Control (VEH) and prenatally-androgenized (PNA) male and female mice were generated using the well-characterized prenatally androgenized (PNA) mouse model protocol (29 (link)–33 (link), 57 (link)). Adult male and female C57BL/6J mice were paired overnight on the day of proestrus. Gestational day 1 was recorded as the following day after overnight mating and the male was removed from the cage. Females were then monitored for signs of pregnancy such as increased body weight and increased belly circumference. From gestational day 16-19, pregnant dams received a daily subcutaneous (s.c.) injection in the nape of the neck of either 100 µL dihydrotestosterone (DHT, 250 µg/100µL) in sesame oil as the PNA treatment or 100 µL of sesame oil only as the vehicle control. This window of prenatal androgen exposure has been shown to lead to PCOS-like features in mice and largely avoid the critical period for the differentiation of external genitalia (29 (link)–33 (link), 57 (link)).The male (M) and female (F) offspring of dams injected with DHT (PNAM or PNAF) and vehicle control (VEH) mice were studied from adulthood (postnatal day (PND) 60 onward) in the following experimental protocols. Oestrous cyclicity of VEH and PNA female mice was assessed to establish the expected loss of oestrous cyclicity in PNA mice by collecting daily vaginal smears over a 20-day period (PND 60–80) (Figure S1
Free full text: Click here
