Cigarette Smoke Extract: A Preclinical Model of Tobacco Dependence

Candice A. Gellner1, Daisy D. Reynaga1, Frances M. Leslie1

1 Department of Pharmacology, University of California, Irvine, Irvine, California
Publication Name:  Current Protocols in Neuroscience
Unit Number:  Unit 9.54
DOI:  10.1002/cpns.14
Online Posting Date:  October, 2016
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Abstract

Animal models are used to study many human diseases, one of which is tobacco addiction. Most preclinical models use nicotine alone, although there are >7000 constituents present in tobacco smoke. The clinical literature suggests that cigarettes have a strong addictive potential, which is not paralleled in preclinical studies using nicotine alone. In order to address the gap between clinical and preclinical literature on tobacco dependence, cigarette smoke extracts containing tobacco constituents have been developed. This unit describes a procedure for producing an aqueous cigarette smoke extract (CSE) which animals readily self‐administer. In addition, we describe how to make the apparatus for producing CSE and how to analyze the solution for nicotine content. © 2016 by John Wiley & Sons, Inc.

Keywords: nicotine; tobacco constituents; self‐administration; smoking

     
 
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Table of Contents

  • Introduction
  • Significance Statement
  • Basic Protocol 1: Preparing Cigarette Smoke Extract (CSE)
  • Support Protocol 1: Constructing the Smoking Apparatus
  • Support Protocol 2: Analyzing Nicotine Content
  • Reagents and Solutions
  • Commentary
  • Literature Cited
  • Figures
     
 
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Materials

Basic Protocol 1: Preparing Cigarette Smoke Extract (CSE)

  Materials
  • Camel unfiltered cigarettes (8)
  • Saline
  • Petroleum jelly
  • 0.1 M, 1 M, and 10 M NaOH
  • 0.1 M, and 1 M HCl
  • Cigarette smoke extract (CSE) apparatus (see protocol 2)
    • Ring stand with 2 three‐prong clamps
    • 50‐ml polypropylene conical tubes (2)
    • 1000‐µl pipet tip
    • 50‐ml glass syringe with glass plunger
  • Aluminum foil
  • Timer
  • Student Adson forceps (12 cm, serrated tip)
  • Black fine tip permanent marker
  • pH test strips

Support Protocol 1: Constructing the Smoking Apparatus

  Materials
  • Plastic repair epoxy (fast setting)
  • Cyanoacrylate (e.g., superglue)
  • 50‐ml polypropylene conical tube cap
  • 10 in. (25 cm) polypropylene tubing (5/16 in. I.D. × 7/16 in. O.D., wall thickness 1/16 in.)
  • 15 in. (38 cm) polypropylene tubing (1/8 in. I.D. × 1/4 in. O.D., wall thickness 1/16 in.)
  • Plastic barbed straight quick‐connect couplers, 3/8 in. × 1/4 in. I.D. (Cole‐Parmer)
  • Cotton‐tipped applicators, 6 in. (15 cm), wood shaft
  • Razor blade
  • Black fine tip permanent marker
  • Ring stand with 2 three‐prong clamps

Support Protocol 2: Analyzing Nicotine Content

  Materials
  • Cigarette smoke extract (CSE; see protocol 1Basic Protocol)
  • (‐)‐Nicotine hydrogen tartrate salt (Glentham Life Sciences)
  • 2 M NaOH (see recipe)
  • 1 M HCl (see recipe)
  • Dichloromethane (DCM; Thermo Fisher Scientific)
  • Magnesium sulfate (anhydrous)
  • Glass vials
  • Vortex mixer
  • Separation funnel
  • Gas chromatography mass spectrophotometer (GC‐MS)
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Figures

Videos

Literature Cited

Literature Cited
  Arnold, M.M., Loughlin, S.E., Belluzzi, J.D., and Leslie, F.M. 2014. Reinforcing and neural activating effects of norharmane, a non‐nicotine tobacco constituent, alone and in combination with nicotine. Neuropharmacology 85:293‐304. doi: 10.1016/j.neuropharm.2014.05.035.
  Belluzzi, J.D., Wang, R., and Leslie, F.M. 2005. Acetaldehyde enhances acquisition of nicotine self‐administration in adolescent rats. Neuropsychopharmacology 30:705‐712. doi: 10.1038/sj.npp.1300586.
  Benowitz, N.L. 1988. Pharmacologic aspects of cigarette smoking and nicotine addiction. N. Engl. J. Med. 319:1318‐1330. doi: 10.1056/NEJM198811173192005.
  Costello, M.R., Reynaga, D.D., Mojica, C.Y., Zaveri, N.T., Belluzzi, J.D., and Leslie, F.M. 2014. Comparison of the reinforcing properties of nicotine and cigarette smoke extract in rats. Neuropsychopharmacology 39:1843‐1851. doi: 10.1038/npp.2014.31.
  Harris, A.C., Mattson, C., Lesage, M.G., Keyler, D.E., and Pentel, P.R. 2010. Comparison of the behavioral effects of cigarette smoke and pure nicotine in rats. Pharmacol. Biochem. Behav. 96:217‐227. doi: 10.1016/j.pbb.2010.05.008.
  Harvey, D.M., Yasar, S., Heishman, S.J., Panlilio, L.V., Henningfield, J.E., and Goldberg, S.R. 2004. Nicotine serves as an effective reinforcer of intravenous drug‐taking behavior in human cigarette smokers. Psychopharmacology (Berl.) 175:134‐142. doi: 10.1007/s00213‐004‐1818‐6.
  Manzardo, A.M., Stein, L., and Belluzzi, J.D. 2002. Rats prefer cocaine over nicotine in a two‐lever self‐administration choice test. Brain. Res. 924:10‐19. doi: 10.1016/s0006‐8993(01)03215‐2.
  Mello, N.K. and Newman, J.L. 2011. Discriminative and reinforcing stimulus effects of nicotine, cocaine, and cocaine + nicotine combinations in rhesus monkeys. Exp. Clin. Psychopharmacol. 19:203‐214. doi: 10.1037/a0023373.
  Small, E., Shah, H.P., Davenport, J.J., Geier, J.E., Yavarovich, K.R., Yamada, H., and Bruijnzeel, A.W. 2010. Tobacco smoke exposure induces nicotine dependence in rats. Psychopharmacology (Berl.) 208:143‐158. doi: 10.1007/s00213‐009‐1716‐z.
  Thomsen, M. and Caine, S.B. 2005. Chronic intravenous drug self‐administration in rats and mice. Curr. Protoc. Neurosci. 32:9.20.1‐9.20.40. doi: 10.1002/0471142301.ns0920s32.
  U.S. Department of Health and Human Services. 2010. How Tobacco Smoke Causes Disease: The Biology and Behavioral Basis for Smoking‐Attributable Disease: A Report of the Surgeon General. Chapter 3. U.S. Department of Health and Human Services, Centers for Disease Control and Prevention, National Center for Chronic Disease Prevention and Health Promotion, Office on Smoking and Health, Atlanta. http://www.ncbi.nlm.nih.gov/books/NBK53017 (Accessed April 1, 2016).
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