Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Protocol
  • Published:

Object recognition test in mice

Abstract

The object recognition test is now among the most commonly used behavioral tests for mice. A mouse is presented with two similar objects during the first session, and then one of the two objects is replaced by a new object during a second session. The amount of time taken to explore the new object provides an index of recognition memory. As more groups have used the protocol, the variability of the procedures used in the object recognition test has increased steadily. This protocol provides a necessary standardization of the procedure. This protocol reduces inter-individual variability with the use of a selection criterion based on a minimal time of exploration for both objects during each session. In this protocol, we describe the three most commonly used variants, containing long (3 d), short (1 d) or no habituation phases. Thus, with a short intersession interval (e.g., 6 h), this procedure can be performed in 4, 2 or 1 d, respectively, according to the duration of the habituation phase. This protocol should allow for the comparison of results from different studies, while permitting adaption of the protocol to the constraints of the experimenter.

This is a preview of subscription content, access via your institution

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

Prices may be subject to local taxes which are calculated during checkout

Figure 1: A photograph of two typical objects (tower of Lego bricks and Falcon tissue culture flask filled with sand) to use in an object recognition protocol.
Figure 2: Experimental design of the three protocols for object recognition.
Figure 3: Typical results showing time mice spent exploring objects.
Figure 4: Typical results showing time to reach the criterion (20 s of total exploration time of both objects) during the two sessions according to the object recognition procedure tested.

Similar content being viewed by others

References

  1. Berlyne, D.E. Novelty and curiosity as determinants of exploratory behaviour. Brit. J. Psychol. 41, 68–80 (1950).

    Google Scholar 

  2. Aggleton, J.P. One-trial object recognition by rats. Q. J. Exp. Psychol. 37, 279–294 (1985).

    Article  Google Scholar 

  3. Ennaceur, A. & Delacour, J. A new one-trial test for neurobiological studies of memory in rats. 1: Behavioral data. Behav. Brain Res. 31, 47–59 (1988).

    Article  CAS  Google Scholar 

  4. Rose, S.A. et al. Rett syndrome: an eye-tracking study of attention and recognition memory. Dev. Med. Child Neurol. 55, 364–371 (2013).

    Article  Google Scholar 

  5. Anderson, M.J. & McGraw, J.J. Novel object recognition in the classroom: establishment of an online video resource for use by instructors in courses on animal learning, memory and behavior. J. Behav. Neurosci. Res. 9, 37–43 (2011).

    Google Scholar 

  6. Stanley, E.M., Wilson, M.A. & Fadel, J.R. Hippocampal neurotransmitter efflux during one-trial novel object recognition in rats. Neurosci. Lett. 511, 38–42 (2012).

    Article  CAS  Google Scholar 

  7. Okuda, S., Roozendaal, B. & McGaugh, J.L. Glucocorticoid effects on object recognition memory require training-associated emotional arousal. Proc. Natl. Acad. Sci. USA 101, 853–858 (2004).

    Article  CAS  Google Scholar 

  8. Barker, G.R. & Warburton, E.C. When is the hippocampus involved in recognition memory? J. Neurosci. 31, 10721–10731 (2011).

    Article  CAS  Google Scholar 

  9. Bouet, V., Freret, T., Dutar, P., Billard, J.M. & Boulouard, M. Continuous enriched environment improves learning and memory in adult NMRI mice through theta burst-related-LTP independent mechanisms but is not efficient in advanced aged animals. Mech. Ageing Dev. 132, 240–248 (2011).

    Article  Google Scholar 

  10. Dodart, J.C., Mathis, C. & Ungerer, A. Scopolamine-induced deficits in a two-trial object recognition task in mice. Neuroreport 8, 1173–1178 (1997).

    Article  CAS  Google Scholar 

  11. Messier, C. Object recognition in mice: improvement of memory by glucose. Neurobiol. Learn. Mem. 67, 172–175 (1997).

    Article  CAS  Google Scholar 

  12. Zhang, R. et al. Novel object recognition as a facile behavior test for evaluating drug effects in AβPP/PS1 Alzheimer's disease mouse model. J. Alzheimers Dis. 31, 801–812 (2012).

    Article  CAS  Google Scholar 

  13. Heyser, C.J. & Chemero, A. Novel object exploration in mice: not all objects are created equal. Behav. Processes 89, 232–238 (2012).

    Article  Google Scholar 

  14. Besheer, J. & Bevins, R.A. The role of environmental familiarization in novel-object preference. Behav. Processes 50, 19–29 (2000).

    Article  CAS  Google Scholar 

  15. Arque, G. et al. Impaired spatial learning strategies and novel object recognition in mice haploinsufficient for the dual specificity tyrosine-regulated kinase-1A (Dyrk1A). PLoS ONE 3, e2575 (2008).

    Article  Google Scholar 

  16. Frick, K.M. & Gresack, J.E. Sex differences in the behavioral response to spatial and object novelty in adult C57BL/6 mice. Behav. Neurosci. 117, 1283–1291 (2003).

