Bombesin-like peptides stimulate somatostatin release from rat fundic D cells in primary culture
In: American journal of physiology. Gastrointestinal and liver physiology, Jg. 36 (1997), Heft 3, S. G686- (10S.)
Online
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Zugriff:
In several species, bombesin-like neuropeptides stimulate somatostatin release in in vitro preparations of gastric mucosa. We sought to determine if this response is due to a direct effect on fundic D cells. Rat fundic mucosal cells were isolated by pronase E (1% D cells). D cells were separated by counterflow elutriation and subsequent density-gradient centrifugation (Nycodenz) (15% D cells) and grown in primary culture for 48 h (46% D cells). Cultured cells were double stained with affinity-purified rabbit-anti-gastrin-releasing peptide (GRP) receptor antibody and mouse monoclonal antibody to human somatostatin. After incubation with rhodamine-labeled antirabbit and fluorescein isothiocyanate-labeled anti-mouse antibodies, reactions were visualized by fluorescence microscopy. All cells positive for somatostatin had GRP receptors, whereas all non-D cells showed no expression in this G cell-free culture system. Somatostatin release from cultured cells was stimulated by sulfated cholecystokinin octapeptide (CCK-8; EC50 3 x 10-10 M) and epinephrine (Ecso 4 x 10-8 M), which are established stimuli for canine fundic D cells. Bombesin (EC50 6 x 10-11 M), its mammalian analog GRP-27, and neuromedin C (GRP-10) (EC50 1 x 10-10 M, for both) were almost equally potent stimuli of somatostatin release, eliciting maximal response at 10-9 M (400-550% above basal). Neuromedin B was less potent and effective (maximal response at 10-8 M, 230% above basal). [D-Phe6]bombesin-(6-13)-OMe, a specific bombesin receptor antagonist, inhibited bombesin-stimulated somatostatin release in a competitive manner (IC50 9 x 10-8 M). Potentiating interactions were observed between bombesin and dibutyryladenosine 3',5'-cyclic monophosphate (DBcAMP) or epinephrine, but not between bombesin and CCK-8. We conclude that bombesin-like peptides directly stimulate somatostatin release by interacting with specific receptors on rat fundic D cells. Bombesin-like peptides appear to induce Ca2+-phospholipid-dependent signal-response transduction, as is indirectly suggested by potentiating interactions with DBcAMP or epinephrine.
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Bombesin-like peptides stimulate somatostatin release from rat fundic D cells in primary culture
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Autor/in / Beteiligte Person: | SCHAFFER, K ; HERRMUTH, H ; MUELLER, J ; COY, D. H ; WONG, H. C ; WALSH, J. H ; CLASSEN, M ; SCHUSDZIARRA, V ; SCHEPP, W |
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Zeitschrift: | American journal of physiology. Gastrointestinal and liver physiology, Jg. 36 (1997), Heft 3, S. G686- (10S.) |
Veröffentlichung: | Bethesda, MD: American Physiological Society, 1997 |
Medientyp: | academicJournal |
Umfang: | print, 42 ref |
ISSN: | 0193-1857 (print) |
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