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Elevated serum vitellogenin and gonadal abnormalities in wild male founder from Tokyo Bay, Japan

日本东京湾野生雄性创始人血清卵黄蛋白原升高和性腺异常

基本信息

批准号：
10680496
负责人：
HASHIMOTO Shinya
金额：
$ 2.18万
依托单位：
Tokyo University of Fisheries
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 1999
项目状态：
已结题

项目摘要

We have investigate the seasonal changes of serum VTG, as a biomarker of the effect of environmental estrogens, in wild male flounder (Pleuronectes yokohamae) collected from Tokyo Bay, and compare these levels to those in male flounder from a reference site along the coast of Japan. Our reference station was located off Shiriuchi in Hokkaido, where sewage and wastewater inputs to the marine environment were considered to be negligible. Pleuronectes yokohamae from Tokyo Bay, aged approximately 1 to 6 yr. were caught between January 1997 and May 1998 (male: n=130, female: n=143), covering an entire reproductive season. Reference specimens were collected off Shiriuchi, in the Strait of Tsugaru, between March 1997 and June 1998 (male: n= 62, female: n=64). Mean serum levels of VTG for female flounder from Tokyo Bay and Hokkaido suggest that no obvious difference (elevated or depressed) of VTG of females was observed between the two sites. At the reference site off Shiriuchi, the mean VTG l … More evels for male flounder was < 50 ng mlィイD1-1ィエD1. In contrast, male flounder in Tokyo Bay showed elevated levels of VTG almost throughout the year (mean: 204 ng mlィイD1-1ィエD1; ranging from < 50 ng ml ィイD1-1ィエD1 to 2200 ng mlィイD1-1ィエD1). The VTG levels in male flounder in Tokyo Bay (n=130) were significantly higher (p <0.001) compared to those of the reference fish from Hokkaido (n=62). The considerable variation in serum VTG concentration observed in individual males from Tokyo Bay could be due to individual differences in sensitivity to or degree of accumulation of estrogenic compounds.The relationship between VTG in male flounder from Tokyo Bay and other factors such as standard length, hepatosomatie index (HSI), 17β-estradiol (EィイD22ィエD2), testosterone (T), and gonadosomatic index (GSI) was investigated. We found no significant relationship between VTG and standard length which indicates that induction of VTG occurs among male flounder of all ages above one year. Furthermore, no relationship was observed between VTG and HSI, EィイD22ィエD2 or T levels. However, high concentrations of serum VTG were observed more often in male flounder whose GSI was lower than 2% in Tokyo Bay. The GSI is therefore an additional factor that should be considered when male flounder is deployed as biomonitor of environmental estrogens.The development of a very sensitive method for the determination of very low levels of plasma VTG in male fish allows, a) detection of trace amounts of estrogenic compounds and mainly, b) analysis of highly diluted serum samples, which can avoid a critical problem of the ELISA technique, the unspecific interference with serum compounds at high concentrations. We developed and optimized a luminometric immunoassay that was able to detect trace amounts of VTG in the serum of male flounder collected from a reference site, located off Shiriuchi in Hokkaido, where sewage and wastewater inputs to the marine environment were considered negligible. This method can be applied successfully to monitor seasonal changes in baseline VTG levels in serum samples obtained from a reference site. Such results will be useful in interpreting the effects of environmental estrogens in ecosystems.Both EィイD22ィエD2 and T in the sera of male and female fish were high during winter, when VTG levels in female fish and the GSI of both sexes were also high. Estrogen has been found in the blood of wild males at levels as high as 10% of those normally seen in mature females in winter. The detectable levels of VTG in the blood of wild male flounder at the reference site in winter may be a consequence of the presence of physiologically active levels of estrogen. Perhaps this species expresses VTG in both sexes in response to "natural" circulating EィイD22ィエD2.Histological analysis revealed that the gonads of about 15% of male flounder (3 out of 20) collected off Haneda in Tokyo Bay contained female gametes (intersexual gonads contained predominately male tissue). The degree of intersexuality was slight, with a few primary oocytes scattered within apparently normal testicular tissue. No abnormalities were seen in the testes of male flounder collected at the reference site in Hokkaido or in the ovaries of females from either site. Less

我们调查了血清VTG的季节性变化，作为环境雌激素的影响的生物标志物，在野生雄性比目鱼（Pleuronectes yokohamae）从东京湾收集，并比较这些水平的男性比目鱼从参考网站沿着日本海岸。我们的基准站位于北海道的Shiriuchi，那里的污水和废水输入海洋环境被认为是微不足道的。东京湾的横滨鲽，年龄约1至6岁。在1997年1月至1998年5月期间捕获的（雄性：n=130，雌性：n=143），覆盖了整个生殖季节。参考标本采集于1997年3月至1998年6月期间，在津轻海峡的Shiriuchi（男性：n= 62，女性：n=64）。东京湾和北海道的雌性比目鱼血清VTG的平均水平表明，两个地点的雌性比目鱼的VTG没有明显的差异（升高或降低）。在Shiriuchi外的参考点，平均VTG为1 ...更多信息雄性比目鱼的水平为< 50 ng ml/kg D1-1 ng/kg D1。相比之下，东京湾的雄性比目鱼几乎全年都显示出VTG水平升高（平均值：204 ng ml VTG D1-1 VTG D1;范围从< 50 ng ml VTG D1-1 VTG D1到2200 ng ml VTG D1-1 VTG D1）。东京湾的雄性比目鱼（n=130）的VTG水平显著高于来自北海道的参考鱼（n=62）（p <0.001）。东京湾雄性比目鱼血清中VTG含量的差异可能是由于个体对雌激素的敏感性或积累程度的差异所致，本文还研究了东京湾雄性比目鱼VTG含量与标准体长、肝体指数（HSI）、17β-雌二醇（17β-E2）等因素的关系（E_（12）D_（22）D_2）、睾酮（T）和性腺指数（GSI）。结果表明，VTG与标准体长之间无显著相关性，表明一龄以上的雄性比目鱼均存在VTG的诱导。此外，VTG与HSI、E β D22、D β D2和T水平之间无相关性。然而，高浓度的血清VTG更常见于雄性比目鱼，其GSI低于2%的东京湾。因此，GSI是一个额外的因素，应该考虑当雄性比目鱼部署作为环境雌激素的biomonitor.一个非常敏感的方法，用于测定非常低水平的血浆VTG在雄鱼的发展允许，a）检测痕量的雌激素化合物，主要是，B）分析高度稀释的血清样品，这可以避免ELISA技术的关键问题，高浓度血清化合物的非特异性干扰。我们开发和优化了一种发光免疫分析，能够检测微量的VTG的血清中收集的雄性比目鱼从参考网站，位于北海道北海道，在那里的污水和废水输入的海洋环境被认为是微不足道的。该方法可以成功地应用于监测从参考站点获得的血清样品中基线VTG水平的季节性变化。冬季雌鱼血清中E_（10）、D_（22）、D_（22）和T均较高，雌鱼VTG和两性GSI也较高。在野生雄性的血液中发现雌激素的水平高达冬季成熟雌性血液中正常水平的10%。在冬季的参考网站的野生雄性比目鱼的血液中的VTG的检测水平可能是生理活性水平的雌激素的存在下的结果。组织学分析表明，在东京湾羽田海域采集的20只雄性比目鱼中，约有15%（3/20）的性腺含有雌性配子（间性性腺主要含有雄性组织）。间性的程度是轻微的，与一些初级卵母细胞分散在明显正常的睾丸组织。在北海道的参考地点采集的雄性比目鱼睾丸或来自两个地点的雌性比目鱼卵巢中均未观察到异常。少