The effects of local administration of cholinergic drugs on the release of acetylcholine in the septo-hippocampal system were investigated using intracerebral microdialysis. Dialysis probes were implanted in the cell-body area of septo-hippocampal neurones in the medial septal area, and in the terminal area of the same neurones in the ventral hippocampus. Drugs were administered locally via the dialysis probe. Administration of the mixed muscarinic/nicotinic receptor agonist carbachol caused a decrease, whereas administration of the muscarinic receptor antagonist methyl-atropine caused an increase in the output of acetylcholine in both the hippocampus and the medial septal area. In contrast, perfusion with the same drugs and the acetylcholine esterase inhibitor neostigmine bromide in the septal area had little or no effect on the output of acetylcholine in hippocampus. The results indicate that acetylcholine autoreceptors are localised on nerve terminals in medial septal area and hippocampus, and exert an inhibitory control over acetylcholine release. However, autoreceptors seem to be sparse or absent on dendrites and cell bodies of septo-hippocampal cholinergic neurones.