Immunocytochemical studies of the distribution of acetylcholine in the crayfish brain

A number of studies indicate that acetylcholine is an important transmitter in most crustacean primary afferents and in at least several central pathways. Little is known, however, regarding the structure or distribution of cholinergio pathways in the central nervous system. The recent introduction of antibodies to choline‐protein conjugates provides a potentially powerful means for localizing putative cholinergic neurons and pathways in the nervous system. Acetylcholine was localized with immunocytochemical procedures in the axons and terminals of cephalic primary afferents and in interneurons of the crayfish brain. The most intensely reactive loci were the primary sensory neuropiles, which contain the terminals of the statocyst afferents (parolfactory lobes) and antennal afferents (antennal lobe). These results are generally in accord with previous findings based upon choline uptake and enzyme assay in lobster cephalic nerves. We also found evidence consistent with the presence of acetylcholine in the globular interneurons of the accessory lobe and in descending interneurons which originate in the dorsal medial and anterior clusters of the protocerebrum. The axons of several neurons in the circumesophageal connective (descending interneurons and primary afferents) are also reactive to the choline antibody.