Characterization of interactions between CCK-33 and CCK receptors in isolated dispersed pancreatic acini.
In isolated dispersed pancreatic acini, we have characterized the interactions between cholecystokinin (CCK) and CCK receptors by simultaneously measuring CCK-33 immunoreactivity and CCK bioactivity. Incubation of acinar cells with CCK-33 at cell density of 0.2-0.3 mg acinar protein per ml resulted in stimulation of amylase release concomitant with significant and time-dependent decrease of the immunoreactive CCK. With L-364,718 (0.1 microM), a specific CCK receptor antagonist, immunoreactive CCK levels in the media were not significantly altered during incubation; however, CCK-stimulated amylase release was almost completely abolished (94% inhibition). Vasoactive intestinal peptide (1 nM) significantly potentiated CCK stimulated amylase release without affecting immunoreactive CCK in the media. Insulin (167 nM) did not affect the CCK stimulated amylase release or immunoreactive CCK in the media. Incubation of acinar cells with CCK-33 at 4 degrees C did not affect the levels of immunoreactive CCK; however, a significant change in levels of immunoreactive CCK were found at 37 degrees C at 90 min. Incubation of cell free medium with CCK-33 in the presence or absence of secreted enzymes revealed no changes in CCK immunoreactivity in the medium at 90 min. Addition of bacitracin in the incubation media did not affect the CCK immunoreactivity or bioactivity. These findings indicate that in isolated rat pancreatic acini, CCK-33 stimulates amylase release through a receptor that is specifically blocked by L-364,718. Specificity of the interactions of CCK-33 with acinar cells in the media appears to be receptor-mediated and time- and temperature-dependent.
D000681:Amylases / Q000378:metabolism; D000818:Animals; D001414:Bacitracin / Q000494:pharmacology; D001570:Benzodiazepinones / Q000494:pharmacology; D002766:Cholecystokinin / Q000037:antagonists & inhibitors / Q000378:metabolism* / Q000494:pharmacology; D020109:Devazepide; D005260:Female; D006652:Histological Techniques; D066298:In Vitro Techniques; D010179:Pancreas / Q000166:cytology / Q000378:metabolism*; D011863:Radioimmunoassay; D051381:Rats; D011949:Receptors, Cholecystokinin / Q000378:metabolism*; D013552:Swine; D014660:Vasoactive Intestinal Peptide / Q000494:pharmacology
D001570:Benzodiazepinones; D011949:Receptors, Cholecystokinin; D001414:Bacitracin; D014660:Vasoactive Intestinal Peptide; D002766:Cholecystokinin; D000681:Amylases; D020109:Devazepide