As described previously [52 (link)], an infusion cannula (IO: 0.3 mm; RWD Life Science.Inc) was inserted into the amygdala through the guide cannula. 0.15 µl of KA (3 mg/ml, Sigma, #420318) was infused at a flow rate of 2 nl/s controlled by microinjector (NanojectIII, Drummond Scientific). The cannula was kept for an additional two mins after completion of infusion and withdrew slowly to minimize reflux along the injection tract. Seizure stages were classified according to the criteria described by Racine [54 (link)] and scored every 5 min by a blinded investigator: stage 0, no seizure; stage 1, arrest and rigid posture; stage 2, head nodding; stage 3, sporadic full-body shaking, spasms; stage 4, chronic full-body spasms; stage 5, jumping, shrieking, falling over; stage 6, violent convulsions or death. Seizures at stage 4–6 that last for ≥ 30 min was defined as SE.
To monitor SE-induced SRSs, diazepam (8 mg/kg, i.p.) was injected 1 h after SE induction to terminate seizures. After a latent period of 2 weeks, mice were video monitored from 8 am to 8 pm each day for 1 week. In some experiments, DPCPX (1 mg/kg, Sigma, #C101) was i.p. injected each day during latent period. SRSs, defined as seizures with score ≥ 4, were counted by a blinded investigator.
To monitor SE-induced SRSs, diazepam (8 mg/kg, i.p.) was injected 1 h after SE induction to terminate seizures. After a latent period of 2 weeks, mice were video monitored from 8 am to 8 pm each day for 1 week. In some experiments, DPCPX (1 mg/kg, Sigma, #C101) was i.p. injected each day during latent period. SRSs, defined as seizures with score ≥ 4, were counted by a blinded investigator.
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