To elucidate the transcriptional profile of the anogenital region of male rats with chemically induced short AGD. This was compared this to the transcriptional profile of control males and control females to investigate whether induction of short AGD resulted in a feminization of the anogenital region at the transcriptional level. Pregnant Sprague Dawley rats were exposed to Finasteride (10mg/kg bw/day) from gestastion day (GD) 7 to GD21 where pups were delivered by cesarean section, their AGD was measured and tissues of the anogenital region were isolated for transcriptional analysis.
Dams were decapitated under CO2/O2-anesthesia at GD21 and fetuses collected by caesarean section. Prior to decapitation of the fetuses, their body weights were recorded and AGD measured by the same, blinded observer using a stereomicroscope with a micrometer eyepiece. The AGD index (AGDi) was calculated by dividing AGD by the cube root of the body weight. The anogenital region was isolated from the fetuses by dissection under a stereomicroscope. The tissue was trimmed to include only the genital tubercle, the perineal tissue and the very base of the tail. The tissue was placed in RNAlater (Qiagen, Hilden, Germany) and stored at -80˚C until further analysis. The animal experiments were carried out at DTU Food (Mørkhøj, Denmark) with ethical approval from the Danish Animal Experiments Inspectorate (license number 2015-15-0201-00553) and by the in-house Animal Welfare committee.