Effects of ammonium ion and ammonia on function and morphology of in vitro frog gastric mucosa

1993 ◽  
Vol 265 (2) ◽  
pp. G277-G288 ◽  
Author(s):  
A. Yanaka ◽  
H. Muto ◽  
S. Ito ◽  
W. Silen
Keyword(s):  
Gastric Mucosa ◽  
Electrical Resistance ◽  
Rana Catesbeiana ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Ammonium Ion ◽  
Gastric Epithelial Cells ◽  
Sudden Drop ◽  
Oxynticopeptic Cells

The effects of ammonium ion (NH+4) and ammonia (NH3) on function and morphology of gastric epithelial cells were studied in intact sheets of in vitro frog (Rana catesbeiana) gastric mucosa. Luminal 115 mM NH4Cl at luminal pH 8.0 (calculated [NH3] 2.7 mM), but not at 5.0 (calculated [NH3] 3 microM) induced 1) an increase in intracellular pH (pHi) in oxynticopeptic cells (OPC) and decreases in transmucosal potential difference (PD) and electrical resistance (R) in resting tissues, 2) a decrease in histamine-stimulated H+ secretion and an increase in H+ backdiffusion after removal of luminal NH4Cl, and 3) augmented acidification of OPC during luminal acidification. Serosal 30 mM NH4Cl at serosal pH 7.2 (calculated [NH3] 0.47 mM) induced 1) an increase in pHi in OPC and inhibition of the alkalinization of OPC after removal of ambient Cl-, 2) a decrease in PD associated with the increase in R and decrease in short-circuit current, effects attenuated by serosal 15 mM K+, accentuated by 0.2 mM Ba2+, and abolished by removal of ambient Cl-, 3) a sudden drop of PD in resting, but not in stimulated tissues, effects prevented by high serosal pH (7.8), serosal HCO3-, or removal of luminal Cl-, 4) a decrease in histamine-stimulated H+ secretion and an increase in H+ backdiffusion after removal of NH4Cl, and 5) augmented acidification of OPC during luminal acidification. These results suggest that 1) luminal NH3, but not NH+4, increases backdiffusion of H+ from the lumen to the mucosa, 2) serosal NH3 and/or NH+4 induces depolarization of OPC and decreases electrogenic Cl- transport, thereby attenuating the activity of the basolateral Cl(-)-HCO3- exchanger in OPC, and 3) both of these effects contribute to the augmented acidification of OPC during exposure to high luminal [H+].

Role of nutrient HCO3(-) in protection of amphibian gastric mucosa

1980 ◽  
Vol 239 (6) ◽  
pp. G536-G542
Author(s):  
R. Schiessel ◽  
A. Merhav ◽  
J. B. Matthews ◽  
L. A. Fleischer ◽  
A. Barzilai ◽  
...  
Keyword(s):  
Gastric Mucosa ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Experimental Conditions ◽  
Slow Decline ◽  
Alkaline Tide ◽  
Artificial Gastric Juice ◽  
Rapid Drop

In in vitro bullfrog fundic mucosa inhibited with 10(-3) M metiamide and exposed to a luminal pH of 2 a progressive slow decline in potential difference (PD) and short-circuit current (Isc) and a rise in resistance (R) were observed when the nutrient solution (N) contained 18 mM HCO3(-), but these changes were restored by an N containing 50 mM HCO3(-). Substitution of PO4(3-) or N-tris(hydroxymethyl)-methyl-2-aminoethanesulfonic acid for NHO3(-) in N caused a rapid drop in PD and Isc in inhibited tissues, changes that could be prevented by 10(-4) M histamine. Ulceration occurred more frequently in metiamide-inhibited gastric sacs exposed to artificial gastric juice with an N of 18 mMHCO3(-) than with 50 mM HCO3(-), but histamine prevented ulceration in the 18 mM HCO3(-) solution. JnetCl approximated Isc under most experimental conditions in inhibited mucosa and was reduced dramatically as were both Jn leads to sCl and Js leads to nCl when HCO3(-) was removed from N. In histamine-stimulated tissues, removal of nutrient HCO3(-) did not influence Cl- transport. Our results are consistent with the proposal that HCO3(-) in N supports normal Cl- flux and that the alkaline tide of actively secreting oxyntic cells can do the same in the absence of ambient HCO3(-).

