Skip to main content
SpringerLink
Log in

The four-plates test–retest paradigm to discriminate anxiolytic effects

  • Original Investigation
  • Published:
Psychopharmacology Aims and scope Submit manuscript

Abstract

Rationale

Animal models of anxiety such as the four-plates test (FPT) enable the detection of an anxiolytic effect not only of benzodiazepines (BZDs) but also of other non-BZD anxiolytic compounds such as the antidepressants paroxetine and venlafaxine. Retesting mice in animal models of anxiety markedly alters the behavioural profile of various drugs.

Objectives

The aim of this study was first to investigate the function of GABAA/BZD receptor and passive avoidance acquisition in the FPT “test–retest”. The second aim of this study was to evaluate the capacity of the FPT to discriminate BZDs from other non-BZD anxiolytics in experienced mice.

Methods

The FPT was performed in naive and experienced mice (submitted to the test 24 h previously). The drugs studied were two BZDs, diazepam (1 mg/kg) and alprazolam (0.25 mg/kg); flumazenil, a GABAA receptor antagonist (8 mg/kg); atropine sulphate, a muscarinic cholinergic receptor antagonist (4 mg/kg) known for its amnesic properties; paroxetine, a selective serotonin reuptake inhibitor (4 and 8 mg/kg); venlafaxine, a serotonin and noradrenalin reuptake inhibitor (4 and 16 mg/kg); and DOI, a 5-HT2A agonist (1 mg/kg).

Results

Our results reveal an increase of anxiety (decrease of punished passages) in saline-experienced mice. Diazepam, alprazolam, paroxetine and venlafaxine did not prevent the increase in anxiety during retest, revealing a passive avoidance acquisition. Flumazenil did not modify the anxiogenic-like behaviour of experienced mice. In contrast, atropine seems to oppose the increase of anxiety; however, its effect is weak and disputable. DOI was the only anxiolytic compound able to oppose the decrease of punished passages of experienced mice.

Conclusion

Anxiogenic behaviour on retesting indicates aversive learning. The protocol test–retest is unable to discriminate between the anxiolytic effect of BZDs from that of paroxetine or venlafaxine. However, this modified model may constitute a new tool to investigate other neural pathways implicated in anxiety.

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

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  • Abi-Saab WM, Bubser M, Roth RH, Deutch AY (1999) 5-HT2 receptor regulation of extracellular GABA levels in the prefrontal cortex. Neuropsychopharmacology 20:92–96

    Article  Google Scholar 

  • Andrews N, File SE (1993) Handling history of rats modifies behavioural effects of drugs in the elevated plus-maze test of anxiety. Eur J Pharmacol 235:109–112

    Article  Google Scholar 

  • Aron C, Simon P, Larousse C, Boissier JM (1971) Evaluation of a rapid technique detecting minor tranquilizers. Neuropharmacology 10:459–469

    Article  Google Scholar 

  • Bertoglio LJ, Carobrez AP (2002a) Prior maze experience required to alter midazolam effects in rats submitted to the elevated plus-maze. Pharmacol Biochem Behav 72:449–455

    Article  Google Scholar 

  • Bertoglio LJ, Carobrez AP (2002b) Anxiolytic effects of ethanol and phenobarbital are abolished in test-experienced rats submitted to the elevated plus maze. Pharmacol Biochem Behav 73:963–969

    Article  Google Scholar 

  • Bertoglio LJ, Carobrez AP (2004) Scopolamine given pre-Trial 1 prevents the one-trial tolerance phenomenon in the elevated plus-maze Trial 2. Behav Pharmacol 15:45–54

    Article  Google Scholar 

  • Biggio G, Concas A, Corda MG, Giorgi O, Sanna E, Serra M (1990) GABAergic and dopaminergic transmission in the rat cerebral cortex: effect of stress, anxiolytic and anxiogenic drugs. Pharmacol Ther 48:121–142

    Article  Google Scholar 

  • Boissier JR, Simon P, Aron C (1968) A new method for rapid screening of minor tranquillizers in mice. Eur J Pharmacol 4:145–151

    Article  CAS  PubMed  Google Scholar 

  • Borsini F, Podhorna J, Marazziti D (2002) Do animal models of anxiety predict anxiolytic-like effects of antidepressants? Psychopharmacology (Berl) 163:121–141

    Article  Google Scholar 

  • Bortolozzi A, Amargos-Bosch M, Adell A, Diaz-Mataix L, Serrats J, Pons S, Artigas F (2003) In vivo modulation of 5-hydroxytryptamine release in mouse prefrontal cortex by local 5-HT(2A) receptors: effect of antipsychotic drugs. Eur J Neurosci 18:1235–1246

