Benzyl benzoate b6630

Females in the estrus phase, as determined by vaginal smears³⁸ , were mated overnight. Wt females were mated with APNtg males and APNtg females were mated with wt males. Pregnancy was confirmed by a distinct weight gain. On GD 16.5 the groups were subdivided and assigned to receive a subcutaneous injection in the interscapular area of vehicle or 250 μg 5α-Androstan-17β-ol-3-one (dihydrotestosterone (DHT), A8380, Sigma-Aldrich, St. Louis, USA) dissolved in 2.5 μl benzyl benzoate (B6630, Sigma-Aldrich) and 47.5 μl sesame oil (S3547, Sigma-Aldrich) for three days. Prenatal androgenization with DHT induces a PCOS-like phenotype in the offspring^{18 (link),39 (link)}. APNtg and wt dams gave birth to both wt and APNtg offspring. Female offspring were weaned at 4 weeks of age and genotyped as previously described^{12 (link)}. The breeding scheme and study design are shown in Fig. 1.

C57Bl/6J mice (3-week-old) were obtained from Janvier Labs (Le Genest-Saint-Isle, France). We generated and phenotype F₀ dams in detail.⁶ (link) To generate prenatal androgen exposed offspring, the CD and HFHS groups were randomly subdivided and injected daily subcutaneously (s.c.) in the inter-scapular area from E16.5 to E18.5 with 50 μL of a solution containing a mixture of 1) a mixture of 5 μL benzyl benzoate (B6630; Sigma-Aldrich) and 45 μL sesame oil (S3547; Sigma-Aldrich, St. Louis, Missouri, USA) i.e. vehicle, or 2) 250 μg DHT (5α androstane-17β-ol-3-one, A8380; Sigma-Aldrich, St. Louis, Missouri, USA) dissolved in a mixture of 5 μL benzyl benzoate and 45 μL sesame oil i.e. PNA by DHT. See key resource table for reagents and resources. Thus, four experimental lineages were studied, i.e., control diet+vehicle (control), control diet+ dihydrotestosterone (androgenized), high-fat, high-sucrose diet+vehicle (obese), and high-fat, high-sucrose diet+DHT (obese and androgenized) lineages.