Single-channel recording in brain slices reveals heterogeneity of nicotinic receptors on individual neurons within the chick lateral spiriform nucleus

Abstract

We examined the functional properties of central nicotinic acetylcholine receptors at the single-channel level using tight-seal, voltage-clamp techniques. Single-channel currents were recorded from cell-attached patches on lateral spiriform neurons in chick brain slices. These neurons are known to express functional nicotinic receptors that are insensitive to the antagonists α-bungarotoxin and κ-bungarotoxin, and which exhibit a high affinity for nicotine and other nicotinic agonists. Single-channel openings were observed in 84% of patches ( n = 118) when the nicotinic agonists acetylcholine (1–100 μM), carbamylcholine (3–100 μM), or nicotine (3–10 μM) were present in the patch pipette. In contrast, single-channels were markedly reduced in number or entirely absent when the nicotinic antagonist dihydro-β-erythroidine was present along with acetylcholine ( n = 7) or when no agonist was present in the pipette ( n = 22). Single-channel openings displayed inward rectification at depolarized potentials, and were dependent on extracellular sodium. Between 1 and 30 μM acetylcholine, a dose-response relationship was observed between agonist concentration and single-channel open probability during the first minute following seal formation. Multiple classes of single nicotinic channels, with calculated mean slope conductances of 15, 31, 40, and ∼ 70 pS, were observed in membrane patches on different neurons within the lateral spiriform nucleus, and even within single patches on individual neurons. We conclude that neurons within the lateral spiriform nucleus express functionally heterogeneous nicotinic receptors and that in some neurons different nicotinic receptor subtypes are present in close proximity to each other on the same cell surface.