Differential expression of vitellogenin and oestrogen receptor genes in the liver of zebrafish, Danio rerio.

Environmental oestrogens are responsible for adverse effects in fish that affect reproduction. Availability of model fish to study the differential effects of endogenous and exogenous oestrogens and to test for oestrogenic activity of chemicals would be advantageous. Zebrafish could provide such a model, but the organisation and expression of vitellogenins (VTGs) and oestrogen receptors (ERs) are not completely understood. VTGs are synthesised in the liver and provide a sensitive biomarker of oestrogenic activity since they are thought to be under the regulation of the ER. There are multiple genes for VTGs and an in silico analysis of their distribution in the Zebrafish genome has identified six genes: VTG-1, VTG-2, VTG-4, VTG-5, VTG-7 located on chromosome 22 and VTG-3 on chromosome 11. VTG-specific, quantitative, real-time, reverse-transcriptase polymerase chain reaction assays were developed and used to measure differential expression in the livers of mature male and female zebrafish. Following normalisation in female fish, relative expression of VTG-5 mRNA is highest and is 1.3x, 1.6x and 2x higher than VTG-4, VTG-2 and VTG-1, respectively, while expression of VTG-3 and VTG-7 is very low. Expression of VTGs in male fish was either undetectable or very low (VTG-4 and VTG-5). ERalpha and ERbeta2 were expressed at higher levels than ERbeta1 in females, but only ERbeta2 was expressed in appreciable quantity in males. Expression of ERalpha in males was significant but only at the limit of detection (<0.1% of female fish), while ERbeta1 could not be detected. The very low level of expression of ERalpha in males raises questions about the accepted mechanism of oestrogenic induction of VTG in male fish.