Retinal amacrine cells can be divided into subgroups on the basis of morphological properties and chemical content. It is likely that these subgroups have specific connections and serve unique functional roles within the inner plexiform layer. In the present study we show that immunoreactivity to neuropeptide Y (NPY) identifies a group of amacrine cells (165,000-170,000) within the adult cat retina. This is the largest group of peptide-containing amacrine cells identified to date in the cat retina. These neurons have small cell bodies and are regularly spaced at all retinal eccentricities examined. The density of NPY-immunoreactive cells, as well as their regular spacing, suggests that these neurons form a specific subgroup of the amacrine cell class and are likely to serve a unique role in the transfer of visual information through the inner plexiform layer.