Development and Application of an Effective Detection Method for Fish Plasma Vitellogenin Induced by Environmental Estrogens.
Development and Application of an Effective Detection Method for Fish Plasma Vitellogenin Induced by Environmental Estrogens.
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环境雌激素诱导的鱼血浆卵黄蛋白原有效检测方法的开发和应用。
DOI：
10.1271/bbb.62.1196
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发表时间：
1998
期刊：
影响因子：
--
通讯作者：
R. Sakakibara
中科院分区：
文献类型：
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作者：
S. Yamanaka;K. Arizono;Y. Matsuda;K. Soyano;H. Urushitani;T. Iguchi;R. Sakakibara
卵黄蛋白原是一种由雌激素诱导的蛋白质，包括具有雌激素活性的环境化学物质。为了检测环境雌激素的影响，我们建立了一种高效、快速的一步法检测和纯化鱼血浆卵黄蛋白原，使用高效阴离子交换色谱柱POOS-HQ。雌二醇处理雄鱼血浆中的卵黄蛋白原以单峰形式洗脱，保留时间为10分钟，经SDS-PAGE鉴定为几乎纯的制剂。卵黄蛋白原的最低检出量为2μg/次。用该方法分析了8月份产卵季节结束时捕获的水产养殖红鱼的血浆卵黄原蛋白水平，从生理上讲，雌性和雄性通常都检测不到卵黄原蛋白。然而，数据显示，除了少数雌性鱼类外，还有一些雄性鱼类合成了卵黄原蛋白，这表明一些具有雌激素活性的化学物质或未知因素诱导了海洋中的鱼类产生卵黄原蛋白。
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.