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Sequential reversal learning: a new touchscreen schedule for assessing cognitive flexibility in mice

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Abstract

Rationale

The widespread deficits in cognitive flexibility observed across psychiatric disorders call for improved rodent tests to understand the biology of cognitive flexibility and development of better psychotherapeutics. Current reversal learning paradigms have a forced-choice setup that challenges the interpretation of results.

Objectives

We aimed at developing a free-choice reversal learning test, where images are presented sequentially and animals are free to move, to enable investigation of the cognitive sub-processes that occur during reversal.

Methods

Behavior in female C57BL/6JOlaHsd mice was characterized using chronic fluoxetine as a reference compound. Additional tests were included to support the interpretation of results and exclude confounding pharmacological effects. Behaviors in vehicle-treated mice were furthermore analyzed for relatedness to deepen the understanding of parameters measured.

Results

We found that exploitation of the previously rewarded image was independent of exploration and acquisition of the new reward contingency and could be differentially modulated by fluoxetine, supporting recent theories that these processes are not mutually exclusive. Specifically, fluoxetine reduced mistake rate, premature and perseverative responses, and promoted conservative strategies during reversal without affecting hit rate. These effects appeared to be most prominent during the late stage of reversal learning, where accuracy was above chance level. Analysis of behaviors in vehicle-treated mice suggested that exploitation was related to an impulsive-like deficit in response inhibition, while exploration was more related to motivation.

Conclusions

This new schedule was feasible, easy to implement, and can provide a deeper understanding of the cognitive sub-processes during reversal.

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References

  • Abuhamdah RM, Hussain MD, Chazot PL, Ennaceur A (2015) Effects of chronic fluoxetine treatment on anxious behaviour of BALB/c mice in a 3-dimensional maze. Stress 18:677–685

    CAS  PubMed  Google Scholar 

  • Alsiö J, Nilsson SR, Gastambide F, Wang RA, Dam SA, Mar AC, Tricklebank M, Robbins TW (2015) The role of 5-HT2C receptors in touchscreen visual reversal learning in the rat: a cross-site study. Psychopharmacology 232:4017–4031

    PubMed  PubMed Central  Google Scholar 

  • Baarendse PJJ, Vanderschuren LJMJ (2012) Dissociable effects of monoamine reuptake inhibitors on distinct forms of impulsive behavior in rats. Psychopharmacology 219:313–326

    CAS  PubMed  Google Scholar 

  • Balleine BW, O’Doherty JP (2010) Human and rodent homologies in action control: corticostriatal determinants of goal-directed and habitual action. Neuropsychopharmacology 35:48–69

    PubMed  Google Scholar 

  • Bari A, Theobald DE, Caprioli D, Mar AC, Aidoo-Micah A, Dalley JW, Robbins TW (2010) Serotonin modulates sensitivity to reward and negative feedback in a probabilistic reversal learning task in rats. Neuropsychopharmacology 35:1290–1301

    CAS  PubMed  PubMed Central  Google Scholar 

  • Bissonette GB, Powell EM (2012) Reversal learning and attentional set-shifting in mice. Neuropharmacology 62:1168–1174

    CAS  PubMed  Google Scholar 

  • Braver TS, Krug MK, Chiew KS, Kool W, Westbrook JA, Clement NJ, Adcock RA, Barch DM, Botvinick MM, Carver CS, Cools R, Custers R, Dickinson A, Dweck CS, Fishbach A, Gollwitzer PM, Hess TM, Isaacowitz DM, Mather M, Murayama K, Pessoa L, Samanez-Larkin GR, Somerville LH, group M (2014) Mechanisms of motivation-cognition interaction: challenges and opportunities. Cogn Affect Behav Neurosci 14:443–472

    PubMed  PubMed Central  Google Scholar 

  • Brigman JL, Graybeal C, Holmes A (2010a) Predictably irrational: assaying cognitive inflexibility in mouse models of schizophrenia. Front Neurosci 4:13

