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Ca²⁺ permeability of mouse and chick nicotinic acetylcholine receptors expressed in transiently transfected human cells

Davide Ragozzino *, Benedetta Barabino *, Sergio Fucile * and Fabrizio Eusebi *¹

1. Combinations of cDNAs encoding mouse and chick nicotinic acetylcholine receptor (nAChR) subunits were transiently transfected into human BOSC 23 cells, and the expressed receptors were studied by simultaneously recording transmembrane currents and fluorescence transients using the whole-cell patch-clamp technique, and confocal microscopy with the Ca²⁺ indicator dye fluo-3.

2. The fractional Ca²⁺ current, P_f, of nAChRs was evaluated as the normalized ratio of nicotine-evoked fluorescence transient over total charge entering the cell (F/Q ratio). Mouse fetal muscle nAChR channels had a P_f, value of 2·1 %. The substitution of the subunit with the subunit resulted in a 2-fold increase in P_f (4·2 %). The difference in Ca²⁺ permeability was confirmed by determination of Ca²⁺/Cs⁺ permeability ratios.

3. Among the chick neuronal nAChRs tested, P_f,34 was 4·6 %, while P_f,44 and P_f,42 were 3·0 % and 2·9 %, respectively.

4. The amplitude of the current elicited by the activation of 34 nAChRs increased as the external Ca²⁺ concentration was raised from 2 to 110 mM, whereas currents flowing through all other nAChRs tested were reduced to various extents.

5. Our findings indicate that the adult-type muscle nAChR () is more permeable to Ca²⁺ than the fetal-type (), while ganglionic-like 34 nAChR is more permeable to Ca²⁺ than the examined 4-containing nAChRs. The functional significance is discussed.

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