Development of a fluorescent transgenic zebrafish biosensor for sensing aquatic heavy metal pollution.
Transgenic Res
; 25(5): 617-27, 2016 10.
Article
em En
| MEDLINE
| ID: mdl-27120052
ABSTRACT
We report a transgenic zebrafish (Danio rerio) designed to respond to heavy metals using a metal-responsive promoter linked to a fluorescent reporter gene (DsRed2). The metallothionein MT-Ia1 promoter containing metal-responsive elements was derived from the Asian green mussel, Perna viridis. The promoter is known to be induced by a broad spectrum of heavy metals. The promoter-reporter cassette cloned into the Tol2 transposon vector was microinjected into zebrafish embryos that were then reared to maturity. A transgene integration rate of 28 % was observed. The confirmed transgenics were mated with wild-type counterparts, and pools of F1 embryos were exposed to sub-lethal doses of Cd(2+), Cu(2+), Hg(2+), Pb(2+) and Zn(2+). The red fluorescence response of zebrafish embryos was observed 8 h post- exposure to these sub-lethal doses of heavy metals using a fluorescence microscope. Reporter expression estimated by real-time PCR revealed eightfold, sixfold and twofold increase on exposure to highest concentrations of Hg(2+), Cd(2+) and Cu(2+), while Pb(2+) and Zn(2+) had no effect. This biosensor could be a first-level screening method for confirming aquatic heavy metal bio-toxicity to eukaryotes.
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MEDLINE
Assunto principal:
Poluentes Químicos da Água
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Animais Geneticamente Modificados
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Técnicas Biossensoriais
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Metais Pesados
Limite:
Animals
Idioma:
En
Ano de publicação:
2016
Tipo de documento:
Article