Acetylcholine (ACh), choline acetyltransferases and cholinesterases occur in cornea, iris-ciliary body complex and retina of several vertebrates. In cornea, ACh may serve as a sensory transmitter as well as a local hormone, the function of which is not delineated. The function of ACh as the parasympathetic neurotransmitter at the iris and ciliary body is well established. The muscarinic receptors on the iris smooth muscle are similar to the muscarinic receptors (M2 type in two way classification) at other smooth muscles towards their interaction with agonists and antagonists. Binding studies using radiolabeled antagonists and their displacement by agonists indicate that muscarinic receptors in membranes of iris-ciliary body complex are heterogeneous indicating more than one subtype of muscarinic receptors. A subtype other than M2 receptors may occur at the presynaptic sites of parasympathetic nerves, which have yet to be investigated using specific agonists and antagonists. Cholinergic markers, choline acetyltransferase and acetylcholinesterase, differ quantitatively and qualitatively in retinas of different species. However, amacrine cells are cholinergic in all vertebrate species. Although they make up 1% of retinal neurons, they influence the activity of a majority of ganglion cells. Cholinergic effects in ganglia are mediated through nicotinic and muscarinic receptors. Both of these types of cholinergic receptors are heterogeneous. They have yet to be investigated for their subtypes using specific agonists and antagonists. Although the role of cholinergic retinal neurons in the processing of visual information is not known, their input to ganglion cells generally increases the rate of spontaneous activity or the number of action potentials in light-evoked responses. Thus, the cholinergic input seems to modify the overall neuronal input to the ganglion cells from the receptive fields. Endothelial cells of blood vessels contain muscarinic receptors, which are activated by ACh to cause relaxation. Although retinal blood vessels provide recognizable characteristic signs in diabetes mellitus and hypertensive disease, no information is available on the muscarinic receptors of these vessels.