Subpial Gene Delivery in the Mouse Spinal Cord to Evaluate Gene Expression

Take an anesthetized mouse, shave its back, and disinfect the skin.

Incise the skin, detach the muscles from the vertebrae, and retract them using clamps.

Drill into the vertebral laminae to create cracks, then remove the bone fragments to expose the dura mater, the outer spinal cord membrane.

Using a needle, create a dural opening and puncture the underlying pia mater to enter the subpial space for direct access to the spinal tissue.

Take a delivery needle containing a suspension of recombinant adeno-associated virus carrying a fluorescent protein-expressing gene and inject the suspension.

Withdraw the needle, suture the muscles and skin, and allow the mouse to recover.

The virus binds to receptors on neurons and glial cells and gets endocytosed.

Once inside the nucleus, the virus triggers gene expression to produce fluorescent proteins.

The subpial gene delivery model is now ready to assess target gene expression in the spinal cord.

Begin by properly anesthetizing the animal for surgery. Isoflurane is used here. Ensure the absence of a paw pinch response before proceeding. Then cover the eyes with ophthalmic ointment. Next, shave the back of the animal with clippers, and clean the skin w