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J Biol Chem, Vol. 275, Issue 18, 13974-13985, May 5, 2000

Amino Acid Determinants of alpha 7 Nicotinic Acetylcholine Receptor Surface Expression*

Kelly T. DineleyDagger and James W. Patrick

From the Division of Neuroscience, Baylor College of Medicine, Houston, Texas 77030

Transient transfection has not been a successful method to express the alpha 7 nicotinic acetylcholine receptor such that these receptors are detected on the cell surface. This is not the case for all ligand-gated ion channels. Transient transfection with the 5-hydroxytryptamine type 3 subunit cDNA results in detectable surface receptor expression. Cell lines stably expressing the alpha 7 nicotinic acetylcholine receptor produce detectable, albeit variable, levels of surface receptor expression. alpha 7 nicotinic acetylcholine receptor surface expression is dependent, at least in part, on cell-specific factors. In addition to factors provided by the cells used for receptor expression, we hypothesize that the surface expression level in transfected cells is an intrinsic property of the receptor protein under study. Employing a set of alpha 7-5-hydroxytryptamine type 3 chimeric receptor subunit cDNAs, we expressed these constructs in a transient transfection system and quantified surface receptor expression. We have identified amino acids that control receptor distribution between surface and intracellular pools; surface receptor expression can be manipulated without affecting the total number of receptors. These determinants function independently of the cell line used for expression and the transfection method employed. How these surface expression determinants in the alpha 7 nicotinic acetylcholine receptor might influence synaptic efficacy is discussed.

* This work was supported by National Institutes of Health Grants F31 DA05694 and NS 13546.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Dagger To whom correspondence should be addressed: Division of Neuroscience, Rm. S603, Baylor College of Medicine, One Baylor Plaza, Houston, TX 77030. Tel.: 713-798-3086; Fax: 713-798-3946; E-mail: kdineley@sensor.neusc.bcm.tmc.edu.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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