, Volume 182, Issue 4, pp 475–484

A comparison of chlordiazepoxide, bretazenil, L838,417 and zolpidem in a validated mouse Vogel conflict test

Original Investigation



GABAA receptors containing an α2 subunit are proposed to mediate the anxiolytic effect of benzodiazepines (BZ) based on studies in transgenic mice using unconditioned models of anxiety. Conditioned models of anxiety were not assessed and are rarely encountered in phenotyping of genetically modified animals. The novel benzodiazepine site ligand L838,417 is a partial agonist at GABAA receptors containing an α2, α3 or α5 subunit and an antagonist at α1 receptors, giving an anxiolytic profile devoid of sedation. However, this compound has not previously been assessed in mice.


(1) Establish the Vogel conflict test (VCT) in C57BL/6J mice and validate it with a range of pharmacological tools and (2) compare the full and partial GABAA receptor positive modulators chlordiazepoxide (CDP) and bretazenil (BRZ), respectively, with the subtype selective ligands zolpidem (ZOL; α1 selective) and L838,417.


(1) enhanced thirst (water deprivation or isoproterenol administration), analgesia (lamotrigine) or cognitive impairment (MK-801) did not generate false positives in the VCT; (2) CDP and BRZ engendered linear dose-related anti-conflict effects and also increased unpunished drinking; (3) L838,417 engendered a bell-shaped anti-conflict effect and did not increase unpunished drinking; (4) the anti-conflict effect of CDP and L838,417 were antagonised by flumazenil, whereas BRZ's effect was insensitive to this antagonist; and (5) ZOL induced motoric deficits and no anti-conflict effect.


We have established the VCT in C57BL/6J mice and validated this test behaviourally, physiologically and pharmacologically. The novel GABAA receptor ligand L838,417 was anxiolytic in this mouse model, and unlike the non-selective compounds, had no effect on unpunished drinking.


Anxiety Benzodiazepine Bretazenil Chlordiazepoxide GABAA Isoproterenol L838,417 Mouse Vogel conflict test Zolpidem 







