Three days after electrode implantation, baseline seizure activity was measured over 3 days. Thereafter, perampanel (2-(2-oxo-1-phenyl-5-pyridin-2-yl-1,2-dihydropyridin-3-yl)benzonitrile; 8 mg/kg, i.p., Eisai Korea Inc., Seoul, Korea), GYKI 52,466 (10 mg/kg, i.p.), or saline (vehicle) was administered daily at PM 6:00 over a 7-day period [31 (link),35 (link),72 (link)]. To select the responders and non-responders, and each dose was chosen as the maximum without adverse effects, based on previous studies [33 (link),34 (link),42 (link),55 (link)]. EEG were detected with a DAM 80 differential amplifier (0.1–3000 Hz bandpass; World Precision Instruments, Sarasota, FL, USA) 2 h a day at the same time over a 7-day period. The data were digitized (1000 Hz) and analyzed using LabChart Pro v7 (ADInstruments, Bella Vista, New South Wales, Australia). Behavioral seizure severity was also evaluated according to Racine’s scale aforementioned. Non-responders were defined as showing no reduction in total seizure occurrence in a 7-day period, as compared with the pre-treatment stage. After recording (18 h after the last treatment), animals were used for Western blot.
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