One‐Step Vital Staining of Presynaptic Terminals and Post‐Synaptic Receptors at Neuromuscular Junctions in Mouse Skeletal Muscle
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Abstract
This protocol describes vital staining of neuromuscular junctions in the mouse triangularis sterni muscle in one incubation step, combining presynaptic, motor nerve terminal staining with the styryl dye FM1?43, which labels recycling synaptic vesicles, and TRITC???bungarotoxin, which labels acetylcholine receptors in the motor endplate membrane. Curr. Protoc. Mouse Biol. 1:489?196 © 2011 by John Wiley & Sons, Inc.
Keywords: neuromuscular junction; fluorescence microscopy; vital staining; styryl dyes; ??bungarotoxin; acetylcholine receptor; synaptic vesicle
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Figure 1. (A ) Low‐power micrograph taken in a dissecting microscope with both transmitted and incident illumination of triangularis sterni muscle preparation. Note the intact muscle fibers oriented vertically and the intramuscular nerve (arrow) running across it. (B ) Examples of intramuscular axons and neuromuscular junctions visualized in a conventional fluorescence microscope. Left panels, upper TRITC‐α‐bungarotoxin staining; lower, FM1‐43; Middle panel, merged channels showing overlay and alignment of motor nerve terminal and motor endplate. (C ) Upper panel, FM1‐43, middle, TRITC‐α‐bungarotoxin; lower, merged channels. Yellow fluorescent region indicates the alignment of presynaptic and post‐synaptic components of the NMJ. View Image
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Literature Cited
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