During the final (fourth) withdrawal cycle, mice were evaluated hourly for handling-induced convulsions (HIC), and were treated with vehicle, LY379268 (0.3, 1, and 3 mg/kg) or MPEP (1, 3, and 10 mg/kg) treatment at 4 and 8 h into withdrawal.
In the light-dark cycle, mice were fed ad libitum for 6 days and then food deprived for 48 hours (starting at ZT12 or ZT16 in 12L:12D or 18L 6D, respectively).
After the end of the treatment cycle, mice remained under observation for additional 3 weeks.
During the light cycle, mice were transferred to the testing room 20 minutes prior to the anxiety tests.
In contrast, during the light cycle, mice experience longer periods of consolidated sleep, which is less likely to favor the generation of SWD (35).
After 10 d in the 12-h LD cycle, mice (male C57BL/6 mice: n = 8/genotype; age 75±± 3 d, mean± sem) received 17 cycles of 3.5 h light (170±2.3 mW/m):3.5 h dark, starting at original light onset.
To detect cells in the S phase of the cell cycle, mice received an intraperitoneal injection of bromodeoxyuridine (BrdU; 100 mg/kg; Sigma-Aldrich, St Louis, MO, USA) 2 h prior to collection of tissues.
