, Volume 226, Issue 2, pp 347–355 | Cite as

Hypothalamic Neuropeptide S receptor blockade decreases discriminative cue-induced reinstatement of cocaine seeking in the rat

  • Marsida Kallupi
  • Giordano de Guglielmo
  • Nazzareno Cannella
  • Hong Wu Li
  • Girolamo Caló
  • Remo Guerrini
  • Massimo Ubaldi
  • John J. Renger
  • Victor N. Uebele
  • Roberto CiccocioppoEmail author
Original Investigation



Previous studies have shown that activation of brain neuropeptide S receptor (NPSR) facilitates reinstatement of cocaine seeking elicited by environmental cues predictive of drug availability. This finding suggests the possibility that blockade of NPSR receptors may be of therapeutic benefit in cocaine addiction. To evaluate this hypothesis, we investigated the effect of two newly synthetized NPSR antagonists, namely the quinolinone-amide derivative NPSR-QA1 and the NPS peptidic analogue [D-Cys(tBu)5]NPS on cocaine self-administration and on discriminative cue-induced relapse to cocaine seeking in the rat.


Separate groups of rats self-administered food and cocaine 0.25 mg/kg/inf in FR1 and FR5 (fixed ratio reinforcement schedules) for 30-min and 2-h sessions per day. After food and cocaine intake reached baseline levels, the effect of NPSR-QA1 was tested on cocaine and food self-administration. The NPSR-QA1 was injected intraperitoneally and its effect on discriminative cue-induced reinstatement was evaluated, while [D-Cys(tBut)5]NPS was injected intracranially, intra-lateral hypothalamus, intra-perifornical area of the hypothalamus, and intra-central amygdala. The effect of the NPSR-QA1 on extinction of cocaine seeking was also assessed.


Intraperitoneal administration of NPSR-QA1 (15–30 mg/kg) did not affect cocaine self-administration. Conversely, NPSR-QA1 (15–30 mg/kg) decreased discriminative cue-induced cocaine relapse. At the lowest dose, this effect was specific, while at the highest dose, NPSR-QA1 also reduced food self-administration. The efficacy of NPSR antagonism on cocaine seeking was confirmed with [D-Cys(tBu)5]NPS (10–30 nmol/rat) as it markedly inhibited relapse behavior following site-specific injection into the lateral hypothalamus and the perifornical area of the hypothalamus but not into the central amygdala.


The identification of the NPS/NPSR system as an important new element involved in the physiopathology of cocaine addiction and the discovery of the anti-addictive properties of NPSR antagonists opens the possibility of exploring a new mechanism for cocaine addiction treatment.


Addiction Drug abuse Neuropeptide S Self-administration Relapse Psychostimulants 



This study was supported by the Compagnia San Paolo Foundation grant (to R.C.). The authors thank Alfredo Fiorelli, Dr. Massimo Nabissi, and Dr. Daniele Tomassoni for expert technical assistance.


