After all injections had been completed, animals were sutured and injected with an analgesic [Rimadyl®; 4 mg/kg (Carprofen), Pfizer, Sandwich, UK; or Metcam®; 0.3 mg/kg (Meloxicam), Boehringer Ingelheim, Germany for replications 1 and 2, respectively].
After the behavioral recordings were completed, animals were perfused under deep thiopental anesthesia with 5 ml of phosphate buffered saline followed by 40 ml of ice-cold 4% paraformaldehyde (PFA) (pH 7.4) (Sigma).
Once the experimental protocol was completed, the animals were killed with intravenous potassium chloride under deep anesthesia.
After the surgical preparation was completed, the animals were allowed to stabilize for 45 to 60 minutes.
After the surgical preparation was completed, the animals were allowed to recover for 30 minutes before the measuring protocol was initiated and the infusion of LR was discontinued.
As the HS operation was completed, the animals were resuscitated for 15 minutes by returning all of the shed blood and then administering a volume of Ringer's solution equal to the volume of the shed blood.
After the behavioral testing was completed, the animals (n = 60) were deeply anesthetized with urethane 40% (1 g/kg, from Sigma Aldrich, France), and then transcardially perfused with saline (0.9%), followed by 4% paraformaldehyde in phosphate-buffered saline (0.2 M).
Once preparation was completed the animal was installed into the PET camera in a supine position.
Once testing was completed on the animals, they were released to the island.
The protocol was completed in 29 animals, comprising 8 in group 1 (sham), 11 in group 2 (oleic acid), and 10 in group 3 (lavage).
