The objective of this research was to identify the psychological and physiological variables that differentiate persons reporting masticatory muscle pain (MMP) from normal controls (NC). This study examined the characteristics of 35 MMP patients in comparison to 35 age-, sex-, and weight-matched NCs. All subjects completed a series of standardized questionnaires prior to undergoing a laboratory evaluation consisting of a psychosocial stressor and pressure pain stimulation at multiple body sites. During the evaluation, subjects' emotional and physiological responses (heart rate, blood pressure, respiration, skin temperature, and muscle activity) were monitored. Results indicated that persons with MMP reported greater fatigue, disturbed sleep, depression, anxiety, menstrual symptoms, and less self-deception (P's<0.05) than matched controls. At rest, MMPs had lower end tidal carbon dioxide levels (P<0.04) and lower diastolic blood pressures than the NCs (P<0.02). During laboratory challenge, both groups responded to the standard stressor with significant physiological activity and emotional responding consistent with an acute stress response (P<0.01), but there were no differences between the MMPs and NCs. Muscle pain patients reported lower pressure pain thresholds than did NCs at the right/left masseter and right temporalis sites (P's<0.05); there were no differences in pressure pain thresholds between MMPs and NCs for the left temporalis (P<0.07) and right/left middle finger sites (P's>0.93). These results are discussed in terms of the psychological and physiological processes that may account for the development of muscle pain in the masticatory system.