Figure 3

(A) Immunofluorescence staining for the accessory (CNXI), C3, and C4 nerves. The sample has been stained with antibodies against NF200 (green) and ChAT (red). (B) The bar graph illustrates the comparison of total axons (NF200+) and cholinergic axons (ChAT+/NF200+) per fascicle for each nerve. (C) A comparison of the detailed proportion of cholinergic (NF200+/ChAT+) and non-cholinergic axons (NF200+/ChAT-) per nerve fascicle in the three nerves. (D) The visualization of myelination of the axons via toluidine blue staining. Myelinated axons (arrows) represent motor or sensory fibers. Some unmyelinated axons (arrowheads) represent sensory fibers (e.g., afferent c-fiber). A cluster of small unmyelinated axons (dashed line) represents sympathetic fibers. (E) Bar graph comparing the somatic and visceral-sympathetic proportions of the three nerves. NF200, neurofilament 200; ChAT, choline acetyltransferase.