Neurons located in the medial septum-nucleus of the diagonal band (vertical limb) area and antidromically activated by electrical stimulation of the fimbria were recorded in urethane anesthetized rats. Forty-five percent of these septo-hippocampal neurons (SHNs) discharged rhythmically in short bursts (mean burst frequency, 4 Hz). They were antidromically driven at short latencies from the fimbria. SHNs driven at long latencies (above 5 ms) were never bursting neurons. Fimbria stimulation also had a powerful inhibitory effect on the spontaneous activity of SHNs. The vast majority of the septo-hippocampal neurons were excited by the iontophoretic application of acetylcholine or cholinergic agonists, carbachol being the most effective. The acetylcholine-induced excitations were readily abolished by atropine. In contrast hexamethonium and mecamylamine were less effective. The rhythmic bursting activity could not be consistently altered by the iontophoretic application of cholinergic agonists or antagonists or of divalent cations. SHNs were also sensitive to various other putative neurotransmitters (substance P, GABA) known to play a role in the medial septal area. Bursting neurons and ACh-sensitive neurons were less frequent among unidentified medial septal neurons. The regulation of the septo-hippocampal cholinergic pathway is therefore not likely to be due to a direct feedback inhibition by locally released acetylcholine. However a strong inhibitory feedback could be exerted by the hippocampo-septal pathway impinging directly or indirectly on septo-hippocampal neurons.