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Research ArticleCELLULAR AND MOLECULAR

Receptor-Mediated Inhibition of Keratinocyte Migration by Nicotine Involves Modulations of Calcium Influx and Intracellular Concentration

Shaheen Zia, Assane Ndoye, Tou X. Lee, Robert J. Webber and Sergei A. Grando
Journal of Pharmacology and Experimental Therapeutics June 2000, 293 (3) 973-981;
Shaheen Zia
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Assane Ndoye
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Tou X. Lee
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Robert J. Webber
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Sergei A. Grando
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Abstract

Early stages of wound healing rely on the ability of keratinocytes (KCs) to move over the denuded dermis to re-epithelialize the defect. The agarose gel keratinocyte outgrowth system (AGKOS) is an in vitro model of skin re-epithelialization designed to study the migratory function of KCs. Endogenously secreted acetylcholine controls crawling locomotion of KCs in AGKOS by binding to the cholinergic receptors of both the nicotinic and the muscarinic classes that are expressed by KCs. In this study, we used AGKOS to elucidate the nicotinic pathway of cholinergic control of keratinocyte migration. Activation of the nicotinic acetylcholine receptors decreased the migration distance of KC in a dose-dependent fashion without altering cell viability. Nicotine also increased in a dose-dependent manner transmembrane influx of 45Ca2+, and caused a transient rise in the concentration of [Ca2+]i. Perfect correlation between concentration responses found in the migration and45Ca2+ influx assays suggested that nicotine-induced inhibition of crawling locomotion relies on modulation of Ca2+ metabolism in KCs. The effects of nicotine could be mediated by the α3- and the α7-containing nicotinic receptors visualized on KCs by immunostaining. Long-term incubation with nicotine up-regulated α7 and down-regulated α3 expression. Thus, nicotine exerts inhibitory effects on keratinocyte migration, and Ca2+ serves as a second messenger in the signaling pathway. These results help explain deleterious effects of nicotine on wound re-epithelialization, and suggest that smoking may delay wound healing via nicotinic receptor-mediated pathway.

Footnotes

  • Send reprint requests to: Sergei A. Grando, M.D., Ph.D., D.Sc., Department of Dermatology, University of California-Davis, UC Davis Medical Center, 4860 Y St., Suite 3400, Sacramento, CA 95817. E-mail: sagrando{at}ucdavis.edu

  • ↵1 This study was supported by National Institutes of Health Grant R29 AR42955 (to S.A.G.).

  • Abbreviations:
    KC
    keratinocyte
    AGKOS
    agarose gel keratinocyte outgrowth system
    ACh
    acetylcholine
    nAChR
    nicotinic acetylcholine receptor
    KGM
    KC growth medium
    IIF
    indirect immunofluorescence
    • Received October 27, 1999.
    • Accepted February 21, 2000.
  • The American Society for Pharmacology and Experimental Therapeutics
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Journal of Pharmacology and Experimental Therapeutics: 293 (3)
Journal of Pharmacology and Experimental Therapeutics
Vol. 293, Issue 3
1 Jun 2000
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Research ArticleCELLULAR AND MOLECULAR

Receptor-Mediated Inhibition of Keratinocyte Migration by Nicotine Involves Modulations of Calcium Influx and Intracellular Concentration

Shaheen Zia, Assane Ndoye, Tou X. Lee, Robert J. Webber and Sergei A. Grando
Journal of Pharmacology and Experimental Therapeutics June 1, 2000, 293 (3) 973-981;

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Research ArticleCELLULAR AND MOLECULAR

Receptor-Mediated Inhibition of Keratinocyte Migration by Nicotine Involves Modulations of Calcium Influx and Intracellular Concentration

Shaheen Zia, Assane Ndoye, Tou X. Lee, Robert J. Webber and Sergei A. Grando
Journal of Pharmacology and Experimental Therapeutics June 1, 2000, 293 (3) 973-981;
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