This article describes a method for inducing brain death in anesthetized mice through the inflation of a balloon catheter within the cranial cavity. The procedure involves careful surgical techniques to avoid damaging brain tissue while increasing intracranial pressure, leading to neuronal death.
Begin with an anesthetized mouse in the prone position, with its head shaved.
Make an incision in the skin over the head.
Drill a small hole in the skull, stopping just before the brain layer or dura mater to avoid damaging brain tissue.
Use blunt forceps to penetrate the final tissue bridge and remove any sharp edges.
Insert a balloon catheter through this opening, ensuring it is fully within the cranial cavity. This catheter is prefilled with saline and free of air.
Infuse additional saline through the catheter to gradually inflate the balloon, increasing intracranial pressure.
As the pressure rises, it compresses the brain's blood vessels, restricting blood flow and oxygen to neurons, leading to neuronal death.
This causes the mouse's tail to stiffen and breathing to stop, indicating brain death.
To induce brain death in the experimental animal, arrange the mouse to the prone position, and holding the skin with forceps, use surgical scissors to remove the skin from the skull. Drill a 1-millimeter caliber borehole, pair immediately above the left parietal cortex, and use blunt forceps to penetrate the final tissue bridge of the skull.
After removing any sharp edges, insert a balloon catheter prefilled with saline with all of the air evacuated. When the catheter is entirely within the cranial cavity, use a syringe pump to begin inflating the catheter at approximately 0.1 millimeters per minute over a period of 10 to 15 minutes. When brain death occurs, stop the inflation of the balloon catheter, and place a heating blanket over the mouse to avoid hypothermia.
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