A Rapid Workflow For Modeling Sleep Deprivation-Induced Recognition Memory Deficits In Mice Using Novel Object...

Inadequate sleep contributes to cognitive decline and may accelerate neurodegenerative processes. This article presents a rapid and reproducible workflow for modeling sleep deprivation-induced recognition memory deficits in mice while linking behavioral outcomes to hippocampal molecular alterations. The protocol uses a 5-h gentle-handling sleep deprivation paradigm administered immediately after a standardized 10-min novel object recognition (NOR) familiarization session. Recognition memory is evaluated using a 5-min NOR test after a precisely defined 5-h retention interval, followed by immediate hippocampal collection from the same animals for quantification of pro-inflammatory cytokines, including interleukin-1 beta (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-α), and oxidative-stress markers, including superoxide dismutase (SOD), glutathione peroxidase (GPx), and malondialdehyde (MDA). The principal methodological advance is not the individual behavioral or biochemical techniques, which are well established, but their operational standardization as one linked workflow. The protocol requires a structured, quantifiable log of gentle-handling interventions, anchors training and deprivation to a fixed circadian time point, preserves animal-level links between NOR recordings and tissue samples, and prespecifies quality-control and analytical criteria. The procedure is designed to minimize physical restraint and forced exercise, can be completed within a short experimental window, and is proposed as a candidate reference workflow that individual laboratories may use to benchmark their own implementation of gentle-handling sleep deprivation, pending independent replication.