    Article  Google Scholar 

  17. Leger, M. et al. Environmental enrichment enhances episodic-like memory in association with a modified neuronal activation profile in adult mice. PLoS ONE 7, e48043 (2012).

    Article  CAS  Google Scholar 

  18. Bevins, R.A. & Besheer, J. Object recognition in rats and mice: a one-trial non-matching-to-sample learning task to study 'recognition memory'. Nat. Protoc. 1, 1306–1311 (2006).

    Article  Google Scholar 

  19. Freret, T. et al. Synergistic effect of acetylcholinesterase inhibition (donepezil) and 5-HT4 receptor activation (RS67333) on object recognition in mice. Behav. Brain Res. 230, 304–308 (2012).

    Article  CAS  Google Scholar 

  20. Sik, A., van Nieuwehuyzen, P., Prickaerts, J. & Blokland, A. Performance of different mouse strains in an object recognition task. Behav. Brain Res. 147, 49–54 (2003).

    Article  Google Scholar 

  21. Ennaceur, A., Michalikova, S., Bradford, A. & Ahmed, S. Detailed analysis of the behavior of Lister and Wistar rats in anxiety, object recognition and object location tasks. Behav. Brain Res. 159, 247–266 (2005).

    Article  CAS  Google Scholar 

  22. Steckler, T., Weis, C., Sauvage, M., Mederer, A. & Holsboer, F. Disrupted allocentric but preserved egocentric spatial learning in transgenic mice with impaired glucocorticoid receptor function. Behav. Brain Res. 100, 77–89 (1999).

    Article  CAS  Google Scholar 

  23. Sheldon, A.B. Preference for familiar versus novel stimuli as a function of the familiarity of the environment. J. Comp. Physiol. Psychol. 67, 516 (1969).

    Article  Google Scholar 

  24. van Goethem, N.P. et al. Object recognition testing: rodent species, strains, housing conditions, and estrous cycle. Behav. Brain Res. 232, 323–334 (2012).

    Article  Google Scholar 

  25. Dellu, F., Fauchey, V., Le Moal, M. & Simon, H. Extension of a new two-trial memory task in the rat: influence of environmental context on recognition processes. Neurobiol. Learn. Mem. 67, 112–120 (1997).

    Article  CAS  Google Scholar 

  26. Kalueff, A.V., Keisala, T., Minasyan, A., Kuuslahti, M. & Tuohimaa, P. Temporal stability of novelty exploration in mice exposed to different open field tests. Behav. Processes 72, 104–112 (2006).

    Article  Google Scholar 

  27. Ennaceur, A. One-trial object recognition in rats and mice: methodological and theoretical issues. Behav. Brain Res. 215, 244–254 (2010).

    Article  CAS  Google Scholar 

  28. Jones, N. & King, S.M. Influence of circadian phase and test illumination on pre-clinical models of anxiety. Physiol. Behav. 72, 99–106 (2001).

    Article  CAS  Google Scholar 

  29. Walf, A.A. & Frye, C.A. The use of the elevated plus maze as an assay of anxiety-related behavior in rodents. Nat. Protoc. 2, 322–328 (2007).

    Article  CAS  Google Scholar 

  30. Gaskin, S. et al. Object familiarization and novel-object preference in rats. Behav. Processes 83, 61–71 (2010).

    Article  Google Scholar 

  31. Wilkinson, J.L., Herrman, L., Palmatier, M.I. & Bevins, R.A. Rats' novel object interaction as a measure of environmental familiarity. Learn. Motiv. 37, 131–148 (2006).

    Article  Google Scholar 

  32. Benice, T.S. & Raber, J. Object recognition analysis in mice using nose-point digital video tracking. J. Neurosci. Methods 168, 422–430 (2008).

    Article  CAS  Google Scholar 

  33. Lad, H.V. et al. Behavioural battery testing: evaluation and behavioural outcomes in 8 inbred mouse strains. Physiol. Behav. 99, 301–316 (2010).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We wish to especially thank G. Bongers (from Noldus Information Technology) for discussions we had on the pros and cons of having experiments automatically analyzed through software and the relevant arguments he put forward. We are grateful to C. Mason for reading the manuscript. A.Q.'s research participation is funded by the French Ministry of Research.

Author information

Authors and Affiliations

Authors

Contributions

A.Q. and M.L. designed and performed the experiments, analyzed the data and drafted the article. T.F. supervised the study. V.B., B.H., M.B. and P.S.-B. revised the manuscript for important intellectual content. All authors discussed the results and commented on the article at all stages.

Corresponding author

Correspondence to Thomas Freret.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Leger, M., Quiedeville, A., Bouet, V. et al. Object recognition test in mice. Nat Protoc 8, 2531–2537 (2013). https://doi.org/10.1038/nprot.2013.155

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nprot.2013.155

This article is cited by

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing