Detection of mercury in aquatic environments using EPRE reporter zebrafish. Mar Biotechnol (NY) 2008;10(6):750-7
Date
06/10/2008Pubmed ID
18537037Pubmed Central ID
PMC8898586DOI
10.1007/s10126-008-9113-xScopus ID
2-s2.0-55949099524 (requires institutional sign-in at Scopus site) 41 CitationsAbstract
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 HgCl(2) 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 HgCl(2). 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 HgCl(2) 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.
Author List
Kusik BW, Carvan MJ 3rd, Udvadia AJAuthor
Ava Udvadia BS,PhD Associate Professor in the Biological Sciences department at University of Wisconsin - MilwaukeeMESH terms used to index this publication - Major topics in bold
AnimalsAnimals, Genetically Modified
Embryo, Nonmammalian
Environmental Monitoring
Gene Expression
Gene Expression Regulation, Developmental
Genes, Reporter
Green Fluorescent Proteins
Luciferases
Mercuric Chloride
Oxidative Stress
Response Elements
Transgenes
Water Pollutants, Chemical
Zebrafish
