The function of the alpha(2C) subclass of adrenergic receptor in the spinal cord is unclear at present. Immunoreactivity for this receptor is found predominantly on axon terminals of the superficial dorsal horn but limited information is available about the properties and origin of these axons. The aim of this study was to determine which classes of neurone give rise to axons that possess this receptor and to investigate the synaptic organisation of these terminals. A series of double-labelling experiments was performed to investigate the relationship between the alpha(2C) receptor and each one of 14 chemical markers that label various types of axon terminal in the dorsal horn. Tissue was examined with two-colour confocal laser scanning microscopy. Quantitative analysis revealed that alpha(2C)-adrenergic receptors are not present on terminals of unmyelinated or peptidergic primary afferents and descending noradrenergic or serotoninergic axons. They were found on a proportion of terminals belonging to a mixed population of excitatory and inhibitory spinal interneurones, including those that contain neurotensin, somatostatin, enkephalin, GABA and neuropeptide Y. However, a greater proportion of terminals originating from excitatory interneurones were found to possess the receptor. Electron microscopic analysis revealed that alpha(2C)-adrenergic receptor immunoreactivity is predominantly associated with axon terminals that are presynaptic to dendrites but a small proportion of immunoreactive terminals formed axo-axonic synaptic arrangements. These studies indicate that noradrenaline can modulate transmission in the dorsal horn by acting through alpha(2C)-adrenergic receptors on terminals of spinal interneurones.