    Article  Google Scholar 

  • Bourin M, Baker GB (1996) The future of antidepressants. Biomed Pharmacother 50:7–12

    Article  Google Scholar 

  • Bourin M, Hascoët M, Mansouri B, Colombel MC, Bradwejn J (1992) Comparison of behavioral effects after single and repeated administrations of four benzodiazepines in three mice behavioral models. J Psychiatry Neurosci 17:72–77

    Google Scholar 

  • Castellano C, Cabib S, Puglisi-Allegra S, Gasbarri A, Sulli A, Pacitti C, Introini-Collison IB, McGaugh JL (1999) Strain-dependent involvement of D1 and D2 dopamine receptors in muscarinic cholinergic influences on memory storage. Behav Brain Res 98:17–26

    Article  Google Scholar 

  • Cook MN, Crounse M, Flaherty L (2002) Anxiety in the elevated zero-maze is augmented in mice after repeated daily exposure. Behav Genet 32:113–118

    Article  Google Scholar 

  • Cruz-Morales SE, Santos NR, Brandao ML (2002) One-trial tolerance to midazolam is due to enhancement of fear and reduction of anxiolytic-sensitive behaviors in the elevated plus-maze retest in the rat. Pharmacol Biochem Behav 72:973–978

    Article  Google Scholar 

  • Dunham NW, Mya TS (1957) A note on a simple apparatus for detecting neurological deficit in rats and mice. J Am Pharm Assoc 46:208–209

    Google Scholar 

  • File SE (1990) One-trial tolerance to the anxiolytic effects of chlordiazepoxide in the plus-maze. Psychopharmacology (Berl) 100:281–282

    Google Scholar 

  • File SE (1993) The interplay of learning and anxiety in the elevated plus-maze. Behav Brain Res 58:199–202

    Article  Google Scholar 

  • File SE (2001) Factors controlling measures of anxiety and responses to novelty in the mouse. Behav Brain Res 125:151–157

    Article  Google Scholar 

  • File SE, Zangrossi H Jr (1993) “One-trial tolerance” to the anxiolytic actions of benzodiazepines in the elevated plus-maze, or the development of a phobic state? Psychopharmacology (Berl) 110:240–244

    Google Scholar 

  • File SE, Mabbutt PS, Hitchcott PK (1990) Characterisation of the phenomenon of “one-trial tolerance” to the anxiolytic effect of chlordiazepoxide in the elevated plus-maze. Psychopharmacology (Berl) 102:98–101

    Google Scholar 

  • File SE, Andrews N, Wu PY, Zharkovsky A, Zangrossi H Jr (1992) Modification of chlordiazepoxide’s behavioural and neurochemical effects by handling and plus-maze experience. Eur J Pharmacol 218:9–14

    Article  Google Scholar 

  • Flood JF, Landry DW, Jarvik ME (1981) Cholinergic receptor interactions and their effects on long-term memory processing. Brain Res 215:177–185

    Article  Google Scholar 

  • Gaggi R, Dall’Olio R, Roncada P (1997) Effect of the selective 5-HT receptor agonists 8-OHDPAT and DOI on behavior and brain biogenic amines of rats. Gen Pharmacol 28:583–587

    Google Scholar 

  • Gobert A, Millan MJ (1999) Serotonin (5-HT)2A receptor activation enhances dialysate levels of dopamine and noradrenaline, but not 5-HT, in the frontal cortex of freely-moving rats. Neuropharmacology 38:315–317

    Article  CAS  PubMed  Google Scholar 

  • Hascoët M, Bourin M, Couetoux du Tertre A (1997) Influence of prior experience on mice behavior using the four-plates test. Pharmacol Biochem Behav 58:1131–1138

    Article  Google Scholar 

  • Hascoët M, Bourin M, Colombel MC, Fiocco AJ, Baker GB (2000) Anxiolytic-like effects of antidepressants after acute administration in a four-plates test in mice. Pharmacol Biochem Behav 65:339–344

    Article  PubMed  Google Scholar 

  • Holmes A, Rodgers RJ (1998) Responses of Swiss–Webster mice to repeated plus-maze experience: further evidence for a qualitative shift in emotional state? Pharmacol Biochem Behav 60:473–488

    Article  Google Scholar 

  • Holmes A, Rodgers RJ (1999) Influence of spatial and temporal manipulations on the anxiolytic efficacy of chlordiazepoxide in mice previously exposed to the elevated plus-maze. Neurosci Biobehav Rev 23:971–980

