Analysis of estrogenic effects by quantification of green fluorescent protein in juvenile fish of a transgenic medaka

Stefan Scholz, Kanta Kurauchi, Masato Kinoshita, Yuji Oshima, Kenjiro Ozato, Kristin Schirmer, Yuko Wakamatsu

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

The ChgH-GFP strain of the teleost medaka contains a regulatory region of the estrogen-responsive choriogenin H (chgH) gene fused to the green fluorescent protein (GFP) gene. The strain was developed for the identification of environmental estrogens by noninvasive analysis of GFP fluorescence. In the present study, a quantification method for GFP by image analysis was established and applied to the analysis of time- and concentration-dependent GFP fluorescence in juvenile fish. Concentration-response analyses were performed with fish exposed for 14 d to 17β-estradiol (0.37-367 pM), genistein (0.37-367 nM), or p-nonylphenol (0.367-1,835 nM). By means of image analysis, it was shown that ChgH-GFP was induced at 183.5 pM or greater 17β-estradiol. Time-course and recovery experiments indicated a strong accumulation of GFP in the liver. Results of reverse transcriptase-polymerase chain reaction analysis of choriogenin H and vitellogenin demonstrated induction of gene expression for the same range of concentrations as that for GFP analysis. Neither expression of these genes nor GFP fluorescence was induced by genistein and p-nonylphenol. Although the ChgH-GFP strain failed to detect these weakly estrogenic compounds, the simplicity of the GFP quantification during early life stages of fish offers promising possibilities for further developments of transgenic strains using different target regulatory sequences.

Original languageEnglish
Pages (from-to)2553-2561
Number of pages9
JournalEnvironmental Toxicology and Chemistry
Volume24
Issue number10
DOIs
Publication statusPublished - Oct 1 2005

Fingerprint

Oryzias
Green Fluorescent Proteins
Fish
Estrogens
Fishes
protein
fish
fluorescence
Genistein
Genes
Fluorescence
image analysis
Image analysis
gene
Estradiol
effect
analysis
estrogenic compound
Vitellogenins
Gene Expression

All Science Journal Classification (ASJC) codes

  • Environmental Chemistry
  • Health, Toxicology and Mutagenesis

Cite this

Analysis of estrogenic effects by quantification of green fluorescent protein in juvenile fish of a transgenic medaka. / Scholz, Stefan; Kurauchi, Kanta; Kinoshita, Masato; Oshima, Yuji; Ozato, Kenjiro; Schirmer, Kristin; Wakamatsu, Yuko.

In: Environmental Toxicology and Chemistry, Vol. 24, No. 10, 01.10.2005, p. 2553-2561.

Research output: Contribution to journalArticle

Scholz, Stefan ; Kurauchi, Kanta ; Kinoshita, Masato ; Oshima, Yuji ; Ozato, Kenjiro ; Schirmer, Kristin ; Wakamatsu, Yuko. / Analysis of estrogenic effects by quantification of green fluorescent protein in juvenile fish of a transgenic medaka. In: Environmental Toxicology and Chemistry. 2005 ; Vol. 24, No. 10. pp. 2553-2561.
@article{d8ce5be4717e45c0803aaceb37a2e68e,
title = "Analysis of estrogenic effects by quantification of green fluorescent protein in juvenile fish of a transgenic medaka",
abstract = "The ChgH-GFP strain of the teleost medaka contains a regulatory region of the estrogen-responsive choriogenin H (chgH) gene fused to the green fluorescent protein (GFP) gene. The strain was developed for the identification of environmental estrogens by noninvasive analysis of GFP fluorescence. In the present study, a quantification method for GFP by image analysis was established and applied to the analysis of time- and concentration-dependent GFP fluorescence in juvenile fish. Concentration-response analyses were performed with fish exposed for 14 d to 17β-estradiol (0.37-367 pM), genistein (0.37-367 nM), or p-nonylphenol (0.367-1,835 nM). By means of image analysis, it was shown that ChgH-GFP was induced at 183.5 pM or greater 17β-estradiol. Time-course and recovery experiments indicated a strong accumulation of GFP in the liver. Results of reverse transcriptase-polymerase chain reaction analysis of choriogenin H and vitellogenin demonstrated induction of gene expression for the same range of concentrations as that for GFP analysis. Neither expression of these genes nor GFP fluorescence was induced by genistein and p-nonylphenol. Although the ChgH-GFP strain failed to detect these weakly estrogenic compounds, the simplicity of the GFP quantification during early life stages of fish offers promising possibilities for further developments of transgenic strains using different target regulatory sequences.",
author = "Stefan Scholz and Kanta Kurauchi and Masato Kinoshita and Yuji Oshima and Kenjiro Ozato and Kristin Schirmer and Yuko Wakamatsu",
year = "2005",
month = "10",
day = "1",
doi = "10.1897/04-525R.1",
language = "English",
volume = "24",
pages = "2553--2561",
journal = "Environmental Toxicology and Chemistry",
issn = "0730-7268",
publisher = "John Wiley and Sons Ltd",
number = "10",

