Functional Properties of the Neuronal Nicotinic Acetylcholine Receptor β3 Promoter in the Developing Central Nervous System*
Abstract
Within the chick central nervous system, expression of the β3 nicotinic acetylcholine receptor gene is restricted to a subset of retinal neurons, the majority of which are ganglion cells. Transient transfection in retinal neurons and in neural and non-neural cells from other regions of the chick embryo allowed the identification of the cis-regulatory domain of the β3 gene. Within this domain, a 75-base pair fragment located immediately upstream of the transcription start site suffices to reproduce the neuron-specific expression pattern of β3. This fragment encompasses an E-box and a CAAT box, both of which are shown to be key positive regulatory elements of the β3 promoter. Co-transfection experiments into retinal, telencephalic, and tectal neurons with plasmid reporters of β3 promoter activity and a number of vectors expressing different neuronal (ASH-1, NeuroM, NeuroD, CTF-4) and non-neuronal (MyoD) basic helix-loop-helix transcription factors indicate that the cis-regulatory domain of β3 has the remarkable property of discriminating accurately between related members of the basic helix-loop-helix protein family. The sequence located immediately 3′ of the E-box participates in this selection, and the E-box acts in concert with the nearby CAAT box.
Footnotes
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↵* This work was supported by the Swiss National Science Foundation and the State of Geneva.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.
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↵‡ To whom correspondence should be addressed: Dept. of Biochemistry, Sciences II, 30 quai Ernest-Ansermet, 1211 Genève 4, Switzerland. Tel.: 41-22-702-6492; Fax: 41-22-702-6483; E-mail:jean-marc.matter{at}biochem.unige.ch.
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↵1 The abbreviations used are: bHLH, basic helix-loop-helix; nAChR, nicotinic acetylcholine receptor; CAT, chloramphenicol acetyltransferase; X-gal, 5-bromo-4-chloro-3-indoyl β-d-galactosidase; CEF, chick embryonic fibroblast; E4–E13, embryonic days 4–13; bp, base pair(s); CNS, central nervous system.
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↵2 J.-M. Matter, unpublished data.
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↵3 T. Roztocil, unpublished observation.
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↵4 L. Matter-Sadzinski, M. Ballivet, and J.-M. Matter, manuscript in preparation.
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- Received October 16, 1997.
- Revision received January 7, 1998.
- The American Society for Biochemistry and Molecular Biology, Inc.