    PubMed  PubMed Central  Google Scholar 

  • Brigman JL, Mathur P, Harvey-White J, Izquierdo A, Saksida LM, Bussey TJ, Fox S, Deneris E, Murphy DL, Holmes A (2010b) Pharmacological or genetic inactivation of the serotonin transporter improves reversal learning in mice. Cereb Cortex (New York, NY: 1991) 20:1955–1963

  • Brown HD, Amodeo DA, Sweeney JA, Ragozzino ME (2012) The selective serotonin reuptake inhibitor, escitalopram, enhances inhibition of prepotent responding and spatial reversal learning. J Psychopharmacol 26:1443–1455

    CAS  PubMed  PubMed Central  Google Scholar 

  • Burke KA, Takahashi YK, Correll J, Brown PL, Schoenbaum G (2009) Orbitofrontal inactivation impairs reversal of Pavlovian learning by interfering with ‘disinhibition’ of responding for previously unrewarded cues. Eur J Neurosci 30:1941–1946

    PubMed  PubMed Central  Google Scholar 

  • Caballero-Puntiverio M, Lerdrup LS, Grupe M, Larsen CW, Dietz AG, Andreasen JT (2019) Effect of ADHD medication in male C57BL/6 J mice performing the rodent continuous performance test. Psychopharmacology 236:1839–1851

    CAS  PubMed  Google Scholar 

  • Carhart-Harris RL, Nutt DJ (2017) Serotonin and brain function: a tale of two receptors. J Psychopharmacol 31:1091–1120

    CAS  PubMed  PubMed Central  Google Scholar 

  • Clark L, Cools R, Robbins TW (2004) The neuropsychology of ventral prefrontal cortex: decision-making and reversal learning. Brain Cogn 55:41–53

    CAS  PubMed  Google Scholar 

  • Clark RA, Shoaib M, Hewitt KN, Stanford SC, Bate ST (2011) A comparison of InVivoStat with other statistical software packages for analysis of data generated from animal experiments. J Psychopharmacol 26:1136–1142

    PubMed  Google Scholar 

  • Clarke HF, Walker SC, Dalley JW, Robbins TW, Roberts AC (2007) Cognitive inflexibility after prefrontal serotonin depletion is behaviorally and neurochemically specific. Cereb Cortex (New York, NY: 1991) 17:18–27

  • Cohen JD, McClure SM, Yu AJ (2007) Should I stay or should I go? How the human brain manages the trade-off between exploitation and exploration. Philos Trans R Soc Lond Ser B Biol Sci 362:933–942

    Google Scholar 

  • Cools R, Roberts AC, Robbins TW (2008) Serotoninergic regulation of emotional and behavioural control processes. Trends Cogn Sci 12:31–40

    PubMed  Google Scholar 

  • D’Cruz A-M, Ragozzino ME, Mosconi MW, Shrestha S, Cook EH, Sweeney JA (2013) Reduced behavioral flexibility in autism spectrum disorders. Neuropsychology 27:152–160

    PubMed  PubMed Central  Google Scholar 

  • Danet M, Lapiz-Bluhm S, Morilak DA (2010) A cognitive deficit induced in rats by chronic intermittent cold stress is reversed by chronic antidepressant treatment. Int J Neuropsychopharmacol 13:997–1009

    CAS  PubMed  Google Scholar 

  • Eskelund A, Li Y, Budac DP, Müller HK, Gulinello M, Sanchez C, Wegener G (2017) Drugs with antidepressant properties affect tryptophan metabolites differently in rodent models with depression-like behavior. J Neurochem 142:118–131

    CAS  PubMed  Google Scholar 

  • Franks GJ, Lattal KA (1976) Antecedent reinforcement schedule training and operant response reinstatement in rats. Anim Learn Behav 4:374–378

    Google Scholar 

  • Hervig ME, Fiddian L, Piilgaard L, Božič T, Blanco-Pozo M, Knudsen C, Olesen SF, Alsiö J, Robbins TW (2020) Dissociable and paradoxical roles of rat medial and lateral orbitofrontal cortex in visual serial reversal learning. Cereb Cortex (New York, NY: 1991) 30:1016–1029

  • Horner AE, Heath CJ, Hvoslef-Eide M, Kent BA, Kim CH, Nilsson SRO, Alsiö J, Oomen CA, Holmes A, Saksida LM, Bussey TJ (2013) The touchscreen operant platform for testing learning and memory in rats and mice. Nat Protoc 8:1961–1984

    CAS  PubMed  PubMed Central  Google Scholar 

  • Hurst JL, West RS (2010) Taming anxiety in laboratory mice. Nat Methods 7:825–826

    CAS  PubMed  Google Scholar 

  • Itami S, Uno H (2002) Orbitofrontal cortex dysfunction in attention-deficit hyperactivity disorder revealed by reversal and extinction tasks. Neuroreport 13:2453–2457

    PubMed  Google Scholar 

  • Iversen SD, Mishkin M (1970) Perseverative interference in monkeys following selective lesions of the inferior prefrontal convexity. Exp Brain Res 11:376–386

    CAS  PubMed  Google Scholar 

  • Izquierdo A, Jentsch J (2011) Reversal learning as a measure of impulsive and compulsive behavior in addictions. Psychopharmacology 219:607–620

    PubMed  PubMed Central  Google Scholar 

  • Izquierdo A, Brigman JL, Radke AK, Rudebeck PH, Holmes A (2017) The neural basis of reversal learning: an updated perspective. Neuroscience 345:12–26

    CAS  PubMed  Google Scholar 

  • Kim CH, Hvoslef-Eide M, Nilsson SRO, Johnson MR, Herbert BR, Robbins TW, Saksida LM, Bussey TJ, Mar AC (2015) The continuous performance test (rCPT) for mice: a novel operant touchscreen test of attentional function. Psychopharmacology 232:3947–3966

    CAS  PubMed  PubMed Central  Google Scholar 

  • Kraeuter AK, Guest PC, Sarnyai Z (2019) The Y-maze for assessment of spatial working and reference memory in mice. Methods Mol Biol 1916:105–111

    CAS  PubMed  Google Scholar 

  • Krebs JR, Kacelnik A, Taylor P (1978) Test of optimal sampling by foraging great tits. Nature 275:27–31

    Google Scholar 

  • Kwon J-W, Armbrust KL (2006) Laboratory persistence and fate of fluoxetine in aquatic environments. Environ Toxicol Chem 25:2561–2568

    CAS  PubMed  Google Scholar 

  • Lucantonio F, Stalnaker TA, Shaham Y, Niv Y, Schoenbaum G (2012) The impact of orbitofrontal dysfunction on cocaine addiction. Nat Neurosci 15:358–366

    CAS  PubMed  PubMed Central  Google Scholar 

  • Mar AC, Horner AE, Nilsson SRO, Alsiö J, Kent BA, Kim CH, Holmes A, Saksida LM, Bussey TJ (2013) The touchscreen operant platform for assessing executive function in rats and mice. Nat Protoc 8:1985–2005

    CAS  PubMed  PubMed Central  Google Scholar 

  • Marazziti D, Consoli G, Picchetti M, Carlini M, Faravelli L (2010) Cognitive impairment in major depression. Eur J Pharmacol 626:83–86

    CAS  PubMed  Google Scholar 

  • Martin KF, Phillips I, Hearson M, Prow MR, Heal DJ (1992) Characterization of 8-OH-DPAT-induced hypothermia in mice as a 5-HT1A autoreceptor response and its evaluation as a model to selectively identify antidepressants. Br J Pharmacol 107:15–21

    CAS  PubMed  PubMed Central  Google Scholar 

  • Matias S, Lottem E, Dugué GP, Mainen ZF (2017) Activity patterns of serotonin neurons underlying cognitive flexibility. eLife 6:e20552

  • McEnaney KW, Butter CM (1969) Perseveration of responding and nonresponding in monkeys with orbital frontal ablations. J Comp Physiol Psychol 68:558–561

