4.3. Neonatal Hypoxic–Ischemic Encephalopathy (HIE) Rat Model

We performed HIE at P9 to reflect human brain development equivalent to a full-term infant [20,59,60]. A modified Rice–Vanucci rat model of hypoxic-ischemic encephalopathy was used as previously described [13]. In brief, P9 rat pups were anesthetized with 2% isoflurane in oxygen. Proper anesthetization was determined by loss of the pedal reflex. Pups were divided into sham and those undergoing HI injury. The right common carotid artery was ligated with a 5.0 silk surgical suture and cut between the ligations. Pups were then returned to dams. After one hour of recovery, pups were placed in a hypoxic chamber at 37 °C with 8% oxygen balanced with 92% nitrogen for 60 min. Pups were then returned to their dams to recover after hypoxic exposure. For the sham-treatment, pups were anesthetized with 2% isoflurane and the left carotid artery was exposed without ligation and without hypoxia treatment. Pups were subsequently returned to dams and allowed to recover.