TABLE 1.

Summary of laboratory studies using anticholinergics with animal models for posttraumatic epilepsy (PTE).

Anticholinergic	Epilepsy model	Protocol	Findings	References
Scopolamine	FPI	Experiment: pre-injury treatment with scopolamine. Adult male Sprague–Dawley rats were assigned to one of four groups: scopolamine 0.1, 1.0, or 10.0 mg/kg, i.p., or an equivalent volume of saline. Drugs were administered 15 min before the injury. Experiment: post-injury treatment with scopolamine. Adult male Sprague–Dawley rats were assigned to one of two groups: scopolamine (1.0 mg/kg, i.p.) or an equivalent volume of saline. Drugs were administered 30 s after injury.	-Scopolamine administrated before and after injury significantly attenuated the duration of suppression of behavioral responses. -Prior administration of scopolamine reduces the incidence of acute seizures and the associated prolonged apnea. -Pretreatment with a 1.0 mg/kg dose of scopolamine reduced mortality by 50%.	Lyeth et al., 1988
	CCI	Adult male Sprague–Dawley rats were trained with the Morris water maze. After five consecutive days of training, injured and uninjured rats were injected with scopolamine (1 mg/kg, i.p.). The drug was administered 15 min before testing in the maze. The next day rats were retested without scopolamine.	-Examined the effects of scopolamine on spatial memory in injured and uninjured rats and determined that when scopolamine was administrated, injured rats showed longer latencies to find the Water-Maze platform compared to sham-injured controls.	Dixon et al., 1994
	CCI	Adult male Sprague–Dawley rats were anesthetized and underwent two craniotomies before receiving a cortical impact through the right craniotomy. Fourteen days, post-injury, rats were surgically prepared for measuring extracellular ACh. Samples were collected every 20 min. After an equilibration period, scopolamine (1 mg/kg, i.p.) was injected. ACh levels were measured using an HPLC system.	-Controlled cortical impact decreases scopolamine-evoked release of ACh in freely moving rats at 14 days following injury.	Dixon et al., 1996
	Pilocarpine	Adult male Wistar EPM-1 rats were injected with pilocarpine (320–350 mg/kg, i.p.) and assigned to one of three groups according to scopolamine doses: 1 or 2 mg/kg, i.p., or an equivalent volume of saline. Scopolamine was injected 2 h after the onset of SE and terminated 17 days later.	-Animals treated with scopolamine had fewer SRS than animals without treatment.	Pereira et al., 2005
	Pilocarpine or kainic acid	Adult male Wistar rats were induced wiyh injections of pilocarpine (350 mg/kg, i.p.) or kainic acid. Approximately 30 min after pilocarpine and 2 h after kainate injection, most animals had reached SE. Animals were assigned to one of four groups according to scopolamine doses: 1, 2, 8 mg/kg, i.p., or an equivalent volume of saline. Scopolamine injections initiated 2 h after the onset of SE. Subsequent injections were administered every 6 h for the following 3 days, followed by administration via an osmotic pump for an additional 14 days. Injections were terminated on the 17th day.	-Scopolamine treatment decreased SRS when it was given in lower doses.	Benassi et al., 2021
	Lithium-pilocarpine	Adult female Sprague–Dawley rats were injected with lithium chloride (127 mg/kg, p.o.) 12–18 h before pilocarpine treatment. Pilocarpine was then administered at a bolus dose of 30 mg/kg, i.p. If needed this was followed by repeated i.p. injection of 10 mg/kg every 30 min until the onset of SE. Treatment with scopolamine (10 mg/kg i.p.) started 2 h after SE onset; the dose was divided by 4 mg/kg administered at 8 a.m. and 6 mg/kg at 6 p.m. over 17 days.	-Compared to the vehicle group, seizure frequency was significantly lower in the scopolamine group at both 8–10 and 24–26 weeks post-SE.	Meller et al., 2021
Pirenzepine	Kindling	Adult male Sprague–Dawley rats underwent bipolar nichrome stimulation and recording electrodes were implanted into the left and right basolateral amygdala and over the cortex. Amygdala stimulation was applied daily with an initial stimulus of 50 μA, and seizure intensity was graded according to Racine’s scale. In experiment 1, animals received saline or pirenzepine at doses of 10, 25, 50, and 100 nmol intracerebroventricularly 1 h before each electrical stimulation. In experiment 2, animals were kindled to the complete stage, and after a recovery period of 3–5 days, a dose of 50 nmol of pirenzepine was administered intracerebroventricularly to kindled animals.	-In experiment 1, the animals pretreated with 50 and 100 nmol doses failed to develop stages 4 or 5 of Racine. Suggesting a role for the M1 receptor in the kindling process. -In experiment 2, the seizure stage and after discharge duration were not affected by pirenzepine in fully kindled animals.	Eşkazan et al., 1999
Biperiden	Pilocarpine	Adult male Wistar rats were induced with injections of pilocarpine (320 mg/kg, i.p.). Biperiden was injected at 8 mg/kg i.p. every 8 h for 10 days, starting 3 h after SE onset.	-Biperiden treatment over 15 to 105 days diminished the frequency of spontaneous epileptic seizures.	Bittencourt et al., 2017
	Pilocarpine or kainic acid	Adult male Wistar rats were induced with injections of pilocarpine (350 mg/kg, i.p.) or kainic acid. Approximately 30 min after pilocarpine and 2 h after kainate injection, most animals had reached SE. Biperiden was injected at 8 mg/kg i.p. every 8 h (starting 3 or 6 h after SE onset) for 5, 10, or 20 days of biperiden administration.	-Animals treated with biperiden for only 5 days showed no effect, but when the treatment was expanded for 20 days, the latency of the first seizure increased, and SRS reduction was observed.	Benassi et al., 2021

FPI, fluid percussion injury; CCI, controlled cortical impact; SRS, spontaneous recurrent seizure; ACh, acetylcholine; i.p., intraperitoneal; p.o., oral administration; h, hours; min, minutes; s, seconds; μA, microampere.