    CAS  PubMed  Google Scholar 

  • McMillan N, Sturdy CB, Spetch ML (2015) When is a choice not a choice? Pigeons fail to inhibit incorrect responses on a go/no-go midsession reversal task. J Exp Psychol Anim Learn Cogn 41:255–265

    PubMed  Google Scholar 

  • Millan MJ, Agid Y, Brüne M, Bullmore ET, Carter CS, Clayton NS, Connor R, Davis S, Deakin B, DeRubeis RJ, Dubois B, Geyer MA, Goodwin GM, Gorwood P, Jay TM, Joëls M, Mansuy IM, Meyer-Lindenberg A, Murphy D, Rolls E, Saletu B, Spedding M, Sweeney J, Whittington M, Young LJ (2012) Cognitive dysfunction in psychiatric disorders: characteristics, causes and the quest for improved therapy. Nat Rev Drug Discov 11:141–168

    CAS  PubMed  Google Scholar 

  • Mosberger AC, de Clauser L, Kasper H, Schwab ME (2016) Motivational state, reward value, and Pavlovian cues differentially affect skilled forelimb grasping in rats. Learn Mem 23:289–302

    PubMed  PubMed Central  Google Scholar 

  • Nonkes LJP, Maes JHR, Homberg JR (2013) Improved cognitive flexibility in serotonin transporter knockout rats is unchanged following chronic cocaine self-administration. Addict Biol 18:434–440

    CAS  PubMed  Google Scholar 

  • Odland AU, Jessen L, Fitzpatrick CM, Andreasen JT (2019a) 8-OH-DPAT Induces compulsive-like deficit in spontaneous alternation behavior: reversal by MDMA but not citalopram. ACS Chem Neurosci 10:3094–3100

  • Odland AU, Jessen L, Kristensen JL, Fitzpatrick CM and Andreasen JT (2019b) The 5-hydroxytryptamine 2A receptor agonists DOI and 25CN-NBOH decrease marble burying and reverse 8-OH-DPAT-induced deficit in spontaneous alternation. Neuropharmacology:107838. https://doi.org/10.1016/j.neuropharm.2019.107838

  • Pattij T, Schoffelmeer ANM (2015) Serotonin and inhibitory response control: focusing on the role of 5-HT1A receptors. Eur J Pharmacol 753:140–145

    CAS  PubMed  Google Scholar 

  • Pejchal T, Foley MA, Kosofsky BE, Waeber C (2002) Chronic fluoxetine treatment selectively uncouples raphe 5-HT(1A) receptors as measured by [(35)S]-GTP gamma S autoradiography. Br J Pharmacol 135:1115–1122

    CAS  PubMed  PubMed Central  Google Scholar 

  • Pho GN, Goard MJ, Woodson J, Crawford B, Sur M (2018) Task-dependent representations of stimulus and choice in mouse parietal cortex. Nat Commun 9:2596

    PubMed  PubMed Central  Google Scholar 

  • Piantadosi PT, Lieberman AG, Pickens CL, Bergstrom HC, Holmes A (2019) A novel multichoice touchscreen paradigm for assessing cognitive flexibility in mice. Learn Mem 26:24–30

    CAS  PubMed  PubMed Central  Google Scholar 

  • Ragozzino ME, Kim J, Hassert D, Minniti N, Kiang C (2003) The contribution of the rat prelimbic-infralimbic areas to different forms of task switching. Behav Neurosci 117:1054–1065

    PubMed  Google Scholar 

  • Runegaard AH, Sørensen AT, Fitzpatrick CM, Jørgensen SH, Petersen AV, Hansen NW, Weikop P, Andreasen JT, Mikkelsen JD, Perrier J-F, Woldbye D, Rickhag M, Wortwein G, Gether U (2018) Locomotor- and reward-enhancing effects of cocaine are differentially regulated by chemogenetic stimulation of Gi-signaling in dopaminergic neurons. eNeuro 5:ENEURO.0345-0317.2018

