PMID- 27585036
OWN - NLM
STAT- MEDLINE
DCOM- 20170811
LR  - 20181202
IS  - 1940-087X (Electronic)
IS  - 1940-087X (Linking)
IP  - 114
DP  - 2016 Aug 11
TI  - The Fibular Nerve Injury Method: A Reliable Assay to Identify and Test Factors 
      That Repair Neuromuscular Junctions.
LID - 10.3791/54186 [doi]
LID - 54186
AB  - The neuromuscular junction (NMJ) undergoes deleterious structural and functional 
      changes as a result of aging, injury and disease. Thus, it is imperative to 
      understand the cellular and molecular changes involved in maintaining and 
      repairing NMJs. For this purpose, we have developed a method to reliably and 
      consistently examine regenerating NMJs in mice. This nerve injury method involves 
      crushing the common fibular nerve as it passes over the lateral head of the 
      gastrocnemius muscle tendon near the knee. Using 70 day old female mice, we 
      demonstrate that motor axons begin to reinnervate previous postsynaptic targets 
      within 7 days post-crush. They completely reoccupy their previous synaptic areas 
      by 12 days. To determine the reliability of this injury method, we compared 
      reinnervation rates between individual 70 day old female mice. We found that the 
      number of reinnervated postsynaptic sites was similar between mice at 7, 9, and 
      12 days post-crush. To determine if this injury assay can also be used to compare 
      molecular changes in muscles, we examined levels of the gamma-subunit of the 
      muscle nicotinic receptor (gamma-AChR) and the muscle-specific kinase (MuSK). The 
      gamma-AChR subunit and MuSK to are highly upregulated following denervation and 
      return to normal levels following reinnervation of NMJs. We found a close 
      relationship between transcript levels for these genes and innervation status of 
      muscles. We believe that this method will accelerate our understanding of the 
      cellular and molecular changes involved in repairing the NMJ and other synapses.
FAU - Dalkin, William
AU  - Dalkin W
AD  - Carilion Research Institute, Virginia Tech; Carilion School of Medicine, Virginia 
      Tech.
FAU - Taetzsch, Thomas
AU  - Taetzsch T
AD  - Carilion Research Institute, Virginia Tech.
FAU - Valdez, Gregorio
AU  - Valdez G
AD  - Carilion Research Institute, Virginia Tech; Carilion School of Medicine, Virginia 
      Tech; Department of Biological Sciences, Virginia Tech; gvaldez1@vtc.vt.edu.
LA  - eng
PT  - Journal Article
PT  - Video-Audio Media
DEP - 20160811
PL  - United States
TA  - J Vis Exp
JT  - Journal of visualized experiments : JoVE
JID - 101313252
SB  - IM
MH  - Animals
MH  - Axons/physiology
MH  - Female
MH  - Mice
MH  - Nerve Crush
MH  - Nerve Regeneration/*physiology
MH  - Neuromuscular Junction/*physiology
MH  - Peroneal Nerve/*injuries
MH  - Reproducibility of Results
MH  - Synapses/physiology
PMC - PMC5091792
EDAT- 2016/09/02 06:00
MHDA- 2017/08/12 06:00
PMCR- 2018/08/11
CRDT- 2016/09/02 06:00
PHST- 2016/09/02 06:00 [entrez]
PHST- 2016/09/02 06:00 [pubmed]
PHST- 2017/08/12 06:00 [medline]
PHST- 2018/08/11 00:00 [pmc-release]
AID - 54186 [pii]
AID - 10.3791/54186 [doi]
PST - epublish
SO  - J Vis Exp. 2016 Aug 11;(114):54186. doi: 10.3791/54186.