Intracellular recordings were made from neurones in laminae I and II of the dorsal horn of a longitudinal, parasagittal spinal cord slice from the neonatal rat. Their responses to peripheral nerve stimulation were first tested. Then the responses to bath application of [Sar(9),Met(O-2)(11)]-substance P and [D-Ala(2), N-MePhe(4), Gly-ol(5)]-enkephalin, neurokinin 1 (NK1) and mu-opioid receptor agonists respectively, were studied. Finally, the structure of each neurone was investigated by injecting neurobiotin intracellularly following recording, and immunocytochemical studies were performed on post-fixed tissues to reveal whether they expressed the NK1 receptor. Nine lamina I neurones where shown to express NK1 receptor and these were depolarised by [Sar(9),Met(O-2)(11)]-substance P. These neurones typically received a powerful C-fibre input that was strongly inhibited, presynaptically, by the mu-opioid receptor agonist. The structure, afferent input, opioid sensitivity and intrinsic properties of these neurones are all consistent with the view that they are a major relay for nociceptive information leading to intense pain. The characteristics of 10 other neurones studied in which the NK1 receptor was not found to be expressed at levels detectable by immunocytochemistry are briefly described for comparison. These results contribute to the emergent view that the large neurones in the most dorsal neuronal layer (lamina 1) of the spinal cord, which express the principal receptor for substance P (NK1) over their entire soma and dendrites, are a major relay for information leading to intense pain. Inhibition of the relay of information by these neurones would be predicted to result in analgesia and hence, a detailed knowledge of their unique neurochemical characteristics is of paramount importance. (C) 2002 IBRO. Published by Elsevier Science Ltd. All rights reserved.