3Rs-based optimization of mice behavioral testing: the habituation/dishabituation olfactory test

Asma Oummadi, Géraldine Meyer-Dilhet, Amandine Béry, Arnaud Aubert, Pascal Barone, Stéphane Mortaud, Gilles J. Guillemin, Arnaud Menuet, Anthony Laugeray*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review


Background: There is clear evidence that most of the paradigms that are used in the field of behavioral neuroscience suffer from a lack of reliability mainly because of oversimplification of both testing procedures and interpretations. In the present study we show how an already existing behavioral test, the olfactory habituation / dishabituation task, can be optimized in such a way that animal number and animal distress could be minimized, number/confidence of behavioral outcomes and number of explored behavioral dimensions could be increased. New method: We used ethologically relevant technical and procedural changes associated with videotracking-based automated quantification of sniffing behavior to validate our new setup. Mainly internal and construct validity were challenged through the implementation of a series of simple experiments. Results: We show that the new version of the test: 1) has very good within and inter laboratory replicability, 2) is sensitive to some environmental / experimental factors while insensitive to others, 3) allows investigating hedonism, both state and trait anxiety, efficacy of anxiolytic molecules, acute stress, mental retardation-related social impairments and learning and memory. 4) We also show that interest for both nonsocial and social odors is stable over time which makes repetitive testing possible. Conclusions: This work paves the way for future studies showing how behavioral tests / procedures may be improved by using ethologically relevant changes, in order to question laboratory animals more adequately. Refining behavioral tests may considerably increase predictivity of preclinical tests and, ultimately, help reinforcing translational research.

Original languageEnglish
Article number108550
Pages (from-to)1-15
Number of pages15
JournalJournal of Neuroscience Methods
Publication statusPublished - 15 Feb 2020


  • 3Rs
  • Behavioral phenotyping
  • Ethology
  • Mice
  • Olfactory system


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