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Characterization of Neuromuscular Junctions in Mice by Combined Confocal and Super-Resolution Microscopy
JoVE Journal
Neuroscience
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JoVE Journal Neuroscience
Characterization of Neuromuscular Junctions in Mice by Combined Confocal and Super-Resolution Microscopy

Characterization of Neuromuscular Junctions in Mice by Combined Confocal and Super-Resolution Microscopy

DOI:
10.3791/63032-v

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11:03 min

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December 08, 2021

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, , , , ,
1Genethon, 91000, Evry, France, 2Université Paris-Saclay, Univ Evry, Inserm, Genethon, Integrare research unit UMR_S951, 91000, Evry, France, 3Ecole Pratique des Hautes Etudes, PSL Research University, 75014, Paris, France

Chapters

  • 00:04Introduction
  • 01:17Dissection of Muscles and Immunostaining
  • 03:21Image Acquisition
  • 03:58Image Analysis to Calculate Postsynaptic NMJ Endplate Volume, MIP Area, and Relative Tortuosity
  • 05:05Image Analysis to Quantify Presynaptic Neurofilament Accumulation and Synaptic Vesicle Glycoprotein 2 Staining
  • 06:04Calculating the Distance Between Acetylcholine Receptor (AChR) Stripes and Their Width
  • 08:22Results: Multi Parameter Postsynaptic NMJ Characterization, STED Imaging, and Morphometric Analysis of Presynaptic Axon Terminal Distribution
  • 10:09Conclusion

Summary

Automatic Translation

Automatic Translation

This protocol describes a method for morphometric analysis of neuromuscular junctions by combined confocal and STED microscopy that is used to quantify pathological changes in mouse models of SMA and ColQ-related CMS.

Tags

Keywords: Neuromuscular JunctionsConfocal MicroscopySuper-resolution MicroscopySTEDMuscle TeasingImmunostainingPresynaptic Axon TerminalsActive ZonesQuantitative Analysis3D StructureAnimal ModelsImage AcquisitionImage Analysis

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