Antagonist pharmacology of desensitizing and non-desensitizing nicotinic acetylcholine receptors in cockroach neurons

• The single-electrode voltage-clamp technique with dual bath and U-tube perfusion was used to measure concentration-response relations for ACh and five nicotinic antagonists on nAChD and nAChN receptors on neurons of acutely dissociated cockroach neurons.
• ACh activated both receptors with the same potency, with an EC50 of 7 μM.
• Dihydro-β-erythroidine, condelphine, phencyclidine and mecamylamine were selective toward nAChD receptors, whereas d-tubocurarine selectively blocked nAChN receptors.

Two α-bungarotoxin-sensitive nicotinic acetylcholine (ACh) receptor subtypes in neurons of the American cockroach have been identified as desensitizing (nAChD) and selectively inhibitable with 100 nM imidacloprid, and non-desensitizing (nAChN) and selectively inhibitable with 100 pM methyllycaconitine. In this paper, the single-electrode voltage-clamp technique was used to measure concentration-response relations for the action of ACh and five antagonists on pharmacologically separated nAChD and nAChN receptors of acutely dissociated neurons from thoracic ganglia of the American cockroach. A dual bath and U-tube perfusion system was used to achieve rapid application of ACh in the continued presence of antagonists, which was essential to accurately measure inhibition by rapidly-reversible antagonists. ACh activated both receptors with an EC50 of 7 μM and the antagonist potencies were (nAChD/nAChN in nM): dihydro-β-erythroidine: 1.0/5.6, d-tubocurarine: 1000/34, condelphine: 0.39/0.65, phencyclidine: 74/980 and mecamylamine 47/1150. While each of these antagonists displayed some subtype selectivity, none are selective enough to be used as subtype-selective tools. These results bring to a total of 16 the number of nicotinic compounds that have been measured on nAChD and nAChN currents. Characterization of these receptors is important for understanding the role of nAChRs in the insect nervous system and the mechanism of action of insecticides.