Kausch Ulf, Alberti Martin, Haindl Stefanie, Budczies Jan, Hock Bertold
Chair of Cell Biology, TU Muenchen, Alte Akademie 12, D-85350, Freising, Germany.
Environ Toxicol. 2008 Feb;23(1):15-24. doi: 10.1002/tox.20306.
Gene expression analyses in male zebrafish (Danio rerio) were carried out using microarray technique and quantitative polymerase chain reaction. Genes responding to the exposure to 17beta-estradiol, bisphenol A and genistein were identified, among them genes involved in metabolism, reproductional and developmental processes. Threshold levels of 17beta-estradiol (200 ng/L), bisphenol A (2000 microg/L), and genistein (5000 microg/L) for the upregulation of the vtg1 gene in short-time exposures (11 days) were determined by qPCR. 14k microarrays were used to generate complete lists of genes regulated by these estrogenic compounds. For this purpose, liver samples from 10 exposed zebrafish and 10 controls were processed. In this case the expressions of 211 genes were significantly regulated by 17beta-estradiol, 47 by bisphenol A and 231 by genistein. Furthermore, it is shown that fish exposed to 17beta-estradiol and genistein have similarities in their gene expression patterns, whereas bisphenol A apparently affected gene expression in a different way. Only genes coding for egg-yolk precursor protein vitellogenin were found to be regulated by all three compounds, which shows that these genes are the only suitable markers for exposure to different estrogenic compounds. The regulated genes were assigned to gene ontology classes. All three estrogenic compounds regulated genes mainly involved in primary and cellular metabolism, but genistein regulated several genes involved in cell cycle-regulation and bisphenol A several genes involved in protein biosynthesis. Genistein also upregulated the expression of four eggshell proteins, which can be used as biomarkers for exposure to this chemical.
利用微阵列技术和定量聚合酶链反应对雄性斑马鱼（Danio rerio）进行了基因表达分析。鉴定出了对17β-雌二醇、双酚A和染料木黄酮暴露有反应的基因，其中包括参与代谢、生殖和发育过程的基因。通过qPCR确定了在短期暴露（11天）中vtg1基因上调的17β-雌二醇（200 ng/L）、双酚A（2000 μg/L）和染料木黄酮（5000 μg/L）的阈值水平。使用14k微阵列生成了由这些雌激素化合物调控的完整基因列表。为此，对10条暴露的斑马鱼和10条对照鱼的肝脏样本进行了处理。在这种情况下，211个基因的表达受到17β-雌二醇的显著调控，47个受双酚A调控，231个受染料木黄酮调控。此外，研究表明，暴露于17β-雌二醇和染料木黄酮的鱼在基因表达模式上有相似之处，而双酚A显然以不同的方式影响基因表达。仅发现编码卵黄前体蛋白卵黄生成素的基因受所有三种化合物调控，这表明这些基因是暴露于不同雌激素化合物的唯一合适标志物。将受调控的基因归类到基因本体类别中。所有三种雌激素化合物调控的基因主要涉及初级和细胞代谢，但染料木黄酮调控了几个参与细胞周期调控的基因，双酚A调控了几个参与蛋白质生物合成的基因。染料木黄酮还上调了四种蛋壳蛋白的表达，这些蛋白可作为暴露于这种化学物质的生物标志物。
