Gene Expression Analysis for Exposure to Estrogenic Substances

Zebrafish (Danio rerio) were exposed with 17β‐estradiol (E2) and nonylphenol (NP) in different concentrations. Gene expression analysis was carried out by two different approaches. First, RT‐PCR experiments were performed for the examination of expression levels of the two marker genes vitellogenin and aromatase. This approach showed a significant increase in the expression of the vitellogenin gene in exposed male fish (500 ng/L 17β‐estradiol and 250 μg/L nonylphenol). This egg yolk protein is usually not synthesized in male vertebrates. A slight decrease of expression of the aromatase gene was observed in exposed female zebrafish. Aromatase is known to catalyze the conversion of androgens to estrogens. Second, DNA microarray experiments were carried out, which allow the simultaneous examination of the expression levels of a great number of marker genes. The microarray experiments resulted in an up‐regulation of vitellogenin up to 850‐fold. In addition, several other genes were identified to be up‐regulated by estrogens, for example the high mobility group box protein ssrp1 (78‐fold) or the chaperonin containing t‐complex polypeptide 1, beta subunit cctb (22‐fold).