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Vol. 302, Issue 3, 889-897, September 2002

Pharmacological Properties of Nicotinic Acetylcholine Receptors Expressed by Guinea Pig Small Intestinal Myenteric Neurons

Xiaoping Zhou, Jim Ren, Erika Brown, David Schneider, Yessi Caraballo-Lopez and James J. Galligan

Department of Pharmacology and Toxicology and the Neuroscience Program, Michigan State University, East Lansing, Michigan

The electrophysiological and pharmacological properties of nicotinic acetylcholine receptors (nAChRs) were studied in guinea pig small intestinal myenteric neurons maintained in culture or in acutely isolated preparations. Acetylcholine and nicotine caused inward currents that desensitized in ~4 s. The current-voltage (I-V) relationship rectified inwardly with a reversal potential near 0 mV. The agonist rank order potency was 1,1-dimethyl-4-phenyl-piperazinium > acetylcholine = nicotine cytisine. Agonist-induced currents were blocked by nAChR antagonists with a rank order potency of mecamylamine > hexamethonium > dihydro-beta -erythroidine (DHbeta E); mecamylamine and DHbeta E exhibit high potency at beta 4 and beta 2 subunit-containing nAChRs, respectively. alpha -Bungarotoxin (0.1 µM) or alpha -methyllycaconitine (0.1 µM), antagonists that block nAChRs containing alpha 7 subunits, did not affect acetylcholine-induced responses. Immunohistochemical studies revealed that nearly every neuron in culture was labeled by an antibody (mAb35) that recognizes nAChR alpha 3 and alpha 5 subunits. Antibodies selective for alpha 3, alpha 5, or beta 2 subunits also stained most neurons, whereas an alpha 7 subunit antibody revealed very few neurons. In neurons in the intact myenteric plexus from newborn and adult guinea pigs, local application of acetylcholine (1 mM) and cytisine (1 mM) caused similar amplitude depolarizations, and these responses were blocked by nAChR antagonists with a rank order potency of mecamylamine > hexamethonium > DHbeta E. These data indicate that myenteric neurons maintained in culture predominately express nAChRs composed of alpha 3, alpha 5, beta 2, and beta 4 subunits. These subunits may be in a homogenous population of receptors with unique pharmacological properties, or multiple receptors of different subunit composition may be expressed by individual neurons.

0022-3565/02/3023-0889$07.00/0
THE JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS
Copyright © 2002 by The American Society for Pharmacology and Experimental Therapeutics


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