In order to determine which types of neuron in laminae I-III of the rat spinal dorsal horn contain the peptide galanin, pre-embedding immunocytochemistry with antiserum to galanin was combined with post-embedding detection of GABA- and glycine-like immunoreactivities. Sixty-eight galanin immunoreactive neurons in laminae I-III selected from four rats were examined, and in each case semi-thin sections through the cell body were tested with a monoclonal antibody to GABA and an antiserum to glycine. All of the 68 galanin-immunoreactive neurons tested were GABA-immunoreactive, while only one of them (in lamina III) was glycine-immunoreactive. This suggests that galanin is contained in inhibitory interneurons, and that (like enkephalin, neuropeptide Y and thyrotropin-releasing hormone) it is mainly restricted to GABAergic neurons which do not use glycine as a co-transmitter.