Modulation of Ca2+ and K+ conductances in an identified insect neurone by the activation of an alpha-bungarotoxin-resistant cholinergic receptor

Abstract

The effects of activation of a population of [alpha]-bungarotoxin ([alpha]-bgt)-insensitive cholinergic receptors on the soma of the cockroach fast coxal depressor motor neurone (Df) have been examined under two-electrode voltage-clamp conditions. Activation of these receptors was achieved by bath-application either of acetylcholine (ACh) in the presence of [alpha]-bgt or of the muscarinic agonist McN-A-343 (McN). Since these receptors have been shown previously to respond to some nicotinic agonists, we refer to them as 'McN-sensitive or mixed pharmacological profile muscarinic receptors' (mMAChRs). Activation of these receptors normally results in a biphasic response consisting of an initial outward current component, which reverses near -70 mV, and a later (delayed) inwardly directed current, which is only observed at membrane potentials more positive than -40 to -20 mV. The initial outwardly directed component of the McN-induced current appears to result from an increase in K+ conductance since it reverses at potentials close to the K+ equilibrium potential (EK) (approximately -70 mV under the experimental conditions used) and is blocked by internal Cs+. This increase in K+ conductance is probably due to an increase in Ca2+-activated K+ current (IK,Ca) which is known to form a large proportion of the outward current observed when this neurone is depolarized. The delayed inwardly directed current induced by McN results from suppression of a Ca2+ current (ICa) which, in turn, causes a decrease in IK,Ca. The net effect is a reduction in outward current, because IK,Ca is considerably larger than ICa. Evidence for an action of McN upon Ca2+ channels is provided by experiments in which K+ currents have been suppressed by internal Cs+ to reveal inward currents produced by the movement of Ba2+ through voltage-dependent Ca2+ channels. Ba2+ currents observed under these conditions are suppressed by bath application of McN. The inwardly directed current component of the McN response is unlikely to involve direct regulation of IK,Ca, since McN has no effect upon this current when it is induced by brief intracellular Ca2+ injections. Both the initial outwardly directed component and the delayed inwardly directed component of the McN-induced current were suppressed by intracellular injection of the Ca2+ chelator BAPTA. These observations suggest that a rise in [Ca2+]i mediates the electrophysiological effects of McN in Df somata.