Gene Transfer into Schwann Cells of the Rodent Sciatic Nerve via Electroporation

Begin with an anesthetized neonatal rat placed on a heating pad with an exposed sciatic nerve.

Lift the distal site of the nerve and inject a plasmid DNA solution mixed with a dye. The dye helps to track plasmid delivery.

Place a tweezer-type electrode around the nerve, maintaining a small gap between the electrode arms.

Introduce saline into this gap for current conduction.

Apply an electric pulse surrounding the nerve fibers that creates transient membrane pores in the Schwann cells. Then, invert the electrode positions and apply a second electric pulse. This enhances plasmid DNA entry through the pores.

After electroporation, the cell membrane reseals and traps the plasmid.

Clean the site, seal the skin incision, disinfect it, and allow the rat to recover.

Later, the plasmid DNA translocates to the nucleus and integrates into the Schwann cell DNA, enabling successful gene transfer.

Gently pull the distal side of the nerve using a needle to lift the exposed nerve. Insert the glass micro-pipette into the distal site on the nerve, and inject the DNA solution by applying slight positive pressure until the nerve appears green. Next, place a tweezer-type platinum electrode at about one to two