Relaxation by vasoactive intestinal polypeptide in the gastric fundus of nitric oxide synthase-deficient mice.
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In many gastrointestinal tissues nitric oxide (NO) and vasoactive intestinal polypeptide (VIP) both play a role as inhibitory non-adrenergic non-cholinergic neurotransmitters. As the mode of interaction between NO and VIP remains controversial, the aim of this study was to investigate the interplay between NO and VIP in the mouse gastric fundus and to evaluate the nitric oxide synthase (NOS) isoform involved in VIP-induced relaxation by using inducible NOS (iNOS), endothelial NOS (eNOS) and neuronal NOS (nNOS) knockout mice. The influence of NOS inhibitors on the relaxant effect of VIP was determined in isolated smooth muscle cells and smooth muscle strips of wild-type and knockout mice. In isolated smooth muscle cells from wild-type, eNOS knockout and nNOS knockout mice, the relaxation induced by VIP (10(-9) M) was inhibited by approximately 70-95 % by both the non-selective NOS inhibitor N(G)-nitro-L-arginine (L-NA; 10(-4) M) and the selective inducible NOS inhibitor N-(3-(aminomethyl)-benzyl)acetamidine (1400W; 10(-6) M). In cells isolated from iNOS knockout mice, VIP still induced full relaxation but it was not influenced by L-NA or 1400W. In smooth muscle strips from wild-type and knockout mice, the concentration-dependent relaxation by VIP (10(-9) to 3 x 10(-7) M) was not influenced by L-NA or 1400W. These results suggest that the experimental method determines the influence of NOS inhibitors on the relaxant effect of VIP. iNOS, probably induced by the isolation procedure, might be involved in the relaxant effect of VIP in isolated smooth muscle cells but not in classic smooth muscle strips. (+info)
Presynaptic modulation of cholinergic neurotransmission in the human proximal stomach.
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1. This study investigates whether the cholinergic neurones, innervating the human proximal stomach, can be modulated by nitric oxide (NO) or vasoactive intestinal polypeptide (VIP), or via presynaptic muscarinic, alpha(2)- or 5-hydroxytryptamine(4) (5-HT(4)-) receptors. 2. Circular muscle strips, without mucosa, were incubated with [(3)H]-choline to incorporate [(3)H]-acetylcholine into the cholinergic transmitter stores. The basal and electrically-induced release of tritium and [(3)H]-acetylcholine were analysed in a medium containing guanethidine (4 x 10(-6) M), hemicholinium-3 (10(-5) M), physostigmine (10(-5) M) and atropine (10(-6) M). Tissues were stimulated twice for 2 min (S(1) and S(2): 40 V, 1 ms, 4 Hz) and drugs were added before S(2). 3. The NO synthase inhibitor L-N(G)-nitroarginine methyl ester (3 x 10(-4) M) and the NO donor sodium nitroprusside (10(-5) M), as well as VIP (10(-7) M) did not influence the basal release nor the electrically-evoked release. 4. The alpha(2)-adrenoceptor agonist UK-14,304 (10(-5) M) significantly inhibited the electrically-evoked release of [(3)H]-acetylcholine, and this was prevented by the alpha(2)-adrenoceptor antagonist rauwolscine (2 x 10(-6) M). 5. The 5-HT(4)-receptor agonist prucalopride (3 x 10(-7) M) significantly enhanced the electrically-evoked release of [(3)H]-acetylcholine, and the 5-HT(4)-receptor antagonist SB204070 (10(-9) M) prevented this. 6. When atropine (10(-6) M) was omitted from the medium and added before the second stimulation, it significantly increased the release of [(3)H]-acetylcholine. 7. These results suggest that the release of acetylcholine from the cholinergic neurones, innervating the circular muscle in the human proximal stomach, can be inhibited via presynaptic muscarinic auto-receptors and alpha(2)-adrenoceptors, and stimulated via presynaptic 5-HT(4)-receptors. No evidence for modulation by NO or VIP was obtained. (+info)
Role of histamine H3 receptors in the regulation of gastric functions.
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The role of central and peripheral histamine H3 receptors in the regulation of gastric acid secretion and gastric mucosal integrity is reviewed. The activation of H3 receptors by peripheral administration of the selective agonist (R)alpha-methylhistamine reduced acid secretion in cats, dogs, rats and rabbits, while increasing it in mice. The antisecretory effects were observed against indirect stimuli that act on vagal pathways or on enterochromaffin-like (ECL) cells, such as 2-deoxy-D-glucose, food or pentagastrin, but not against histamine or dimaprit. Inhibitory effects on acid production were observed in rats after central administration of histamine or of H3 receptor agonists. In the conscious rat intragastric administration of (R)alpha-methylhistamine caused gastroprotective effects against the damage induced by absolute ethanol, HCl, aspirin and stress. The mechanism involved seems to be related to the increased mucus production, via nitric oxide-independent mechanisms. Gastroprotective effects against ethanol were also observed after central administration of histamine or its metabolite N(alpha)-methylhistamine, suggesting that brain receptors participate the histamine-mediated effects on gastric functions. (+info)
Molecular and functional characterization of ERG, KCNQ, and KCNE subtypes in rat stomach smooth muscle.
