*2.2.1 Middle cerebral artery occlusion*

Ischemia and reperfusion were simulated in rats using the middle cerebral artery occlusion (MCAO) technique [46], by placing a thread in the left cerebral artery to reduce blood flow in the left hemisphere, and withdrawing it to re-establish circulation. Briefly, adult female Long-Evans rats (250–300 g) were anesthetized with isoflurane (5% induction and 2% for maintenance). The pterygopalatine artery and branches of the carotid artery were cauterized on the left side, after which a 3 centimeter length of 3.0 Dermalon suture (blunt tip) was introduced in a retrograde direction into the external carotid artery. It was advanced cranially in the internal carotid artery for 23 mm, as measured from the bifurcation of the common carotid artery. This model reduced cerebral blood flow to about 10% of control in the core of the ischemic area. Reperfusion was achieved by withdrawing the thread into the external carotid artery.
