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High prevalence of chronic orofacial pain in women and its relationship with ovarian states suggest that ovarian hormones may be involved in the control of orofacial nociception. Since the interaction between ovarian hormones and nociception seems more evident in the orofacial area than in many other parts of the body, a possible site specificity of an ovarian hormone effect on nociception was tested in rats. Two nociceptive tests were applied to three groups of male rats ( n =46) and three groups of female rats ( n =46), that were gonadectomised ( n =17), sham-operated ( n =15) or intact ( n =14). Each rat in each group received a local subcutaneous injection of formalin in the upper lip and in the hindpaw. Upper lip injection resulted in an increased occurrence of upper lip rubbing for more than 45 min and hindpaw injection resulted in an increased occurrence of hindpaw licking for about 1 h. The duration of the nociceptive behaviours was measured at 3 months after surgery. No significant difference was found between intact and sham-operated animals. A significant increase (54%) in the upper lip rubbing but not the hindpaw licking was observed in gonadectomised females. No difference was observed in castrated males for upper lip rubbing, but a tendency towards an increased duration (102%) of hindpaw licking was noted. The depletion in gonadal hormones was confirmed 3 months after gonadectomy and after the sacrifice of the animals, by the observed decline in the bone mineral density measured on the femur of 40 rats belonging to the six groups. A role of ovarian hormones was also suggested after immunostaining of oestrogen receptors in the lamina II of Caudalis subnucleus of the trigeminal sensory complex and cervical (C1–C2) spinal dorsal horn. The number of cells expressing oestrogen receptors displayed a small (13.6%) but significant ( P =0.037) increase in ovariectomised compared with sham-operated rats. These results suggest that the lack of ovarian hormones induces a site-specific increase in the sensitivity to orofacial nociceptive stimulation, and that an up-regulation of oestrogen receptors in the Caudalis subnucleus and C1-C2 dorsal horn may be one of the factors involved in this effect.