  1. Camarda V, Rizzi A, Ruzza C, Zucchini S, Marzola G, Marzola E, Guerrini R, Salvadori S, Reinscheid RK, Regoli D, Calo G (2009) In vitro and in vivo pharmacological characterization of the neuropeptide S receptor antagonist [D-Cys(tBu)5]neuropeptide S. J Pharmacol Exp Ther 328:549–555PubMedCrossRefGoogle Scholar
  2. Cannella N, Economidou D, Kallupi M, Stopponi S, Heilig M, Massi M, Ciccocioppo R (2009) Persistent increase of alcohol-seeking evoked by neuropeptide S: an effect mediated by the hypothalamic hypocretin system. Neuropsychopharmacology 34:2125–2134PubMedCrossRefGoogle Scholar
  3. Ciccocioppo R, Sanna PP, Weiss F (2001) Cocaine-predictive stimulus induces drug-seeking behavior and neural activation in limbic brain regions after multiple months of abstinence: reversal by D(1) antagonists. Proc Natl Acad Sci U S A 98:1976–1981PubMedCrossRefGoogle Scholar
  4. Cifani C, Micioni Di Bonaventura MV, Cannella N, Fedeli A, Guerrini R, Calo G, Ciccocioppo R, Ubaldi M (2011) Effect of neuropeptide S receptor antagonists and partial agonists on palatable food consumption in the rat. Peptides 32:44–50PubMedCrossRefGoogle Scholar
  5. Clark SD, Duangdao DM, Schulz S, Zhang L, Liu X, Xu YL, Reinscheid RK (2011) Anatomical characterization of the neuropeptide S system in the mouse brain by in situ hybridization and immunohistochemistry. J Comp Neurol 519:1867–1893PubMedCrossRefGoogle Scholar
  6. Fedeli A, Braconi S, Economidou D, Cannella N, Kallupi M, Guerrini R, Calo G, Cifani C, Massi M, Ciccocioppo R (2009) The paraventricular nucleus of the hypothalamus is a neuroanatomical substrate for the inhibition of palatable food intake by neuropeptide S. Eur J Neurosci 30:1594–1602PubMedCrossRefGoogle Scholar
  7. Guerrini R, Camarda V, Trapella C, Calo G, Rizzi A, Ruzza C, Fiorini S, Marzola E, Reinscheid RK, Regoli D, Salvadori S (2009) Further studies at neuropeptide s position 5: discovery of novel neuropeptide S receptor antagonists. J Med Chem 52:4068–4071PubMedCrossRefGoogle Scholar
  8. Jungling K, Seidenbecher T, Sosulina L, Lesting J, Sangha S, Clark SD, Okamura N, Duangdao DM, Xu Y-L, Reinscheid RK, Pape H-C (2008) Neuropeptide S-mediated control of fear expression and extinction: role of intercalated GABAergic neurons in the amygdala. Neuron 59:298–310PubMedCrossRefGoogle Scholar
  9. Kallupi M, Cannella N, Economidou D, Ubaldi M, Ruggeri B, Weiss F, Massi M, Marugan J, Heilig M, Bonnavion P, de Lecea L, Ciccocioppo R (2010) Neuropeptide S facilitates cue-induced relapse to cocaine seeking through activation of the hypothalamic hypocretin system. Proc Natl Acad Sci U S A 107:19567–19572PubMedCrossRefGoogle Scholar
  10. Leonard SK, Ring RH (2011) Immunohistochemical localization of the neuropeptide S receptor in the rat central nervous system. Neuroscience 172:153–163PubMedCrossRefGoogle Scholar
  11. Leonard SK, Dwyer JM, Sukoff Rizzo SJ, Platt B, Logue SF, Neal SJ, Malberg JE, Beyer CE, Schechter LE, Rosenzweig-Lipson S, Ring RH (2008) Pharmacology of neuropeptide S in mice: therapeutic relevance to anxiety disorders. Psychopharmacology (Berl) 197:601–611CrossRefGoogle Scholar
  12. Liu X, Zeng J, Zhou A, Theodorsson E, Fahrenkrug J, Reinscheid RK (2011) Molecular fingerprint of neuropeptide S-producing neurons in the mouse brain. J Comp Neurol 519:1847–1866PubMedCrossRefGoogle Scholar
  13. Meis S, Bergado-Acosta JR, Yanagawa Y, Obata K, Stork O, Munsch T (2008) Identification of a neuropeptide S responsive circuitry shaping amygdala activity via the endopiriform nucleus. PLoS One 3(7):e2695PubMedCrossRefGoogle Scholar
  14. Melamed JY, Zartman AE, Kett NR, Gotter AL, Uebele VN, Reiss DR, Condra CL, Fandozzi C, Lubbers LS, Rowe BA, McGaughey GB, Henault M, Stocco R, Renger JJ, Hartman GD, Bilodeau MT, Trotter BW (2010) Synthesis and evaluation of a new series of neuropeptide S receptor antagonists. Bioorg Med Chem Lett 20:4700–4703PubMedCrossRefGoogle Scholar
  15. O'Brien CP, Childress AR, Ehrman R, Robbins SJ (1998) Conditioning factors in drug abuse: can they explain compulsion? J Psychopharmacol 12:15–22PubMedCrossRefGoogle Scholar
  16. Okamura N, Habay SA, Zeng J, Chamberlin AR, Reinscheid RK (2008) Synthesis and pharmacological in vitro and in vivo profile of 3-oxo-1,1-diphenyl-tetrahydro-oxazolo[3,4-a]pyrazine-7-carboxylic acid 4-fluoro-benzylamide (SHA 68), a selective antagonist of the neuropeptide S receptor. J Pharmacol Exp Ther 325:893–901PubMedCrossRefGoogle Scholar
