The neuropeptide Antho-RFamide is extremely abundant in Renilla koellikeri (sea pansy), a representative of the cnidarians (octocorallians) considered to be closest to the stem ancestors of metazoans with nervous systems. Therefore, a knowledge of the distribution of Antho-RFamide-containing neurons in this species would contribute to our understanding of the early evolution of nervous systems. Using antisera raised against RFamide and FMRFamide, we detected immunostaining in numerous neurons throughout the nervous system of the sea pansy. The antisera revealed ectodermal nerve-nets on the upper and lower sides of the colony and on the oral side of tentacles, in the oral disk, and in the pharynx of feeding polyps. Neurons were immunostained also in the mesogleal nerve-net of feeding polyps and in the through-conducting mesogleal nerve-net of the colonial mass. Varying densities of stained neurons were observed in the different compartments of the endoderm: muscular walls of the feeding and water circulation polyps, mesenteric filaments and their derived follicles containing either ovocytes or spermatophores, in the endodermal channels connecting the different compartments of the colony, and in circular muscle of the peduncle. The distribution of immunostained neurons suggests that they play important roles in feeding, reproduction, neuromuscular transmission, and in neuro-neuronal transmission coordinating the different parts of the colony.