Characterization of the Nicotinic Acetylcholine Receptor 3 Gene

doi:10.1074/jbc.270.7.3224

Characterization of the Nicotinic Acetylcholine Receptor 3 Gene

ITS REGULATION WITHIN THE AVIAN NERVOUS SYSTEM IS EFFECTED BY A PROMOTER 143 BASE PAIRS IN LENGTH (*)

From the Department of Biochemistry, Sciences II, University of Geneva, 1211 Geneva 4, Switzerland

** To whom correspondence should be addressed:
Dept. of Biochemistry, Sciences II, 30 Quai Ernest Ansermet, 1211 Geneva 4, Switzerland.
Tel.: 41-22-7026494; Fax: 41-22-7026483.

↵^§ Present address: Brain Tumor Research Center, The Preuss Laboratory, University of California, San Francisco, CA 94143-0520.
↵^¶ Present address: Dept. of Cellular and Molecular Medicine, University of California at San Diego, La Jolla, CA 92093-0648.

Abstract

Genomic and cDNA clones encoding the chicken neuronal nicotinic acetylcholine receptor β3 subunit were isolated and sequenced. The β3 gene consists of six protein-encoding exons and the deduced protein has the structural features found in all other members of the neuronal nicotinic acetylcholine receptor subunit family. Although they are undetectable in most brain compartments, β3 mRNAs are relatively abundant in the developing retina and in the trigeminal ganglion. In situ hybridization and immunohistochemical analysis demonstrated that in retina, β3 transcripts and protein are confined to subpopulations of cells in the inner nuclear and ganglion cell layers. β3 is expressed in the proximal and distal regions of the developing trigeminal ganglion, i.e. in both placode- and neural crest-derived neurons. Transient transfection assays in cells freshly dissociated from selected regions of the central nervous system at different developmental stages allowed the identification of genetic elements involved in the neuronal-selective expression of the β3 gene. A promoter fragment 143 base pairs in length and containing TATA, CAAT, and other consensus sequences is sufficient to restrict reporter gene expression to a subpopulation of retinal neurons. This promoter is totally inactive upon transfection into neuronal and non-neuronal cells from other regions of the central nervous system.

Footnotes

↵* This research was funded by the State of Geneva and by grants from the Swiss National Science Foundation. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore by hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBank™/EMBL Data Bank with accession number(s) X83739 [GenBank]and X83740[GenBank].
↵¹ The abbreviations used are:

nAChR(s)

nicotinic acetylcholine receptor(s)

GCL

ganglion cell layer

INL

inner nuclear layer

IPL

inner plexiform layer

HLH

helix-loop-helix

CAT

chloramphenicol acetyltransferase

β-gal

β-galactosidase

CEF(s)

chick embryonic fibroblast-like cell(s)

CHO

Chinese hamster ovary

MOPS

3-morpholinepropanesulfonic acid

PBS

phosphate-buffered saline

PIPES

1,4-piperazinediethanesulfonic acid

X-gal

5-bromo-4-chloro-3-indoyl β-D-galactoside

E

embryonic day

bp

base pair

kbp

kilobase pair

CNS

central nervous system.
↵² L. Erkman, M. Gomez, J.-M. Matter, and M. Ballivet, submitted for publication.
↵³ M.-C. Hernandez, L. Erkman, L. Matter-Sadzinski, T. Roztocil, M. Ballivet, and J.-M. Matter, unpublished results.
↵⁴ S. Bertrand, N. Hussy, and D. Bertrand, unpublished results.
↵⁵ J.-M. Matter, L. Matter-Sadzinski, and M. Ballivet, submitted for publication.
- Received August 19, 1994.
- Revision received November 3, 1994.