Detection of environmental androgens: A novel method based on enzyme‐linked immunosorbent assay of spiggin, the stickleback (Gasterosteus aculeatus) glue protein
Detection of environmental androgens: A novel method based on enzyme‐linked immunosorbent assay of spiggin, the stickleback (Gasterosteus aculeatus) glue protein
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环境雄激素的检测：一种基于刺鱼（Gasterosteus aculeatus）胶蛋白 spiggin 酶联免疫吸附测定的新方法
DOI：
10.1002/etc.5620210924
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发表时间：
2002
影响因子：
4.1
通讯作者：
I. Mayer
中科院分区：
文献类型：
--
作者：
Ioanna Katsiadaki;A. Scott;M. Hurst;P. Matthiessen;I. Mayer
我们报告了一种新的水性雄激素体内生物标志物测试的开发和验证。在繁殖过程中，雄性棘鱼（Gasterosteus aculeatus）在肾脏中制造一种胶蛋白，spiggin，用于筑巢。Spiggin的产生受雄激素的控制。然而，到目前为止，只能通过测量肾脏上皮细胞的高度来量化其产量。在本研究中，我们报告了spiggin的酶联免疫吸附测定（ELISA）的发展，并证明了其应用于测量已暴露于水中雄激素的雌性棘鱼肾脏中的spiggin。来自ELISA程序的结果显示与肾上皮细胞高度的测量的强相关性（r2 = 0.93）。然而，ELISA要快得多，并且具有相当高的响应范围（100，000倍对四倍）。通过将完整雌性动物暴露于浓度递增的17α-甲基睾酮和5α-二氢睾酮3周试验期，获得了spiggin产生的清晰、分级反应。这两种类固醇的最低有效浓度分别为100 ng/L和3 μg/L。雌性刺鱼暴露在纸浆厂的废水中，肾脏中也会产生spiggin。雌性棘鱼拥有雄激素调节蛋白，使其成为检测水生环境中雄激素污染的独特生物测定生物。
We report the development and validation of a novel in vivo biomarker test for waterborne androgens. During breeding, male sticklebacks (Gasterosteus aculeatus) manufacture a glue protein, spiggin, in their kidneys that they use to build their nests. Spiggin production is under the control of androgens. Until now, however, it has only been possible to quantify its production by measurement of the height of kidney epithelial cells. In the present study, we report the development of an enzyme‐linked immunosorbent assay (ELISA) for spiggin and demonstrate its application to the measurement of spiggin in the kidneys of female sticklebacks that have been exposed to androgens in water. Results from the ELISA procedure revealed a strong correlation with measurement of kidney epithelial cell height (r2 = 0.93). However, the ELISA was much quicker and had a considerably higher response range (100,000‐fold vs fourfold). Clear, graded responses in spiggin production were obtained by exposing intact females to increasing concentrations of 17α‐methyltestosterone and 5α‐dihydrotestosterone over three‐week test periods. The lowest effective concentrations for these two steroids were 100 ng/L and 3 μg/L, respectively. Female sticklebacks that were exposed to pulp mill effluent also produced spiggin in their kidneys. Possession of an androgen‐regulated protein by the female stickleback makes it a unique bioassay organism for detecting androgenic contamination in the aquatic environment.
影响因子：
4.8
作者：
T. Sperry;Peter Thomas
通讯作者：
T. Sperry;Peter Thomas