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Murray, L., Gillingwater, T. H., Kothary, R. Dissection of the Transversus Abdominis Muscle for Whole-mount Neuromuscular Junction Analysis. J. Vis. Exp. (83), e51162, doi:10.3791/51162 (2014).

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Abstract

Materials

Name Company Catalog Number Comments
Paraformaldehyde Aqueous Solution (16% ) Electron Micropscopy Sciences 15700
Dumont #5 Forceps Fine Science Tools 11251-10
Angled Sprung Scissors Fine Science Tools 15006-09
Fine Scissors - ToughCut Fine Science Tools 14058-09
SYLGARD 184 Silicone Elastomer Kit Dow Corning dependant on local supplier Use this to make dissection dish for pinning out muscle
Minutien Pins Fine science tools 26002-20
α-Bungarotoxin, Alexa Fluor 488 Conjugate Invitrogen B-13422
Albumin from Bovine Serum Sigma Aldrich A4503
Neurofilament Primary antibody (2H3), Supernatant Developmental Studies Hybridoma Bank
SV2 Primary antibody (SV2), Supernatant Developmental Studies Hybridoma Bank
Goat Anti-Mouse IgG (H+L) Jackson ImmunoResearch 115-166-003
Fluorescence Mounting Medium Dako S3023
Slides (Superfrost Plus; White) Fisher 12-550-15
Coverslips Fisher
Triton X-100 Sigma Aldrich T8787
CD1/C57Bl6 mouse Jackson Labs

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References

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  2. Dupuis, L., Loeffler, J. P. Neuromuscular junction destruction during amyotrophic lateral sclerosis: insights from transgenic models. Curr. Opin. Pharmacol. 9, 341-346 (2009).
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  4. Murray, L. M., et al. Selective vulnerability of motor neurons and dissociation of pre- and post-synaptic pathology at the neuromuscular junction in mouse models of spinal muscular atrophy. Hum. Mol. Genet. 17, 949-962 (2008).
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  15. Brill, M. S., Marinkovic, P., Misgeld, T. Sequential photo-bleaching to delineate single Schwann cells at the neuromuscular junction. J. Vis. Exp. (4460), (2013).
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  17. Bowerman, M., Murray, L. M., Beauvais, A., Pinheiro, B., Kothary, R. A critical smn threshold in mice dictates onset of an intermediate spinal muscular atrophy phenotype associated with a distinct neuromuscular junction pathology. Neuromuscul. Disord. 22, 263-276 (2012).
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