FUNCTIONAL EXPRESSION OF NICOTINIC ACETYLCHOLINE-RECEPTORS CONTAINING RAT ALPHA-7 SUBUNITS IN HUMAN SH-SY5Y NEUROBLASTOMA-CELLS

被引:93
|
作者
PUCHACZ, E [1 ]
BUISSON, B [1 ]
BERTRAND, D [1 ]
LUKAS, RJ [1 ]
机构
[1] UNIV GENEVA, CTR MED, DEPT PHYSIOL, CH-1211 GENEVA 4, SWITZERLAND
来源
FEBS LETTERS | 1994年 / 354卷 / 02期
关键词
ACETYLCHOLINE; NICOTINE; NICOTINIC RECEPTOR; ALPHA-BUNGAROTOXIN; SH-SY5Y CELL LINE;
D O I
10.1016/0014-5793(94)01108-7
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Neuronal nicotinic acetylcholine receptors (nAChR) are made from different combinations of subunits encoded by a diverse family of genes. However, the recently cloned alpha 7 gene codes for subunits that can form homooligomeric nAChR complexes when expressed in Xenopus oocytes. Electrophysiological studies reveal that these alpha 7-nAChR function as alpha-bungarotoxin (Bgt)-sensitive, quickly activating/inactivating ion channels with a unique pharmacological profile and an unusually high permeability to calcium ions. Although similar observations have been made in studies of Bgt-sensitive, functional nAChR subtypes that are naturally expressed in neuronal cells, all attempts until now to reconstitute functional alpha 7-nAChR in cell lines have failed. Here we report the successful use of SH-SY5Y human neuroblastoma cells, which naturally express low levels of endogenous alpha 7 transcripts, to stably overexpress heterologous rat nAChR alpha 7 transgenes. These transgenes are expressed as the appropriately-sized alpha 7 messages and protein, and stably transfected SH-SY5Y cells have over 30-times higher levels of specific Bgt binding sites than do wild-type cells. Whole cell current recordings confirm that transfected cells express functional nAChR that are sensitive to blockade by Bgt and display the typical physiological and pharmacological profiles of alpha 7-nAChR. We conclude that stable, functional expression of alpha 7 transgenes in a mammalian cell line has been achieved for the first time.
引用
收藏
页码:155 / 159
页数:5
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