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Free access | 10.1172/JCI110091
Unité de Recherche de Diabétologie et d'Etudes Radio-Immunologiques des Hormones Protéiques (Institut National de la Santé et de Recherche Medical U 55, Centre National de la Recherche Scientifique, European Research Associates 494), Hôpital Saint-Antoine, Paris 12, France
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Unité de Recherche de Diabétologie et d'Etudes Radio-Immunologiques des Hormones Protéiques (Institut National de la Santé et de Recherche Medical U 55, Centre National de la Recherche Scientifique, European Research Associates 494), Hôpital Saint-Antoine, Paris 12, France
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Unité de Recherche de Diabétologie et d'Etudes Radio-Immunologiques des Hormones Protéiques (Institut National de la Santé et de Recherche Medical U 55, Centre National de la Recherche Scientifique, European Research Associates 494), Hôpital Saint-Antoine, Paris 12, France
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Unité de Recherche de Diabétologie et d'Etudes Radio-Immunologiques des Hormones Protéiques (Institut National de la Santé et de Recherche Medical U 55, Centre National de la Recherche Scientifique, European Research Associates 494), Hôpital Saint-Antoine, Paris 12, France
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Unité de Recherche de Diabétologie et d'Etudes Radio-Immunologiques des Hormones Protéiques (Institut National de la Santé et de Recherche Medical U 55, Centre National de la Recherche Scientifique, European Research Associates 494), Hôpital Saint-Antoine, Paris 12, France
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Unité de Recherche de Diabétologie et d'Etudes Radio-Immunologiques des Hormones Protéiques (Institut National de la Santé et de Recherche Medical U 55, Centre National de la Recherche Scientifique, European Research Associates 494), Hôpital Saint-Antoine, Paris 12, France
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Published March 1, 1981 - More info
An EDTA procedure was used to prepare isolated epithelial cells of human gallbladder devoid of endogenous vasoactive intestinal peptide (VIP) as measured by radioimmunoassay. Specific binding sites for VIP were characterized in these cells. At 37°C, the binding of 125I-labeled VIP reached a peak within 20 min and then declined rapidly. At 15°C, binding was stable between 90 and 180 min of incubation. Binding of the labeled peptide was inhibited by concentrations of native VIP of 30 pM-0.1 μM. Half-maximal inhibition was observed at 2 nM. Scatchard analysis indicated two functionally independent classes of receptor sites: 62,000 high affinity sites/cell with a dissociation constant (Kd) of 1.3 nM, and 510,000 low affinity sites/cell with a Kd of 16.2 nM. Secretin inhibited tracer binding but with a 1,000 times lower potency than native VIP. VIP strongly stimulated adenosine 3′:5′ monophosphate (cyclic AMP) production in human gallbladder epithelial cells. At 37°C, 0.1 nM and 10 nM VIP raised cyclic AMP levels 44 and 100 times above the basal level, respectively. Maximal values remained constant between 60 and 90 min at 15°C. The importance of the VIP-induced cyclic AMP rise was related, at least in part, to a low phosphodiesterase activity in human gallbladder epithelial cells. At equilibrium, during a 60-min incubation at 15°C, cyclic AMP production was noted at concentrations of VIP as low as 3 pM. Maximal and half-maximal stimulations were observed at 10 nM and 0.2 nM VIP, respectively. Secretin also stimulated cyclic AMP production but with a 10,000 lower potency than VIP.
In the guinea pig, VIP and secretin were equipotent stimulators of cyclic AMP in gallbladder epithelial cells. This particular feature was shown to be due to receptors specific for each peptide that were present in these cells.
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