Transport of H plus and other electrolytes across isolated gastric mucosa of the rabbit.

The relationship of transmural and cellular phenomena to the rate of spontaneous luminal HH+ secretion by short-circuited gastric mucosa bathed in HCO3- and CO2-free Ringer solution was studied by the pH-stat technique. The bulk of luminal acidification can be accounted for by the appearance of H+ and not by organic acids. Acid secretion requires the presence of O2 and exogenous substrate but is not limited by endogenous CO2 production. The rate of H+ secretion is matched by the serosal appearance of alkali. The greater appearance of CO2 on the serosal side of the mucosa is consistent with hydroxylation of CO2 and greater permeability of the serosal border to HCO3-. Luminal changes in H+ concentrations affect H+ secretion but serosal changes do not, suggesting that the gradient produced by H+ secretion is at the luminal border. Steady-state isotopic Na+ and Cl- fluxes indicate net secretion of both ions, but net K+ transport is negligible.     

pH stat studies on bicarbonate secretion in the isolated mouse ileum

Bicarbonate secretion occurs in almost all segments of the gastrointestinal tract. This study examined HCO(3)(-) secretion in the ileum, since it is less understood than HCO(3)(-) secretion in other intestinal segments. Mouse ileal mucosa was mounted in vitro in Ussing chambers, and the mucosal alkalinization rate (J(OH)) was determined by pH stat titration, while the mucosal side was bathed with a buffer-free solution (100% O(2)) and the serosal side with a HCO(3)(-)/CO(2)-buffered solution. The transmural potential difference (PD) was recorded. The mucosal alkalinization rate (J(OH)) was higher in the presence of mucosal Cl(-) than in its absence. Forskolin, an activator of adenylate cyclase, enhanced J(OH) and PD in both the presence and absence of mucosal Cl(-). Mucosal SO(4)(2-) also caused an increase in J(OH), although the magnitude was smaller than that induced by Cl(-). Mucosal Cl(-)-dependent J(OH) was partially inhibited by acetazolamide, 5-nitro-2-(3-phenylpropylamino)-benzoic acid (NPPB), tenidap and probably also by niflumic acid, but not by glibenclamide, DIDS or bumetanide. The forskolin-induced J(OH) value and PD were both inhibited by NPPB and probably also by tenidap. It is concluded that HCO(3)(-)- secretion in the ileum follows a mucosal Cl(-)-dependent pathway and a cAMP-activated pathway, each being distinct from each other. The Cl(-)-dependent pathway is probably mediated by the slc26a6 anion exchanger, and possibly also by the slc26a3 anion exchanger. The cAMP-activated HCO(3)(-) secretion is probably mediated by the cystic fibrosis transmembrane conductance regulator.     

Effects of bicarbonate and pH on chloride transport by gastric mucosa

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.     

Intracellular pH regulation in guinea-pig caecal and colonic enterocytes during and after loading with short-chain fatty acids and ammonia

Absorption of short-chain fatty acids (SCFA) and ammonia implies considerable fluxes of protons across the epithelium of the large intestine. Efficient regulation of intracellular pH (pH(i)) is therefore essential in these cells. The aim of the present study was to examine the effects of SCFA and of ammonia on pH(i), on pH(i) regulation and to characterize the mechanisms involved in pH(i) regulation in surface enterocytes of the guinea-pig caecal and colonic mucosa. Intact epithelia from the caecum and the distal colon were mounted in a microperfusion chamber. pH(i) was measured by fluorescence microscopy using 2',7'-bis(2-carboxyethyl)-5(6)-carboxyfluorescein (BCECF). Addition of SCFA or ammonia to the serosal side changed the enterocyte pH(i) markedly, whereby ammonia caused larger changes in pH(i) than SCFA. In contrast, addition of SCFA to the mucosal solution had no effect on pH(i) and ammonia increased pH(i) only slightly. Basolaterally located pH(i) regulation mechanisms, Na(+)-H(+) exchange and Cl(-)-HCO(3)(-) exchange, are involved mainly in returning pH(i) to normal values. It is concluded that, due to apparently lower permeability of the apical membranes, the caecal and colonic epithelium is protected against pH(i) disturbances caused by the naturally high luminal SCFA and NH(3) concentrations. The major regulation mechanisms of pH(i) are located in the basolateral membrane of the enterocytes.     

The effect of carbon dioxide on the intracellular pH and buffering power of snail neurones.

