Ragozzino D, Fucile S, Giovannelli A, Grassi F, Mileo A M, Ballivet M, Alemà S, Eusebi F
Istituto Pasteur-Fondazione Cenci Bolognetti, Dipartimento di Medicina Sperimentale e Patologia, Università di Roma, Italy.
Eur J Neurosci. 1997 Mar;9(3):480-8. doi: 10.1111/j.1460-9568.1997.tb01625.x.
To study how subunit composition affects the functional properties of neuronal nicotinic acetylcholine receptors (nAChRs), we examined the behaviour of acetylcholine (ACh)-induced single-channel currents in human BOSC 23 cells transiently transfected with various subunit cDNA combinations. For all nAChRs examined (chick and rat alpha 3 beta 4, chick alpha 3 beta 2, alpha 4 beta 2, alpha 7 and alpha 8), expression levels were high enough to allow measurements of acetylcholine-evoked whole-cell currents and nicotine-elicited Ca2+ transients as well as the functional characterization of nAChR channels. Unitary acetylcholine-evoked events of alpha 8 nAChR had a slope conductance of 23 pS, whereas two conductance classes (19-23 and 32-45 pS) were identified for all other nAChR channels. The mean channel open times were significantly longer for homomeric alpha 7 and alpha 8 nAChRs (6-7 ms) than for heteromeric nAChRs (1-3 ms), with the exception of alpha 3 beta 4 nAChRs (8.4 ms for rat, 7 ms for chick). At least two species of heterologously expressed nAChRs (alpha 3 beta 4 and alpha 3 beta 2) exhibited single-channel characteristics similar to those reported for native receptors. The variety of nAChR channel conductance and kinetic properties encountered in human cells transfected with nAChR subunits contributes to the functional diversity of nAChRs in nerve cells.
为了研究亚基组成如何影响神经元烟碱型乙酰胆碱受体(nAChRs)的功能特性,我们检测了用各种亚基cDNA组合瞬时转染的人BOSC 23细胞中乙酰胆碱(ACh)诱导的单通道电流行为。对于所有检测的nAChRs(鸡和大鼠的α3β4、鸡α3β2、α4β2、α7和α8),表达水平足够高,足以测量乙酰胆碱诱发的全细胞电流和尼古丁引发的Ca2+瞬变以及nAChR通道的功能特性。α8 nAChR的单一乙酰胆碱诱发事件的斜率电导为23 pS,而所有其他nAChR通道则鉴定出两种电导类别(19 - 23和32 - 45 pS)。同聚体α7和α8 nAChRs的平均通道开放时间(6 - 7毫秒)显著长于异聚体nAChRs(1 - 3毫秒),大鼠α3β4 nAChRs除外(大鼠为8.4毫秒,鸡为7毫秒)。至少两种异源表达的nAChRs(α3β4和α3β2)表现出与天然受体报道的单通道特性相似。用nAChR亚基转染的人细胞中遇到的nAChR通道电导和动力学特性的多样性有助于神经细胞中nAChRs的功能多样性。