Facilitation by acetylcholine of tetrodotoxin-resistant spikes in rat hippocampal pyramidal cells

The electrical activity of hippocampal pyramidal cells was studied in slice cultures during blockade of the regenerative Na currents. In the presence of tetrodotoxin, these neurones had a mean resting potential of -68 mV, a membrane input resistance of 87 M omega and displayed marked non-linearities in their current voltage relationship. In response to depolarizing stimuli, pyramidal cells generated action potentials of small amplitude, slow rise and long duration. These tetrodotoxin-resistant spikes were abolished by calcium conductance blockers such as cobalt and cadmium ions. Acetylcholine applied to the bath or by iontophoresis depolarized pyramidal cells, elicited spontaneous tetrodotoxin-resistant spikes and facilitated spiking evoked by depolarizing rectangular current pulses or a current ramp. The effects of acetylcholine were not only slow in onset, but also prolonged; they were completely reversible and sensitive to atropine and calcium-antagonists such as cadmium and cobalt ions which, respectively, reduced and abolished these effects. After hyperpolarizations following injection of depolarizing current pulses were suppressed by acetylcholine and often transformed into depolarizing afterpotentials. Acetylcholine had no effect on voltage-independent conductances as determined by application of hyperpolarizing current pulses. These results could be explained by inhibition of the voltage-dependent K+-current, i.e. the M current (blockade of the calcium current could remove any depolarizing influence resulting from M current inhibition) or by a direct activation of a voltage-dependent calcium current by muscarinic agonists.