Acetylcholine-evoked afterdischarge in Aplysia bag cell neurons.

A brief synaptic input to the bag cell neurons of Aplysia evokes a lengthy afterdischarge and the secretion of peptide hormones that trigger ovulation. The input transmitter is unknown, although prior work has shown that afterdischarges are prevented by strychnine. Because molluscan excitatory cholinergic synapses are blocked by strychnine, we tested the hypothesis that acetylcholine acts on an ionotropic receptor to initiate the afterdischarge. In cultured bag cell neurons, acetylcholine induced a short burst of action potentials followed by either return to near baseline or, like a true afterdischarge, transition to continuous firing. The current underlying the acetylcholine-induced depolarization was dose dependent, associated with increased membrane conductance, and sensitive to the nicotinic antagonists hexamethonium, mecamylamine, and α-conotoxin ImI. Whereas nicotine, choline, carbachol, and glycine did not mimic acetylcholine, tetramethylammonium did produce a similar current. Consistent with an ionotropic receptor, the response was not altered by intracellular dialysis with the G protein blocker guanosine 5'-(β-thio)diphosphate. Recording from the intact bag cell neuron cluster showed acetylcholine to evoke prominent depolarization, which often led to extended bursting, but only in the presence of the acetylcholinesterase inhibitor neostigmine. Extracellular recording confirmed that exogenous acetylcholine caused genuine afterdischarges, which, as per those generated synaptically, rendered the cluster refractory to further stimulation. Finally, treatment with a combination of mecamylamine and α-conotoxin ImI blocked synaptically induced afterdischarges in the intact bag cell neuron cluster. Acetylcholine appears to elicit the afterdischarge through an ionotropic receptor. This represents an expedient means for transient stimulation to elicit prolonged firing in the absence of ongoing synaptic input.