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• W2153537269 abstract "Background & AimsGastrin is a key regulator of gastric acid secretion. We aimed to isolate pure G cells to identify the mechanistic basis of luminal- and strain-mediated regulation.MethodsUsing gradient centrifugation and fluorescence-activated cell sorting, rat G cells were prepared and luminal, neural, hormonal, and mechanical activation of secretion and signaling pathways studied.ResultsPure G-cell preparations (>97%) were isolated. Reverse-transcription polymerase chain reaction identified neural, hormonal, bacterial, and luminal G protein–coupled receptors, and immunostaining visualized specific sweet/bitter receptors and the tastant-associated G protein α-gustducin. Gastrin release was stimulated by forskolin (adenosine 3′,5′-cyclic monophosphate [cAMP] inducer, 10 μmol/L; >3-fold), potentiated by 3-isobutyl-1-methylxanthine (IBMX; phosphodiesterase type 5 inhibitor and adenosine antagonist, 10 μmol/L) and phorbol myristate acetate (phorbol ester, 10 μmol/L), and inhibited by H-89 (protein kinase A inhibitor, 10 μmol/L), PD98059 (MEK1 inhibitor, 0.1 μmol/L), and wortmannin (phosphatidylinositol 3-kinase inhibitor, 1 nmol/L). Gastrin release was stimulated by neuronal G protein-coupled receptor ligands, pituitary adenylate cyclase-activating protein (20 pmol/L, >8-fold) and bombesin (0.1 μmol/L, 8-fold) through cAMP signaling. The tastants sucralose, glucose, caffeine, denatonium, and the vanilloid receptor activator capsaicin all stimulated secretion (>3-fold), as did bacterial lipopolysaccharides Salmonella enteritidis (0.24 nmol/L, 5-fold) greater than Helicobacter pylori (0.57 μmol/L, 3-fold). Secretion was associated with elevated cAMP levels (∼2-fold) and could be inhibited by H-89 and PD98059 and potentiated by IBMX and cholera toxin (250 μg/mL). Bacterially mediated secretion also involved activation of nuclear factor κB and the c-Jun-N-terminal kinase pathway. Mechanical strain stimulated (2-fold to 8-fold) gastrin release, and decreasing pH from 7.4 to 5.5 inhibited release. The adenosine receptor 2B antagonist MRS1754 inhibited mechanically induced gastrin release.ConclusionsG cells are luminal sampling chemomechanosensory cells whose secretion is regulated by neural, hormonal, luminal, and mechanical factors through protein kinase A activation, cAMP signaling, and mitogen-activated protein kinase phosphorylation. Gastrin is a key regulator of gastric acid secretion. We aimed to isolate pure G cells to identify the mechanistic basis of luminal- and strain-mediated regulation. Using gradient centrifugation and fluorescence-activated cell sorting, rat G cells were prepared and luminal, neural, hormonal, and mechanical activation of secretion and signaling pathways studied. Pure G-cell preparations (>97%) were isolated. Reverse-transcription polymerase chain reaction identified neural, hormonal, bacterial, and luminal G protein–coupled receptors, and immunostaining visualized specific sweet/bitter receptors and the tastant-associated G protein α-gustducin. Gastrin release was stimulated by forskolin (adenosine 3′,5′-cyclic monophosphate [cAMP] inducer, 10 μmol/L; >3-fold), potentiated by 3-isobutyl-1-methylxanthine (IBMX; phosphodiesterase type 5 inhibitor and adenosine antagonist, 10 μmol/L) and phorbol myristate acetate (phorbol ester, 10 μmol/L), and inhibited by H-89 (protein kinase A inhibitor, 10 μmol/L), PD98059 (MEK1 inhibitor, 0.1 μmol/L), and wortmannin (phosphatidylinositol 3-kinase inhibitor, 1 nmol/L). Gastrin release was stimulated by neuronal G protein-coupled receptor ligands, pituitary adenylate cyclase-activating protein (20 pmol/L, >8-fold) and bombesin (0.1 μmol/L, 8-fold) through cAMP signaling. The tastants sucralose, glucose, caffeine, denatonium, and the vanilloid receptor activator capsaicin all stimulated secretion (>3-fold), as did bacterial lipopolysaccharides Salmonella enteritidis (0.24 nmol/L, 5-fold) greater than Helicobacter pylori (0.57 μmol/L, 3-fold). Secretion was associated with elevated cAMP levels (∼2-fold) and could be inhibited by H-89 and PD98059 and potentiated by IBMX and cholera toxin (250 μg/mL). Bacterially mediated secretion also involved activation of nuclear factor κB and the c-Jun-N-terminal kinase pathway. Mechanical strain stimulated (2-fold to 8-fold) gastrin release, and decreasing pH from 7.4 to 5.5 inhibited release. The adenosine receptor 2B antagonist MRS1754 inhibited mechanically induced gastrin release. G cells are luminal sampling chemomechanosensory cells whose secretion is regulated by neural, hormonal, luminal, and mechanical factors through protein kinase A activation, cAMP signaling, and mitogen-activated protein kinase phosphorylation." @default.
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• W2153537269 date "2009-07-01" @default.
• W2153537269 modified "2023-10-18" @default.
• W2153537269 title "Delineation of the Chemomechanosensory Regulation of Gastrin Secretion Using Pure Rodent G Cells" @default.
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• W2153537269 doi "https://doi.org/10.1053/j.gastro.2009.01.005" @default.
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