Environmental estrogens: dose–response relationships for vitellogenin formation and reproductive toxicity in male rainbow trout

The appearance of the egg-yolk protein vitellogenin (Vg) in plasma of male fish is a sensitive indicator of exposure to estrogenic compounds. We have been studying the kinetics of Vg formation and excretion in rainbow trout with a goal towards developing an integrated pharmacokinetic–pharmcodynamic (PK–PD) model to quantitatively relate cumulative estrogenic exposure of fish to the expression and appearance of Vg in plasma. We administered graded doses of ethynylestradiol (EE2), o,p-DDT, DDD and DDE and octylphenol 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 the xenobiotics and Vg were simultaneously quantified using ELISA and GC–MS or GC–ECD. In separate experiments, sexually mature trout were exposed to graded water concentrations of EE2 for 3 months and various parameters indicative of the functional status of the male reproductive system determined. These parameters included tissue-somatic indices, histopathological evaluation, spermatocrit, sperm motility (quantified using computer-assisted-motion analysis) and viability of semen based on fertilization assays using eggs harvested from untreated trout. Results from fertilization assays indicated that 12 week exposure to EE2 concentrations of 10 and 100 ng/l caused a 50% reduction in the fertilization rate of semen harvested from exposed trout. PK–PD modeling strategies proved valuable tools for linking chemical exposures to Vg formation.