This article describes a protocol for inducing status epilepticus (SE) in mice using pilocarpine, along with the administration of scopolamine and terbutaline to minimize side effects. The study outlines the monitoring of behavioral changes and the recovery process following SE induction.
To begin, administer an injection of scopolamine and terbutaline solution to minimize the peripheral side effects of pilocarpine. Next, administer an injection of pilocarpine solution to induce status epilepticus or SE. Pilocarpine will alter the brain's neuronal excitability.
Immediately after that, place the mouse in a warm incubator and monitor for induction of SE. Look for head nodding and rhythmic movements of the front legs or any behavior of stage 3 or higher on the Racine's scale. Then, place the mouse into a room temperature incubator and continue to monitor the animal's behavior.
Next, administer an injection of diazepam solution to end SE. Lastly, administer an injection of dextrose solution to aid with the animal's recovery. Following SE induction, the animal enters a seizure-free latent period until the onset of spontaneous recurrent seizures.
Begin by weighing each 8-week-old male mouse. Make 2 milligrams per kilogram of scopolamine and terbutaline as well as 280 milligrams per kilogram of pilocarpine solution. Then, inject the solution intraperitoneally into each mouse and return the mice to their cages.
30 minutes after the scopolamine and terbutaline administration, inject pilocarpine solution into each mouse. Immediately after pilocarpine injection, place the mice in an incubator with 28 to 30 degrees Celsius for observation. Next, monitor the behavior of the mice until status epilepticus or SE is induced. If limbic motor seizures that correspond to stage 3 or higher according to Racine's scale are detected, record the time and monitor the mice.
Once continuous motor seizures last more than 2 minutes, place the mouse in a new cage at room temperature and keep monitoring for 3 hours to determine whether their convulsive seizures continue and SE is induced. Then, terminate behavioral acute seizures at 3 hours after SE onset by injecting 10 milligrams per kilogram of diazepam solution. After diazepam injection, administer 1 milliliter of 5% dextrose per individual mouse to help with recovery.
At day 1 after SE induction, weigh the mice and keep them in the incubator for one extra day. At day 2 after SE induction, weigh the mice and return them to their home cage. Provide moist food to facilitate recovery. Finally, measure daily body weight of the animals until 7 days post pilocarpine, and if the body weight has not increased, inject the mice with 5% dextrose.
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