An immunocytochemical study of choline acetyltransferase-containing neurons and axon terminals in normal and partially deafferented hippocampal formation.

Monoclonal antibodies to the acetylcholine synthesizing enzyme, choline acetyltransferase (ChAT), have been used to study putative cholinergic structures in immunocytochemical preparations of normal rat hippocampal formation and of hippocampal formation deprived of its septal innervation. Small numbers of ChAT-positive (ChAT+) neuronal somata were observed scattered throughout the septotemporal extent of the normal hippocampal formation. They were most common in stratum lacunosum-moleculare of regio superior, but were also found in various layers of the dentate gyrus and occasionally in the remaining hippocampal laminae. In addition, light microscopy demonstrated that ChAT+ terminal fields in normal hippocampal formation were organized in discrete bands and laminae. Pronounced dense bands were observed: immediately superficial to stratum granulosum; deep to stratum pyramidale; and at the border between stratum radiatum and stratum lacunosum-moleculare. In the dentate gyrus, ChAT+ staining was pronounced in the hilus at temporal levels, but only moderate staining occurred in the anterior hilus and throughout the molecular layer. A close correspondence was observed in the density and distribution of ChAT+ immunoreactivity and acetylcholinesterase staining. Electrolytic lesions of the medial septal nucleus/diagonal band complex had no effect on the occurrence of ChAT+ somata, but virtually abolished the ChAT+ laminar staining pattern and eliminated all but occasional small patches of ChAT+ terminals. These results confirm that the vast majority of hippocampal cholinergic terminals originate either from neurons of the medial septum/diagonal band complex or from fibers of passage. The newly observed intrinsic hippocampal neurons can account for at least some of the ChAT activity remaining after septal lesions, and they apparently contribute to the cholinergic innervation of the hippocampal formation.