Failure of 16,16-dimethyl PGE2 to prevent inhibitory effect of ethanol on sodium transport in canine gastric mucosa

1985 ◽  
Vol 248 (3) ◽  
pp. G299-G306
Author(s):  
T. A. Miller ◽  
J. M. Henagan ◽  
Y. J. Kuo ◽  
L. L. Shanbour
Keyword(s):  
Gastric Mucosa ◽  
Sodium Transport ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Inhibitory Effect ◽  
Gastric Epithelium ◽  
Bathing Solution ◽  
Inhibitory Effects ◽  
Stimulation Of

By use of an in vitro canine gastric mucosal preparation, we evaluated the effects of ethanol (2, 4, 6, and 8%, vol/vol) and indomethacin (2.2 X 10(-4)M), with and without 16,16-dimethyl PGE2 pretreatment, on net sodium transport (JNanet) (mucosal to serosal) across gastric epithelium. Although administration of 2 or 4% ethanol to the mucosal bathing solution had no appreciable inhibitory effects on sodium transport, 6 and 8% ethanol and indomethacin significantly inhibited JNanet when compared with untreated control mucosa. This effect was accompanied by inhibition of transmucosal potential difference (PD) and short-circuit current (Isc). In other mucosae exposed to dimethyl PGE2 (8 X 10(-6) M) in the serosal bathing solution, significant increases in JNanet, PD, and Isc were noted when compared with control mucosa. Addition of 6 or 8% ethanol to the mucosal solution of dimethyl PGE2-pretreated tissue resulted in significant decreases in PD, Isc, and JNanet below control values that were not significantly different from mucosa exposed to 6 and 8% ethanol without PG pretreatment. When indomethacin was added to the mucosal solution following dimethyl PGE2 pretreatment, only slight decreases in PD and Isc below control levels were observed, and the inhibitory effects on JNanet induced by indomethacin without such treatment were abolished. These findings suggest that stimulation of JNanet by prostaglandin may play a role in its ability to prevent indomethacin damage to gastric epithelium but does not appear to be of importance in mediating protection against ethanol damage.

H(+)-K(+)-ATPase contributes to regulation of pHi in frog oxynticopeptic cells

1991 ◽  
Vol 261 (5) ◽  
pp. G781-G789 ◽  
Author(s):  
A. Yanaka ◽  
K. J. Carter ◽  
P. J. Goddard ◽  
M. C. Heissenberg ◽  
W. Silen
Keyword(s):  
Steady State ◽  
Gastric Mucosa ◽  
Histamine Release ◽  
Rana Catesbeiana ◽  
Intracellular Acidosis ◽  
Endogenous Histamine ◽  
Decreasing Ph ◽  
Oxynticopeptic Cells

The effect of intracellular acidosis on luminal H+ secretion and the role of H(+)-K(+)-ATPase in regulation of intracellular pH (pHi) in oxynticopeptic cells (OPC) (measured with a pH-sensitive fluorescent dye) were examined in intact sheets of in vitro frog (Rana catesbeiana) gastric mucosa. Intracellular acidosis of OPC induced by decreasing pH in the serosal solution (pHs) from 7.2 to 6.0 reversibly increased forskolin-stimulated H+ secretion without increasing endogenous histamine release. The observed increase in H+ secretion was unaffected by either 1 mM cimetidine or 1 mM histamine, but was accentuated by 1 mM amiloride, an effect abolished by 0.3 mM omeprazole. Steady-state pHi values in stimulated or resting OPC at pHs 7.2 were not significantly different. However, pHi in OPC was significantly higher in stimulated than in resting tissues at pHs 6.9, a difference accentuated by decreasing pHs to 6.4 or by 1 mM amiloride. Amiloride completely prevented recovery from intracellular acidosis induced by pHs 6.4 or 6.9 in omeprazole-treated tissues, but only partially mitigated recovery in cimetidine- or forskolin-treated tissues. At pHs 6.4, high luminal [K+] (100 mM) increased H+ secretion and hastened recovery of pHi in cimetidine-treated tissues in the presence of amiloride. These results suggest that, in intact sheets of in vitro frog gastric mucosa, 1) intracellular acidosis stimulates luminal H+ secretion via histamine-independent mechanisms and 2) H(+)-K(+)-ATPase contributes to the recovery of OPC from intracellular acidosis.

Influence of Cl- on pH(i) in oxynticopeptic cells of in vitro frog gastric mucosa

1990 ◽  
Vol 258 (5) ◽  
pp. G815-G824 ◽  
Author(s):  
A. Yanaka ◽  
K. J. Carter ◽  
H. H. Lee ◽  
W. Silen
Keyword(s):  
Gastric Mucosa ◽  
Intracellular Ph ◽  
Rana Catesbeiana ◽  
Basolateral Membrane ◽  
Fluorescent Dye ◽  
Disulfonic Acid ◽  
Fundic Mucosa ◽  
Ethanesulfonic Acid ◽  
Oxynticopeptic Cells

