In vivo visual reporter system for detection of estrogen-like substances by transgenic medaka.
Detection of endocrine disrupting chemicals, in particular, environmental estrogens with living organisms, has many advantages if compared to chemical analysis. The screening of novel pollutants with meaningful endpoints, the integration of uptake, bioconcentration, and excretion as well as the evaluation of endocrine disrupting effects with respect to toxicity require in vivo biotests for estrogen-like substances (ELSs). Critical disadvantages of whole organism biotests are their low sensitivity and the need for laborious and time-consuming work. To overcome these problems, we have developed a transgenic medaka strain harboring the green fluorescence protein (GFP) gene driven by choriogenin H gene regulatory elements. Choriogenin H is an egg envelope protein induced by estrogens in the liver. With yolk sac larvae of this strain, GFP induction in liver was observed 24 h after onset of aqueous exposure to 0.63 nM 17beta-estradiol (E2), 0.34 nM ethynylestradiol, or 14.8 nM estrone. Furthermore, concentrated sewage treatment effluent induced GFP expression. Comparison of E2 equivalents estimated by GFP-induction in transgenic medaka, a YES assay, and GC/MS showed detection limits in the same order of magnitude. These results indicated that the sensitivity of the transgenic medaka strain was sufficient for application as an alternative model in monitoring environmental water samples for ELSs.
Kurauchi
Kanta
K
Graduate School of Agriculture, Kyoto University, Sakyo, Kyoto 606-8502, Japan.
Nakaguchi
Yoshitsugu
Y
Tsutsumi
Makiko
M
Hori
Hirosi
H
Kurihara
Ryo
R
Hashimoto
Shinya
S
Ohnuma
Ryoko
R
Yamamoto
Yoshikazu
Y
Matsuoka
Sumiko
S
Kawai
Shin'Ichiro
S
Hirata
Takashi
T
Kinoshita
Masato
M
D000818:Animals; D030801:Animals, Genetically Modified; D001483:Base Sequence; D001681:Biological Assay / Q000379:methods*; D015415:Biomarkers / Q000032:analysis; D004527:Egg Proteins / Q000235:genetics; D004703:Endocrine System / Q000187:drug effects; D004967:Estrogens / Q000032:analysis* / Q000633:toxicity; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000032:analysis; D008099:Liver / Q000378:metabolism; D011498:Protein Precursors / Q000235:genetics; D012722:Sewage / Q000032:analysis* / Q000737:chemistry; D012733:Sex Differentiation / Q000235:genetics; D013997:Time Factors; D014874:Water Pollutants, Chemical / Q000032:analysis* / Q000633:toxicity
D015415:Biomarkers; D004527:Egg Proteins; D004967:Estrogens; D011498:Protein Precursors; D012722:Sewage; D014874:Water Pollutants, Chemical; C104950:choriogenin H; D049452:Green Fluorescent Proteins