Ganglion cells within the cat retina have been traditionally grouped by morphological criteria into three major classes: alpha, beta, and gamma. The gamma-type cells have been least well characterized, but the available evidence indicates that this class comprises a relatively heterogeneous population of neurons. In the present study we demonstrate that an antibody for neuropeptide Y (NPY) recognizes a subpopulation of about 2,000 gamma-type ganglion cells. The NPY-immunoreactive (IR) neurons project to the superior colliculus and to the C layers of the lateral geniculate nucleus as demonstrated by retrograde labeling with fluorescent tracers (fluorogold or rhodamine latex microspheres). Virtually all of these cells disappear following lesions of the optic nerve. The NPY-IR ganglion cells were identified as gamma cells on the basis of soma size and dendritic branching patterns. The somas of these neurons are small (8-22 microns in diameter), and each cell is characterized by sparsely branching dendritic processes, usually extending into the middle third of the inner plexiform layer, the physiologically defined ON sublamina. These neurons are distributed across the entire retina, with the highest density at the area centralis. Within local regions of the retina, however, there was no indication that the NPY-IR gamma cells are arrayed in a regular mosaic pattern. These results provide the first evidence that the gamma class of ganglion cells of the cat retina can be subdivided on the basis of immunocytochemical properties.