Mice were injected with CFA or 20 μl of saline as control in the morning. Morphine (10 mg/kg; Professional Compounding Centers of America Corp., London, Ontario, Canada) or 0.9% saline (Sigma) was injected intraperitoneally 1 hour after CFA and then again in the afternoon. Morphine injections were repeated twice daily, for two more consecutive days (day 1 and day 2). On day 3, mechanical nociceptive threshold was determined by the simplified up-and-down method using von Frey filament (63) in response to increasing doses of Morphine injected either intrathecally (at doses of 0, 1, 2.5, 5, 10, and 15 μg) or locally (in the ankle, at doses of 0, 2.5, 5, 10, 25, 30, and 40 μg) in the inflamed paw.

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