Analysis of variance










gamma-aminobutyric acid








not significant


Nmethyl-d-asparic acid




Vogel conflict test




  1. Agmo A, Pruneda R, Guzman M, Gutierrez M (1991) GABAergic drugs and conflict behavior in the rat: lack of similarities with the actions of benzodiazepines. Naunyn-Schmiedeberg's Arch Pharmacol 344:314–322CrossRefGoogle Scholar
  2. Anagnostopoulos AV, Mobraaten LE, Sharp JJ, Davisson MT (2001) Transgenic and knockout databases: behavioral profiles of mouse mutants. Physiol Behav 73:675–689CrossRefPubMedGoogle Scholar
  3. Benke D, Michel C, Mohler H (1997) GABA(A) receptors containing the alpha4-subunit: prevalence, distribution, pharmacology, and subunit architecture in situ. J Neurochem 69:806–814PubMedCrossRefGoogle Scholar
  4. Blackburn-Munro G, Ericksen HK (2004) Antiepileptics in the treatment of neuropathic pain: evidence from animal models. Curr Pharm Des (in press)Google Scholar
  5. Boissier JR, Simon P, Aron C (1968) A new method for rapid screening of minor tranquillizers in mice. Eur J Pharmacol 4:145–151CrossRefPubMedGoogle Scholar
  6. Borsini F, Podhorna J, Marazziti D (2002) Do animal models of anxiety predict anxiolytic-like effects of antidepressants? Psychopharmacology (Berl) 163:121–141CrossRefGoogle Scholar
  7. Brauer HR, Nowicki PW, Catalano G, Catalano MC (2002) Panic attacks associated with citalopram. South Med J 95:1088–1089PubMedGoogle Scholar
  8. Brocco MJ, Koek W, Degryse AD, Colpaert FC (1990) Comparative studies on the anti-punishment effects of chlordiazepoxide, buspirone and ritanserin in the pigeon, Geller–Seifter and Vogel conflict procedures. Behav Pharmacol 1:403–418PubMedCrossRefGoogle Scholar
  9. Burghardt NS, Sullivan GM, McEwen BS, Gorman JM, LeDoux JE (2004) The selective serotonin reuptake inhibitor citalopram increases fear after acute treatment but reduces fear with chronic treatment: a comparison with tianeptine. Biol Psychiatry 55:1171–1178CrossRefPubMedGoogle Scholar
  10. Carli M, Samanin R (1982) Evidence that agents increasing water consumption do not necessarily generate “false positives” in conflict procedures using water as a reinforcer. Pharmacol Biochem Behav 17:1–3CrossRefPubMedGoogle Scholar
  11. Conti LH, Maciver CR, Ferkany JW, Abreu ME (1990) Footshock-induced freezing behavior in rats as a model for assessing anxiolytics. Psychopharmacology (Berl) 102:492–497CrossRefGoogle Scholar
  12. Cooper SJ (1982) Benzodiazepine mechanisms and drinking in the water-deprived rat. Neuropharmacology 21:775–780CrossRefPubMedGoogle Scholar
  13. Cooper SJ (1991) Ingestional responses following benzodiazepine receptor ligands, selective 5-HT1A agonists and selective 5-HT3 receptor antagonists. In: Rodgers RJ, Cooper SJ (eds) 5-HT1A agonists, 5-HT3 antagonists and benzodiazepines. Their comparative pharmacology. Wiley, Chichester, pp 233–265Google Scholar
  14. Crawley JN, Belknap JK, Collins A, Crabbe JC, Frankel W, Henderson N, Hitzemann RJ, Maxson SC, Miner LL, Silva AJ, Wehner JM, Wynshaw-Boris A, Paylor R (1997) Behavioral phenotypes of inbred mouse strains: implications and recommendations for molecular studies. Psychopharmacology (Berl) 132:107–124CrossRefGoogle Scholar
  15. Crestani F, Martin JR, Mohler H, Rudolph U (2000) Mechanism of action of the hypnotic zolpidem in vivo. Br J Pharmacol 131:1251–1254CrossRefPubMedGoogle Scholar
  16. Crestani F, Mohler H, Rudolph U (2001) Anxiolytic action of diazepam: mediated by GABAA receptors containing the α2 subunit. Trends Pharmacol Sci 22:403CrossRefGoogle Scholar
  17. Czech DA, Vander Zanden JM (1991) Drinking behavior in the spiny mouse (Acomys cahirinus) following putative dipsogenic challenges. Pharmacol Biochem Behav 38:913–916CrossRefPubMedGoogle Scholar
