Canadian Rivers Institute, Department of Biology, University of Prince Edward Island, Charlottetown, Prince Edward Island, Canada.
Environ Toxicol Chem. 2012 Jun;31(6):1391-5. doi: 10.1002/etc.1815. Epub 2012 Apr 18.
Previous research demonstrated that exposure to exogenous androgens and effluents with androgenic activity can induce spiggin mRNA production in the kidney of the three-spined stickleback (Gasterosteus aculeatus). In the present study, we determine whether a short-term exposure to a known antiandrogenic pesticide, linuron (LN), suppresses spiggin mRNA in male stickleback and in androgenized female stickleback. Primers were designed from previously characterized sequences for each androgen receptor (AR) isoform in stickleback, arα and arβ, to assess whether these receptors are differentially regulated by androgen or antiandrogen exposure. Fish were exposed for 72 h to one of four treatments: control, LN (250 µg/L), 17α-methyltestosterone (MT, 500 ng/L), and an LN-MT mixture at those same concentrations. There was no effect of LN on spiggin and arβ mRNA levels in male kidney, while levels of arα were significantly increased twofold. Exposure to LN significantly inhibited MT-induced spiggin RNA production in female kidney with no effect on expression of arα and arβ. The present study is the first to demonstrate the antiandrogenic effect of LN at the transcript level and to examine androgenic/antiandrogenic responsiveness of the two ARs in the stickleback. From the present study, it was determined that measurement of spiggin RNA is a reliable and sensitive screening tool for the detection of both androgenic and antiandrogenic compounds.
先前的研究表明,外源性雄激素和具有雄激素活性的污水会导致三刺鱼(Gasterosteus aculeatus)肾脏中 spiggin mRNA 的产生。在本研究中,我们确定短期暴露于已知的抗雄激素农药——利谷隆(LN)是否会抑制雄性和雄性化雌性三刺鱼的 spiggin mRNA。我们从已鉴定的三刺鱼 AR 同工型(arα 和 arβ)的序列中设计了引物,以评估这些受体是否受到雄激素或抗雄激素暴露的差异调节。鱼在 72 小时内分别暴露于以下四种处理中的一种:对照、LN(250μg/L)、17α-甲基睾酮(MT,500ng/L)和相同浓度的 LN-MT 混合物。LN 对雄性肾脏中的 spiggin 和 arβ mRNA 水平没有影响,而 arα 水平则显著增加了两倍。LN 暴露显著抑制了 MT 诱导的雌性肾脏中的 spiggin RNA 产生,而对 arα 和 arβ 的表达没有影响。本研究首次证明了 LN 在转录水平上的抗雄激素作用,并研究了两种 AR 在三刺鱼中的雄激素/抗雄激素反应性。本研究确定,spiggin RNA 的测量是检测雄激素和抗雄激素化合物的可靠和敏感的筛选工具。