Effects of cold storage on relaxation responses in the rat oesophageal tunica muscularis mucosae

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Abstract
Tetrodotoxin-resistant relaxations produced by electrical field stimulation, 5-hydroxytryptamine (5-HT), and A23187 (calimycin) were investigated in the rat isolated oesophageal tunica muscularis mucosae subjected to cold storage of varied duration. Cold storage for 2 days abolished cholinergic nerve mediated contractions to field stimulation; however, field stimulation evoked relaxations and muscarinic agonist evoked contractions persisted undiminished. After 5 days of cold storage, field-stimulated relaxations, as well as tension generating capacity of the tissue, were significantly reduced. Proximodistal differences were observed in tunica muscularis mucosae sensitivity to the relaxant effects of serotonin and A23187 and these were exaggerated after cold storage. In the distal segment, cold storage for 2 days unmasked ketanserin-sensitive 5-HT receptors mediating contractions. Similarly, A23187 induced contractions rather than relaxation in cold-stored distal tunica muscularis mucosae; however, this effect was resistant to ketanserin. Immunohistochemical staining by means of the peroxidase–antiperoxidase technique revealed 5-HT-like immunoreactive "mast cells" within the tunica muscularis mucosae. These cells appeared to be associated with the smooth muscle rather than the vasculature. It is concluded that (i) field-stimulated relaxation is not dependent on intramural nerves, and (ii) it is unlikely that the release of 5-HT from mast cells or other cells mediates field-stimulated relaxation. However, confirmation awaits the provision of an antagonist against the novel 5-HT receptor that mediates 5-HT-induced relaxations.