Butyrylcholinesterase (BChE) is closely related to acetylcholinesterase (AChE), but its function in nervous system development or physiology is unclear. Here, the distribution of BChE was investigated by immunohistochemical methods in the developing chick retina. Using a specific anti-BChE antibody, we detected immunoreactivity associated with different cell types in two nuclear layers and in plexiform layers of the retina. At embryonic day 10 (E10), a transient BChE staining is detected in the inner plexiform layer (IPL) and in radial cells, the latter possibly representing Müller glia. At E12, a subpopulation of amacrine cells appeared, followed by cells in the middle and outer half of the inner nuclear layer. These cells at locations of amacrine, bipolar and horizontal cells represented the predominant three cell types persisting until hatching. The BChE+ amacrine cells were studied in more detail. Their distribution was not significantly different in the central and peripheral retina. Double labelling experiments revealed that BChE+ amacrine cells did not express choline acetyltransferase (ChAT), and, thus, are non-cholinergic. Only a minority of them coexpressed AChE. On the other hand, the majority of them colocalized with anti-GABA immunoreactivity. Taken together, these data support a hitherto unsuspected role of BChE in non-cholinergic cells, possibly in conjunction with GABA.