The treatment paradigm previously established in our lab was followed [11 (link),17 (link)]. Briefly, female (64–70 days of age) Sprague Dawley rats were treated with oxycodone HCl (Sigma Aldrich, St. Louis, MO, USA) dissolved in a saline vehicle via oral gavage. An ascending dosing procedure was used where doses of 10 mg/kg/day of oxy were orally-gavaged for 5 days followed by a 0.5 mg/kg/day escalation for 10 days until reaching a final dose of 15 mg/kg/day, after which females were mated with proven male breeders. The treatment regimen continued throughout mating, gestation, and parturition until weaning on post-natal day (P) 21. For the PNO paradigm, dams were orally-gavaged with 15 mg/kg/day of oxy only after parturition until weaning. The solo control group received no treatment in order to mimic a real-life scenario, i.e., normal drug-free mothers. For this study, offspring were sacrificed at P14, and brains were removed and stored at −80 °C.
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