FIG. 5.

Rabbit anti-NF-200 (polyclonal 1:80, Sigma), which reacts with the 200-kDa neurofilament protein, was used to visualize axonal sprouting from the transplanted nerve at the nerve/muscle junctions. In the phase-contrast image of the transplantation group at 12 weeks, axonal sprouts from the proximal terminal axon grew prominently, with an even distribution of axon fibers (A and B). Alexa Fluor^® 488-conjugated alpha-bungarotoxin (1:400, Invitrogen/Molecular Probes) was used to stain the acetylcholine receptor (AchR). (C) A double stained image of normal muscle shows an intact neuromuscular junction. In the denervation group, remnant axon filaments had disappeared almost completely, although staining for the AchR remained even at 12 weeks (D–G). In the transplantation group, the integration of axon and AchR improved with time (H–K).