This article details a surgical procedure to induce hypoxia and ischemia in a mouse model, simulating conditions that lead to thrombotic stroke. The methodology involves surgical exposure of the carotid artery and subsequent manipulation to restrict blood flow.
Begin with an anesthetized and secured mouse.
Remove hair from the mouse's neck and disinfect the surgical site.
Make an incision on the neck and retract the underlying muscle and trachea to expose the right common carotid artery.
The carotid artery supplies the oxygenated blood to the brain.
Tie two knots around the artery to restrict the upward blood flow.
Suture the incision and transfer the mouse to a low-oxygen chamber. Insert a rectal probe to monitor body temperature, then adjust the heat lamp to maintain the appropriate temperature.
The low oxygen in the chamber reduces the mouse's blood oxygen levels, creating a hypoxia condition.
With the carotid artery sutured, the blood and oxygen supply to the brain are further reduced. This leads to ischemia, which causes neuronal damage.
Gradually, blood components adhere to the vessel wall, forming a clot or thrombus.
This obstructs the blood flow, increasing neuronal damage and inducing a thrombotic stroke.
Place a 10 to 13-week-old, 22 to 30-gram male C57 black 6 mouse onto the surgical bed. After confirming anesthesia by toe pinch, apply ointment to the animal's eyes. Stretch the four limbs along the neck roll and secure them with medical tape.
Then, using a depilatory cream, remove the hair on the animal's neck and clean the surgical site. Now move the surgical bed under a dissecting microscope and make a 0.5-centimeter right cervical incision in the skin along the midline. Using a pair of fine forceps, pull apart the trachea and muscle to expose the right common carotid artery and place two pre-cut 5-0 silk sutures underneath the artery. Be careful to separate the carotid artery from the vagus nerve.
Live knot to pre-cut releasable 5-0 silk sutures on the RCCA. Then use 4-0 nylon monofilament sutures to close the skin, and quickly transfer the mouse to the hypoxia system.
Place the face mask over the animal's nose and mouth and a rectal probe, so that a 37.5 degrees Celsius body temperature is maintained throughout the hypoxia delivery. Adjust the heat lamps.
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