The effect of Cl- on intracellular pH (pH(i)) was studied using sheets of frog (Rana catesbeiana) fundic mucosa in which oxynticopeptic cells were selectively loaded with the acetomethoxy ester form of the pH-sensitive fluorescent dye 2',7'-bis(2-carboxyethyl)-5(6)-carboxyfluorescein (BCECF/AM). Before the measurement of pH(i), tissues were exposed to either 10(-5) M forskolin in the serosal solution (stimulated tissues) or 3 x 10(-4) omeprazole in the serosal solution (inhibited tissues). In HCO3- and N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (HEPES) buffers, pH(i) increased significantly after removal of Cl- from serosal and luminal solution, both in stimulated and inhibited tissues. The presence of Cl- in the luminal solution prevented this rise in pHi, an effect abolished by serosal 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS, 3 x 10(-4) M) but not by serosal amiloride (10(-3)M). In the presence of serosal Cl-, pH(i) increased after exposure to serosal DIDS, more prominently in the stimulated than in the inhibited tissues. These results confirm the presence of a Cl(-)-HCO3-exchanger in the basolateral membrane of oxynticopeptic cells in intact sheets of mucosa and suggest that luminal Cl- contributes to the regulation of pH(i) in oxynticopeptic cells.

Action of 2-deoxy-D-glucose on frog gastric mucosa

1965 ◽  
Vol 209 (3) ◽  
pp. 461-466 ◽  
Author(s):  
George Sachs ◽  
R. Shoemaker ◽  
B. I. Hirschowitz
Keyword(s):  
Gastric Mucosa ◽  
Acid Secretion ◽  
Glucose Uptake ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Potential Difference ◽  
Atp Levels

2-Deoxyglucose (2-DG) has been found to inhibit chloride and acid secretion by the in vitro frog mucosa, with a fall in short-circuit current and potential difference and a rise in resistance. The ATP levels and phosphohexoisomerase activity were essentially unchanged following 2-DG treatment. 3-Methyl-O-glucose uptake was reduced by about 50% following preincubation with 2-DG. The O2 consumption was only slightly reduced with 10 mmoles 2-DG, but the CO2 ratio from glucose-6-C14/glucose-1-C14 fell from 0.98 to 0.37, indicating activation of the hexosemonophosphate (HMP) shunt.

Effect of imidazoles on active transport by gastric mucosa and urinary bladder

1965 ◽  
Vol 208 (6) ◽  
pp. 1183-1190 ◽  
Author(s):  
Darwin Alonso ◽  
Richard Rynes ◽  
John B. Harris
Keyword(s):  
Gastric Mucosa ◽  
Urinary Bladder ◽  
Competitive Inhibition ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Buffering Capacity ◽  
Toad Urinary Bladder ◽  
Inhibitory Effects ◽  
Active Sodium

Studies were undertaken to determine whether the stimulatory effect of histamine and the methyl xanthines on the frog gastric mucosa in vitro could be ascribed to the imidazole component of these compounds. The actively transported ions of gastric secretion appear in the form of hydrochloric acid and of a nonacid chloride component that is responsible for the short-circuit current (Isc). At 10–2 m, imidazole reduced acid secretion (Jh) by 80%, which was somewhat greater than the reduction produced by thiocyanate. N-methylimidazole, 2-methylimidazole, 4(5)-methylimidazole, and benzimidazole abolished Jh. The Isc tended to rise as Jh fell. In stimulated mucosae, imidazole and N-methylimidazole promptly reduced oxygen consumption (qO2) by 25%; thiocyanate had a delayed and lesser effect. Pyrazole, 4-hydroxymethylimidazole, and tris had little or no effect on Jh, Isc, or qO2. Imidazole and N-methylimidazole reduced active sodium transport by the toad urinary bladder by 50%. The results indicate that the inhibitory effects of the imidazoles cannot be ascribed to their buffering capacity or to competitive inhibition of histamine. Instead the effects may result from depletion of tissue content of adenosine-3',5'-phosphate or from interference with protein-bound phosphohistidine in oxidative phosphorylation.

Acid secretion, potential difference, and resistance of elasmobranch stomach

1962 ◽  
Vol 203 (6) ◽  
pp. 1091-1093 ◽  
Author(s):  
Warren S. Rehm
Keyword(s):  
Gastric Mucosa ◽  
Acid Secretion ◽  
Present Report ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Potential Difference ◽  
Secretory Rate ◽  
Negaprion Brevirostris ◽  
Ion Secretion

The present report is concerned with in vitro studies on gastric mucosa of the skate, Raja eglentaria, the electric ray, Narcine braziliensis, and the shark, Negaprion brevirostris. Maximum secretory rates of from 0.65 to 2.7 µEq hr–1 cm–2 were found. An increase in the secretory rate from an initial low level was associated with an increase in potential difference (PD), a decrease in resistance, and an increase in the calculated short-circuit current. The average PD and resistance before the increase in the secretory rate was 2.2 mv (nutrient positive) and 268 ohms cm2. After the increase they were 6.4 mv and 199 ohms cm2. Thiocyanate (10–2 m) to nutrient resulted in a decrease of secretory rate to zero and an increase in PD and resistance. The results can be explained on the basis of the separate mechanisms theory of H+ and Cl– ion secretion on the assumption that the resistance in the Cl– ion limb of the circuit is considerably lower than that in the H+ ion limb of the circuit.

