Bombesin-like peptides stimulate somatostatin release from rat fundic D cells in primary culture

被引:15
作者
Schaffer, K
Herrmuth, H
Mueller, J
Coy, DH
Wong, HC
Walsh, JH
Classen, M
Schusdziarra, V
Schepp, W
机构
[1] TECH UNIV MUNICH, DEPT MED 2, D-81675 MUNICH, GERMANY
[2] TECH UNIV MUNICH, DEPT PATHOL, D-81675 MUNICH, GERMANY
[3] TULANE UNIV, MED CTR, DEPT MED, NEW ORLEANS, LA 70112 USA
[4] GASTROENTER BIOL CTR, CURE, LOS ANGELES, CA 90073 USA
来源
AMERICAN JOURNAL OF PHYSIOLOGY-GASTROINTESTINAL AND LIVER PHYSIOLOGY | 1997年 / 273卷 / 03期
关键词
gastrin-releasing peptide receptor; gastrin-releasing peptide receptor antagonist; gastric cell culture; staurosporine;
D O I
10.1152/ajpgi.1997.273.3.G686
中图分类号
R57 [消化系及腹部疾病];
学科分类号
摘要
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 growth 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 anti-rabbit 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 (EC50 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-Phe(6)]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 DBcCAMP or epinephrine.
引用
收藏
页码:G686 / G695
页数:10
相关论文
共 42 条
[1]   DIFFERENTIAL EFFECT OF BOMBESIN ON INTRALUMINAL AND INTRAVASCULAR RELEASE OF GASTRIC GASTRIN AND SOMATOSTATIN IN ANESTHETIZED RATS [J].
ALINO, SF ;
UVNASMOBERG, K .
ACTA PHYSIOLOGICA SCANDINAVICA, 1989, 135 (04) :565-571
[2]   MOLECULAR-CLONING OF THE BOMBESIN GASTRIN-RELEASING PEPTIDE RECEPTOR FROM SWISS 3T3 CELLS [J].
BATTEY, JF ;
WAY, JM ;
CORJAY, MH ;
SHAPIRA, H ;
KUSANO, K ;
HARKINS, R ;
WU, JM ;
SLATTERY, T ;
MANN, E ;
FELDMAN, RI .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1991, 88 (02) :395-399
[3]  
BERTACCINI G, 1973, BRIT J PHARMACOL, V49, P437, DOI 10.1111/j.1476-5381.1973.tb17254.x
[4]   ACTION OF SECRETAGOGUES ON AMYLASE RELEASE FROM DOG PANCREATIC ACINI [J].
BOMMELAER, G ;
ROZENTAL, G ;
BERNIER, C ;
VAYSSE, N ;
RIBET, A .
DIGESTION, 1981, 21 (05) :248-254
[5]   INHIBITION OF BOMBESIN-STIMULATED GASTRIN-RELEASE FROM ISOLATED HUMAN G-CELLS BY BOMBESIN ANALOGS [J].
BUCHAN, AMJ ;
MELOCHE, M ;
COY, DH .
PHARMACOLOGY, 1990, 41 (05) :237-245
[6]   SIGNAL-TRANSDUCTION EVENTS INVOLVED IN BOMBESIN-STIMULATED GASTRIN-RELEASE FROM HUMAN G-CELLS IN CULTURE [J].
BUCHAN, AMJ ;
MELOCHE, RM .
CANADIAN JOURNAL OF PHYSIOLOGY AND PHARMACOLOGY, 1994, 72 (09) :1060-1065
[7]   RELEASE OF SOMATOSTATIN IMMUNOREACTIVITY FROM HUMAN ANTRAL D-CELLS IN CULTURE [J].
BUCHAN, AMJ ;
CURTIS, SB ;
MELOCHE, RM .
GASTROENTEROLOGY, 1990, 99 (03) :690-696
[8]   EFFECTS OF VARIOUS GASTROINTESTINAL PEPTIDES ON GASTRIC SOMATOSTATIN RELEASE [J].
CHIBA, T ;
TAMINATO, T ;
KADOWAKI, S ;
INOUE, Y ;
MORI, K ;
SEINO, Y ;
ABE, H ;
CHIHARA, K ;
MATSUKURA, S ;
FUJITA, T ;
GOTO, Y .
ENDOCRINOLOGY, 1980, 106 (01) :145-149
[9]  
CHIBA T, 1988, SOMATOSTATIN BASIC C, P229
[10]  
DELLEFAVE G, 1985, LIFE SCI, V27, P993