OECD validation of phase-3 Hershberger assay using the stimulated weanling male rat in Korea

Abstract

The OECD has proposed a new, validated test guideline with the stimulated weanling male Hershberger assay to avoid the surgical castration step. In the present study, we assessed the relevance and reliability of the stimulated weanling Hershberger assay in four stages. All chemicals except for testosterone propionate (TP) were orally administered to sexually immature male rats of 22 days old for 10 days. The weights of four mandatory accessory sex organs, two additional reproductive tissues and optional systemic organs were evaluated. At the first two stages, TP, as reference androgen, significantly increased the weights of epididymides and accessory sex organs (ASO) at 1.0 mg kg(-1) and flutamide (FLU), as a positive anti-androgen control, decreased the TP-stimulated organ weights at 3.0 mg kg(-1). At stage 3, trenbolone (40 mg kg(-1)), an anabolic steroid, significantly increased ASO weights, and weak anti-androgens (DDE and linuron) decreased the TP-stimulated ASO weights at each high dose. The above results were confirmed in a blind test with coded substances provided by OECD. Compared with results from our previous castrated male assay, the intact weanling version is less sensitive than the castrated male version, in terms of a smaller response at the reference dose of TP or FLU. However, this study suggests that the stimulated weanling Hershberger assay can detect the effects of both potent and weak anti-androgens on androgen-producing and androgen-dependent tissues.