  18. Dalvi A, Rodgers RJ (1999) Behavioral effects of diazepam in the murine plus-maze: flumazenil antagonism of enhanced head dipping but not the disinhibition of open-arm avoidance. Pharmacol Biochem Behav 62:727–734CrossRefPubMedGoogle Scholar
  19. Davis M (1979) Diazepam and flurazepam: effects on conditioned fear as measured with the potentiated startle paradigm. Psychopharmacology (Berl) 62:1–7CrossRefGoogle Scholar
  20. Dekeyne A, Brocco M, Adhumeau A, Gobert A, Millan MJ (2000) The selective serotonin (5-HT)1A receptor ligand, S15535, displays anxiolytic-like effects in the social interaction and Vogel models and suppresses dialysate levels of 5-HT in the dorsal hippocampus of freely-moving rats. A comparison with other anxiolytic agents. Psychopharmacology (Berl) 152:55–66CrossRefGoogle Scholar
  21. Depoortere H, Zivkovic B, Lloyd KG, Sanger DJ, Perrault G, Langer SZ, Bartholini G (1986) Zolpidem, a novel nonbenzodiazepine hypnotic. I. Neuropharmacological and behavioral effects. J Pharmacol Exp Ther 237:649–658PubMedGoogle Scholar
  22. De Vry J, Benz U, Schreiber R, Traber J (1993) Shock-induced ultrasonic vocalization in young adult rats: a model for testing putative anti-anxiety drugs. Eur J Pharmacol 249:331–339CrossRefPubMedGoogle Scholar
  23. Di Scala G, Oberling P, Rocha B, Sandner G (1992) Conditioned place preference induced by Ro 16-6028, a benzodiazepine receptor partial agonist. Pharmacol Biochem Behav 41:859–862CrossRefPubMedGoogle Scholar
  24. File SE, Kenny PJ, Ouagazzal AM (1998) Bimodal modulation by nicotine of anxiety in the social interaction test: role of the dorsal hippocampus. Behav Neurosci 112:1423–1429CrossRefPubMedGoogle Scholar
  25. Flores P, Pellon R (2000) Antipunishment effects of diazepam on two levels of suppression of schedule-induced drinking in rats. Pharmacol Biochem Behav 67:207–214CrossRefPubMedGoogle Scholar
  26. Gonzalez LE, File SE (1997) A five minute experience in the elevated plus-maze alters the state of the benzodiazepine receptor in the dorsal raphe nucleus. J Neurosci 17:1505–1511PubMedGoogle Scholar
  27. Griebel G, Perrault G, Tan S, Schoemaker H, Sanger DJ (1999) Comparison of the pharmacological properties of classical and novel BZ-omega receptor ligands. Behav Pharmacol 10:483–495PubMedCrossRefGoogle Scholar
  28. Griebel G, Belzung C, Perrault G, Sanger DJ (2000) Differences in anxiety-related behaviours and in sensitivity to diazepam in inbred and outbred strains of mice. Psychopharmacology (Berl) 148:164–170CrossRefGoogle Scholar
  29. Griebel G, Simiand J, Serradeil-Le Gal C, Wagnon J, Pascal M, Scatton B, Maffrand JP, Soubrie P (2002) Anxiolytic- and antidepressant-like effects of the non-peptide vasopressin V1b receptor antagonist, SSR149415, suggest an innovative approach for the treatment of stress-related disorders. Proc Natl Acad Sci U S A 99:6370–6375CrossRefPubMedGoogle Scholar
  30. Itoh J, Nabeshima T, Kameyama T (1991) Utility of an elevated plus-maze for dissociation of amnesic and behavioral effects of drugs in mice. Eur J Pharmacol 194:71–76CrossRefPubMedGoogle Scholar
  31. Kennett GA, Trail B, Bright F (1998) Anxiolytic-like actions of BW 723C86 in the rat Vogel conflict test are 5-HT2B receptor mediated. Neuropharmacology 37:1603–1610CrossRefPubMedGoogle Scholar
  32. Korsgaard MGP, Hartz BP, Brown WD, Ahring PK, Strøbæk D, Mirza NR (2005) Kv7 channel modulators: novel anxiolytics. J Pharmacol Exp Ther 314:282–292CrossRefPubMedGoogle Scholar
  33. Kuribara H, Haraguchi H, Tadokoro S (1989) Anticonflict effect of caffeine: investigation by punishment and hypertonic NaCl solution procedures in mice. Arukoru Kenkyu to Yakubutsu Izon 24:144–153PubMedGoogle Scholar
  34. Kuribara H, Fujiwara S, Yasuda H, Tadokoro S (1990) The anticonflict effect of MK-801, an NMDA antagonist: investigation by punishment procedure in mice. Jpn J Pharmacol 54:250–252PubMedCrossRefGoogle Scholar
