Spatiotemporal Pavlovian head-fixed reversal learning task for mice

Kohei Yamamoto, Kota Yamada, Saya Yatagai, Yusuke Ujihara, Koji Toda

Research output: Contribution to journalArticlepeer-review

1 Citation (Scopus)


Our world is full of uncertainty. Animals, including humans, need to behave flexibly to adjust to ever-changing environments. Reversal learning tasks have been used to assess behavioral flexibility in many species. However, there are some limitations in the traditional free-moving methodology, including (1) sessions to train the animals, (2) within-session number of trials associated with reversals, (3) factors of physical movement unrelated to the task in the maze or operant box, and (4) incompatibility with techniques, such as two-photon imaging. Therefore, to address these limitations, we established a novel spatiotemporal Pavlovian head-fixed reversal learning task for mice. Six experimentally naive adult C57BL/6J mice were used in this study. First, we trained head-fixed mice on a fixed-time schedule task. Sucrose solution was delivered every 10 s with a single drinking spout placed within the licking distance of the mice. After the mice showed anticipatory licking toward the timing of sucrose solution delivery, we began training the mice on the fixed-time schedule reversal learning task with two licking spouts. In this task, sucrose solution was delivered through one of the two drinking spouts. The rewarding spout was switched every 10 trials. Mice quickly learned to switch anticipatory licking to the rewarding side of the spouts, suggesting that they learned this head-fixed reversal learning task. Using the head-fixed experimental design, behavioral measures can be simplified by eliminating the complex behavioral sequences observed in free-moving animals. This novel head-fixed reversal learning task is a useful assay for studying the neurobiological mechanism of behavioral flexibility that is impaired in various psychopathological conditions.

Original languageEnglish
Article number78
JournalMolecular brain
Issue number1
Publication statusPublished - 2022 Dec


  • Behavioral flexibility
  • Head-fixed
  • Mice
  • Pavlovian conditioning
  • Reversal learning

ASJC Scopus subject areas

  • Molecular Biology
  • Cellular and Molecular Neuroscience


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