Detection of Mercury in Aquatic Environments Using EPRE Reporter Zebrafish |
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Authors: | Brandon W Kusik Michael J Carvan III Ava J Udvadia |
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Institution: | (1) Department of Biological Sciences, University of Wisconsin–Milwaukee, Milwaukee, WI 53204, USA;(2) Great Lakes WATER Institute, University of Wisconsin–Milwaukee, 600 E. Greenfield Ave., Milwaukee, WI 53204, USA |
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Abstract: | It has been proposed that transgenic zebrafish could be designed to detect low levels of chemical contaminants that cause
oxidative stress in aquatic environments, such as heavy metals or pesticides. In this paper, we describe such a transgenic
zebrafish that produces a luciferase–green fluorescent protein (LUC–GFP) fusion protein under conditions of oxidative stress.
The reporter gene expression is under the regulation of the electrophile responsive element (EPRE), which activates gene expression
in response to oxidative stressors. The GFP component of this fusion protein allows us to visually detect reporter gene activity
in live animals to determine if activity is localized to a particular tissue. The luciferase component is capable of returning
a quantitative assessment of reporter gene activity that allows us to determine if reporter gene activity is directly correlated
to the concentration of the chemical inducer. We have tested this reporter construct in both transient and stable transgenic
fish after exposure to a range of HgCl2 concentrations. GFP expression from the EPRE–LUC–GFP construct was inducible in transient assays but was below the limit
of detection in stable lines. In contrast, we observed inducible luciferase activity in both transient assays and stable lines
treated with HgCl2. We conclude that the EPRE is capable of driving reporter gene expression in a whole animal assay under conditions of oxidative
stress. Furthermore, expression was induced at HgCl2 concentrations that do not result in obvious morphological defects, making this approach useful for the detection of low
levels of oxidative contaminants in aquatic environments. |
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Keywords: | Transgenic zebrafish Oxidative stress Electrophile response element Antioxidant response element EPRE ARE |
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