Distraction interventions are used clinically to relieve pain. Exposure to distracting stimuli causes withdrawal of attention from the painful stimulus and reduces perceived pain. However, the neurobiological mechanisms mediating distraction-induced analgesia are poorly understood due, in part, to a paucity of animal studies modelling this phenomenon. The present study investigated the effects of three distracting stimuli on formalin-evoked nociceptive behaviour and brain tissue monoamine levels in rats. The three distractors were: exposure to a novel environment, exposure to a novel object, and exposure, without contact, to another rat (conspecific). A control group, habituated to the test arena, was also included. Formalin-evoked nociceptive behaviour was significantly reduced in rats exposed to the novel object or novel arena, but not those exposed to the conspecific, compared with controls. Antinociception resulting from exposure to the novel object was of longer duration than that resulting from exposure to the novel arena. Failure to detect any distractor-induced effects on plasma corticosterone levels or aversive behaviours suggests that the stimuli used were non-stressful. HPLC analysis revealed that there was a significant reduction in serotonin and dopamine metabolites in the medial prefrontal cortex in animals exposed to the novel object. These results indicate that exposure to a novel object or arena reduces nociceptive behaviour in rats, effects accompanied by discrete alterations in serotonin and dopamine metabolites in the medial prefrontal cortex.