Seo Jung Soo, Lee Young-Mi, Jung Sang-Oun, Kim Il-Chan, Yoon Yong-Dal, Lee Jae-Seong
Department of Molecular and Environmental Bioscience, The National Research Lab of Marine Molecular and Environmental Bioscience, Hanyang University, Seoul 133-791, South Korea.
Biochem Biophys Res Commun. 2006 Jul 21;346(1):213-23. doi: 10.1016/j.bbrc.2006.05.123. Epub 2006 May 30.
To uncover the effect of estrogenic chemicals [4-nonylphenol (NP) and bisphenol A (BisA)] on the expression of androgen receptor (AR) and estrogen receptors (ERalpha and ERbeta) in the hermaphroditic fish Rivulus marmoratus, we cloned the full length of the cDNAs encoding AR, ERalpha, and ERbeta from gonadal tissue of R. marmoratus and analyzed the modulation of expression of these genes following exposure to estrogenic chemicals using real-time RT-PCR. R. marmoratus AR, ERalpha, and ERbeta genes showed a high similarity to the relevant fish species on amino acid residues, respectively. Rm-ERalpha and Rm-ERbeta cDNAs included a serine-rich region when compared to other teleost fish ER genes. Tissue-specific expression of Rm-AR and Rm-ERbeta mRNAs in adult hermaphrodite R. marmoratus was high in the gonad, while Rm-ERalpha mRNA was high in the liver based on real-time RT-PCR. In addition, Rm-AR and Rm-ERalpha mRNAs increased along with developmental stage from stage 3 (5 dpf) to hatching, while Rm-ERbeta mRNA increased from stage 2 (2 dpf). To uncover the effect of estrogenic chemicals on R. marmoratus, we exposed the fish to NP (300 microg/l) and BisA (600 microg/l) for 96 h. Significant down-regulation of Rm-AR, Rm-ERalpha, and Rm-ERbeta mRNA was observed in gonadal tissue after exposure to NP but not BisA. In the liver, there were gender differences in gene expression after EDC exposure. These results demonstrate that expression patterns of the Rm-AR, Rm-ERalpha, and Rm-ERbeta genes in the hermaphroditic fish, R. marmoratus, vary according to tissue and developmental stage as well as the specificity of environmental estrogenic chemicals. These genes can be useful as molecular biomarkers in assessing the potential impact of estrogenic compounds using this species as a model system.
为了揭示雌激素类化学物质[4-壬基酚(NP)和双酚A(BisA)]对雌雄同体鱼类花鳉(Rivulus marmoratus)雄激素受体(AR)和雌激素受体(ERα和ERβ)表达的影响,我们从花鳉的性腺组织中克隆了编码AR、ERα和ERβ的cDNA全长,并使用实时逆转录聚合酶链反应(RT-PCR)分析了暴露于雌激素类化学物质后这些基因表达的调控情况。花鳉的AR、ERα和ERβ基因在氨基酸残基上分别与相关鱼类物种具有高度相似性。与其他硬骨鱼类的ER基因相比,Rm-ERα和Rm-ERβ cDNA包含一个富含丝氨酸的区域。基于实时RT-PCR,成年雌雄同体花鳉中Rm-AR和Rm-ERβ mRNA的组织特异性表达在性腺中较高,而Rm-ERα mRNA在肝脏中较高。此外,从第3阶段(5日龄仔鱼)到孵化,Rm-AR和Rm-ERα mRNA随着发育阶段而增加,而Rm-ERβ mRNA从第2阶段(2日龄仔鱼)开始增加。为了揭示雌激素类化学物质对花鳉的影响,我们将鱼暴露于NP(300微克/升)和BisA(600微克/升)中96小时。暴露于NP后,性腺组织中观察到Rm-AR、Rm-ERα和Rm-ERβ mRNA显著下调,但暴露于BisA后未观察到。在肝脏中,暴露于环境内分泌干扰物(EDC)后基因表达存在性别差异。这些结果表明,雌雄同体鱼类花鳉中Rm-AR、Rm-ERα和Rm-ERβ基因的表达模式根据组织、发育阶段以及环境雌激素类化学物质的特异性而有所不同。这些基因可作为分子生物标志物,用于以该物种为模型系统评估雌激素化合物的潜在影响。