Reproduction Abstracts

Introduction: Androgens: testosterone, dihydrotestosterone (DHT), and a fungicide with antiandrogenic activity – vinclozolin (Vnz) served to study agonism and antagonism of the androgen receptor.

Material and methods: Granulosa cells (GC) isolated from pig follicles were cultured for 48 h with testosterone (10⁻⁷ M), DHT (10⁻⁷ M), and Vnz (1.4×10⁻⁵ M) separately or in combinations. Cells were then assessed for viability, cytotoxicity, and caspase activation events with ApoTox-Glo Triplex assay. Additionally, to determine the mechanism of GC cell death induced by Vnz, Apoptosis Antibody Array Kit has been used.

Results and discussion: Supplementation with Vnz resulted in a dramatical reduction in the viability of GC but suprisingly without an increase in cytotoxicity. This might result from the ability of Vnz to alter normal cell division (cell-cycle arrest) without producing membrane integrity changes. Moreover, caspase-3 activity was suppressed in cultures treated with a combination of Vnz and testosterone. Such an antiapoptotic effect of Vnz applied together with high concentration of androgens might be explained by the suppression of two intra-cellular proapoptotic proteins bad and bax. It seems that selective destruction of porcine follicles is a serious consequence of their exposure to Vnz leading to premature ovarian failure in the affected organisms.

Supported by grant 2013/09/N/NZ4/00596.