Cholinergic neurons have been identified in the rabbit retina by an immunocytochemical technique for the localization of choline acetyltransferase. The dendritic processes of these cells form two strata in the inner plexiform layer; these strata show almost no overlap with GABAergic strata (identified with glutamate decarboxylase immunocytochemistry) or with acetylcholinesterase (AChE) containing strata (identified with AChE cytochemistry), but the latter two overlap almost exactly. Implications of these data for the possible mechanisms of directional selectivity are discussed.