A nicotinic acetylcholine receptor-like α-bungarotoxin-binding site on outer hair cells

Abstract

Acetylcholine (ACh) appears to be the major neurotransmitter liberated from olivocochlear efferents terminating on outer hair cells (OHC). Recently, cholinergic receptor epitopes were visualized at the basal pole of the OHCs. To evaluate the ACh receptor type at OHC we performed binding studies with [ 125I]-labelled α-bungarotoxin (α-bgtx), a close to irreversibly acting blocker of the nicotinic acetylcholine receptor (nAChR) of skeletal muscle and of electrocytes of Torpedo and Electrophorus. An irreversible and saturable binding (80 nM) of the radiolabelled compound to OHCs was observed. The number of α-bgtx sensitive binding sites present on each OHC was calculated to be about 2×10 −17 mol/OHC, which would amount to about 10 7 binding sites/cell. Preincubation with the reversibly acting cholinergic ligands, carbamylcholine (1 mM), nicotine (0.1 mM) and d-tubocurarine (1–100 μM) was found to inhibit α-bgtx binding to a varying degree. Atropine (0.05 mM), a muscarinic antagonist, had no influence on the binding of α-bgtx to OHCs. [ 3H]-QNB, a specific marker and antagonist for muscarinic AChR, and [ 125I]-κ-toxin, known to react with neuronal and ganglionic nAChR, showed no specific binding to OHCs. The data indicate that a peripheral type nAChR is present on OHCs mediating ACh-induced modulation of the biomechanics of the cochlea by influencing OHC motility.