, exquisitely sensitive, and inexpensive assay for monitoring aquatic pollution. We have already initiated studies using sentinel zebrafish to monitor a public drinking water source.
Genes / Markers
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Engineered Foreign Genes