The cross-generational THC animal model has been described in detail previously (Szutorisz et al., 2014; Watson et al., 2015) and is illustrated in Fig. 1. Briefly, for the F0 breeding parents, 21-day old male and female Long-Evans rats were purchased from Charles River Laboratories, Inc. (Wilmington, MA) and housed in same sex groups. After one week of acclimation in the facility, the rats were administered THC (1.5 mg/kg intraperitoneally, every 3rd day from postnatal day 28-49) or VEH. After birth at ~PND2, mixed litters were established combining an approximately equal number (12-14) of pups from THC- and VEH-exposed parents with a balanced proportion of males and females in each litter. The litters were cross-fostered to drug-naïve surrogates, which were used as nursing mothers. F1 offspring were weaned at ~PND24 and groups of 3-4 animals were maintained without any drug treatment or testing on normal 12-h light/dark cycle with ad libitum access to food and water until adolescence (~PND35) or adulthood (~PND62). Animal care and handling were performed by technicians unfamiliar with parental treatment history. Animals were anaesthetized with CO2, decapitated, brains were frozen in isopentane, and stored at -80°C until subsequent experiments.
