Middle cerebral artery occlusion (MCAO) surgery in mice

The protocol of MCAO surgery for mice is basically conformed to the Guide for the Care and Use of Laboratory Animals (NIH publication no. 85–23, 1996) and was approved by IACUC of TMU. For the MCAO surgery, mice were anesthetized with a mixture containing 75% air and 3% isoflurane maintained in 25% of oxygen. Mice were placed on a 37 °C heating pad throughout and after the surgery for another two hour for maintaining the body temperature at 37 °C ± 0.5 °C. For the procedure of transient MCAO, the right common carotid artery (CCA) was exposed and a 6–0 monofilament nylon suture (20 mm) coated with silicon (3 mm) was inserted from the external to the internal carotid artery until the tip occluded the right middle cerebral artery (MCA) which is approximately 10 mm from the surgical incision site [18]. Then, the filament suture was kept for the occlusion of MCA for thirty- or sixty-minutes and then gently withdrawal to regain the blood reperfusion for the mice.