  35. Liao JF, Hung WY, Chen CF (2003) Anxiolytic-like effects of baicalein and baicalin in the Vogel conflict test in mice. Eur J Pharmacol 464:141–146CrossRefPubMedGoogle Scholar
  36. Loiseau F, Le Bihan C, Hamon M, Thiebot MH (2003) Distinct effects of diazepam and NK1 receptor antagonists in two conflict procedures in rats. Behav Pharmacol 14:447–455PubMedGoogle Scholar
  37. Low K, Crestani F, Keist R, Benke D, Brunig I, Benson JA, Fritschy JM, Rulicke T, Bluethmann H, Mohler H, Rudolph U (2000) Molecular and neuronal substrate for the selective attenuation of anxiety. Science 290:131–134CrossRefPubMedGoogle Scholar
  38. Maickel RP, Maloney GJ (1973) Effects of various depressant drugs on deprivation-induced water consumption. Neuropharmacology 12:777–782CrossRefPubMedGoogle Scholar
  39. Maruyama Y, Kuribara H, Kishi E, Weintraub ST, Ito Y (2001) Confirmation of the anxiolytic-like effect of dihydrohonokiol following behavioural and biochemical assessments. J Pharm Pharmacol 53:721–725CrossRefPubMedGoogle Scholar
  40. McKernan RM, Rosahl TW, Reynolds DS, Sur C, Wafford KA, Atack JR, Farrar S, Myers J, Cook G, Ferris P, Garrett L, Bristow L, Marshall G, Macaulay A, Brown N, Howell O, Moore KW, Carling RW, Street LJ, Castro JL, Ragan CI, Dawson GR, Whiting PJ (2000) Sedative but not anxiolytic properties of benzodiazepines are mediated by the GABA(A) receptor alpha1 subtype. Nat Neurosci 3:587–592CrossRefPubMedGoogle Scholar
  41. Millan MJ, Brocco M (2003) The Vogel conflict test: procedural aspects, gamma-aminobutyric acid, glutamate and monoamines. Eur J Pharmacol 463:67–96CrossRefPubMedGoogle Scholar
  42. Mirza NR, Bright JL, Stanhope KJ, Wyatt A, Harrington NR (2005) Lamotrigine has an anxiolytic-like profile in the rat conditioned emotional response test of anxiety: a potential role for sodium channels? Psychopharmacology (Berl) 180:159–168CrossRefGoogle Scholar
  43. Nazar M, Jessa M, Plaznik A (1997) Benzodiazepine-GABAA receptor complex ligands in two models of anxiety. J Neural Transm 104:733–746CrossRefPubMedGoogle Scholar
  44. Patel JB, Malick JB (1980) Effects of isoproterenol and chlordiazepoxide on drinking and conflict behaviors in rats. Pharmacol Biochem Behav 12:819–821CrossRefPubMedGoogle Scholar
  45. Plaznik A, Palejko W, Nazar M, Jessa M (1994) Effects of antagonists at the NMDA receptor complex in two models of anxiety. Eur Neuropsychopharmacol 4:503–512CrossRefPubMedGoogle Scholar
  46. Rodgers RJ (1997) Animal models of ‘anxiety’: where next? Behav Pharmacol 8:477–496PubMedCrossRefGoogle Scholar
  47. Rodgers RJ, Davies B, Shore R (2002) Absence of anxiolytic response to chlordiazepoxide in two common background strains exposed to the elevated plus-maze: importance and implications of behavioural baseline. Genes Brain Behav 1:242–251CrossRefPubMedGoogle Scholar
  48. Rudolph U, Crestani F, Benke D, Brunig I, Benson JA, Fritschy JM, Martin JR, Bluethmann H, Mohler H (1999) Benzodiazepine actions mediated by specific gamma-aminobutyric acid(A) receptor subtypes. Nature 401:796–800CrossRefPubMedGoogle Scholar
  49. Shekhar A, McCann UD, Meaney MJ, Blanchard DC, Davis M, Frey KA, Liberzon I, Overall KL, Shear MK, Tecott LH, Winsky L (2001) Summary of a National Institute of Mental Health workshop: developing animal models of anxiety disorders. Psychopharmacology (Berl) 157:327–339CrossRefGoogle Scholar
  50. Sieghart W (1995) Structure and pharmacology of gamma-aminobutyric acidA receptor subtypes. Pharmacol Rev 47:181–234PubMedGoogle Scholar
  51. Soderpalm A, Blomqvist O, Soderpalm B (1995a) The yohimbine-induced anticonflict effect in the rat, Part I. Involvement of noradrenergic, serotonergic and endozepinergic(?) mechanisms. J Neural Transm Gen Sect 100:175–189CrossRefPubMedGoogle Scholar
  52. Soderpalm AK, Blomqvist O, Engel JA, Soderpalm B (1995b) Characterization of the anticonflict effect of MK-801, a non-competitive NMDA antagonist. Pharmacol Toxicol 76:122–127PubMedGoogle Scholar
