Tactile cues are important to environmental novelty during repeated open field tests

The open field test (OFT) is a commonly used protocol to measure anxiety-like behaviors in rodents. Exploration in the central area of the open field and rearing frequency are often readouts of anxiety measurement. However, concerns about carry-over effects associated with repeated assessments limit its application, with the underlying mechanisms of this phenomenon still to be fully described. Here, we showed that repeated OFTs in the same mice led to reductions in the percentage of time spent in the central area and frequency of rearing. This effect reduced with an increase in the intervals between test. The decay caused by repeated OFTs was due to habituation, rather than frequent handling of the experimenter, since novel environments could prevent decay from repeated OFTs. Our results also indicated that tactile cues of the environment played important roles in the habituation of repeated OFTs. Furthermore, the decay of central area activity and rearing behavior during repeated OFTs would be blocked if the hippocampal CA1 was lesioned, suggesting that CA1 is a crucial region for habituation of the OFT in mice. Taken together, our study uncovers the important roles of tactile cues and hippocampal CA1 during repeated OFTs in mice.