Physical restraint mouse models to assess immune responses under stress with or without habituation.

Physical confinement, or restraint, is a psychological stressor used in rodent studies. A single restraint episode elevates blood corticosterone levels, a hallmark of stress responses. Repeated restraint results in habituation (or desensitization), whereas chronic exposure to unpredictable stressors fails to induce habituation. Here, we provide our protocols and guidelines in using three mouse restraint models, namely prolonged restraint stress, repeated restraint stress, and chronic variable stress, to examine immunological homeostasis/competence, or lack thereof, under stress with or without habituation. For complete information on the generation and use of these protocols, please refer to Rudak et al. (2021).