The distribution of calcitonin (CT) binding sites in serial sections of the rat brain and spinal cord has been examined by an 'in vitro' autoradiographic technique using a radioisotope-sensitive sheet film and [125]salmon CT. Autoradiograms of the diencephalic region had the highest grain density throughout the entire hypothalamus, with the exception of the nuclei ventromedialis, posterior and mammillaris, which were not labeled at all. In the brainstem, large amounts of grains were found in the ventrolateral division of the periaqueductal gray, in the locus coeruleus, in the nucleus tractus spinalis nervi trigemini and in the raphe obscurus, pallidus and magnus, while a widespread and lower grain density was observed in the reticular formation. In the spinal cord the labeling was discretely localized in laminae IV, V and VI of the dorsal horn. The observed distribution of CT binding sites is closely related to the neuroendocrine and analgesic effects of exogenous CT and reinforces the concept of a possible neuromodulatory role proposed for the peptide at brain level.