The aims of this project were to investigate whether inflammation in the orofacial muscle alters mu opioid receptor (MOR) mRNA and protein expressions in trigeminal ganglia (TG), and to assess the contribution of peripheral MORs under acute and inflammatory muscle pain conditions. mRNA and protein levels for MOR were quantified by reverse-transcription-polymerase chain reaction (RT-PCR) and Western blot, respectively, from the TG of naïve rats, and compared with those from the rats treated with complete Freund's adjuvant (CFA) in the masseter. TG was found to express mRNA and protein for MOR, and CFA significantly up-regulated both MOR mRNA and protein by 3 days following the inflammation. The MOR protein up-regulation persisted to day 7 and returned to the baseline level by day 14. We then investigated whether peripheral application of a MOR agonist, D-Ala2, N-Me-Phe4, Gly5-ol-enkephalin acetate salt (DAMGO), attenuates masseter nociception induced by masseteric infusion of hypertonic saline (HS) in lightly anesthetized rats. DAMGO (1, 5, 10 microg) or vehicle was administered directly into the masseter 5-10 min prior to the HS infusion. The DAMGO effects were assessed on mean peak counts (MPC) and overall magnitude as calculated by the area under the curve (AUC) of the HS-evoked behavioral responses. Under this condition, only the highest dose of DAMGO (10 microg) significantly reduced MPC, which was prevented when H-D-Phe-Cys-Tyr-D-Trp-Arg-Thr-Pen-Thr-NH2 (CTAP), a selective MOR antagonist, was co-administered. DAMGO pre-treatment in the contralateral masseter did not attenuate MPC. The same doses of DAMGO administered into CFA-inflamed rats, however, produced a greater attenuation of both MPC and AUC of HS-evoked nocifensive responses. These results demonstrated that activation of peripheral MOR provides greater anti-nociception in inflamed muscle, and that the enhanced MOR effect can be partly explained by significant up-regulation of MOR expression in TG.