Abstract The septohippocampal pathway plays an important role in learning and memory. It projects from the medial septum-vertical limb of the diagonal band of Broca (MSDB) to the hippocampus and provides the latter with its main cholinergic innervation. To assess the importance of cholinergic selectivity and timing of MSDB stimulation in modulating learning and memory, we directly compared the effects of several MSDB stimulation strategies in healthy rats. We evaluated the effects of DREADD-mediated selective cholinergic neuronal MSDB stimulation and nonselective neuronal MSDB stimulation on spatial learning and memory in the appetitive radial arm maze and on resting-state brain networks using resting-state functional MRI. DREADDs were activated with the novel DREADD agonist J60. Selective cholinergic MSDB stimulation during – but not after – radial arm maze training improved spatial learning compared with J60-treated sham rats and had no effect on working memory or reversal learning. J60-treated sham rats had a worse working memory than saline-treated sham rats during the reversal phase of the radial arm maze task, suggesting an adverse effect of chronic use of J60. Nonselective MSDB stimulation during training resulted in a loss of appetite and exclusion from the radial arm maze training. Acute selective cholinergic and nonselective MSDB stimulation induced decreased functional connectivity (FC) in the default mode-like network. In addition, acute nonselective MSDB stimulation resulted in increased intrahippocampal FC, while selective cholinergic MSDB stimulation led to globally increased FC with the nucleus accumbens. While the combined effect of radial arm maze learning and the necessary chronic food restriction with or without chronic MSDB stimulation had no observable effect on resting-state networks, chronic food restriction alone globally increased FC in the brain.

Selective cholinergic stimulation of the medial septum-diagonal band of Broca via DREADDs improves spatial learning in healthy rats