Objective To construct a fluorescent transgenic zebrafish model sensitive to 17α-ethynylestradiol (EE2) to directly monitor environmental estrogens (EEs) pollution in aquatic system.
Methods Tg(ere-vtg:egfp) recombined plasmid was constructed, linearized and microinjected into 1-2 cell zebrafish embryos. The injected embryos were exposed to 10 ng/L EE2 to induce green fluorescent protein (GFP), and further raised till sexually mature to build transgenic zebrafish line. The sensitivity was tested with protocols using homozygote larvae and embryos exposed to different concentrations of EE2. The specificity was tested using several environmental chemicals with similar activity or structure.
Results Tg(ere-vtg:egfp) transgenic zebrafishes was successfully generated after microinjection with Tg(ere-vtg:egfp) plasmid and genetic screening. GFP expression in Tg(ere-vtg:egfp) transgenic zebrafish line was related to the concentration and time of EE2 exposure. GFP expression was induced at 0.1 ng/L EE2 (131 hpf) in zebrafish embryos. Tg(ere-vtg:egfp) transgenic zebrafish larvae expressed GFP after exposure to estrogens with varied structures but not after exposure to non-estrogen chemicals.
Conclusion Using Tg(ere-vtg:egfp) transgenic zebrafish can monitor EEs pollution dynamically.
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