2.3. Mouse pilocarpine model of SE

Adult male mice (8‐10 weeks) were injected with methylscopolamine and terbutaline (2 mg/kg each in saline, ip) to minimize the unwanted effects of pilocarpine in the periphery. Thirty minutes later, pilocarpine (280 mg/kg in saline, freshly prepared, ip) was injected to induce seizures in mice. Control mice received methylscopolamine and terbutaline, followed by saline injection instead of pilocarpine. Seizures were classified as we previously described. ²² , ²³ 0: normal behavior—walking, exploring, sniffing, and grooming; 1: immobile, staring, jumpy, and curled‐up posture; 2: automatisms—repetitive blinking, chewing, head bobbing, vibrissae twitching, scratching, face‐washing, and “star‐gazing”; 3: partial body clonus, occasional myoclonic jerks, and shivering; 4: whole body clonus, “corkscrew” turning and flipping, loss of posture, rearing, and falling; 5: (SE onset): nonintermittent seizure activity; 6: wild running, bouncing, and tonic seizures; and 7: death. SE was defined by nonintermittent seizure activity, which was indicated by continuous generalized clonic seizures without returning to lower‐stage seizures. The SE was allowed to proceed for 1 hr and terminated by sodium pentobarbital (30 mg/kg, ip). The animal mortality rate after pilocarpine‐induced SE was about 40% in this study.