Localization of amino acids, neuropeptides and cholinergic markers in neurons of the septum-diagonal band complex projecting to the retrosplenial granular cortex of the rat.

Retrograde labelling was combined with immunohistochemistry to localize neurons containing choline acetyltransferase, gamma-aminobutyric acid (GABA), glutamate, leu-enkephalin, neurotensin, and substance P-like immunoreactivity in the projection pathways from the septum-diagonal band complex to the retrosplenial granular cortex in the rat. Injections of horseradish peroxidase conjugated to subunit B of cholera toxin (CT-HRP) into the retrosplenial granular cortex resulted in retrogradely labelled neurons in the ipsilateral nuclei of the diagonal band of Broca, especially in the horizonatal nucleus of the diagonal band, and small numbers of CT-HRP-labelled neurons were also found in the medial septal nucleus. In the horizontal and vertical nuclei of the diagonal band of Broca, 90-95% of CT-HRP-labelled neurons (35-45 per section) were immunoreactive for choline acetyltransferase and small numbers of retrogradely labelled neurons (2 to 4-5 per section) were also immunoreactive for GABA, glutamate, neurotensin, leu-enkephalin, or substance P. In the medial septal nucleus approximately 75-80% of the retrogradely labelled neurons (8-10 per section) were immunoreactive for choline acetyltransferase and up to 25% of the CT-HRP labelled neurons (1-3 per section) in the medial septal nucleus also displayed GABA-, glutamate-, neurotensin-, leu-enkephalin-, or substance P-immunoreactivity. These results suggest that the complexity of the neurotransmitter(s)/neuromodulator(s) of septum-diagonal band complex projections to the retrosplenial granular cortex should be taken into account when considering the mechanisms of cortical activation.