Electrophysiology of the frog gastric mucosa with sufficient CO2

1975 ◽  
Vol 228 (3) ◽  
pp. 928-933 ◽  
Author(s):  
Kidder GW
Keyword(s):  
Gastric Mucosa ◽  
Acid Secretion ◽  
Rate Increase ◽  
Short Circuit ◽  
Potential Drop ◽  
Short Circuit Current ◽  
Secretory Rate ◽  
Sudden Drop ◽  
Tissue Resistance ◽  
Per Se

Changing from 5% CO2 in the serosal solution only to 10% CO2 on both sides doubles the acid secretory rate of the bullfrog stomach (as shown previously) and raises the transmural potential difference, although the short-circuit current is not changed. The secretory overshoot on reoxygenation after anaerobiosis, observed in 5% CO2, is virtually eliminated in 10% CO2, as predicted by the diffusion model which explains the secretory rate increase. It was found that the rate of rise of the secretory rate during anoxia was identical in the 2 conditions, which explains the increased secretory lag in 10% CO2 and suggests an interesting limitation on the rate with which acid secretion can be activated. During anoxia in 10% CO2, but not in 5% CO2, there occurs a sudden drop in PD to a slightly negative value associated with a fall in tissue resistance. This effect occurs under SCN-inhibition and thus seems unrelated to H+ transport per se. A working model is presented which can explain some of the events surrounding the sudden potential drop.

In vitro primate gastric mucosa: electrical characteristics

1980 ◽  
Vol 239 (2) ◽  
pp. G77-G82 ◽  
Author(s):  
S. Tripathi ◽  
P. K. Rangachari
Keyword(s):  
Gastric Mucosa ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Passive Permeability ◽  
Significant Edge ◽  
Sodium Dependent ◽  
Edge Damage ◽  
Net Fluxes ◽  
High Conductance

Ion transport by the resting, isolated, rhesus gastric mucosa was assessed under conditions of minimal diffusion limitation to oxygen by 1) the substitution of Na+ and Cl- of the bathing solutions with less permeant ions, 2) the drugs amiloride and ouabain, and 3) estimation of net fluxes of 22Na by methods designed to circumvent the problem of poorly matched tissues. The mucosae developed potential differences of 51.3 +/ 3.5 mV, serosal side positive and had conductances of 5.56 +/- 0.30 mS x cm-2. The permeabilities of the tissues to D-mannitol were between 7.80 x 10(-7) and 3.15 x 10(-7) cm x s-1. The relatively high conductance of this epithelium in the absence of significant edge damage and a low (32%) paracellular conductance stems mainly from a passive permeability to Cl-; active absorption of Na+ and active secretion of Cl- contribute equally to the short-circuit current. The mucosal entry step for Na+ is amiloride sensitive, whereas the serosal exit step can be inhibited by ouabain. The entry step for Cl- at the serosal membrane is possibly sodium dependent.

Effects of bicarbonate and pH on chloride transport by gastric mucosa

1982 ◽  
Vol 243 (1) ◽  
pp. G60-G68 ◽  
Author(s):  
E. C. Manning ◽  
T. E. Machen
Keyword(s):  
Gastric Mucosa ◽  
Intracellular Ph ◽  
Chloride Transport ◽  
Short Circuit ◽  
Ph Dependence ◽  
Short Circuit Current ◽  
Extracellular Ph ◽  
Solution Ph ◽  
Ph Changes

HCO3 and pH dependence of net Cl transport (JClnet) by resting (metiamide-treated) frog gastric mucosa has been investigated in vitro by measuring short-circuit current (Isc = JClnet) and transepithelial conductance (G). With either 100% O2 or 95% O2-5% CO2 gassing, HCO3-free solutions caused large (greater than 50%) reductions in Isc and G. Increases in [HCO3] of the serosal, but not mucosal, solution caused increases in Isc and G. At least part of the effect appeared to be due specifically to the HCO3 moiety, as opposed to the pH changes that also occurred. In HCO3-free solutions (100% O2), increasing serosal solution pH above 7 with either permeable or impermeable buffers caused Isc and G to increase; permeable buffers were somewhat more effective than impermeable buffers. Measurements of intracellular pH (pHc) with [14C]DMO or [14C]methylamine showed that increases in extracellular pH (pHo) caused increases in cellular pH (pHc), and these changes in pHc were independent of buffer type. We conclude that HCO3 and/or high pHo stimulate Isc and G and that buffer permeability and cellular concentration can also affect transport.

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