Staining of acetylcholine receptors at the adult mouse and embryonic chick neuromuscular junctions using erabutoxin b and the antibody against acetylcholine receptors.

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Abstract
Acetylcholine receptors (AChRs) at the adult mouse and embryonic chick neuromuscular junctions were visualized by using erabutoxin b (Eb), one of the short-chain neurotoxins from Laticauda semifasciata, and antibody against AChRs of the electric organ from Narke japonica. Visualization of AChRs was performed by the following three methods: (1) staining with rhodaminelabeled Eb (TMR-Eb), (2) staining with horseradish peroxidase-labeled Eb (HRP-Eb), and (3) indirect immunofluorescence microscopy using the antibody against purified AChRs. At the light microscopic level, TMR-Eb offered the most simple and rapid procedure for producing clear images. The HRP-Eb method was suitable for viewing the distribution of AChRs at the ultrastructural level. These procedures revealed that adult neuromuscular junctions had a nearly uniform distribution of AChRs at their postsynaptic membrane, whereas embryonic ones possessed aggregations of small discrete regions of high AChR concentration. Similar regions of AChRs were observed on the nonjunctional myotube surface.

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