Figure 1

Double fluorescence labeling for N-CAM (red) and AChR (green) in longitudinal sections of soleus muscles in 14-day embryonic rats, 14-day postnatal rats and adult ratsRed and green stainnings obtained simultaneously were superimposed to show apposition, which appears yellow between N-CAM and postsynaptic AChR plaques.A-C: Intense immunoreactivity for N-CAM is found throughout the axons and axon terminals in embryonic rats. The cholinesterase staining in the endplates with α-BTX shows being distributed in a characteristic curved band with smooth edges.D-F: Immunoreactivity for N-CAM is reduced distinctly from axon to terminal in 14-day postnatal rats, and the area of contact cholinesterase is decreased too.G-I: Immunoreactivity for N-CAM disappeared in the neuromuscular junction except AChR site where the N-CAM staining shows very faint. The ring-shaped AChR plaques in normal adult rats comprise multiple AChR-rich branches where AChR are uniformly distributed. Scale bar: 20 μm."

Figure 2

Confocal laser scanning photomicrographs of double stainning for N-CAM and AchR in the untreated groups and the Yiqi Huayu Recipe-treated groupsA: Untreated group on the 10th day after L5 nerve root compression. The intensity of N-CAM staining is very weak at neuromuscular junction, and the AChR plaques display apparent granular or loss of uniformity of edge thickening (arrowheads).B: Yiqi Huayu Recipe-treated group on the 10th day after L5 nerve root compression. The intensity of N-CAM staining in this picture is stronger than that of the untreated group. The sprouts from endplate (arrows) and a few contact sites (arrowheads) of N-CAM with AChR can be seen.C: Untreated group on the 20th day after L5 nerve root compression. There are some AChR plaques of morphological abnormality (arrows), and no contact sites of N-CAM with AChR although the intensity and scope of N-CAM staining is increased.D: Yiqi Huayu Recipe-treated group on the 20th day after L5 nerve root compression. All the AChR sites are reoccupied by N-CAM and the morphology of AChR are restored. E: Untreated group on the 30th day after lumbar nerve root compression. AChR plaques display irregularity in shape, with a few round or spindle shaped cupulae. Meanwhile, some N-CAM stainings are still shown to be around the AChR. F: Yiqi Huayu Recipe-treated group on the 30th day after L5 nerve root compression. The shape of AChR plaques is close to the normal and the staining of N-CAM has nearly disappeared. Scale bar: 20 μm."

Figure 3

Overlap areas of N-CAM and AChR sites in neuromuscular junctions of rats in different groupThe intensity of N-CAM, which is very strong at embryonic stage of rats, is gradually decreased during the developing stage and nearly disappears until adult stage. The N-CAM reappears after nerve root compression and is gradually decreased according to the regeneration condition in neuromuscular junction. The effect of Yiqi Huayu Recipe on degeneration and regeneration is better than that of the untreated group (*P＜0.05)."