Hepatic and extrahepatic expression of estrogen-responsive genes in male adult zebrafish (Danio rerio) as biomarkers of short-term exposure to 17β-estradiol

Abstract

Growing concern over reproductive hormones in the environment demands sensitive and efficient methods by use of molecular biomarkers to detect these contaminants in oviparous vertebrates. In this study, a real-time quantitative RT-PCR was adopted to investigate the expressions of three estrogen-responsive genes, Vtg I, Vtg II and ERalpha, in hepatic and extrahepatic tissues of male adult zebrafish exposed to varying concentrations of 17beta-estradiol (E2) for selected periods. Without exposure to E2, all the genes were expressed in the tissues of male controls with their levels being much lower than those in the respective tissues of female controls. The expressions of hepatic Vtg I and II mRNAs were induced significantly after 1-day exposure to E2 at as low as 0.25 microg L(-1). Significant induction in the expression of hepatic ERalpha mRNA required a higher E2 concentration (> or = 0.5 microg L(-1)) and a longer exposure (> or = 2 days), suggesting that Vtg I and Vtg II are more sensitive to E2 exposure. The induction of Vtg mRNA in the skin of zebrafish was also significant following a short exposure (1 day) to low E2 concentration (0.25 microg L(-1)), with the levels of Vtg I and Vtg II mRNA being increased by 25 and 5 times, respectively. These results suggest that Vtg I mRNA is a highly sensitive biomarker for determining the estrogenic effects of E2 and that the skin of zebrafish may be an appropriate substitute for liver for such a determination.