Protocol to induce memory generalization and stress-like behaviors without memory strengthening using contextual fear conditioning in mice

Over-generalization of negative memory is common in psychiatric disorders, including depression and anxiety. Here, we present a protocol to dissociate memory generalization from memory strengthening and induce lasting stress-related behavioral alterations using contextual fear conditioning in adult mice. We describe the steps for repeated conditioning in identical or distinct contexts, followed by memory testing and behavioral assays of stress-related phenotypes. This protocol enables the investigation of the behavioral and biological mechanisms underlying pathological memory generalization and its consequences. For complete details on the use and execution of this protocol, please refer to Cheng et al. 1