Inducing Fibular Nerve Injury in a Mouse to Study Neuromuscular Junction Regeneration

Place an anesthetized mouse with a shaved hind leg under a surgical microscope.

Incise the skin on the shaved leg, then cut through the connective tissue layers to expose the underlying muscles.

Using retractors, separate the muscles to expose the common fibular nerve.

The nerve contains motor neurons, whose axon terminals connect with muscle fibers to form neuromuscular junctions or NMJs.

Locate the nerve's intersection with the tendon of the gastrocnemius muscle in the knee.

Hold the nerve with forceps and apply pressure to induce injury.

A translucent injury site indicates axonal damage, which leads to NMJ impairment.

Ensure the nerve's outer epineurium layer is intact to guide axonal regeneration.

Remove the retractors and realign the muscles. Suture the incision site and allow the mouse to recover.

Signals from the damaged axons recruit macrophages to clear axonal debris, which supports axonal regeneration and the reestablishment of NMJs.

Place the mouse on the surgical board and confirm a lack of response to toe pinch. Align the limbs within the restraints, with the target hind limb in an anatomically natural position and the knee joint slightly extended without internal or external r