  17. Paneda C, Huitron-Resendiz S, Frago LM, Chowen JA, Picetti R, de Lecea L, Roberts AJ (2009) Neuropeptide S reinstates cocaine-seeking behavior and increases locomotor activity through corticotropin-releasing factor receptor 1 in mice. J Neurosci 29:4155–4161PubMedCrossRefGoogle Scholar
  18. Paxinos G, Watson C (1998) The rat brain in stereotaxic coordinates, 4th edn. Academic, San DiegoGoogle Scholar
  19. Peng Y-L, Han R-W, Chang M, Zhang L, Zhang R-S, Li W, Han Y-F, Wang R (2010) Central Neuropeptide S inhibits food intake in mice through activation of neuropeptide S receptor. Peptides 31:2259–2263PubMedCrossRefGoogle Scholar
  20. 'Rizzi A, Vergura R, Marzola G, Ruzza C, Guerrini R, Salvadori S, Regoli D, Calo G (2008) Neuropeptide S is a stimulatory anxiolytic agent: a behavioural study in mice. Br J Pharmacol 154:471–479PubMedCrossRefGoogle Scholar
  21. Roth AL, Marzola E, Rizzi A, Arduin M, Trapella C, Corti C, Vergura R, Martinelli P, Salvadori S, Regoli D, Corsi M, Cavanni P, Calo G, Guerrini R (2006) Structure-activity studies on neuropeptide S: identification of the amino acid residues crucial for receptor activation. J Biol Chem 281:20809–20816PubMedCrossRefGoogle Scholar
  22. Ruzza C, Rizzi A, Trapella C, Pela M, Camarda V, Ruggieri V, Filaferro M, Cifani C, Reinscheid RK, Vitale G, Ciccocioppo R, Salvadori S, Guerrini R, Calo G (2010) Further studies on the pharmacological profile of the neuropeptide S receptor antagonist SHA 68. Peptides 31:915–925PubMedCrossRefGoogle Scholar
  23. Smith KL, Patterson M, Dhillo WS, Patel SR, Semjonous NM, Gardiner JV, Ghatei MA, Bloom SR (2006) Neuropeptide S stimulates the hypothalamo-pituitary-adrenal axis and inhibits food intake. Endocrinology 147:3510–3518PubMedCrossRefGoogle Scholar
  24. Somaini L, Donnini C, Raggi MA, Amore M, Ciccocioppo R, Saracino MA, Kalluppi M, Malagoli M, Gerra ML, Gerra G (2011) Promising medications for cocaine dependence treatment. Recent Patents CNS Drug Discov 6:146–160CrossRefGoogle Scholar
  25. Tancredi T, Guerrini R, Marzola E, Trapella C, Calo G, Regoli D, Reinscheid RK, Camarda V, Salvadori S, Temussi PA (2007) Conformation–activity relationship of neuropeptide S and some structural mutants: helicity affects their interaction with the receptor. J Med Chem 50:4501–4508PubMedCrossRefGoogle Scholar
  26. Trotter BW, Nanda KK, Manley PJ, Uebele VN, Condra CL, Gotter AL, Menzel K, Henault M, Stocco R, Renger JJ, Hartman GD, Bilodeau MT (2010) Tricyclic imidazole antagonists of the neuropeptide S receptor. Bioorg Med Chem Lett 20:4704–4708PubMedCrossRefGoogle Scholar
  27. Vitale G, Filaferro M, Ruggieri V, Pennella S, Frigeri C, Rizzi A, Guerrini R, Calo G (2008) Anxiolytic-like effect of neuropeptide S in the rat defensive burying. Peptides 29:2286–2291PubMedCrossRefGoogle Scholar
  28. Xu Y-L, Reinscheid RK, Huitron-Resendiz S, Clark SD, Wang Z, Lin SH, Brucher FA, Zeng J, Ly NK, Henriksen SJ, de Lecea L, Civelli O (2004) Neuropeptide S: a neuropeptide promoting arousal and anxiolytic-like effects. Neuron 43:487–497PubMedCrossRefGoogle Scholar
  29. Xu Y-L, Gall CM, Jackson VR, Civelli O, Reinscheid RK (2007) Distribution of neuropeptide S receptor mRNA and neurochemical characteristics of neuropeptide S-expressing neurons in the rat brain. J Comp Neurol 500:84–8102PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2012

Authors and Affiliations

  • Marsida Kallupi
    • 1
  • Giordano de Guglielmo
    • 1
  • Nazzareno Cannella
    • 1
  • Hong Wu Li
    • 1
  • Girolamo Caló
    • 2
  • Remo Guerrini
    • 3
  • Massimo Ubaldi
    • 1
  • John J. Renger
    • 4
  • Victor N. Uebele
    • 4
  • Roberto Ciccocioppo
    • 1
    Email author
  1. 1.School of Pharmacy (Pharmacology Unit)University of CamerinoCamerinoItaly
  2. 2.Department of Experimental and Clinical Medicine, Section of Pharmacology and National Institute of NeuroscienceUniversity of FerraraFerraraItaly
  3. 3.Department of Pharmaceutical Sciences and LTTA (Laboratorio per le Tecnologie delle Terapie Avanzate)University of FerraraFerraraItaly
  4. 4.Department of NeuroscienceMerck Research LaboratoriesWest PointUSA

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