  53. Sorbera LA, Leeson PA, Silvestre J, Castaner J (2001) Pagoclone-anxiolytic GABA-A/BZD site partial agonist. Drugs Future 26:651–657CrossRefGoogle Scholar
  54. Soubrie P, de Angelis L, Boissier JR (1976) Effects of antianxiety drugs on the water intake in trained and untrained rats and mice (author's translation). Psychopharmacology (Berl) 50:41–45CrossRefGoogle Scholar
  55. Stanhope KJ, Dourish CT (1996) Effects of 5-HT1A receptor agonists, partial agonists and a silent antagonist on the performance of the conditioned emotional response test in the rat. Psychopharmacology (Berl) 128:293–303CrossRefGoogle Scholar
  56. Stocker SD, Sved AF, Stricker EM (2000) Role of renin-angiotensin system in hypotension-evoked thirst: studies with hydralazine. Am J Physiol Regul Integr Comp Physiol 279:R576–R585PubMedGoogle Scholar
  57. Tam SW, Worcel M, Wyllie M (2001) Yohimbine: a clinical review. Pharmacol Ther 91:215–243CrossRefPubMedGoogle Scholar
  58. Teloken C, Rhoden EL, Sogari P, Dambros M, Souto CA (1998) Therapeutic effects of high dose yohimbine hydrochloride on organic erectile dysfunction. J Urol 159:122–124PubMedCrossRefGoogle Scholar
  59. Treit D (1985) Animal models for the study of anti-anxiety agents: a review. Neurosci Biobehav Rev 9:203–222CrossRefPubMedGoogle Scholar
  60. Trullas R, Skolnick P (1993) Differences in fear motivated behaviors among inbred mouse strains. Psychopharmacology (Berl) 111:323–331CrossRefGoogle Scholar
  61. Umezu T (1995) Assessment of anxiolytics (5)-Vogel-type conflict task in mice. Nihon Shinkei Seishin Yakurigaku Zasshi 15:305–314PubMedGoogle Scholar
  62. Umezu T (1999) Effects of psychoactive drugs in the Vogel conflict test in mice. Jpn J Pharmacol 80:111–118CrossRefPubMedGoogle Scholar
  63. Uyeno ET, Davies MF, Pryor GT, Loew GH (1990) Selective effect on punished versus unpunished responding in a conflict test as the criterion for anxiogenic activity. Life Sci 47:1375–1382CrossRefPubMedGoogle Scholar
  64. van Gaalen MM, Steckler T (2000) Behavioural analysis of four mouse strains in an anxiety test battery. Behav Brain Res 115:95–106CrossRefPubMedGoogle Scholar
  65. Vanover KE, Robledo S, Huber M, Carter RB (1999) Pharmacological evaluation of a modified conflict procedure: punished drinking in non-water-deprived rats. Psychopharmacology (Berl) 145:333–341CrossRefGoogle Scholar
  66. Vogel JR, Beer B, Clody DE (1971) A simple and reliable conflict procedure for testing anti-anxiety agents. Psychopharmacologia 21:1–7CrossRefPubMedGoogle Scholar
  67. Voikar V, Koks S, Vasar E, Rauvala H (2001) Strain and gender differences in the behavior of mouse lines commonly used in transgenic studies. Physiol Behav 72:271–281CrossRefPubMedGoogle Scholar
  68. Wafford KA, Thompson SA, Thomas D, Sikela J, Wilcox AS, Whiting PJ (1996) Functional characterization of human gamma-aminobutyric acidA receptors containing the alpha 4 subunit. Mol Pharmacol 50:670–678PubMedGoogle Scholar
  69. Whitwam JG, Amrein R (1995) Pharmacology of flumazenil. Acta Anaesthesiol Scand Suppl 108:3–14PubMedCrossRefGoogle Scholar
  70. Witkin JM, Acri JB, Gleeson S, Barrett JE (1997) Blockade of behavioral effects of bretazenil by flumazenil and ZK 93,426 in pigeons. Pharmacol Biochem Behav 56:1–7CrossRefPubMedGoogle Scholar
  71. Witkin JM, Morrow D, Li X (2004) A rapid punishment procedure for detection of anxiolytic compounds in mice. Psychopharmacology (Berl) 172:52–57CrossRefGoogle Scholar
  72. Xie ZC, Buckner E, Commissaris RL (1995) Anticonflict effect of MK-801 in rats: time course and chronic treatment studies. Pharmacol Biochem Behav 51:635–640CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  1. 1.Department of in-vivo PharmacologyNeuroSearch A/SBallerupDenmark

Personalised recommendations