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Assessment of environmental endocrine disruption with plasma concentration of vitellogenin in Koi carp of rivers and wetland sites.

通过河流和湿地锦鲤血浆卵黄蛋白原浓度评估环境内分泌干扰。

基本信息

批准号：
11839007
负责人：
KERA Yoshio
金额：
$ 2.3万
依托单位：
Nagaoka University of Technology
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2000
项目状态：
已结题

项目摘要

Vitellogenin (Vtg) is a female specific plasma protein and the major precursor of egg yolk protein in oviparous vertebrates. The protein can serve as a useful biomarker for environmental endocrine disruption. The present study was carried out to assess environmental endocrine disruption with plasma Vtg concentration in Koi carp (Cyprinus carpio) of rivers and wetland sites.We purified Koi carp Vtg to homogeneity, raised specific polyclonal antiserum to the purified Koi carp Vtg and established a single radial immunodiffusion assay and a homologous enzyme-linked immunosorbent assay (ELISA) to measure plasma Vtg concentration.To assess environmental endocrine disruption, wild Koi carp were collected in the wetland site Sakata and Toyanogata and the Shinano River. Two of 24 males captured in the Shinano River in the Ojiya City area and 5 of 15 males captured in the Shinano River in Niigata City area possessed slightly higher plasma Vtg concentrations (>1 μg/ml).Plasma Vtg levels in juvenile male carp, which were exposed to effluent from sewage treatment plant for 10-14 days, were higher than those in the unexposed male fish, suggesting that the effluent contained estrogenic compounds at a biological active level. The analyses of the effluent with GC/MS revealed that estradiol- 17β in the effluent is one of the main compounds to increase plasma Vtg level in Koi carp.

卵黄原蛋白（Vitellogenin, Vtg）是雌性特有的血浆蛋白，是卵生脊椎动物卵黄蛋白的主要前体。该蛋白可作为环境内分泌干扰的有用生物标志物。本研究旨在探讨河流和湿地环境中锦鲤血浆Vtg浓度对环境内分泌的影响。将锦鲤Vtg纯化至均匀性，对纯化的锦鲤Vtg建立特异性多克隆抗血清，并建立单径向免疫扩散法和同源酶联免疫吸附法（ELISA）检测血浆Vtg浓度。为了评估环境内分泌干扰，在坂田、丰口和信野河湿地采集了野生锦鲤。在Ojiya市新野河地区捕获的24只雄鱼中有2只和在新潟市新野河地区捕获的15只雄鱼中有5只血浆中Vtg浓度略高（bb0.1 μg/ml）。暴露于污水处理厂出水10 ~ 14天的幼鱼血浆Vtg水平高于未暴露于污水处理厂出水10 ~ 14天的幼鱼血浆Vtg水平，表明污水中含有具有生物活性的雌激素化合物。气相色谱质谱分析表明，出水中的雌二醇- 17β是提高锦鲤血浆Vtg水平的主要化合物之一。