Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Attenuation of nicotine taking and seeking in rats by the stoichiometry-selective alpha4beta2 nicotinic acetylcholine receptor positive allosteric modulator NS9283

  • 378 Accesses

  • 8 Citations



The rewarding and reinforcing effects of nicotine are produced, in large part, by activation of neuronal α4β2* nicotinic acetylcholine receptors (nAChRs), pentameric protein complexes comprised of different stoichiometries of α4 and β2 subunits. However, little is known about the functional role of distinct subtypes of α4β2* nAChRs in nicotine addiction.


NS9283 represents a new class of stoichiometry-selective positive allosteric modulators (PAMs) that selectively bind to α4β2 nAChRs containing three α4 and two β2 subunits (3(α4)2(β2) nAChRs). The present experiments were designed to determine the effects of NS9283 on nicotine self-administration and the reinstatement of nicotine-seeking behavior, an animal model of smoking relapse. Parallel studies of sucrose self-administration and reinstatement were conducted in separate cohorts of rats to determine if the effects of NS9283 generalized to other reinforced behaviors.


Acute and repeated administration of NS9283 dose-dependently reduced nicotine self-administration and reinstatement in male Sprague Dawley rats. These effects were reinforcer specific as no effects of NS9283 on sucrose self-administration and reinstatement were noted. NS9283 also failed to substitute for nicotine in supporting self-administration behavior suggesting that, at the doses tested, NS9283 alone is not reinforcing.


Taken together, these results provide compelling evidence that stoichiometry-selective PAMs of 3(α4)2(β2) nAChRs attenuate nicotine taking and seeking in rats and suggest that targeting 3(α4)2(β2) nAChRs may represent a promising therapeutic strategy for preventing smoking relapse.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3.
Fig. 4
Fig. 5


  1. Anderson DJ, Malysz J, Gronlien JH, El Kouhen R, Hakerud M, Wetterstrand C, Briggs CA, Gopalakrishnan M (2009) Stimulation of dopamine release by nicotinic acetylcholine receptor ligands in rat brain slices correlates with the profile of high, but not low, sensitivity alpha4beta2 subunit combination. Biochem Pharmacol 78:844–851

  2. Ashare RL, Schmidt HD (2014) Optimizing treatments for nicotine dependence by increasing cognitive performance during withdrawal. Expert Opin Drug Discov 9:579–594

  3. Ashare RL, Kimmey BA, Rupprecht LE, Bowers ME, Hayes MR, Schmidt HD (2016) Repeated administration of an acetylcholinesterase inhibitor attenuates nicotine taking in rats and smoking behavior in human smokers. Transl Psychiatry 6:e713

  4. CDC CfDCaP (2015) Tobacco use among middle and high school students—United States, 2011–2014. Morb Mortal Wkly Rep 64:381–385

  5. CDCP CfDCaP (2015) Current cigarette smoking among adults—United States, 2005–2014. Morb Mortal Wkly Rep 64:1244–1240

  6. Changeux JP (2010) Nicotine addiction and nicotinic receptors: lessons from genetically modified mice. Nat Rev Neurosci 11:389–401

  7. Chaudhri N, Caggiula AR, Donny EC, Palmatier MI, Liu X, Sved AF (2006) Complex interactions between nicotine and nonpharmacological stimuli reveal multiple roles for nicotine in reinforcement. Psychopharmacology 184:353–366

  8. Coe JW, Brooks PR, Vetelino MG, Wirtz MC, Arnold EP, Huang J, Sands SB, Davis TI, Lebel LA, Fox CB, Shrikhande A, Heym JH, Schaeffer E, Rollema H, Lu Y, Mansbach RS, Chambers LK, Rovetti CC, Schulz DW, Tingley FD 3rd, O’Neill BT (2005) Varenicline: an alpha4beta2 nicotinic receptor partial agonist for smoking cessation. J Med Chem 48:3474–3477

  9. Grupe M, Jensen AA, Ahring PK, Christensen JK, Grunnet M (2013a) Unravelling the mechanism of action of NS9283, a positive allosteric modulator of (alpha4)3(beta2)2 nicotinic ACh receptors. Br J Pharmacol 168:2000–2010

  10. Grupe M, Paolone G, Jensen AA, Sandager-Nielsen K, Sarter M, Grunnet M (2013b) Selective potentiation of (alpha4)3(beta2)2 nicotinic acetylcholine receptors augments amplitudes of prefrontal acetylcholine- and nicotine-evoked glutamatergic transients in rats. Biochem Pharmacol 86:1487–1496

