A Population of Supramammillary Area Calretinin Neurons Terminating on Medial Septal Area Cholinergic and Lateral Septal Area Calbindin-Containing Cells Are Aspartate/Glutamatergic

Abstract

The excitatory amino acid, aspartate/glutamate content of septal complex calretinin (CR)-, choline acetyltransferase plus substance P-, and Leu-enkephalin (Leu-enk)-containing extrinsic afferents was examined. Experiments were carried out using the transmitter-specific [3H]-D-aspartate retrograde tracer technique in combination with immunostaining for CR, choline acetyltransferase, and Leu-enk. The extrinsic and intrinsic CR innervation of the same brain areas were elucidated on control rats and on animals in which the septum was surgically separated from its ventral afferents. Correlated light and electron microscopic double-immunostaining experiments were used to determine the synaptic connections between CR axon terminals and lateral septal area calbindin (CB)- and medial septal area choline acetyltransferase-immunoreactive neurons. Furthermore, to determine the synaptic power of supramammilloseptal aspartate/glutamatergic neurons on the septal complex, semiquantitative analyses were performed in the supramammillary area on retrogradely (1) [3H]-D-aspartate-radiolabeled and (2) HRP-labeled material. The results demonstrated that a population of the extrinsic CR axons originating in the supramammillary area are aspartate/glutamatergic. These fibers forming asymmetric synaptic contacts terminate on both CB and cholinergic neurons. Intraseptal CR neurons, which establish symmetric synapses, innervate only lateral septal area neurons, including the CB-containing cells. These observations, together with other published data, raise the possibility of a hippocampus-lateral septal (GABAergic CB-containing neurons)-supramammillary area (aspartate/glutamatergic cells)-medial septal (cholinergic neurons)-hippocampus signal loop, which might be involved in the generation and regulation of hippocampal theta rhythm activity.