1. Intracellular pH (pHi) was measured using pH-sensitive glass micro-electrodes. The effects on pHi of CO2 applied externally and HCO3-, H+ and NH4+ injected iontophoretically, were investigated. 2. The transport numbers for iontophoretic injection into aqueous micro-droples were found by potentiometric titration to be 0-3 for HCO3- and 0-94 for H+. 3. Exposure to Ringer, pH 7-5, equilibrated with 2-2% CO2 caused a rapid, but only transient, fall in pHi. Within 1 or 2 min pHi began to return exponentially to normal, with a time constant of about 5 min. 4. When external CO2 was removed, pHi rapidly increased, and then slowly returned to normal. The pHi changes with CO2 application or removal gave a calculated intracellular buffer value of about 30 m-equiv H+/pH unit per litre. 5. Injection of HCO3- caused a rise in pHi very similar to that seen on removal of external CO2. 6. The pHi responses to CO2 application, CO2 removal and HCO3- injection were slowed by the carbonic anhydrase inhibitor acetazolamide. 7. H+ injection caused a transient fall in pHi. In CO2 Ringer pHi fell less and recovered faster than in CO2-free Ringer. Calculation of the internal buffer value from the pHi responses to H+ and HCO3- injection gave very similar values. 8. The internal buffer value (measured by H+ injection) was greatly increased by exposure to CO2 Ringer. Acetazolamide reduced this effect of CO2, suggesting that the function of intracellular carbonic anhydrase may be to maximize the internal buffering power in CO2. 9. It was concluded that the internal HCO3- was determined primarily by the CO2 level and pHi, that internal HCO3- made a large contribution to the buffering power, and that after internal acidfication pHi was restored to normal by active transport of H+, OH- or HCO3- across the cell membrane. The active transport was much faster in CO2 than in CO2-free Ringer.     

Influence of extracellular bicarbonate on the short-circuit current and intracellular free calcium of human cultured sweat duct cells.

Transepithelial short-circuit current (Iscc) and intracellular free Ca2+ (Ca2+i) was studied in monolayers of cultured human sweat duct cells (CSDCs) in the presence or absence of HCO3- (and CO2) in the bathing solutions. Addition of HCO3- (and CO2) increased the control Iscc by more than 50%. The effect of HCO3- (and CO2) on Iscc was confined to the serosal bath. The HCO3- (and CO2) effect was also studied during stimulation with the cholinergic agonist methacholine (MCh), which in CSDC induces a complex response consisting of an initial Iscc and Ca2+i spike, which is independent of extracellular Ca2+, followed by regular Iscc and Ca2+i oscillations, which are absent during Ca(2+)-free bathing conditions. The sustained Iscc and Ca2+i oscillations, but not the initial Iscc and Ca2+i spike were abolished by the removal of extracellular HCO3- (and CO2). It is concluded that the Ca2+ influx and the Iscc in CSDCs are critically influenced by the presence of extracellular HCO3- (and CO2) in the bathing solutions.     

HCO3 – potentiates the cAMP-dependent secretory response of the human distal colon through a DIDS-sensitive pathway

We used the Ussing short-circuit technique to investigate the role of HCO3- in the adenosine 3',5'-cyclic monophosphate (cAMP)-dependent secretory response of the human distal colon. In HCO3(-)-free 4-(2-hydroxyethyl)-1-piperazineethanesulphonic acid (HEPES)-Ringer's, forskolin (10 mumol l-1 mucosal and serosal) evoked a sustained increase in short-circuit current (Isc) (delta Isc = 24 +/- 3 microA cm-2, n = 57). However, this was only 25% of the forskolin-stimulated Isc response in HCO3(-)-Ringer's (delta Isc = 84 +/- 8 microA cm-2, n = 57). The reduced response to forskolin in HCO3(-)-free HEPES-Ringer's was not due to inhibition of the secretory mechanism by HEPES, as replacing HCO3- with a different buffer, N-tris[hydroxymethyl)methyl-2-aminoethanesulphonic acid (TES), had a similar effect and inclusion of HEPES in the HCO3(-)-Ringer's did not reduce the secretory response. Similarly, it was not due to an indirect modulation of electrogenic Cl- secretion, as the forskolin-stimulated bumetanide-sensitive Isc was comparable in the two Ringer's. Rather it was due to the activation of a HCO3(-)-dependent Isc which was inhibited by serosal 4,4'-diisothiocyano-stilbene-2,2'-disulphonate (DIDS). This DIDS-sensitive Isc was not inhibited by acetazolamide, but it was inhibited by the replacement of bathing solution Cl- with gluconate, suggesting a role for a Na(+)-dependent Cl-/HCO3- exchanger in the cAMP-dependent secretory response of the human distal colon.     

