Regulation of mucus secretion by cells isolated from the rat gastric mucosa

1. A suspension of cells, containing about 30% mucous cells, was isolated from the rat fundic mucosa, and was pre-incubated with D-[6-3H]glucosamine. 3H-labelled material subsequently released into the medium was separated by Fast Protein Liquid Chromatography on a Superose 6 column. 2. A sharp peak of labelled high molecular weight material eluted from the column close to the void volume. This material was identified as mucous glycoprotein by its similar chromatographic behaviour to partially purified rat gastric mucous glycoprotein, by its resistance to complete degradation by papain and by its behaviour on treatment with dithiothreitol. On a caesium chloride density gradient the labelled material was virtually all located between densities of 1.35 and 1.53 g/ml. with the main peak at 1.40 g/ml. 3. A broad peak of lower molecular weight material was also eluted from the column. The release of this unidentified material did not seem to be closely associated with the release of mucous glycoprotein from the cells. 4. Release of mucous glycoprotein was stimulated by secretin (half-maximally effective concentration 2.3 nM, 84% stimulation above basal release at 100 nM), and by isoprenaline (half-maximally effective concentration 34 nM, 33% stimulation at 1 microM). Carbachol (0.5 nM) produced a significant (18-29%) stimulation of mucus secretion, but gastrin (100 nM), histamine (0.5 mM) and epidermal growth factor (200 nM) were without effect. 5. The preparation should prove useful in the identification of the agents which regulate gastric mucus secretion.