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Contribution of K(+) channels derived from the expression of ERG, KCNQ, and KCNE subtypes, which are responsible for rapidly and slowly activating delayed rectifier K(+) currents (I(Kr) and I(Ks), respectively) in cardiac myocytes, to membrane currents was examined in stomach circular smooth muscle cells (SMCs). The region-qualified multicell RT-PCR showed that ERG1/KCNE2 transcripts were expressed in rat stomach fundus and antrum SMCs and that KCNQ1/KCNE1 transcripts were expressed in antrum but not fundus. Western blotting and immunocytochemical analyses indicate that ERG1 proteins were substantially expressed in both regions, whereas KCNE1 proteins were faintly expressed in antrum and not in fundus. Both I(Kr)- and I(Ks)-like currents susceptible to E-4031 and indapamide, respectively, were identified in circular SMCs of antrum but only I(Kr)-like current was identified in fundus. It is strongly suggested that I(Kr)- and I(Ks)-like currents functionally identified in rat stomach SMCs are attributable to the expression of ERG1/KCNE2 and KCNQ1/KCNE1, respectively. The membrane depolarization by 1 microM E-4031 indicates the contribution of K(+) channels encoded by ERG1/KCNE2 to the resting membrane potential in stomach SMCs. (+info)
Effects of glucagon-like peptide-1(7-36)amide on motility and sensation of the proximal stomach in humans.
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BACKGROUND: Glucagon-like peptide-1(7-36)amide (GLP-1) retards gastric emptying, reduces food intake, and inhibits antroduodenal and stimulates pyloric motility. AIMS: To assess the effects of synthetic GLP-1 on fundus tone and volume waves, gastric compliance, and perception of gastric distension. SUBJECTS: Eleven healthy male volunteers. METHODS: Background infusions were saline, or GLP-1 at 0.3 or 0.9 pmol/ kg/min on separate days in random order. Interdigestive fundus motility was recorded by barostat (maximum capacity of intragastric bag 1200 ml) during basal and peptide periods of 60 minutes each. Thereafter stepwise isobaric distensions were performed with ongoing peptide infusion, and gastric sensation was scored. RESULTS: Low and high loads of GLP-1 induced physiological and supraphysiological plasma immunoreactivities, respectively. GLP-1 dose dependently diminished fundus tone (162.9 (15.0) and 259.5 (17.2) v 121.1 (6.0) ml with saline; p<0.0001). It greatly reduced volume waves and total volume displaced by these events (p<0.0001). Gastric compliance derived from isobaric distension rose in a dose related manner (42.6 (5.5) and 63.6 (7.7) v 27.0 (3.5) ml/mm Hg; p=0.0004) with a concomitant reduction of the pressure at half maximum bag volume (6.4 (0.4) and 5.5 (0.4) v 7.2 (0.1) mm Hg; p<0.0001). GLP-1 did not change perception of isobaric distension but reduced the perception score related to corresponding bag volume (p<0.0001). CONCLUSIONS: GLP-1 is a candidate physiological inhibitory regulator of fundus motility. It allows the stomach to afford a larger volume without increase in sensation. (+info)
Influence of antioxidant depletion on nitrergic relaxation in the pig gastric fundus.
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1. The hypothesis that endogenous tissue antioxidants might explain the inability of the superoxide generators 6-anilino-5,8-quinolinedione (LY83583) and hydroquinone (HQ) and of the NO-scavengers hydroxocobalamin (HC) and 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (c-PTIO) to affect nitrergic neurotransmission in the porcine gastric fundus was tested by selective pharmacological depletion of respectively Cu/Zn superoxide dismutase (Cu/Zn SOD) and reduced glutathione (GSH) in circular smooth muscle preparations. 2. Diethyldithiocarbamate (DETCA; 3x10(-3) M), which almost completely abolished tissue Cu/Zn SOD activity, had no effect per se on nitrergic relaxations induced by either electrical field stimulation (EFS; 4 Hz, 10 s) or exogenous nitric oxide (NO; 10(-5) M). In these DETCA-treated tissues however, electrically-induced nitrergic relaxations became sensitive to inhibition by LY83583 (10(-5) M) or HC (10(-4) M), but not by HQ (10(-4) M) or c-PTIO (10(-4) M); only for the combination of DETCA plus LY83583, this inhibition was partially reversed by exogenous Cu/Zn SOD (1000 u ml(-1)). 3. Immunohistochemical analysis of porcine gastric fundus revealed a 100% colocalization of Cu/Zn SOD and neuronal nitric oxide synthase (nNOS) in the intrinsic neurons of the myenteric plexus. 4. Buthionine sulphoximine (BSO; 10(-3) M) in the absence or presence of LY83583 (10(-5) M) or HC (10(-4) M) did not alter nitrergic relaxations, although it reduced per se the tissue GSH content to 62% of control. 5. Pharmacological depletion studies, corroborated by immunohistochemical data, thus suggest a role for Cu/Zn SOD but not for GSH in nitrergic neurotransmission in the porcine gastric fundus. (+info)
Characterization of pre- and postsynaptic muscarinic receptors in circular muscle of pig gastric fundus.
