Abstract
A method was devised for the simultaneous localization of neuropeptides and acetylcholine (ACh) -like cation at ultrastructural level in a paraformaldehyde fixed nervous tissue (myenteric plexus). In order to minimize the loss of tissual ACh, no membrane permeabilization was made before the immunocytochemical reaction. Conventional pre-embedding immunoperoxidase reaction for neuropeptides detection [substance P and vasoactive intestinal polypeptide (VIP)] was followed by cytochemical ionic fixation of ACh-like cation. All the substance P- or VIP-immunoreactive nerve endings possessed punctiform precipitates of ACh-like cations situated in the small clear synaptic vesicles (30-50 nm). In the vicinity of substance P- and VIP-immunoreactive nerve terminals, some other nerve terminals containing ACh-like cations did not contain any peptide immunoreactivity.
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