title: Development and Application of an Effective Detection Method for Fish Plasma Vitellogenin Induced by Environmental Estrogens.
abstract: Vitellogenin is a protein induced by estrogens, including environmental chemicals with estrogenic activity. To measure the effects of environmental estogens, we developed an effective and rapid one-step method of detecting and purifying fish plasma vitellogenin using a high-performance anion-exchange chromatography column, POROS-HQ. Vitellogenin in a plasma of estradiol-treated male fish (mummichog and red sea bream) was eluted as a single peak with a retention time of 10 minutes from the column, which gives an almost pure preparation as assessed by SDS-PAGE. The lowest detectable amount of vitellogenin was 2 μg per assay. The method was used to analyze the plasma vitellogenin level of aquacultured red sea breams caught in August, when the spawning season is over, and usually no vitellogenin is detected in either females or males, physiologically. However, the data showed that in addition to a few females, some male fish synthesized vitellogenin, suggesting that some chemicals or unknown factors with estrogenic activity have induced fish in the ocean to produce vitellogenin.
authors.affiliation: a School of Pharmaceutical Sciences, Nagasaki University.
authors.forename: S
authors.initials: S
authors.lastname: Yamanaka
authors.affiliation: b Faculties of Environmental Studies, Nagasaki University.
authors.forename: K
authors.initials: K
authors.lastname: Arizono
authors.affiliation: b Faculties of Environmental Studies, Nagasaki University.
authors.forename: Y
authors.initials: Y
authors.lastname: Matsuda
authors.affiliation: c Faculties of Fisheries of Nagasaki University.
authors.forename: K
authors.initials: K
authors.lastname: Soyano
authors.affiliation: d Faculty of Sciences of Yokohama City University.
authors.forename: H
authors.initials: H
authors.lastname: Urushitani
authors.affiliation: d Faculty of Sciences of Yokohama City University.
authors.forename: T
authors.initials: T
authors.lastname: Iguchi
authors.affiliation: a School of Pharmaceutical Sciences, Nagasaki University.
authors.forename: R
authors.initials: R
authors.lastname: Sakakibara
keywords: HPLC; endocrine disrupters; environmental estrogens; red sea bream; vitellogenin