Title

Functional Expression Of Nicotinic Acetylcholine Receptors Containing Rat α7 Subunits In Human Sh-Sy5Y Neuroblastoma Cells

Department

neurobiology

Document Type

Article

Abstract

Neuronal nicotinic acetylcholine receptors (nAChR) are made from different combinations of subunits encoded by a diverse family of genes. However, the recently cloned α7 gene codes for subunits that can form homooligomeric nAChR complexes when expressed in Xenopus oocytes. Electrophysiological studies reveal that these α7-nAChR function as α-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 α7-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 α7 transcripts, to stably overexpress heterologous rat nAChR α7 transgenes. These transgenes are expressed as the appropriately-sized α7 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 α7-nAChR. We conclude that stable, functional expression of α7 transgenes in a mammalian cell line has been achieved for the first time. © 1994.

Publication Date

11-7-1994

Publication Title

FEBS Letters

ISSN

00145793

Volume

354

Issue

2

First Page

155

Last Page

159

Digital Object Identifier (DOI)

10.1016/0014-5793(94)01108-7

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