Vitellogenin expression in zebrafish Danio rerio: evaluation by histochemistry, immunohistochemistry, and in situ mRNA hybridisation.

Several histological methods were tested for their potential to detect the in vivo induction of vitellogenin in zebrafish, after exposure to 17beta-oestradiol (E2), and validated by correlating semi-quantitative measurements on digital images to vitellogenin plasma values measured by ELISA and morphological criteria. All methods, except for vitellogenin-specific immunohistochemistry on liver, detected vitellogenin production in male zebrafish at the exposure level of 1 nM E2/l, and correlated well to each other and to ELISA results on plasma, thus indicating their specificity. The level of sensitivity is in the range of the induction of clinical (histopathological) effects, although slightly below the level of sensitivity of the plasma ELISA. Vitellogenin specific in situ mRNA hybridisation on liver appeared laborious and not applicable on routinely prepared material. Vitellogenin specific immunohistochemistry on plasma and basophilia of male liver are cost- and effort-effective detection methods of vitellogenin production, and can be applied routinely on standard histological sections. These methods are, therefore, suitable to evaluate vitellogenin production as an indicator of exposure to compounds with estrogenic activity, at the level of induction of clinical effects. They are a useful tool for hazard identification of endocrine disruption, especially when combined with routine histopathology.