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Ca 2+ permeability of human heteromeric nAChRs expressed by transfection in human cells

Cell Calcium, ISSN: 0143-4160, Vol: 32, Issue: 2, Page: 53-58
2002
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Article Description

The Ca 2+ permeability of the human heteromeric α3β4, α4β2 and α4β4 neuronal nicotinic acetylcholine receptors (nAChRs) was estimated by measuring the fractional Ca 2+ current ( P f ) flowing through the ligand-activated receptor-channels. Simultaneous recordings of transmembrane currents and fluorescence transients, using the whole-cell patch-clamp technique combined with fura-2 fluorescence microscopy, were performed in transiently transfected human cells. The human α4β2 nAChR showed a P f value of 2.6%, while the human α3β4 nAChR showed a similar P f value of 2.7%. Conversely, α4β4 nAChR exhibited a P f value (1.5%) significantly smaller than those of both α4β2 and α3β4 nAChRs. In test experiments performed in HEK 293 cells stably expressing rat GluR1 AMPA receptor subunit, we repeated the determination of P f, whose value (3.2%) has previously been reported by others using the same fluorescent dye; and we found a very similar P f value (3.5%). In further test experiments, we found that P f values of chick α3β4 (4.4%) and α4β4 (2.1%) matched those previously reported by us using confocal fluorescence microscopy. Thus, our findings are consistent with those elsewhere reported even using different experimental procedures, giving a strong support to the following sequence of Ca 2+ permeability: h-α3β4>h-α4β2>h-α4β4.

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