Ionic mechanisms of a two-component cholinergic inhibition in Aplysia neurones.

1. A two-component inhibition, consisting of a rapid and slow i.p.s.p., has been observed in the medial cells of the pleural ganglion of Aplysia. Each i.p.s.p. has been shown to be mediated by a distinct cholinergic receptor. The ionic mechanisms of the two components of the inhibitory response (whether elicited synaptically or by ACh injection) are analysed in this paper.2. The inversion potential (typically -60 mV) of the rapid i.p.s.p. and of the rapid response to ACh injection is selectively altered by an intracellular injection of chloride or by partial substitution of the external chloride by impermeant anions. The shift caused by this last procedure is similar to that predicted for the chloride equilibrium potential (E(Cl)) by the Nernst equation.3. The slow i.p.s.p. and the slow response to ACh injection (both of which invert around -80 mV) are insensitive to changes in either internal or external chloride concentrations; on the contrary, with alterations of the concentration of potassium in the external medium, the inversion potential of the slow responses is altered in a way similar to that expected for the potassium equilibrium potential (E(K)).4. It is concluded that the rapid i.p.s.p. and the corresponding ACh potential are due to a change in chloride permeability of the post-synaptic membrane, whereas the slow responses are due to a selective change in potassium permeability.5. Additional data suggest that the fast, ;chloride' channel is impermeable to sulphate and methylsulphate, but slightly permeable to propionate and isethionate. The slow, ;potassium' channel is impermeable to caesium ions, whereas its permeability to rubidium ions is half that to potassium.6. The potassium permeability of both the non-synaptic and synaptic membrane is markedly reduced by an intracellular injection of either tetraethylammonium (TEA) or caesium. These ions not only block the cholinergic potassium currents (whether inward or outward) but likewise block the potassium currents activated in the same cells by an iontophoretic injection of dopamine.7. The potassium dependent synaptic potentials are also selectively affected by manipulations known to block the electrogenic sodium pump. In the presence of ouabain or in sea water in which sodium has been replaced by lithium, there is an apparent reduction of these potentials which was shown to be simply a reflexion of the movement of E(K) towards a less polarized level. This shift in inversion potential was not seen for the potassium dependent response to ACh iontophoretic injection. These results are interpreted in terms of accumulation of potassium ions assumed to occur in the extracellular spaces of the neuropile, but not in the thoroughly dissected somatic region.8. Cooling was shown to eliminate, selectively, the synaptic and ACh potential changes caused by an increase in potassium permeability.