  • Schoenbaum G, Setlow B, Nugent SL, Saddoris MP, Gallagher M (2003) Lesions of orbitofrontal cortex and basolateral amygdala complex disrupt acquisition of odor-guided discriminations and reversals. Learn Mem 10:129–140

    PubMed  PubMed Central  Google Scholar 

  • Schreiner DC, Gremel CM (2018) Orbital frontal cortex projections to secondary motor cortex mediate exploitation of learned rules. Sci Rep 8:10979

    PubMed  PubMed Central  Google Scholar 

  • Sharma RK, Singh T, Mishra A, Goel RK (2017) Relative safety of different antidepressants for treatment of depression in chronic epileptic animals associated with depression. J Epilepsy Res 7:25–32

    PubMed  PubMed Central  Google Scholar 

  • Sokolic L, McGregor IS (2007) Benzodiazepines impair the acquisition and reversal of olfactory go/no-go discriminations in rats. Behav Neurosci 121:527–534

    CAS  PubMed  Google Scholar 

  • Thomas A, Burant A, Bui N, Graham D, Yuva-Paylor LA, Paylor R (2009) Marble burying reflects a repetitive and perseverative behavior more than novelty-induced anxiety. Psychopharmacology 204:361–373

    CAS  PubMed  PubMed Central  Google Scholar 

  • Wallace A, Pehrson AL, Sánchez C, Morilak DA (2014) Vortioxetine restores reversal learning impaired by 5-HT depletion or chronic intermittent cold stress in rats. Int J Neuropsychopharmacol 17:1695–1706

    CAS  PubMed  Google Scholar 

  • Wobrock T, Ecker UK, Scherk H, Schneider-Axmann T, Falkai P, Gruber O (2009) Cognitive impairment of executive function as a core symptom of schizophrenia. World J Biol Psychiatry 10:442–451

    PubMed  Google Scholar 

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Acknowledgments

The authors wish to thank Dr. Simon Bate, the creator of the program InVivoStat, for his helpful statistical advice.

Funding

This research was supported by Lundbeckfonden (grant R263-2017-3000) for the PhD project of A.U.O. The funding source had no influence on the design, analysis, or reporting of the study.

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Authors and Affiliations

  1. Department of Drug Design and Pharmacology, University of Copenhagen, Universitetsparken 2, 2100, Copenhagen, Denmark

    Anna U. Odland, Rune Sandahl & Jesper T. Andreasen

Authors
  1. Anna U. Odland
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  2. Rune Sandahl
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  3. Jesper T. Andreasen
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Contributions

A.U.O. and J.T.A. together conceptualized the research, designed model parameters, and directed analyses. A.U.O. did the final data analysis, completed the first draft of the manuscript, and contributed to experimental work. R.S. did most of the experimental work, collected raw data, and contributed to writing the manuscript. J.T.A. provided critical feedback that helped shape the direction of research and contributed to writing the manuscript.

Corresponding author

Correspondence to Jesper T. Andreasen.

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Highlights

• New sequential reversal learning paradigm that separates exploration from exploitation.

• Exploitation of previous reward contingencies was independent of exploration of new reward contingencies during reversal.

• Exploitation was related to an impulsive-like phenotype, and exploration to motivation.

Supplementary materials

Supplementary material I

ARRIVE guidelines checklist with references to sections in this manuscript (PDF 1003 kb)

Supplementary material II

Pearson’s correlation analysis results (PDF 523 kb)

Supplementary material III

Principal component analysis results (PDF 434 kb)

Supplementary material IV

Touchscreen training results (PDF 603 kb)

Supplementary material V

Supplementary correlations of hit rate and spontaneous alternation behavior (PDF 506 kb)

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Odland, A.U., Sandahl, R. & Andreasen, J.T. Sequential reversal learning: a new touchscreen schedule for assessing cognitive flexibility in mice. Psychopharmacology 238, 383–397 (2021). https://doi.org/10.1007/s00213-020-05687-6

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  • DOI: https://doi.org/10.1007/s00213-020-05687-6

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