Coexistence of vasoactive intestinal polypeptide and neuropeptide Y immunoreactivity within axon terminals in the canine and human lower esophageal sphincter: electron microscopy by a double immunogold labeling procedure.

Subcellular localization of vasoactive intestinal polypeptide (VIP) and neuropeptide Y (NPY) immunoreactivity within nerves was investigated in the lower esophageal sphincter of dogs and humans by double immunogold staining of sections prepared for electron microscopy. Coexistence of VIP and NPY immunoreactivity was clearly demonstrated in the large granular vesicles (LGVs) in axon terminals that were closely associated with the smooth muscle cells, as well as in the LGVs within the perikarya of neurons located in the myenteric plexus. Some LGVs appeared immunopositive only for VIP or NPY. This phenomenon might have been partly due to the fact that the double-labeling procedure with immunogold particles of different sizes was performed on both faces of each section. The results obtained in this study suggest that VIP and NPY are synthesized in the same neuron, stored in the same axon terminal, and released together to act on sphincter muscle cells.