Research Paper Volume 14, Issue 2 pp 728—746

Figure 1. Schematic diagram of the experimental paradigm for sepsis induction, daily chronic stress (DCS), and tissue collection. Young adult (~4 months) and old (~20 months) male and female mice were purchase from Jackson Laboratory (JAX Bar Harbor, ME). Prior to initiation of the experiment, mice were acclimated to a 12-hour light-dark cycle for a minimum of 14 days. Sepsis was induced by employing cecal ligation and puncture (CLP) under isoflurane anesthesia. DCS was conducted by placing mice in weighted plexiglass animal restraint holders (Kent Scientific; Torrington, CT) for 2 hours daily commencing the day after CLP. Mice were euthanized for tissue collection either 24 or 96 hours post CLP+DCS. The hippocampus was dissected, flash frozen, and stored at −80, for miR isolation and sequencing.