Ultrastructural co-localization of neuropeptide Y and vasoactive intestinal polypeptide in neurosecretory vesicles of submucous neurons in the rat jejunum

The localization of vasoactive intestinal polypeptide and neuropeptide Y in submucous nerves of rat jejunum was studied using both single-label pre-embedding immunocytochemistry and post-embedding double-label immunogold techniques. Vasoactive intestinal polypeptide-immunoreactive fibres and cell bodies were regularly observed in submucous plexus and a similar distribution was seen for neuropeptide Y. Varicose fibres were observed in single-label studies and when areas of specific interest were subjected to double-label immunogold protocols these immunoreactive profiles exhibited vesicles clearly stained for both vasoactive intestinal polypeptide and neuropeptide Y. Synaptic vesicles in immunopositive fibres observed close to the mucosa (and elsewhere in the submucosa) were dense-cored with an average diameter of 80 nm. Nerves associated with vascular elements only stained for neuropeptide Y, not for vasoactive intestinal polypeptide. These findings suggest that these two unrelated enteric peptides are co-released in the vicinity of the mucosal lining and the likely implications of such co-release are discussed.