SUMMARY Memory traces are believed to be ensembles of cells used to store memories. To visualize memory traces, we created a transgenic line that allows for the comparison between cells activated during encoding and expression of a memory. Mice re-exposed to a fear-inducing context froze more and had a greater percentage of reactivated cells in the dentate gyrus (DG) and CA3 than mice exposed to a novel context. Over time, these differences disappeared, in keeping with the observation that memories become generalized. Optogenetically silencing DG or CA3 cells that were recruited during encoding of a fear-inducing context prevented expression of the corresponding memory. Mice with reduced neurogenesis displayed less contextual memory and less reactivation in CA3 but, surprisingly, normal reactivation in the DG. These studies suggest that distinct memory traces are located in the DG and in CA3 but that the strength of the memory is related to reactivation in CA3.

Recent studies have focused on the identification and manipulation of memory traces in rodent models. The two main mouse models utilized are either a CreERT2/loxP tamoxifen (TAM)- or a tetracycline transactivator (tTA)/tetracycline-response element (TRE) doxycycline (DOX)-inducible system. These systems, however, could be improved to label a more specific population of activated neurons corresponding to behavior. Here, we sought to identify an improved selective estrogen receptor (ER) modulator (SERM) in which we could label an individual memory trace in ArcCreERT2 mice. We found that 4-hydroxytamoxifen (4-OHT) is the most selective SERM in the ArcCreERT2 x ROSA26-CAG-stopflox-channelrhodospin (ChR2)-enhanced yellow fluorescent protein (eYFP) mice. The half-life of 4-OHT is also shorter than TAM, allowing for more specificity of memory trace labeling. Furthermore, 4-OHT allowed for context-specific labeling in the dentate gyrus (DG) and CA3. In summary, we believe 4-OHT improves the specificity of memory trace labeling and will allow for refined memory trace studies in the future.