    Article  Google Scholar 

  • Holmes A, Rodgers RJ (2003) Prior exposure to the elevated plus-maze sensitizes mice to the acute behavioral effects of fluoxetine and phenelzine. Eur J Pharmacol 459:221–230

    Article  CAS  PubMed  Google Scholar 

  • Holmes A, Iles JP, Mayell SJ, Rodgers RJ (2001) Prior test experience compromises the anxiolytic efficacy of chlordiazepoxide in the mouse light/dark exploration test. Behav Brain Res 122:159–167

    Article  Google Scholar 

  • Ichikawa J, Dai J, Meltzer HY (2001) DOI, a 5-HT2A/2C receptor agonist, attenuates clozapine-induced cortical dopamine release. Brain Res 907:151–155

    Article  Google Scholar 

  • Ichikawa J, Ishii H, Bonaccorso S, Fowler WL, O’Laughlin IA, Meltzer HY (2001) 5-HT(2A) and D(2) receptor blockade increases cortical DA release via 5-HT(1A) receptor activation: a possible mechanism of atypical antipsychotic-induced cortical dopamine release. J Neurochem 76:1521–1531

    Article  CAS  PubMed  Google Scholar 

  • Jacob JJ, Tremblay EC, Colombel MC (1974) Enhancement of nociceptive reactions by naloxone in mice and rats. Psychopharmacologia 37:217–223

    Article  Google Scholar 

  • Lamprea MR, Cardenas FP, Silveira R, Morato S, Walsh TJ (2000) Dissociation of memory and anxiety in a repeated elevated plus maze paradigm: forebrain cholinergic mechanisms. Behav Brain Res 117:97–105

    Article  Google Scholar 

  • Liao JF, Hung WY, Chen CF (2003) Anxiolytic-like effects of baicalein and baicalin in the Vogel conflict test in mice. Eur J Pharmacol 464:141–146

    Article  Google Scholar 

  • Lister RG (1987) The use of a plus-maze to measure anxiety in the mouse. Psychopharmacology (Berl) 92:180–185

    Google Scholar 

  • McGregor IS, Dielenberg RA (1999) Differential anxiolytic efficacy of a benzodiazepine on first versus second exposure to a predatory odor in rats. Psychopharmacology (Berl) 147:174–181

    Article  Google Scholar 

  • Meneses A (2002) Involvement of 5-HT(2A/2B/2C) receptors on memory formation: simple agonism, antagonism, or inverse agonism? Cell Mol Neurobiol 22:675–688

    Article  Google Scholar 

  • Meneses A, Hong E (1999) 5-HT1A receptors modulate the consolidation of learning in normal and cognitively impaired rats. Neurobiol Learn Mem 71:207–218

    Article  Google Scholar 

  • Nic Dhonnchadha BA, Bourin M, Hascoet M (2003a) Anxiolytic-like effects of 5-HT2 ligands on three mouse models of anxiety. Behav Brain Res 140:203–214

    Article  CAS  PubMed  Google Scholar 

  • Nic Dhonnchadha BA, Hascoët M, Jolliet P, Bourin M (2003b) Evidence for a 5-HT2A receptor mode of action in the anxiolytic-like properties of DOI in mice. Behav Brain Res 147:175–184

    Article  Google Scholar 

  • Nic Dhonnchadha BA, Ripoll N, Hascoët M, Bourin M (in press) Implication of the 5-HT2 receptor subtypes in the mechanism of action of antidepressants in the mouse four plates test

  • Nutt DJ, Malizia AL (2001) New insights into the role of the GABA(A)-benzodiazepine receptor in psychiatric disorder. Br J Psychiatry 179:390–396

    Article  Google Scholar 

  • Pan SY, Han YF (2000) Learning deficits induced by 4 belladonna alkaloids are preferentially attenuated by tacrine. Acta Pharmacol Sin 21:124–130

    Google Scholar 

  • Pattij T, Groenink L, Oosting RS, van der Gugten J, Maes RA, Olivier B (2002) GABA(A)-benzodiazepine receptor complex sensitivity in 5-HT(1A) receptor knockout mice on a 129/Sv background. Eur J Pharmacol 447:67–74

    Article  CAS  PubMed  Google Scholar 

  • Pehek EA, McFarlane HG, Maguschak K, Price B, Pluto CP (2001) M100,907, a selective 5-HT(2A) antagonist, attenuates dopamine release in the rat medial prefrontal cortex. Brain Res 888:51–59

    Article  Google Scholar 

  • Pierce PA, Peroutka SJ (1989) Hallucinogenic drug interactions with neurotransmitter receptor binding sites in human cortex. Psychopharmacology 97:118–122