Functional evidence for intracellular acid extruders in human ventricular myocardium

Intracellular pH (pH(i)) is a major homeostatic system within the cell. Changes in pH(i) exert great influence on cardiac contractility and rhythm. Both the housekeeping Na+ - H+ exchanger (NHE) and the Na+ - HCO3- symporter (NHS) have been confirmed as major transporters for the active acid extrusion mechanism in animal cardiomyocytes. However, whether the NHE and NHS functionally coexist in human ventricular cardiomyocytes remains unclear. We therefore examined the mechanism of pH(i) recovery following an NH4Cl-induced intracellular acidosis in the human ventricular myocardium. The pH(i) was monitored by microspectrofluorimetry by the use of intracellular 2',7'-bis(2-carboxyethyl)-5(6)-carboxy-fluorescein (BCECF)-fluorescence. HOE 694 (30 microM), a specific NHE inhibitor could block pH(i) recovery from induced intracellular acidosis completely in nominally HCO3- -free HEPES Tyrode solution, but it only partially inhibited the pH(i) recovery in 5% CO2/HCO3- Tyrode solution. In 5% CO2/HCO3- Tyrode solution, the addition of HOE 694 together with DIDS (an NHS inhibitor) or the removal of [Na+](o) could entirely inhibit the acid extrusion. We conclude for the first time that two different acid extruders, HCO3- -independent and -dependent, were most likely the NHE and NHS, respectively, that functionally coexisted in the human ventricular cardiomyocytes.     

Effects of CO2/HCO3- in perilymph on the endocochlear potential in guinea pigs

The effect of CO(2)/HCO(3)(-) on the endocochlear potential (EP) was examined by using both ion-selective and conventional microelectrodes and the endolymphatic or perilymphatic perfusion technique. The main findings were as follows: (i) A decrease in the EP from approximately +75 to approximately +35 mV was produced by perilymphatic perfusion with CO(2)/HCO(3)(-)-free solution, which decrease was accompanied by an increase in the endolymphatic pH (DeltapH(e), approximately 0.4). (ii) Perilymphatic perfusion with a solution containing 20 mM NH(4)Cl produced a decrease in the EP (DeltaEP, approximately 20 mV) with an increase in the pH(e) (DeltapH(e), approximately 0.2), whereas switching the perfusion solution from the NH(4)Cl solution to a 5% CO(2)/25 mM HCO(3)(-) solution produced a gradual increase in the EP to the control level with the concomitant recovery of the pH(e). (iii) The perfusion with a solution of high or low HCO(3)(-) with a constant CO(2) level within 10 min produced no significant changes in the EP. (iv) Perfusion of the perilymph with 10 microg/ml nifedipine suppressed the transient asphyxia-induced decrease in EP slightly, but not significantly. (v) By contrast, the administration of 1 microg/ml nifedipine via the endolymph inhibited significantly the reduction in the EP induced by transient asphyxia or perilymphatic perfusion with CO(2)/HCO(3)(-)-free or 20 mM NH(4)Cl solution. These findings suggest that the effect of CO(2) removal from perilymphatic perfusion solution on the EP may be mediated by an increase in cytosolic Ca(2+) concentration induced by an elevation of cytosolic pH in endolymphatic surface cells.     

Chloride-dependent bicarbonate secretion in the mouse large intestine

The gastrointestinal HCO(3)(-) secretion functions to limit the mucosal acid damage due to HCl secreted in the stomach or organic acids produced in the large intestine. We studied HCO(3)(-) secretion in the mouse large intestine with isolated tissues mounted in chambers by using pH stat method. Addition of Cl(-) to the mucosal side caused an increase in HCO(3)(-) secretion in the cecum and distal colon but had little, if any, effect in the proximal colon. In agreement with this, mucosal surface pH was higher in the cecum and distal colon than in the proximal colon. The Cl(-)-induced HCO(3)(-) secretion in the cecum was inhibited by 5-nitro-2-(3-phenylpropylamino)benzoic acid (NPPB, mucosal addition), but not by DIDS (mucosal or serosal), acetazolamide, amiloride (serosal) or glibenclamide (mucosal). Removal of Na(+) or addition of propionate had hardly any effect on the Cl(-)-induced HCO(3)(-) secretion. These results suggest that a NPPB-sensitive, DIDS-resistant Cl(-)/ HCO(3)(-) exchanger is present in the apical membrane, and mediates Cl(-)-dependent HCO(3)(-) secretion. This process is probably mainly responsible for the formation of the high pH at the mucosal surface.     

Intracellular pH transients in rat diaphragm muscle measured with DMO.

