Necropsy of Mature Male Offspring
Female and male pups were necropsied separately. Beginning on PCD 147, males were necropsied in a randomized block design in which one male per litter per treatment block was selected at a time. One male was selected from each of the other treatment blocks before an additional male was selected from a given block. This process was continued until all males in the study were necropsied. Androgen-dependent organ weights were measured in each treatment block until three animals per litter were examined.
This design ensured that differences in age at necropsy were accounted for across litters and treatments. Animals were anesthetized with CO2 before decapitation. All males were examined for vaginal pouches, the presence of nipples, and for phallus abnormalities including hypospadias, cleft phallus, or incomplete PPS. Androgen-dependent organs that were weighed included ventral prostate, dorsolateral prostate, seminal vesicles (with seminal fluid), testes, epididymides, levator ani plus bulbocav-ernosus muscles (combined), and glans penis. Kidney weights and liver weights were taken as diagnostics for general toxicological effects.