The Presence of a Shelter in an Open Field Test Has Differential Effects on the Behavior and Stress Response of Two Mouse Species

The open field test (OFT) is frequently used in research to assess anxiety-like behavior and locomotor activity. Its simple design can lead to the misconception that it is a standardized procedure comparable between laboratories. However, some modifications in the setup can cause changes in behavior. Different species might also react differently to the modifications introduced. There is thus need for a better understanding of the impact of modifications and their value for the species in question. Here, we tested two closely related mouse species, Mus musculus and Mus spretus, in an OFT with and without the presence of a shelter. We assessed mouse exploratory behavior through the analysis of multiple behavioral traits, and stress response through the measurement of circulating cortisol levels. Both species had elevated cortisol levels during the OFT in contrast to control animals which were not exposed to the OFT. While the presence of a shelter in the OFT increased the exploratory behavior in both mouse species, M. spretus, but not M. musculus, showed a reduction in cortisol levels. Also, other measured behaviors show a rather proactive coping strategy of the commensal M. musculus in contrast to a reactive strategy of the non-commensal M. spretus. Our study revealed a strong species-specific influence of the OFT design on the resulting behavior and stress levels of mice, illustrating the importance of OFT designs to account for the characteristics of the species under study. The addition of a shelter might be considered to improve experimental results by promoting animal welfare.