MCAO

Rodent models of focal cerebral ischemia have been developed to mimic human ischemic stroke, using the procedure of intraluminal suture occlusion of the middle cerebral artery [34]. This MCAO mouse model has been widely used to study stroke-induced pathophysiology such as cell death and changes in synaptic structures [35–37], and to design new prophylactic, neuroprotective, and therapeutic agents [38]. The mice were anesthetized with pentobarbital sodium (7 mg/kg, i.p.) and body temperature was maintained at 38 °C during surgery. A midline incision was made at the neck and the left common carotid artery (CCA), external carotid artery (ECA), and internal carotid artery (ICA) were identified and ligated. For MCAO, a silicone-coated round-tipped MCAO suture (MSMC21B120PK50, RWD Life Science Co.) was gently inserted from the ECA stump to the ICA, up to ~10 mm, stopping at the MCA, following the previously reported method [39]. After 90, 60, or 0 min of occlusion, the MCAO suture and ligation were withdrawn. The neck skin was sewn back after blood reperfusion was confirmed.