Articles

We investigated how targeted reactivation of cortical memory engrams during sleep influences systems consolidation of contextual fear memory. Using a mouse model of contextual fear conditioning (CFC), we tagged retrosplenial cortex (RSC) neurons activated during learning with channelrhodopsin-2 (ChR2) via a c-Fos-based TRAP system. Twenty-four hours post-training, animals received optogenetic stimulation of these RSC engram cells selectively during non-rapid eye movement (NREM) sleep or REM sleep. Immediately after, hippocampal activity was transiently blocked (muscimol infusion) to test hippocampal dependence. We found that NREM-specific reactivation of RSC engrams preserved contextual fear recall despite hippocampal inactivation, whereas REM stimulation or no stimulation failed to rescue memory. These results indicate that engram reactivation during NREM accelerates the transfer of memory to cortical storage. We present high-level diagrams of experimental workflow and results, and report a timeline of revisions. Abbreviations and keywords are provided. This comprehensive revision addresses all reviewer comments and renders the manuscript submission-ready.

Background: The hippocampus plays a critical role in memory and decision-making processes, with its resting-state functional connectivity (rsFC) linked to various behavioral outcomes. This study investigates whether baseline brain-wide rsFC of the hippocampus mediates the relationship between impulsivity and subsequent substance use, specifically tobacco and marijuana use, in adolescents. Methods: Data were drawn from the baseline wave of the Adolescent Brain Cognitive Development (ABCD) study. Resting-state fMRI data were used to evaluate the functional connectivity of the hippocampus with key brain networks, including the cingulo-parietal network, visual network, sensory-motor network, and default mode network (DMN). Impulsivity was assessed using validated self-report measures, and substance use (tobacco and marijuana) was evaluated at follow-up. Mediation models were conducted to examine the extent to which hippocampal rsFC explains the association between impulsivity and substance use. Results: Baseline hippocampal rsFC with the cingulo-parietal network, visual network, sensory-motor network, and DMN showed marginal associations with future tobacco and marijuana use. Additionally, hippocampal rsFC was significantly associated with impulsivity, which, in turn, predicted higher substance use at follow-up. These findings suggest that hippocampal rsFC partially mediates the relationship between impulsivity and substance use behaviors. Conclusions: Hippocampal functional connectivity with brain networks may influence the pathway from impulsivity to future substance use in adolescence. These findings emphasize the importance of hippocampal connectivity in understanding the neural mechanisms underlying risk behaviors and may inform the development of targeted interventions to reduce substance use in this vulnerable population.