Abstract The substance Preceptor neurokinin-1 is expressed by a subset of neurons in the rat spinal cord. We have combined immunostaining for Fos, a marker of noxious peripheral stimulation, and neurokinin-1 to examine whether nociceptive signals from particular peripheral tissues (skin, muscle or knee joint) or activity generated by nerve injury or formalin-induced inflammation are preferentially modulated by substance P. Our results indicate that superficial and deep spinal neurokinin-l-positive neurons process nociceptive information in markedly different ways. In lamina I, the number of double-labelled neurons was positively correlated with the intensity of the stimulus (defined by the total Fos count) and was not directly related to any particular peripheral target. However, in the deeper layers of the spinal cord (V–X), there was no such correlation, and stimulation of joint nociceptors and formalin-induced inflammation produced the greatest proportion of Fos/neurokinin-1 co-localization, suggesting a particular role for substance P in the mediation of joint pain and inflammatory hyperalgesia. Thus, lamina I neurokinin-I receptor-bearing neurons appear to be involved in intensity discriminative aspects of pain, whereas the deep neurokinin-1 cells are involved in spatial localization or the detection of particular nociceptive submodalities.