Several lines of evidence indicate that 5‐HT7 receptors are involved in pain control at the level of the spinal cord, although their mechanism of action is poorly understood. To provide a morphological basis for understanding the action of 5‐HT on this receptor, we performed an immunocytochemical study of 5‐HT7 receptor distribution at the lumbar level. 5‐HT7 immunolabelling is localized mainly in the two superficial laminae of the dorsal horn and in small and medium‐sized dorsal root ganglion cells, which is consistent with a predominant role in nociception. In addition, moderate labelling is found in the lumbar dorsolateral nucleus (Onuf's nucleus), suggesting involvement in the control of pelvic floor muscles. Electron microscopic examination of the dorsal horn revealed three main localizations: 1) a postsynaptic localization on peptidergic cell bodies in laminae I–III and in numerous dendrites; 2) a presynaptic localization on unmyelinated and thin myelinated peptidergic fibers (two types of axon terminals are observed, large ones, presumably of primary afferent origin, and smaller ones partially from intrinsic cells; this presynaptic labelling represents 60% and 22% of total labelling in laminae I and II, respectively); and 3) 16.9% of labelling in lamina I and 19.8% in lamina II are observed in astrocytes. Labeled astrocytes are either intermingled with neuronal elements or make astrocytic “feet” on blood vessels. In dendrites, the labelling is localized on synaptic differentiations, suggesting that 5‐HT may act synaptically on the 5‐HT7 receptor. This localization is compared with other 5‐HT receptor localizations, and their physiological consequences are discussed. J. Comp. Neurol. 490:256–269, 2005. © 2005 Wiley‐Liss, Inc.