Under anaesthesia (a mixture medetomidine 0.25 mg/kg [Pfizer] and ketamine 60 mg/kg [Boehringer-Ingelheim]), a small laminectomy was performed at the sixth thoracic vertebra and a flexible cannula was inserted under the dura mater and glued in place, such that the tip is rested at the lumbar enlargement of the spinal cord. The opposite end of the cannula was placed subcutaneously, and an osmotic minipump (ALZET, Charles River Laboratories) was connected to the cannula [9 (link), 11 (link)]. For the first pharmacological experiment, the P2X7 receptor antagonist A-438079 (50 μg/12 μL/day) or saline was delivered for 8 days (from 1 day before to 7 days post-immunization). For the second experiment, rats received the irreversible CatS inhibitor LHVS (30 nmol/12 μL/day) or vehicle (20 % Cremophor EL/saline) for 14 days beginning 1 day before and until day 13 post-immunization. Doses of A-438079 and LHVS were based on previous studies ([29 (link), 30 (link)], respectively).
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