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

Abstract

Source: Dalkin, W., et al. The Fibular Nerve Injury Method: A Reliable Assay to Identify and Test Factors That Repair Neuromuscular Junctions. J. Vis. Exp. (2016)

This video demonstrates a surgical procedure to induce fibular nerve injury in a mouse to study neuromuscular junction (NMJ) regeneration. The procedure involves exposing the fibular nerve, applying controlled pressure to induce injury, and ensuring the epineurium remains intact to support axonal regeneration. Over time, macrophages clear axonal debris, facilitating nerve repair and the reestablishment of NMJs.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Preparing Animals for Surgery

1. Anesthetize mice with a mixture of ketamine (90 mg/kg) and xylazine (10 mg/kg) via subcutaneous inguinal injection with a sterile 1 ml insulin syringe. The carrier solution contains a mixture of 0.9% saline, 17.4 mg/ml ketamine, and 2.6 mg/ml xylazine. Place animals back in cages while waiting for medication to take effect.
   NOTE: If the loading dose does not provide sufficient anesthesia for the duration of the procedure, an additional 25% of the loading dose....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Ketamine	VetOne	501072
Xylazine	Lloyd Inc.	3437
Buprenorphine	Zoopharm	1Z-73000-150910
Nair	Nair
Kim-wipes	Kimtech	34155
Electric Razor	Braintree Scientific	CLP-64800
80% EtOH/H₂O
10% Proviodine
1 ml Insulin Syringe
Spring Scissors	Vannas	91500-09
No. 15 scalpel	Braintree Scientific	SSS 15
#5 Forceps	Dumont	11252-00
6-0 silk suture on reverse cutting needle	Suture Express	752B
Rodent Heating Pad	Braintree Scientific	AP-R-18.5
Alexa 555 conjugated alpha-BTX	Molecular Probes	B35451
Vectashield	Vector Labs	H-1000
Olympus Stereo Zoom Microscope	Olympus	562037192
Zeiss 700 Confocal Microscope	Zeiss
Variable-flow peristaltic perfusion pump	Fisher Scientific	13-876-3