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).     

Effects of Anthropogenic Estrogens Nonylphenol and 17α‐Ethinylestradiol in Aquatic Model Ecosystems

Microcosm tests were conducted to investigate the effects of the estrogenic substances nonylphenol (NP) and 17α‐ethinylestradiol (EE) on aquatic ecosystems. Maximum concentrations of 9 to 120 μg L^—1 NP resp. 49 to 724 ng L^—1 EE were induced by controlled release. The controlled release method allows the establishment of a continuous concentration course. The microcosms proved to run robustly with abiotic conditions close to natural. They developed biocenosis with similar characteristics as in natural ecosystems and, considering their given level of complexity, they can be used to describe possible risks for the environment. Both tested chemicals unveiled the potential to affect the plankton communities in the tested concentration range. NP exposure caused a reduction of Cladocera and Copepoda abundances and disturbed the phytoplankton structure. A NOEC_community of 30 μg L^—1 was calculated. In the first EE study, a flood in the lake where the microcosm water was collected caused additional stress and thereby a high variability, both between the microcosms and in each microcosm over time. Probably therefore the only effect found was a reduction of Copepoda abundance. In a second EE study Cladocera and Copepoda abundances were reduced, from which the phytoplankton benefited. Although a final interpretation is difficult for results of microcosm tests, there are indications that the found effects of EE and perhaps also NP may be caused at least partially by endocrine disruptive activity.