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1. This study investigated the subtype of muscarinic receptors on the cholinergic neurones and smooth muscle in the circular muscle of the pig gastric fundus. 2. Muscarinic antagonists, except MT-3, concentration-dependently inhibited the contractions induced by a given concentration of acetylcholine. Concentration-response curves by acetylcholine were shifted rightwards in a parallel manner without depression of the maximum by the muscarinic antagonists, except by MT-3 that induced a leftward shift. Correlation of the pIC(50) and pA(2) values with published pK(i) values for the five muscarinic receptor subtypes suggests that the muscarinic receptors on pig gastric fundus circular muscle belong to the M(3) subtype. 3. Electrically-evoked contractions (40 V, 4 Hz, 0.25 ms, 2 min) were concentration-dependently inhibited by the muscarinic antagonists except for methoctramine and AF-DX 116, that increased the amplitude of the electrically-induced contractions in lower concentrations. MT-3 tended to increase the electrically-induced contractions. 4. The antagonists, except MT-3, concentration-dependently increased the electrically-induced tritium outflow (40 V, 4 Hz, 0.25 ms, 2 min) after incubation of the tissues with [(3)H]-choline. MT-3 (3 x 10(-8) and 10(-7) M) decreased the electrically-induced tritium release. Correlation of the pIC(50) values with published pK(i) values for the different muscarinic receptor subtypes yielded a significant and comparable correlation for M(1), M(3), M(4) and M(5) receptors. 5. These results suggest that the postsynaptic receptors in circular muscle of the pig gastric fundus belong to the M(3) subtype. However, the presynaptic receptor could not be clearly defined, although it does certainly not belong to the M(2) subtype. (+info)
Structure-activity relationship studies of (+/-)-terbutaline and (+/-)-fenoterol on beta3-adrenoceptors in the guinea pig gastric fundus.
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(+/-)-Terbutaline and (+/-)-fenoterol are both arylethanolamine analogs that have tertbutyl and aryliso-propyl substituents respectively at the a position on the nitrogen of the ethanolamine side chain. In the present study, we have investigated the structure-activity relationships of (+/-)-terbutaline and (+/-)-fenoterol as beta3-adrenoceptor agonists in the guinea pig gastric fundus. (+/-)-Terbutaline and (+/-)-fenoterol induced concentration-dependent relaxation of the precontracted gastric fundus with pD2 values of 4.45+/-0.10 and 5.90+/-0.09, and intrinsic activities of 1.00+/-0.03 and 0.99+/-0.01 respectively. The combination of the selective beta1-adrenoceptor antagonist (+/-)-atenolol (100 microM), and the selective beta2-adrenoceptor antagonist (+/-)-butoxamine (100 microM), produced a 2 and 6 fold rightward shift of the concentration-response curves for (+/-)-terbutaline and (+/-)-fenoterol respectively, without depressing the maximal responses. The order of potency of these agonists was (pD2 value): (+/-)-fenoterol (5.09+/-0.10) > (+/-)-terbutaline (4.13+/-0.08). In the presence of (+/-)-atenolol and (+/-)-butoxamine, however, the non-selective beta1, beta2- and beta3-adrenoceptor antagonist (+/-)-bupranolol caused a concentration-dependent rightward shift of the concentration-response curves for (+/-)-terbutaline and (+/-)-fenoterol. Schild plot analyses of the effects of (+/-)-bupranolol against these agonists gave pA2 values of 6.21+/-0.07 ((+/-)-terbutaline) and 6.37+/-0.06 ((+/-)-fenoterol) respectively, and the slopes of the Schild plot were not significantly different from unity (p>0.05). These results suggest that the relaxant responses to (+/-)-terbutaline and (+/-)-fenoterol are mainly mediated through beta3-adrenoceptors in the guinea pig gastric fundus. The beta3-adrenoceptor agonist potencies of arylethanolamine analogs depend on the size of the end of the alkylamine side chain. (+info)