A lack of knowledge persists concerning the combination of kinetics on protein and mRNA levels of the most commonly used biomarker for estrogenic influences-vitellogenin (VTG). Consequently, male fathead minnows were exposed to 17alpha-ethinylestradiol (EE(2)) for 35 days, followed by an equally long depuration period in a flow-through system. VTG mRNA levels reached a plateau after 3 days of exposure, which remained stable until 3 days after EE(2) removal. Control levels were re-attained within 7 days of the depuration phase. VTG protein accumulated in the plasma following a two-phased model. The first phase depicting an exponential increase lasted 15 days and was followed by a saturation phase approaching a plateau of approximately 47 mg VTG/ml plasma. Clearance kinetics could be described by a two-compartment open model, with half-lives of 2.17 and 21.32 days for the alpha- and beta-phases, respectively. In addition, a high VTG protein synthesis rate seemed to adversely affect fitness and mortality of the fish.