Segment-specific modulation of the electrophysiological activity of leech Retzius neurons by acetylcholine

1. The acetylcholine responses of Retzius neurons were electrophysiologically and pharmacologically characterized in situ and in culture. Single-electrode voltage-clamp was used to record currents from leech Retzius neurons from standard segments [Rz(X)] and from reproductive segments [Rz(5,6)]. 2. A 1 s pressure pulse of acetylcholine (ACh) produced a fast inward current followed by a slower outward current in Rz(X) neurons, whereas it produced only an outward current in Rz(5,6) neurons. These segment-specific responses were maintained when the two types of Retzius neurons were isolated in culture for up to 12 days. 3. The inward current of Rz(X) reversed at around -25 mV and was partially carried by Na+. This cationic current desensitized rapidly. The outward current of Rz(X) and Rz(5,6) neurons reversed at around -65 mV and was carried by Cl-. This anionic current desensitized very slowly upon prolonged applications of ACh. 4. The expression of the ACh-induced outward current in Rz(X) was season-dependent and was recorded in a larger proportion of Rz(X) neurons during the summer than during the winter. The expression of the ACh-induced outward current in Rz(5,6) did not show any seasonal pattern. 5. The fast inward current of Rz(X) was also elicited by nicotine; it was blocked by d-tubocurarine, hexamethonium and mecamylamine, but was not affected by alpha-bungarotoxin. The outward current of Rz(X) and Rz(5,6) was also elicited by nicotine and by 4-[N-(3-chlorophenyl)carbamoxyloxy]2-butynyltrimethylammonium chloride (a muscarinic agonist); it was blocked by d-tubocurarine and by alpha-bungarotoxin, but it was not affected by hexamethonium or mecamylamine. 6. The results show that the serotonergic Retzius neurons of the leech could be tonically inhibited by ACh. In addition, the Retzius neurons from standard segments could also be phasically excited by ACh. The receptors responsible for the excitation fit into the classification of neuronal nicotinic receptors, whereas the receptors mediating the inhibition are closer in type to the muscular nicotinic receptor.