Dose–response relationships and pharmacokinetics of vitellogenin in rainbow trout after intravascular administration of 17α-ethynylestradiol

Abstract

A commonly used endpoint in bioassays testing the estrogenicity of chemicals is the induction of the egg yolk precursor vitellogenin (VTG) in male fish. However, relatively little is known about the kinetics of induction and elimination of VTG in fish exposed to xenoestrogens. In this study, we administered graded intra-arterial doses (0.001, 0.1, 1.0 and 10.0 mg/kg) of 17α-ethynylestradiol (EE 2) to male rainbow trout via a dorsal aortic cannula which allowed repetitive blood sampling from individual fish for up to 48 days after injection. The plasma concentrations of VTG was quantified using an enzyme-linked immunosorbent assay procedure and the simultaneous concentrations of EE 2 were determined by gas chromatography-mass spectrometry. The pattern of VTG induction was similar for all doses of EE 2, with a 12-h lag-time before increase from basal levels (0.006–0.008 μg/ml), then increasing sharply to maximum levels within 7–9 days ( C max=0.05, 711, 1521 and 2547 μg/ml VTG for the 0.001, 0.1, 1.0 and 10.0 mg/kg doses, respectively). After induction by EE 2, VTG declined mono-exponentially with an elimination half-life of 42–49 h. The half-life of VTG increased to 145 h in the 10 mg/kg treated fish. The pharmacokinetics of EE 2 were distinctly nonlinear with substantial increases in the elimination half-life with increasing dose. The plasma concentration–time profiles of EE 2 were influenced by enterohepatic recirculation that caused multiple or secondary peaks in the profiles. In a separate experiment, the pharmacokinetics of purified VTG was characterized after intra-arterial injection in trout. After direct injection of VTG, plasma levels declined tri-exponentially with an apparent steady-state volume of distribution of 837 ml/kg; total body clearance was 31.1 ml/h per kg, and the elimination half-life was 43.7 h.