title: Evaluation of Estrogenic Activity of Wastewater: Comparison Among In Vitro ERα Reporter Gene Assay, In Vivo Vitellogenin Induction, and Chemical Analysis.
abstract: The in vitro estrogen receptor (ER) reporter gene assay has long been used to measure estrogenic activity in wastewater. In a previous study, we demonstrated that the assay represents net estrogenic activity in the balance between estrogenic and antiestrogenic activities in wastewater. However, it remained unclear whether the net estrogenic activity measured by the in vitro ERα reporter gene assay can predict the in vivo estrogenic effect of wastewater. To determine this, we measured the following: estrogenic and antiestrogenic activities of wastewater and reclaimed water by the in vitro ERα reporter gene assay, expression of vitellogenin-1 (vtg1) and choriogenin-H (chgH) in male medaka (Oryzias latipes) by quantitative real-time PCR, and estrone, 17β-estradiol, estriol, and 17α-ethynylestradiol concentrations chemically to predict estrogenic activity. The net estrogenic activity measured by the in vitro medaka ERα reporter gene assay predicted the in vivo vtg1/chgH expression in male medaka more accurately than the concentrations of estrogens. These results also mean that in vivo vtg1/chgH expression in male medaka is determined by the balance between estrogenic and antiestrogenic activities. The in vitro medaka ERα reporter gene assay also predicted in vivo vtg1/chgH expression on male medaka better than the human ERα reporter gene assay.
authors.affiliation: †Research Center for Environmental Quality Management, Graduate School of Engineering, Kyoto University, 1-2 Yumihama, Otsu, Shiga 520-0811, Japan.
authors.forename: Masaru
authors.initials: M
authors.lastname: Ihara
authors.affiliation: ‡Public Works Research Institute, 1-6 Minamihara, Tsukuba, Ibaraki 305-8516, Japan.
authors.forename: Tomokazu
authors.initials: T
authors.lastname: Kitamura
authors.affiliation: †Research Center for Environmental Quality Management, Graduate School of Engineering, Kyoto University, 1-2 Yumihama, Otsu, Shiga 520-0811, Japan.|§University of South Bohemia in Ceske Budejovice, Faculty of Fisheries and Protection of Waters, South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses, Research Institute of Fish Culture and Hydrobiology, Zátiší 728/II, 389 25 Vodňany, Czech Republic.
authors.forename: Vimal
authors.initials: V
authors.lastname: Kumar
authors.affiliation: ‡Public Works Research Institute, 1-6 Minamihara, Tsukuba, Ibaraki 305-8516, Japan.|∥Environment and Bio Group, Korea Institute of Science and Technology Europe, 66123 Saarbrücken, Germany.
authors.forename: Chang-Beom
authors.initials: CB
authors.lastname: Park
authors.affiliation: †Research Center for Environmental Quality Management, Graduate School of Engineering, Kyoto University, 1-2 Yumihama, Otsu, Shiga 520-0811, Japan.
authors.forename: Mariko O
authors.initials: MO
authors.lastname: Ihara
authors.affiliation: †Research Center for Environmental Quality Management, Graduate School of Engineering, Kyoto University, 1-2 Yumihama, Otsu, Shiga 520-0811, Japan.
authors.forename: Sang-Jung
authors.initials: SJ
authors.lastname: Lee
authors.affiliation: †Research Center for Environmental Quality Management, Graduate School of Engineering, Kyoto University, 1-2 Yumihama, Otsu, Shiga 520-0811, Japan.
authors.forename: Naoyuki
authors.initials: N
authors.lastname: Yamashita
authors.affiliation: ⊥Okazaki Institute for Integrative Bioscience, National Institute for Basic Biology, National Institutes of Natural Sciences, and Department of Basic Biology, Faculty of Life Science, Graduate University for Advanced Studies, SOKENDAI, Higashiyama 5-1, Myodaiji, Okazaki, Aichi 444-8787, Japan.
authors.forename: Shinichi
authors.initials: S
authors.lastname: Miyagawa
authors.affiliation: ⊥Okazaki Institute for Integrative Bioscience, National Institute for Basic Biology, National Institutes of Natural Sciences, and Department of Basic Biology, Faculty of Life Science, Graduate University for Advanced Studies, SOKENDAI, Higashiyama 5-1, Myodaiji, Okazaki, Aichi 444-8787, Japan.
authors.forename: Taisen
authors.initials: T
authors.lastname: Iguchi
authors.affiliation: ‡Public Works Research Institute, 1-6 Minamihara, Tsukuba, Ibaraki 305-8516, Japan.
authors.forename: Seiichiro
authors.initials: S
authors.lastname: Okamoto
authors.affiliation: ‡Public Works Research Institute, 1-6 Minamihara, Tsukuba, Ibaraki 305-8516, Japan.
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Suzuki
authors.affiliation: †Research Center for Environmental Quality Management, Graduate School of Engineering, Kyoto University, 1-2 Yumihama, Otsu, Shiga 520-0811, Japan.
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Tanaka
mesh_terms: D000818:Animals; D001681:Biological Assay / Q000379:methods*; D047628:Estrogen Receptor alpha / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D004967:Estrogens / Q000032:analysis / Q000737:chemistry / Q000633:toxicity*; D015870:Gene Expression / Q000187:drug effects*; D008297:Male; D009990:Oryzias; D014819:Vitellogenins / Q000032:analysis* / Q000378:metabolism; D014874:Water Pollutants, Chemical / Q000032:analysis / Q000737:chemistry / Q000633:toxicity*
chemical_list: D047628:Estrogen Receptor alpha; D004967:Estrogens; D014819:Vitellogenins; D014874:Water Pollutants, Chemical