Atropine methylbromide

KA (Milestone Pharmatech, New Brunswick, NJ), atropine methylbromide
(Sigma-Aldrich, St. Louis, MO) and pilocarpine hydrochloride (Sigma-Aldrich)
were dissolved in 0.9% NaCl. Raloxifene hydrochloride
(Sigma-Aldrich) was dissolved in dimethylsulfoxide (Sigma-Aldrich).
Pentobarbital was purchased as a solution (50 mg/ml; Nembutal; Henry Schein
Inc., Melville, NY), as was diazepam (5 mg/ml; Baxter Healthcare; New
Providence, NJ) and stored at room temperature. ICI 182,780 (Tocris
Bioscience, Bristol, UK) was dissolved in corn oil (vehicle; Mazola, ACH
Food Companies, Summit, IL) as a 10 mM stock solution and stored at room
temperature in the dark until use.

Animal procedures were performed using protocols approved by the University Committee on Use and Care of Animals of the University of Michigan. Animals were purchased from Charles River and kept under a constant 12 hour light/dark cycle with access to food and water ad libitum. Epileptic animals (n = 32) and sham controls (n = 11) were generated as described previously (Kron et al 2010 (link)). Briefly, eight week-old male Sprague Dawley rats were pretreated with atropine methylbromide (5 mg/kg i.p.; Sigma-Aldrich, St. Louis, MO) 20 minutes prior to induction of SE with pilocarpine hydrochloride (340 mg/kg i.p.; Sigma-Aldrich, St. Louis, MO) for epileptic animals, or an equivalent volume of 0.9% saline for sham animals. After 90 minutes of SE, seizures were terminated with diazepam (10 mg/kg i.p; Hospira Inc, Lake Forest, IL). Sham controls were treated with diazepam two hours after the saline injection.