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(T/EH033) Protein electrophoresis and RT-PCR as tools to detect endocrine disruption in zebrafish (Danio rerio). Versonnen, Bram1, Janssen, Colin1, 1 ABSTRACT- The increasing concern about endocrine disruptive effects of many chemicals has led to a growing need for standardized, robust and short-term assays. Zebrafish (Danio rerio) is an internationally accepted test species for aquatic toxicity tests and existing standard procedures with this species can be adapted, to include endpoints for the detection of endocrine disruption. In this research, the female specific protein vitellogenin was used as an endpoint to detect endocrine disruption in estradiol-exposed male and female fish. Based on previous research aimed at isolating the VTG mRNA of zebrafish, the present study examined the influence of ethinylestradiol on VTG content. Adult zebrafish were exposed for 7 to 21 days to different concentrations of ethinylestradiol. Next to condition factors and gonadosomatic indices, induction of VTG mRNA in the liver (RT-PCR) and in blood (electrophoresis) were used as endpoints. With RT-PCR, VTG was detected in the liver of adult zebrafish exposed to EE and not in unexposed fish. These results were confirmed by protein electrophoresis of blood plasma: significant differences in VTG content in the plasma were found between exposed and control fish. It may be concluded that VTG analysis through RT-PCR and protein electrophoresis are suitable tools for the detection of effects of xeno-estrogens and that effects short-term exposures (7 days) are detectable in adult zebrafish. Key words: endocrine disruption, zebrafish, vitellogenin, RT-PCR |