  11. Han ZY, Le Novere N, Zoli M, Hill JA Jr, Champtiaux N, Changeux JP (2000) Localization of nAChR subunit mRNAs in the brain of Macaca mulatta. Eur J Neurosci 12:3664–3674

  12. Hodos W (1961) Progressive ratio as a measure of reward strength. Science 134:943–944

  13. Hopkins TJ, Rupprecht LE, Hayes MR, Blendy JA, Schmidt HD (2012) Galantamine, an acetylcholinesterase inhibitor and positive allosteric modulator of nicotinic acetylcholine receptors, attenuates nicotine taking and seeking in rats. Neuropsychopharmacology : official publication of the American College of Neuropsychopharmacology 37:2310–2321

  14. Ishii K, Wong JK, Sumikawa K (2005) Comparison of alpha2 nicotinic acetylcholine receptor subunit mRNA expression in the central nervous system of rats and mice. J Comp Neurol 493:241–260

  15. Kimmey BA, Rupprecht LE, Hayes MR, Schmidt HD (2012) Donepezil, an acetylcholinesterase inhibitor, attenuates nicotine self-administration and reinstatement of nicotine seeking in rats. Addiction biology.

  16. Lee AM, Arreola AC, Kimmey BA, Schmidt HD (2014) Administration of the nicotinic acetylcholine receptor agonists ABT-089 and ABT-107 attenuates the reinstatement of nicotine-seeking behavior in rats. Behav Brain Res 274:168–175

  17. Lerman C, LeSage MG, Perkins KA, O’Malley SS, Siegel SJ, Benowitz NL, Corrigall WA (2007) Translational research in medication development for nicotine dependence. Nat Rev Drug Discov 6:746–762

  18. Levin ED, Johnson JE, Slade S, Wells C, Cauley M, Petro A, Rose JE (2011) Lorcaserin, a 5-HT2C agonist, decreases nicotine self-administration in female rats. J Pharmacol Exp Ther 338:890–896

  19. Liu X (2013) Positive allosteric modulation of alpha4beta2 nicotinic acetylcholine receptors as a new approach to smoking reduction: evidence from a rat model of nicotine self-administration. Psychopharmacology 230:203–213

  20. Lubbers BR, van Mourik Y, Schetters D, Smit AB, De Vries TJ, Spijker S (2014) Prefrontal gamma-aminobutyric acid type a receptor insertion controls cue-induced relapse to nicotine seeking. Biol Psychiatry 76:750–758

  21. Mohler EG, Franklin SR, Rueter LE (2014) Discriminative-stimulus effects of NS9283, a nicotinic alpha4beta2* positive allosteric modulator, in nicotine-discriminating rats. Psychopharmacology 231:67–74

  22. Moroni M, Zwart R, Sher E, Cassels BK, Bermudez I (2006) alpha4beta2 nicotinic receptors with high and low acetylcholine sensitivity: pharmacology, stoichiometry, and sensitivity to long-term exposure to nicotine. Mol Pharmacol 70:755–768

  23. Nelson ME, Kuryatov A, Choi CH, Zhou Y, Lindstrom J (2003) Alternate stoichiometries of alpha4beta2 nicotinic acetylcholine receptors. Mol Pharmacol 63:332–341

  24. O’Connor EC, Chapman K, Butler P, Mead AN (2011) The predictive validity of the rat self-administration model for abuse liability. Neurosci Biobehav Rev 35:912–938

  25. Picciotto MR, Zoli M, Rimondini R, Lena C, Marubio LM, Pich EM, Fuxe K, Changeux JP (1998) Acetylcholine receptors containing the beta2 subunit are involved in the reinforcing properties of nicotine. Nature 391:173–177

  26. Richardson NR, Roberts DC (1996) Progressive ratio schedules in drug self-administration studies in rats: a method to evaluate reinforcing efficacy. J Neurosci Methods 66:1–11

  27. Rode F, Munro G, Holst D, Nielsen EO, Troelsen KB, Timmermann DB, Ronn LC, Grunnet M (2012) Positive allosteric modulation of alpha4beta2 nAChR agonist induced behaviour. Brain Res 1458:67–75

