Concentration- and Time-dependent Effects of the Synthetic Estrogen, 17α-ethinylestradiol, on Reproductive Capabilities of the Zebrafish, Danio rerio

Abstract

Partial or full life-cycle tests are needed to assess the potential of endocrine-disrupting compounds (EDCs) to adversely affect development and reproduction of fish. Small fish species such as zebrafish, Danio rerio, are under consideration as model organisms for appropriate test protocols. The present study examines how reproductive effects resulting from exposure of zebrafish to the synthetic estrogen 17α-ethinylestradiol (EE₂) vary with concentration (0.05 to 10 ng EE₂ L⁻¹, nominal), and with timing/duration of exposure (partial life-cycle, full life-cycle, and two-generation exposure). Partial life-cycle exposure of the parental (F1) generation until completion of gonad differentiation (0–75 d postfertilization, dpf) impaired juvenile growth, time to sexual maturity, adult fecundity (egg production/female/day), and adult fertilization success at 1.1 ng EE₂ L⁻¹ and higher. Lifelong exposure of the F1 generation until 177 dpf resulted in lowest observed effect concentrations (LOECs) for time to sexual maturity, fecundity, and fertilization success identical to those of the developmental test (0–75 dpf), but the slope of the concentration-response curve was steeper. Reproduction of zebrafish was completely inhibited at 9.3 ng EE₂ L⁻¹, and this was essentially irreversible as a 3-mo depuration restored fertilization success to only a very low rate. Accordingly, elevated endogenous vitellogenin (VTG) synthesis and degenerative changes in gonad morphology persisted in depurated zebrafish. Full life-cycle exposure of the filial (F2) generation until 162 dpf impaired growth, delayed onset of spawning and reduced fecundity and fertilization success at 2.0 ng EE₂ L⁻¹. In conclusion, results show that the impact of estrogenic agents on zebrafish sexual development and reproductive functions as well as the reversibility of effects, varies with exposure concentration (reversibility at ≤ 1.1 ng EE2 L⁻¹ and irreversibility at 9.3 ng EE2 L⁻¹), and between partial and full life-cycle exposure (exposure to 10 ng EE2 L⁻¹ during critical period exerted no permanent effect on sexual differentiation, but life-cycle exposure did).

This work was funded by the European Commission (Contract ENV4-CT97-0509). The authors are grateful to Uwe Boshof, Josef Greve, Eva Leu, and Kerstin Hutter for excellent technical assistance. The chemical analyses were performed by Walter Böhmer and Britta Kurzawa.