Ontogeny of substance P receptors in rat spinal cord: Quantitative changes in receptor number and differential expression in specific loci

Abstract

Anatomic distribution and functional studies of substance P (SP) and its binding sites show a role for the peptide in sensory (nociception), autonomic and somatic motor control. These physiologic functions show postnatal developmental changes, which, if mediated by SP, suggest that the receptors for the peptide may also undergo postnatal changes. This hypothesis was tested by using light microscopic autoradiography and membrane homogenate binding of 125I-Bolton-Hunter-SP ( 125I-BH-SP) to study SP binding sites in the spinal cord of rats of different ages. In cervicothoracic segments of rat spinal cord, the autoradiographs showed that specific binding of 125I-BH-SP occurred predominantly in the grey matter and varied inversely to age. In pups, up to about 15 days old, binding sites were diffusely distributed over the grey matter, and became progressively more defined in specific nuclei as the rats aged. A novel nucleus which is located in the ventrolateral ventral horn of caudal cervical segments and contained a high density of SP binding sites has been identified. High densities of SP binding sites in this nucleus and the intermediolateral cell column were visualized from the first postnatal day; however, those in the phrenic motor nucleus and in the dorsal horn were not fully expressed until after the 8th postnatal day. The age-related binding was confirmed in a membrane homogenate binding study of whole spinal cord which showed that the ratio for the concentration (cpm/mg protein) of specific binding was 106:12:4:1, for rats 11 (26 g), 38 (145 g), 90 (329 g) and 260 (553 g) days old. The ratio for the specific binding to the spinal cord (uncorrected for tissue weight) for the same groups of rats was 6:3:2:1. These data suggest that SP receptors decreased as a function of age. Furthermore, the decrease in SP receptors was not entirely due to growth of the spinal cord.