Ultrastructural localization of VIP-immunoreactivity in canine distal oesophagus

The localization of vasoactive intestinal polypeptide (VIP) immunoreactivity in canine distal oesophagus was studied using different fixation and embedding procedures and labelling with protein A-gold. The retention of immunoreactivity and the preservation of ultrastructure was best after fixation with a mixture of 0.1% glutaraldehyde and 4% paraformaldehyde, and embedding in 'LR White' resin using the 'cold-cured' method. VIP immunoreactivity was localized exclusively over large granular vesicles in the myenteric plexus and in nerves of the circular muscle. Most varicose profiles in circular muscle were labelled, but some large granular vesicles in the same profiles which contained labelled vesicles as well as some varicose profiles with large granular vesicles were unlabelled. From these data it was uncertain whether unlabelled large granular vesicles contained VIP or other neuropeptides. A striking finding was the dense and close innervation of the interstitial cells of Cajal with nerve containing VIP-labelled large granular vesicles. This finding is consistent with earlier suggestions that VIP-immunoreactive nerves may innervate these cells which are in gap junction contact with smooth muscle and that this arrangement may be involved in the transmission of non-adrenergic, non-cholinergic nerve activity in distal oesophagus.