Schauwecker1 project protocol

Drug study: Pilocarpine-induced status epilepticus in 11 inbred strains of mice   (2012)

Schauwecker PE
With: Kornacki M, Lorenzana A, Chancer Z, McCord M




Schauwecker1_protocol

Project protocol - Contents

Workflow and sampling

Step
Procedure performed
Data collected
1
 Mice injected with pilocarpine (repeated low-dose treatment) until onset of status epilepticus (SE)
Dose required to induce SE
2
 Mice monitored for a minimum of 5 h
Latency to stage 4 seizures, duration of stage 4 seizures, survival

Equipment and supplies

  • Syringes and needles

Reagents and solutions

  • Methyl scopolamine (Sigma Aldrich, St. Louis MO)
  • Pilocarpine (Sigma Aldrich, St. Louis MO)
  • 0.9% NaCl

Acclimation to test conditions

Mice were allowed to adapt to laboratory conditions for at least 1 wk. Experiments were performed between 0800 and 1200h.

Procedure: Pilocarpine administration and assessment of treatment

    1. Mice are injected with methyl scopolamine (i.p.) 15-30 min before pilocarpine injection; at a dose of 1 mg/kg body weight (in 0.9% NaCl) [methyl scopolamine minimizes peripheral cholinergic side effets of pilocarpine].
    2. Mice are injected with pilocarpine (i.p.) using a repeated low-dose treatment of 100 mg/kg body weight (in 0.9% NaCl) every 30-45 min until onset of status epilepticus (SE); final doses are recorded.
    3. Mice are monitored for a minimum of 5 h to assess the severity and length of seizures. See Schauwecker (2012) for more details. Briefly, seizures are classified according to the following stages:
      1. rigid posture or immobility
      2. stiffened, extended and often arched tail
      3. partial body clonus, including forelimb or hindlimb clonus or head bobbing
      4. whole body continuous clonic seizures with rearing
      5. severe whole body continuous clonic seizures with rearing and falling
      6. tonic-clonic seizures with loss of posture or jumping

Definitions and calculations

  • SE: Status epilepticus (defined by continuous or intermittent limbic seizure activity for at least 45 min without full recovery between seizures, as described in humans)

Data collected by investigator

  • pilocarpine dose required to induce seizures
  • latency to induce stage 4 seizures
  • duration of stage 4 seizures
  • status epilepticus seizure mortality