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J. Biol. Chem., Vol. 278, Issue 18, 15633-15640, May 2, 2003
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From the Division of Neurobiology, Barrow Neurological Institute,
Phoenix, Arizona 85013
Nicotine exposure can have long lasting effects
on nervous system function, some of which must contribute to nicotine
dependence. Up-regulation, an increase in numbers of
radioligand-binding nicotinic acetylcholine receptors (nAChR), occurs
on exposure to nicotine at high concentrations. To determine whether
altered gene expression might account for long term changes and
up-regulation following nicotine exposure, we assessed effects of
1 h of 1 mM nicotine exposure on alteration of
gene expression in the neuron-like SH-SY5Y neuroblastoma clonal line.
Repeat and cross-controlled microarray analyses yielded a list of 17 genes from the initially screened ~5,000 whose expression was
consistently altered following nicotine treatment. Subsequent
quantitative, real time reverse transcriptase PCR analyses
confirmed altered expression in 14 of 16 genes tested. Further, the
general nAChR antagonist, d-tubocurarine, blocked all but two of the
observed changes in gene expression, indicating that these changes are
dependent on nAChR activation. Use of other antagonists revealed that
nAChR subtypes can differentially affect gene expression. The genes
affected code for proteins that may be broadly categorized into four
groups: transcription factors, protein processing factors, RNA-binding
proteins, and plasma membrane-associated proteins. Our results suggest
that nicotinic activation of nAChR may have a broad role in affecting
cellular physiology through modulating gene expression.
Nicotine Modulates the Expression of a Diverse Set of Genes in
the Neuronal SH-SY5Y Cell Line*
*
This work was supported in part by endowment and/or
capitalization funds from the Men's and Women's Boards of the Barrow
Neurological Foundation, by the Robert and Gloria Wallace Foundation,
by National Institutes of Health Grant R01-NS40417, by Arizona Disease
Control Research Commission Grant 10011, and by Philip Morris Inc.
through an External Research Program postdoctoral fellowship (to
T. D.), and was conducted in part in the Charlotte and Harold Simensky Neurochemistry of Alzheimer's Disease Laboratory.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.
To whom correspondence should be addressed: Div. of Neurobiology,
Barrow Neurological Inst., 350 West Thomas Rd., Phoenix, AZ 85013. Tel.: 602-406-3398; Fax: 602-406-4172; E-mail:
rlukas@chw.edu.
Copyright © 2003 by The American Society for Biochemistry and Molecular Biology, Inc.
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