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Various synthetic chemicals released to the environment can interfere with the endocrine system of vertebrates. Many of these endocrine disrupting compounds (EDCs) exhibit estrogenic activity and can interfere with sexual development and reproductive physiology. More recently, also chemicals with different modes of action (MOAs), such as antiestrogenic, androgenic and antiandrogenic EDCs, have been shown to be present in the environment. However, to date EDC-research primarily focuses on exposure to EDCs with just one MOA, while studies examining the effects of simultaneous exposure to EDCs with different MOAs are rare, although they would reflect more real, natural exposure situations. In the present study the combined effects of estrogenic and antiestrogenic EDCs were assessed by analyzing the calling behavior of short-term exposed male Xenopus laevis. The estrogenic 17α-ethinylestradiol (EE2), and the antiestrogenic EDCs tamoxifen (TAM) and fulvestrant (ICI) were used as model substances. As previously demonstrated, sole EE2 exposure (10-10 M) resulted in significant alterations of the male calling behavior, including altered temporal and spectral parameters of the advertisement calls. Sole TAM (10-7 M, 10-8 M, 10-10 M) or ICI (10-7 M) exposure, on the other hand, did not affect any of the measured parameters. If frogs were co-exposed to EE2 (10-10 M) and TAM (10-7 M) the effects of EE2 on some parameters were abolished, but co-exposure to EE2 and ICI (10-7 M) neutralized all estrogenic effects. Thus, although EDCs with antiestrogenic MOA might not exhibit any effects per se, they can alter the estrogenic effects of EE2. Our observations demonstrate that there is need to further investigate the combined effects of EDCs with various, not only opposing, MOAs as this would reflect realistic wildlife situations.
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???displayArticle.pmcLink???PMC3445530 ???displayArticle.link???PLoS One
Figure 2. Effects of various EDC on proportions of the call type rasping produced by male Xenopus laevis.Median (IQR) (nâ=â10 per treatment) of (A) percentages of rasping calls in each of the four recorded nights and (B) median percentages of rasping calls produced by male Xenopus laevis exposed to 17α-ethinylestradiol (EE2), a mixture of EE2 and tamoxifen (EE2+TAM) or EE2 and fulvestrant (EE2+ICI). Statistical differences were determined using General Linear Mixed models. Significant differences from solvent control (CTRL) + human chorionic gonadotropin (hCG) treatment are marked by asterisks (* pâ¤0.05; ** pâ¤0.01; *** pâ¤0.001).
Figure 3. Effects of various EDC on the number of accentuated clicks within advertisement calls produced by male Xenopus laevis.Median (IQR) (nâ=â10 per treatment) of (A) the number of accentuated clicks in each of the four recorded nights and (B) the median number of accentuated clicks produced by male Xenopus laevis exposed to 17α-ethinylestradiol (EE2), a mixture of EE2 and tamoxifen (EE2+TAM) or EE2 and fulvestrant (EE2+ICI). Statistical differences were determined using General Linear Mixed models. Significant differences from solvent control (CTRL) + human chorionic gonadotropin (hCG) treatment are marked by asterisks (* pâ¤0.05; ** pâ¤0.01; *** pâ¤0.001).
Figure 4. Effects of various EDC on the duration of clicks within advertisement calls produced by male Xenopus laevis.Median (IQR) (nâ=â10 per treatment) of (A) the duration of clicks of male advertisement calls in each of the four recorded nights and (B) the median duration of clicks of male advertisement calls produced by male Xenopus laevis exposed to 17α-ethinylestradiol (EE2) or a mixture of EE2 and tamoxifen (EE2+TAM) or EE2 and fulvestrant (EE2+ICI). Statistical differences were determined using General Linear Mixed models. Significant differences from solvent control (CTRL) + human chorionic gonadotropin (hCG) treatment are marked by asterisks (* pâ¤0.05; ** pâ¤0.01; *** pâ¤0.001).
Figure 1. Effects of various EDC on proportions of advertisement calls produced by male Xenopus laevis.Median (IQR) (nâ=â10 per treatment) of (A) percentages of advertisement calls in each of the four recorded nights and (B) median percentages of advertisement calls produced by male Xenopus laevis exposed to 17α-ethinylestradiol (EE2), a mixture of EE2 and tamoxifen (EE2+TAM) or EE2 and fulvestrant (EE2+ICI). Statistical differences were determined using General Linear Mixed models. Significant differences from solvent control (CTRL) + human chorionic gonadotropin (hCG) treatment are marked by asterisks (* pâ¤0.05; ** pâ¤0.01; *** pâ¤0.001).
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