We examine some of the biological and physiological properties of the avian alpha(6) neuronal nicotinic acetylcholine receptor (nAChR) subunit, We show here that, beginning at embryonic day 5, alpha(6) mRNA is abundantly expressed in the developing chick neuroretina, where it coexists with other nicotinic receptor subunit mRNAs such as alpha(3), beta(2) and beta(4). In contrast, alpha(6) mRNA is absent from the optic tectum and from the peripheral ganglia, Despite numerous efforts, the alpha(6) subunit has long failed the critical test of functional reconstitution, Here we use patch-clamp techniques and confocal laser microscopy to measure ACh-activated currents and nicotine-elicited Ca2+ transients in human BOSC 23 cells transfected with chick alpha(6) in combination with other chick nAChR neuronal subunits. Heterologously expressed alpha(6) and beta(4) subunits form functional heteromeric nAChRs, which are permeable to Ca2+ ions and blocked by the nicotinic antagonist methyllycaconitine (10 mu M). Likewise, ACh elicits measurable currents in cells transfected with alpha(6) and beta(2). Hill analysis of the dose-response curves in cells transfected with alpha(3), beta(4) and alpha(6) cDNAs, suggests the assembly of functional alpha(3) beta(4) alpha(6) receptor, with an apparent affinity for ACh threefold lower than alpha(3) beta(4). Our results indicate that alpha(6)-containing nAChRs assemble in heterologous expression systems and are probably present in retinal cells.
The neuronal alpha(6) subunit forms functional heteromeric acetylcholine receptors in human transfected cells
1998
Abstract
We examine some of the biological and physiological properties of the avian alpha(6) neuronal nicotinic acetylcholine receptor (nAChR) subunit, We show here that, beginning at embryonic day 5, alpha(6) mRNA is abundantly expressed in the developing chick neuroretina, where it coexists with other nicotinic receptor subunit mRNAs such as alpha(3), beta(2) and beta(4). In contrast, alpha(6) mRNA is absent from the optic tectum and from the peripheral ganglia, Despite numerous efforts, the alpha(6) subunit has long failed the critical test of functional reconstitution, Here we use patch-clamp techniques and confocal laser microscopy to measure ACh-activated currents and nicotine-elicited Ca2+ transients in human BOSC 23 cells transfected with chick alpha(6) in combination with other chick nAChR neuronal subunits. Heterologously expressed alpha(6) and beta(4) subunits form functional heteromeric nAChRs, which are permeable to Ca2+ ions and blocked by the nicotinic antagonist methyllycaconitine (10 mu M). Likewise, ACh elicits measurable currents in cells transfected with alpha(6) and beta(2). Hill analysis of the dose-response curves in cells transfected with alpha(3), beta(4) and alpha(6) cDNAs, suggests the assembly of functional alpha(3) beta(4) alpha(6) receptor, with an apparent affinity for ACh threefold lower than alpha(3) beta(4). Our results indicate that alpha(6)-containing nAChRs assemble in heterologous expression systems and are probably present in retinal cells.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