    Article  Google Scholar 

  • Porter RHP, Benwell KR, Lamb H, Malcolm CS, Allen NH, Revell DF et al (1999) Functional characterisation of agonists at recombinant human 5-HT2A, 5-HT2B, and 5-HT2C receptors in CHO-K1 cells. Br J Pharmacol 128:13–20

    CAS  PubMed  Google Scholar 

  • Prado-Alcala RA, Solana-Figueroa R, Galindo LE, Medina AC, Quirarte GL (2003) Blockade of striatal 5-HT2 receptors produces retrograde amnesia in rats. Life Sci 74:481–488

    Article  Google Scholar 

  • Ripoll N, Hascoët M, Bourin M. The four-plate test: anxiolytic paradigm or analgesic one? submitted in Behav Brain Res

  • Rodgers RJ, Shepherd JK (1993) Influence of prior maze experience on behaviour and response to diazepam in the elevated plus-maze and light/dark tests of anxiety in mice. Psychopharmacology (Berl) 113:237–242

    Google Scholar 

  • Rodgers RJ, Lee C, Shepherd JK (1992) Effects of diazepam on behavioural and antinociceptive responses to the elevated plus-maze in male mice depend upon treatment regimen and prior maze experience. Psychopharmacology (Berl) 106:102–110

    Google Scholar 

  • Rodgers RJ, Johnson NJ, Cole JC, Dewar CV, Kidd GR, Kimpson PH (1996) Plus-maze retest profile in mice: importance of initial stages of trail 1 and response to post-trail cholinergic receptor blockade. Pharmacol Biochem Behav 54:41–50

    Article  Google Scholar 

  • Sanchez C, Hyttel J (1999) Comparison of the effects of antidepressants and their metabolites on reuptake of biogenic amines and on receptor binding. Cell Mol Neurobiol 19:467–489

    Article  Google Scholar 

  • Shimada T, Matsumoto K, Osanai M, Matsuda H, Terasawa K, Watanabe H (1995) The modified light/dark transition test in mice: evaluation of classic and putative anxiolytic and anxiogenic drugs. Gen Pharmacol 26:205–210

    Google Scholar 

  • Steenbergen HL, Heinsbroek RP, Van Hest A, Van de Poll NE (1990) Sex-dependent effects of inescapable shock administration on shuttlebox-escape performance and elevated plus-maze behavior. Physiol Behav 48:571–576

    Article  Google Scholar 

  • Titeler M, Lyon RA, Glennon RA (1988) Radioligand binding evidence implicates the brain 5-HT2 receptor as a site of action for LSD and phenylisopropylamine hallucinogens. Psychopharmacology 94:213–216

    CAS  PubMed  Google Scholar 

  • Vaswani M, Linda FK, Ramesh S (2003) Role of selective serotonin reuptake inhibitors in psychiatric disorders: a comprehensive review. Prog Neuro-Psychopharmacol Biol Psychiatry 27:85–102

    Article  Google Scholar 

  • Williams GV, Rao SG, Goldman-Rakic PS (2002) The physiological role of 5-HT2A receptors in working memory. J Neurosci 22:2843–2854

    Google Scholar 

  • Zangrossi H Jr, File SE (1992) Behavioral consequences in animal tests of anxiety and exploration of exposure to cat odor. Brain Res Bull 29:381–388

    Article  Google Scholar 

Download references

Acknowledgements

The authors thank Marie-Claude Colombel, Marie-Noëlle Hervé, Fabienne Massé, Sophie Drinkwater and Florence Clénet for their help.

Author information

Authors and Affiliations

  1. Neurobiologie de l’anxiété et de la dépression, Faculté de Médecine, EA 3256, 1 rue Gaston Veil, BP 53508, 44035, Nantes cedex 01, France

    Nadège Ripoll, Bríd Áine Nic Dhonnchadha, Michel Bourin & Martine Hascoët

  2. Laboratoire de Biomathématiques et Biostatistiques, Faculté de Pharmacie, 1 rue Gaston Veil, 44035, Nantes cedex 01, France

    Véronique Sébille

Authors
  1. Nadège Ripoll
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Bríd Áine Nic Dhonnchadha
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Véronique Sébille
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Michel Bourin
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Martine Hascoët
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Michel Bourin.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ripoll, N., Nic Dhonnchadha, B.Á., Sébille, V. et al. The four-plates test–retest paradigm to discriminate anxiolytic effects. Psychopharmacology 180, 73–83 (2005). https://doi.org/10.1007/s00213-004-2130-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00213-004-2130-1

Keywords

Search

Navigation