Transient focal ischemia was induced by occlusion of the middle cerebral artery as previously described [79 (link),80 (link)], with slight modifications [47 (link),48 (link)]. This model of brain ischemia is considered one of the best models to mimic human ischemic stroke [81 (link)], representing an appropriate in vivo procedure to define the protective role of Ngb-NP against stroke.
Briefly, the middle cerebral artery is occluded using a nylon filament suture with a 3–4 mm coating (Doccol Corporation, Redlands, CA, USA), inserted through the right common carotid artery, and advanced until the origin of the middle cerebral artery. A laser-Doppler flow probe (tip diameter 1 mm) attached to a flowmeter (moorVMS-LDF1, Moor Instruments, Axminster, UK) is located over the thinned skull in the MCAO territory (4 mm lateral to bregma) to obtain a continuous measure of relative cerebral brain flow during the surgery. Only animals with a cerebral blood flow reduction over 60% were included in the study. After surgery, anesthesia was discontinued, and rats were returned to their home cages. Animals in the three groups studied were sacrificed 24 h after MCAO procedure.
Briefly, the middle cerebral artery is occluded using a nylon filament suture with a 3–4 mm coating (Doccol Corporation, Redlands, CA, USA), inserted through the right common carotid artery, and advanced until the origin of the middle cerebral artery. A laser-Doppler flow probe (tip diameter 1 mm) attached to a flowmeter (moorVMS-LDF1, Moor Instruments, Axminster, UK) is located over the thinned skull in the MCAO territory (4 mm lateral to bregma) to obtain a continuous measure of relative cerebral brain flow during the surgery. Only animals with a cerebral blood flow reduction over 60% were included in the study. After surgery, anesthesia was discontinued, and rats were returned to their home cages. Animals in the three groups studied were sacrificed 24 h after MCAO procedure.
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