Immunohistochemical and retrograde tracing techniques were combined to demonstrate the occurrence of displaced ganglion cells (DGCs) exhibiting substance P-like immunoreactivity (SP-LI) in the pigeon retina. Following injections of rhodamine-labeled latex microspheres into the nucleus of the basal optic root (accessory optic system), about 5200 DGCs were observed to contain rhodamine fluorescence in the contralateral retina. Approximately 26% of the retrogradely labeled DGCs also contained SP-LI. The soma sizes of the doubly labeled DGCs ranged from 12 to 24-mu-m, and their distribution mirrored the overall distribution of DGCs projecting to the nucleus of the basal optic root. The density of doubly labeled DGCs ranged from 2 to 15 cells/mm2, with density peaks occurring in the superior-nasal and inferior-temporal retinal quadrants. Larger DGCs projecting to the nBOR (25-32-mu-m) were never seen to contain SP-LI. Together with previous results of enucleation experiments, these data indicate the existence of a subpopulation of SP-LI DGCs which are connected with the accessory optic system in the pigeon. The present results also contribute information on the heterogeneity of retinal ganglion cells transmitters and modulators.