Quantal analysis of action of hemicholinium-3 studied at a central cholinergic synapse of Aplysia

The effects of hemicholinium-3 (HC-3) on cholinergic transmission were studied on central identified inhibitory (H-type post-synaptic cell, Cl- channels) and on excitatory (D-type post-synaptic cell, cationic channels) synapses of Aplysia californica. In the H-type post-synaptic cell, the amplitude and the decay time of miniature post-synaptic currents (m.p.s.c.s.) were calculated by statistical analysis of long duration induced post-synaptic current (l.d.i.p.s.c.) due to 3 s depolarizations of the presynaptic neurone in the presence of tetrodotoxin. On H-type receptors, with respect to acetylcholine (ACh), HC-3 acted as an agonist and a blocker whereas on D-type receptors, it acted only as a blocker. At low concentration of bath-applied HC-3, in the H-type synapse, the decay time of the evoked inhibitory post-synaptic current (i.p.s.c.) as well as that of the m.p.s.c. was lengthened. These changes were rapidly reversible by wash. The decay time of excitatory post-synaptic current (e.p.s.c.) at the D-type synapse was not affected. On the inhibitory synapse, HC-3 applied in the bath at the concentration of 10(-5) M, reduced considerably the size of the m.p.s.c.s whereas the evoked i.p.s.c.s and the l.d.i.p.s.c.s were only slightly affected pointing to an increase of the quantal content of both responses. After wash, both i.p.s.c.s and l.d.i.p.s.c.s showed a clear facilitation which persisted for several tens of minutes. The presence of presynaptic receptors was considered. Similar facilitation of e.p.s.c.s by HC-3 was observed at the D-type synapse. The comparison of the degree of depression by HC-3 of the m.p.s.c.s and of the responses to ionophoretically applied ACh, indicated that the size of the quantum was not changed. Intracellular injection of HC-3 into the presynaptic neurone of the H-type synapse led to a decrease of transmitter release which affected solely the quantal content of the responses. As the synaptic transmission could not be restored by injection of exogenous ACh into the presynaptic neurone, it was concluded that the depression of transmission was not due to a decrease of ACh synthesis.