}

TY - JOUR

T1 - Analysis of estrogenic effects by quantification of green fluorescent protein in juvenile fish of a transgenic medaka

AU - Scholz, Stefan

AU - Kurauchi, Kanta

AU - Kinoshita, Masato

AU - Oshima, Yuji

AU - Ozato, Kenjiro

AU - Schirmer, Kristin

AU - Wakamatsu, Yuko

PY - 2005/10/1

Y1 - 2005/10/1

N2 - The ChgH-GFP strain of the teleost medaka contains a regulatory region of the estrogen-responsive choriogenin H (chgH) gene fused to the green fluorescent protein (GFP) gene. The strain was developed for the identification of environmental estrogens by noninvasive analysis of GFP fluorescence. In the present study, a quantification method for GFP by image analysis was established and applied to the analysis of time- and concentration-dependent GFP fluorescence in juvenile fish. Concentration-response analyses were performed with fish exposed for 14 d to 17β-estradiol (0.37-367 pM), genistein (0.37-367 nM), or p-nonylphenol (0.367-1,835 nM). By means of image analysis, it was shown that ChgH-GFP was induced at 183.5 pM or greater 17β-estradiol. Time-course and recovery experiments indicated a strong accumulation of GFP in the liver. Results of reverse transcriptase-polymerase chain reaction analysis of choriogenin H and vitellogenin demonstrated induction of gene expression for the same range of concentrations as that for GFP analysis. Neither expression of these genes nor GFP fluorescence was induced by genistein and p-nonylphenol. Although the ChgH-GFP strain failed to detect these weakly estrogenic compounds, the simplicity of the GFP quantification during early life stages of fish offers promising possibilities for further developments of transgenic strains using different target regulatory sequences.

AB - The ChgH-GFP strain of the teleost medaka contains a regulatory region of the estrogen-responsive choriogenin H (chgH) gene fused to the green fluorescent protein (GFP) gene. The strain was developed for the identification of environmental estrogens by noninvasive analysis of GFP fluorescence. In the present study, a quantification method for GFP by image analysis was established and applied to the analysis of time- and concentration-dependent GFP fluorescence in juvenile fish. Concentration-response analyses were performed with fish exposed for 14 d to 17β-estradiol (0.37-367 pM), genistein (0.37-367 nM), or p-nonylphenol (0.367-1,835 nM). By means of image analysis, it was shown that ChgH-GFP was induced at 183.5 pM or greater 17β-estradiol. Time-course and recovery experiments indicated a strong accumulation of GFP in the liver. Results of reverse transcriptase-polymerase chain reaction analysis of choriogenin H and vitellogenin demonstrated induction of gene expression for the same range of concentrations as that for GFP analysis. Neither expression of these genes nor GFP fluorescence was induced by genistein and p-nonylphenol. Although the ChgH-GFP strain failed to detect these weakly estrogenic compounds, the simplicity of the GFP quantification during early life stages of fish offers promising possibilities for further developments of transgenic strains using different target regulatory sequences.

UR - http://www.scopus.com/inward/record.url?scp=26844546168&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=26844546168&partnerID=8YFLogxK

U2 - 10.1897/04-525R.1

DO - 10.1897/04-525R.1

M3 - Article

C2 - 16268157

AN - SCOPUS:26844546168

VL - 24

SP - 2553

EP - 2561

JO - Environmental Toxicology and Chemistry

JF - Environmental Toxicology and Chemistry

SN - 0730-7268

IS - 10

ER -