Ultrastructural localization of VIP-like immunoreactivity in large dense-core vesicles of ‘cholinergic-type’ nerve terminals in cat exocrine glands

Abstract

Exocrine glands of the cat were analysed with the peroxidase-antiperoxidase method and routine electron microscopy. Vasoactive intestinal polypeptide (VIP)-like immunoreactivity was observed in certain nerve endings in the submandibular salivary gland, lacrimal gland and Harderian gland. The distribution of the VIP immunoreactive nerve fibres agreed well with earlier light microscopic findings. At the electron microscopic level electron-dense precipitates representing VIP-like immunoreactivity were seen in so-called large dense-core vesicles (median diameter about 990A˚) in nerve fibres and varicosities also containing many small (‘immunonegative’) agranular vesicles. In conventional electron microscopy, the small agranular vesicles outnumbered the large dense-core vesicles by about 9 to 1. Immunoreactive fibres and varicosities could be seen close to the secretory acini (distance less than 400A˚) and more distant (1500A˚or more) to e.g. demilunes, ducts and blood vessels of the glands. The number and distribution of immunoreactive nerve fibres were not affected by sympathectomy. Furthermore, no typical ‘p-type’ bouton profiles, which are dominated by large opaque vesicles (dia. 800–2000A˚), could be seen in the ultrastructural analysis of conventional preparations of the glands. The morphological features of the VIP immunoreactive nerve endings could not be distinguished from those often described as representing cholinergic fibres. These findings are in agreement with earlier suggestions of a possible coexistence of acetylcholine and VIP in neurons innervating exocrine glands and indicate possible functions for VIP in the roles of these nerves in evoking vasodilation and exocrine secretion.