  28. Schmidt HD, Mietlicki-Baase EG, Ige KY, Maurer JJ, Reiner DJ, Zimmer DJ, Van Nest DS, Guercio LA, Wimmer ME, Olivos DR, De Jonghe BC, Hayes MR (2016) Glucagon-like peptide-1 receptor activation in the ventral tegmental area decreases the reinforcing efficacy of cocaine. Neuropsychopharmacology : official publication of the American College of Neuropsychopharmacology 41:1917–1928

  29. Stolerman IP, Jarvis MJ (1995) The scientific case that nicotine is addictive. Psychopharmacology 117:2–10 discussion 14-20

  30. Taly A, Corringer PJ, Guedin D, Lestage P, Changeux JP (2009) Nicotinic receptors: allosteric transitions and therapeutic targets in the nervous system. Nat Rev Drug Discov 8:733–750

  31. Timmermann DB, Sandager-Nielsen K, Dyhring T, Smith M, Jacobsen AM, Nielsen EO, Grunnet M, Christensen JK, Peters D, Kohlhaas K, Olsen GM, Ahring PK (2012) Augmentation of cognitive function by NS9283, a stoichiometry-dependent positive allosteric modulator of alpha2- and alpha4-containing nicotinic acetylcholine receptors. Br J Pharmacol 167:164–182

  32. Tuesta LM, Fowler CD, Kenny PJ (2011) Recent advances in understanding nicotinic receptor signaling mechanisms that regulate drug self-administration behavior. Biochem Pharmacol 82:984–995

  33. Uteshev VV (2014) The therapeutic promise of positive allosteric modulation of nicotinic receptors. Eur J Pharmacol 727:181–185

  34. Wada E, Wada K, Boulter J, Deneris E, Heinemann S, Patrick J, Swanson LW (1989) Distribution of alpha 2, alpha 3, alpha 4, and beta 2 neuronal nicotinic receptor subunit mRNAs in the central nervous system: a hybridization histochemical study in the rat. J Comp Neurol 284:314–335

  35. Wang F, Chen H, Sharp BM (2008) Neuroadaptive changes in the mesocortical glutamatergic system during chronic nicotine self-administration and after extinction in rats. J Neurochem 106:943–956

  36. Wang J, Kuryatov A, Sriram A, Jin Z, Kamenecka TM, Kenny PJ, Lindstrom J (2015) An accessory agonist binding site promotes activation of alpha4beta2* nicotinic acetylcholine receptors. J Biol Chem 290:13907–13918

  37. Williams DK, Wang J, Papke RL (2011) Positive allosteric modulators as an approach to nicotinic acetylcholine receptor-targeted therapeutics: advantages and limitations. Biochem Pharmacol 82:915–930

  38. Winzer-Serhan UH, Leslie FM (1997) Codistribution of nicotinic acetylcholine receptor subunit alpha3 and beta4 mRNAs during rat brain development. J Comp Neurol 386:540–554

Download references


HDS was supported in part by a K01 training grant (DA030445), an R01 (DA037897) and an R21 (DA039393) from the National Institutes of Health (NIH), a pilot grant (P50-CA-143187) from the Center for Interdisciplinary Research on Nicotine Addiction (CIRNA) at UPenn, and an Institutional Research Grant (IRG-78-002-31) from the American Cancer Society and the Abramson Cancer Center at UPenn. The authors would like to thank Aniona for generously providing NS9283 for these studies. The authors also extend a note of gratitude to Adrian Arreola and Duncan Van Nest for their technical assistance.

Author information

Correspondence to Heath D. Schmidt.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Authors’ contribution

HDS was responsible for the study concept and design, supervised and contributed to the acquisition of data, analyzed the data, and drafted the manuscript. JJM contributed to the acquisition of animal data, analyzed the data, and helped draft the manuscript. KS-N also provided critical revision of the manuscript for important intellectual content. All authors critically reviewed content and approved the final version for publication.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Maurer, J.J., Sandager-Nielsen, K. & Schmidt, H.D. Attenuation of nicotine taking and seeking in rats by the stoichiometry-selective alpha4beta2 nicotinic acetylcholine receptor positive allosteric modulator NS9283. Psychopharmacology 234, 475–484 (2017).

Download citation


  • Nicotine addiction
  • Relapse
  • Self-administration
  • Smoking
  • Reinstatement
  • Positive allosteric modulator
  • E-cigarette