Alkaline secretion by Necturus proximal duodenal mucosa

Proximal duodenum from the amphibian Necturus was stripped of muscle layers and the mucosa was mounted as a tube for studies of alkali transport or as a flat sheet for intracellular impalement by voltage-sensitive glass micro-electrodes. The mucosa alkalinized the unbuffered luminal perfusate at a high rate (3.4 muequiv. cm-1 h-1) and developed a transepithelial electric potential difference of 5.7 mV (lumen negative). Transport was inhibited by 2,4-dinitrophenol (10(-4) M) and by furosemide (10(-3) M) and SITS (10(-3) M) on the seros but not on the mucosal side, indicating dependence on tissue metabolism and on serosal membrane Cl-/HCO3- exchange. Prostaglandin E2 (10(-7)-10(-5) M) and dibutyryl cyclic AMP (10(-6)-10(-4) M) had no effects on the secretion or transepithelial electrical potential difference. removal of serosal HCO3- decreased luminal alkalinization by 75%, indicating a contribution by passive migration of HCO3- and/or a dependence of transcellular transport on the nutrient supply of this ion. Administration of HCO3- (17.8 mM) to the luminal perfusate affected neither the transepithelial nor transmembrane electrical potential differences nor the resistance ratio. It is thus unlikely that the luminal membrane possesses any major HCO3- conductance.     

Inhibition of H+ secretion in frog gastric mucosa by somatostatin.

Somatostatin added to the serosal bathing solution of the isolated gastric mucosa of Rana pipiens significantly inhibited pentagastrin- and histamine-stimulated H+ secretion. The decrease in H+ secretion rate was accompanied by an increase in the transmucosal potential difference and resistance. Somatostatin (10(-5) M) had no effect on the N6,O2-dibutyryl adenosine 3'-5'-cyclic monophosphate (DBcAMP)-stimulated H+ secretion rate. The mucosa exposed to somatostatin secreted H+ on stimulation by DBcAMP or histamine, but did not respond to 2.8 X 10(-7) M pentagastrin. However, pentagastrin added to the serosal solution stimulated H+ secretion after the somatostatin was washed away. Calcium inophore (3 X 10(-5) M) alone or 10(-2) M Ca2+ plus calcium ionophore temporarily increased the H+ secretion rate inhibited by somatostatin. The data suggest that somatostatin has a direct effect on the oxyntic cells in the gastric mucosa.     

Inhibition of H+ secretion in the frog gastric mucosa by ATP and related compounds.

Addition of adenosine triphosphate (ATP) to the nutrient (submucosal-facing) solution of the histamine-stimulated in vitro frog (Rana pipiens) gastric mucosa produces a marked reduction in the H+ secretory rate and an increase in transmucosal potential difference (PD) and resistance in both Cl- and Cl-free media. The effects are reversible upon removal of ATP. The threshold concentration is between 1 and 2 mM, and 5 mM produce maximal inhibition. It is shown that the effects of ATP are not due to a change in pH or osmolarity of the nutrient fluid, or to a decrease in the Ca2+ and/or Mg2+ activities of the nutrient fluid. It is also shown that the inhibitory action of ATP is not dependent on a chelation complex between Ca2+ or Mg2+. Adenosine diphosphate also produces effects essentially the same as ATP whereas 5'-adenosine monophosphate and adenosine produce relatively little or no change.     

H+ production by isolated cells from human gastric mucosa

Gastric mucosal cells were isolated from human mucosa obtained at surgery. H⁺ production was indirectly estimated by¹⁴C aminopyrine (AP) uptake. The maximal response to histamine occurred after 30 min of incubation whereas intrinsic factor (IF) secretion was maximal after only 7.5 to 15 min. According to the concentration response curve 10^–4 mol/l histamine proved to be the most effective concentration, the response to which was completely inhibited by ranitidine. Carbachol, dibutyryl cAMP and IMX also enhanced AP uptake, IMX being even more powerful than histamine. Carbachol and IMX failed to potentiate the response to histamine. Parietal cell fractions potentiate the response to histamine. Parietal cell fractions enriched by a Percoll^® density gradient revealed a pronounced background stimulation so that additional stimulation by test agents was less effective than in non-fractionized cells.     

Acid secretion by isolated primate gastric mucosa.

Pentagastrin, histamine, and acetylcholine stimulated acid secretion in the isolated rhesus (Macaca mulatta) gastric mucosa. The potencies were in the order pentagastrin greater than histamine greater than acetylcholine. However, histamine was more effective, the calculated maximal response being twice as great as for the other two agonists. Metiamide inhibited acid secretion induced by both histamine and pentagastrin. Whereas the inhibition of histamine by metiamide was surmountable, that of pentagastrin was not. Atropine inhibited the response to acetylcholine. Even in higher doses, atropine had no effect on histamine. With stimulation, transmucosal potential differences and resistances decreased; the changes were reversed on inhibition.     

