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J Neurophysiol 84: 1314-1329, 2000;
0022-3077/00 $5.00
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The Journal of Neurophysiology Vol. 84 No. 3 September 2000, pp. 1314-1329
Copyright ©2000 by the American Physiological Society

Expression and Channel Properties of alpha -Bungarotoxin-Sensitive Acetylcholine Receptors on Chick Ciliary and Choroid Neurons

Mary Ellen McNerney, Desiree Pardi, Phyllis C. Pugh, Qiang Nai, and Joseph F. Margiotta

Department of Anatomy and Neurobiology, Medical College of Ohio, Toledo, Ohio 43614-5804

McNerney, Mary Ellen, Desiree Pardi, Phyllis C. Pugh, Qiang Nai, and Joseph F. Margiotta. Expression and Channel Properties of alpha -Bungarotoxin-Sensitive Acetylcholine Receptors on Chick Ciliary and Choroid Neurons. J. Neurophysiol. 84: 1314-1329, 2000. Cell-specific expression of nicotinic acetylcholine receptors (AChRs) was examined using ciliary and choroid neurons isolated from chick ciliary ganglia. At embryonic days 13 and 14 (E13,14) the neurons can be distinguished by size, with ciliary neuron soma diameters exceeding those of choroid neurons by about twofold. Both neuronal populations are known to express two major AChR types: alpha 3*-AChRs recognized by mAb35, that contain alpha 3, alpha 5, beta 4, and occasionally beta 2 subunits, and alpha -bungarotoxin (alpha Bgt)-AChRs recognized and blocked by alpha Bgt, that contain alpha 7 subunits. We found that maximal whole cell current densities (I/Cm) mediated by alpha Bgt-AChRs were threefold larger for choroid compared with ciliary neurons, while alpha 3*-AChR current densities were similar in the two populations. Different densities of total cell-surface alpha Bgt-AChRs could not explain the distinct alpha Bgt-AChR response densities associated with ciliary and choroid neurons. Ciliary ganglion neurons display abundant [125I]-alpha Bgt binding (approx 106 sites/neuron), but digital fluorescence measurements revealed equivalent site densities on both populations. AChR channel classes having single-channel conductances of approx 30, 40, 60, and 80 pS were present in patches excised from both ciliary and choroid neurons. Treating the neurons with alpha Bgt selectively abolished the 60- and 80-pS events, identifying them as arising from alpha Bgt-AChRs. Kinetic measurements revealed brief open and long closed durations for alpha Bgt-AChR channel currents, predicting a very low probability of being open (po) when compared with 30- or 40-pS alpha 3*-AChR channels. None of the channel parameters associated with the 60- and 80-pS alpha Bgt-AChRs differed detectably, however, between choroid and ciliary neurons. Instead calculations based on the combined whole cell and single-channel results indicate that choroid neurons express approximately threefold larger numbers of functional alpha Bgt-AChRs (NF) per unit area than do ciliary neurons. Comparison with total surface [125I]-alpha Bgt-AChR sites (NT), reveals that NF/NT 1 for both neuron populations, suggesting that "silent" alpha Bgt-AChRs predominate. Choroid neurons may therefore express a higher density of functional alpha Bgt-AChRs by recruiting a larger fraction of receptors from the silent pool than do ciliary neurons.




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