Visualisation of the gastrin receptor within rat mucosa using a biotinylated gastrin antagonist.
We have previously demonstrated that the peptide Boc-L-Trp-L-Leu-beta-Ala is a potent and specific antagonist of pentagastrin-stimulated acid secretion in both the rat and the dog. Using conventional solution phase methodology, the analogue biotinyl-L-Trp-L-Leu-beta-Ala was prepared in reasonable yield and purity and applied to cryostat sections of rat intestinal and other tissues. The sections were exposed to 5-10 micrograms of peptide and the bound analogue was visualised using streptavidin-fluorescein. The binding of the analogue was demonstrated in sections from fundus, duodenum, ileum, colon, and lung. However, the analogue failed to bind to tissue from the pancreas, heart, kidney, or liver. The binding of the probe was greatly reduced or completely inhibited by preincubation with Boc-L-Trp-L-Leu-beta-Ala, pentagastrin, or gastrin 1-17. The distribution of the cells recognised by the probe was consistent with the distribution of histamine-containing enterochromaffin-like cells. The results of this study may have some bearing on current theories of the mechanism of gastrin-stimulated acid release.
Douglas
A J
AJ
Department of Biochemistry, Queen's University, Belfast, Ireland.
Walker
B
B
Johnston
C F
CF
Murphy
R F
RF
D000595:Amino Acid Sequence; D000818:Animals; D005629:Frozen Sections; D005755:Gastrins / Q000037:antagonists & inhibitors*; D007413:Intestinal Mucosa / Q000378:metabolism*; D008297:Male; D008969:Molecular Sequence Data; D009842:Oligopeptides; D051381:Rats; D011919:Rats, Inbred Strains; D011949:Receptors, Cholecystokinin / Q000378:metabolism*; D014018:Tissue Distribution
D005755:Gastrins; D009842:Oligopeptides; D011949:Receptors, Cholecystokinin; C064296:biotinyl-tryptophyl-leucyl-beta-alanine