Improved intraepithelial two-dimensional cable analysis with application to Necturus gastric antral mucosa

 Previously 2D cable analysis has been performed with two microelectrodes, one for passing intraepithelial current and the other for measuring the voltage response in multiple cells along the distance scale. This requires that the epithelium must be in a stable state for a considerably long period because of the multiple impalements. To follow changes of intraepithelial resistances in chambered Necturus antral mucosa with good temporal resolution, four/five electrodes were used to impale cells in the same preparation and the intraepithelial current (6.5–20 nA) was conducted sequentially to three/four of the electrodes, one at a time, to obtain six/ten independent voltage response measurements along the distance scale. The solution to the 2D cable equations was fitted to results and apical, basolateral and shunt resistances were calculated. It was found that an incorrect distance configuration can ruin the precision of the measurement. The distance configuration can, however, be optimized. The resistance values calculated with the 2D cable analysis were very close to those obtained by the amiloride exposure technique in the same tissues. The improvement gained with this work is better temporal resolution (even <10 s) when measuring epithelial resistances. Received: 15 January 1998 / Accepted: 3 June 1998     

Effect of sodium taurocholate on secretion by amphibian gastric mucosa in vitro

Effects of sodium taurocholate on the electrical and secretory activity of amphibian gastric mucosa have been studied in vitro. Exposure of the luminal surface of fundic mucosa to high concentrations (5 X 10(-2) M) at low pH (2.0 and 3.0) produced a marked fall in potential difference and electrical resistance. At lower concentrations (10(-3) to 10(-4) M) and higher pH (7.4), taurocholate did not alter the electrical properties but significantly increased net acidification from 1.39 +/- 0.27 to 2.01 +/- 0.18 mueq . cm-2 . h-1 (means +/- SE; P less than 0.01). Pretreatment of fundic mucosa with cimetidine resulted in net alkaline secretion (0.27 +/- 0.07 mueq . cm-2 . h-1), and addition of taurocholate (10(-4) M) to the luminal surface at pH 7.4 converted net alkalinization to net acidification (0.94 +/- 0.28 mueq . cm-2 . h-1). This response was not inhibited by atropine (10(-5) M) or somatostatin (10(-6) M) but exhibited marked tachyphylaxis. Taurocholate (10(-4) M) inhibited alkaline secretion in thiocyanate-treated fundic mucosa (0.63 +/- 0.04 to 0.14 +/- 0.09 mueq . cm-2 . h-1; P less than 0.001) and in spontaneously alkaline-secreting antral mucosa (0.36 +/- 0.12 to 0.09 +/- 0.06 mueq . cm-2 . h-1; P less than 0.05), but acidification did not occur. Apparent stimulation of acid secretion and simultaneous inhibition of alkaline secretion of sodium taurocholate may play a role in the pathogenesis of mucosal damage by bile.     

Effects of Ca2+ ions on gastric acid secretion by the rat isolated stomach

The role of Ca²⁺ in the stimulatory action of histamine has been evaluated in the isolated gastric fundus from immature rats, by changing the concentration of calcium ions in the bathing solutions. Lowering Ca²⁺ to 1.2 mM greatly enhanced the secretory response to histamine, while leaving unaffected that to the H₂-receptor agonist, dimaprit. The effect of histamine was competitively antagonized by ranitidine (pA₂=6.78) in normal solutions; conversely in 1.2 mM Ca²⁺, the antagonism by ranitidine became unsurmountable. Basal rates of acid secretion did not change in low Ca²⁺ solutions, whereas they were reduced approximately by 50% in Ca²⁺-free media. Finally, the secretory response to theophylline was significantly lower in low Ca²⁺ solutions in comparison with that in control conditions. From the above results it may be concluded that changes in the concentration of Ca²⁺ ions caused different changes in the secretory response of the rat stomach in the various experimental conditions. The marked enhancement of the response to histamine observed in low Ca²⁺ is unlikely to be connected with H₂-receptors, as suggested by the lack of interference in the response to dimaprit, but it could be related to intracellular mechanisms (H⁺/K⁺-ATPase, carbonic anhydrase activation etc.).     

Effects of increased O2 and CO2 on acid secretion by dogfish gastric mucosa in vitro.

The gastric mucosa of the dogfish (Squalus acanthias), as usually prepared for in vitro chambered experiments, shows a secretory rate (JH) of about 2 mueq/cm2-h, but a potential difference (PD) of zero. Raising PCO2 from 0.05 to 0.1 atm increases JH by 40% and causes the development of a PD of about 2 mV, mucosal surface positive. Increasing PO2 from 0.9 to 1.9 atm in a hyperbaric chamber (at constant PCO2 = 0.1 atm) doubles JH and increases PD to 5 mV. Transepithelial resistance falls by 20% at high PO2. It appears that the dogfish gastric mucosa, like that of the frog, is rate limited by CO2 diffusion into the tissue from the usual 5% mixture and is also rate limited by the usual O2 levels (unlike the frog), presumably due to its thicker structure and higher O2 consumption. The mucosal-positive PD, which is reversed from all other mucosae studied, is readily explained by separate electrogenic H+ and Cl- pumps, but less readily by schemes embodying a neutral HCl pump. It is not yet known whether the hyperbaric conditions are sufficient to ensure O2 sufficiency.