Changes of the intracellular pH of rat diaphragm muscle were monitored at 30-min intervals with the weak acid DMO (5,5-dimethyl-2,4-oxazolidinedione). Transferring the muscle from a CO2-containing to a CO2-free solution caused intracellular pH (pHi) to rise by an average of 0.18 during the first 30 min and then to level off at a slightly lower value over the next 60-90 min. Transferring the muscle from a CO2-free to a CO2-containing solution caused pHi to fall by 0.18 during the first 30 min and then to recover by 0.05 over the next 90 min. Subsequent return to the CO2-free solution caused pHi to overshoot the control value by 0.10. Both the recovery and the overshoot can be accounted for by an acid-extruding pump. Intracellular acid loading with 118 mM DMO similarly caused pHi to fall initially, to recover slowly during the acid loading, and then to overshoot the control pHi on removal of the acid load. In the absence of HCO3-/CO2, acid extrusion was reduced by about a fifth. SITS (4-acetamido-4'-isothiocyanostilbene-2,2'-disulfonic acid) had no effect. The absence of either Na+ or Cl- from HCO3-/CO2- free solution reduced acid extrusion by about a half.     

Propionate-induced epithelial K+ and Cl?/HCO 3 ? secretion and free fatty acid receptor 2 (FFA2, GPR43) expression in the guinea pig distal colon

Propionate, a fermented product in the lumen of the large intestine, is a short-chain fatty acid (SCFA) known to have a variety of localized physiological and pathophysiological functions (e.g., luminal fluid secretion and anti-inflammatory response). In the present study, we investigated propionate-induced transepithelial ion transport and the expression of SCFA receptor, free fatty acid receptor 2 (FFA2, otherwise known as GPR43) in the guinea pig distal colon utilizing the Ussing chamber technique and immunohistochemistry. The addition of propionate to the luminal bathing solution concentration-dependently induced transient K(+) and Cl(-) and/or bicarbonate secretion within approximately 30?s and long-lasting Cl(-) secretion for approximately 60?min was first identified in the present study. The transient anion secretion was tetrodotoxin (TTX)-sensitive and mediated through the cholinergic (both nicotinic and muscarinic) neural pathway, but the transient K(+) and long-lasting Cl(-) secretion were due to TTX-insensitive mechanism. Immunohistochemistry studies showed that some chromogranin A-immunoreactive enteroendocrine cells were also immunoreactive for FFA2 but not colocalized with 5-hydroxytryptamine. In conclusion, the propionate-induced secretion consisted of the neural and non-neural three-phase secretory manner possibly mediated by the stimulation of FFA2 expressed by enteroendocrine cells.     

Concentration dependence of bicarbonate-induced calcium current modulation

High-voltage-activated calcium currents (HVA) of CA1 neurons are prominently attenuated following a switch from HEPES-buffered solution to one buffered with CO(2)/HCO(3)(-). In the present study we investigated whether bicarbonate ions or the dissolved CO(2) induce this alteration in current characteristic. The study was carried out on freshly isolated CA1 neurons using the whole cell patch-clamp technique. Maximal calcium conductance and the mean peak amplitude of the currents showed a concentration-dependent decrease when cells were consecutively bathed in solutions containing increasing amounts of bicarbonate and CO(2). This decrease is best described by the Hill equation, yielding a maximal attenuation of 69%, a half-maximal concentration (EC(50)) of 7.4 mM HCO(3-), and a Hill coefficient of 1.8. In parallel, the potentials of half-maximal activation (V(h,a)) and inactivation (V(h,i)) were linearly shifted in hyperpolarizing direction with a maximal shift, in the 10% CO(2)/37 mM HCO(3)(-) containing solution of 10 +/- 1 mV for V(h,a) (n = 23) and 17 +/- 1.4 mV for V(h,i) (n = 18). When currents were evoked in solutions containing equal concentrations of bicarbonate but different amounts of CO(2), only nonsignificant changes were observed, while marked alterations of the currents were induced when bicarbonate was changed and CO(2) held stable. The experiments suggest that bicarbonate is the modulating agent and not CO(2). This bicarbonate-induced modulation may be of critical relevance for the excitation level of the CNS under pathological situation with altered concentration of this ion, such as hyperventilation and metabolic acidosis.     

Secretion of fluid and amylase in the perfused rat pancreas.

1. The isolated rat pancreas was perfused with physiological salt solutions of varying composition. Flow of pancreatic juice and output of amylase during rest and after stimulation with pure secretin, pure cholecystokinin-pancreozymin (CCK-PZ), caerulein or acetylcholine (ACh) were measured. 2. Basal fluid secretion was abolished replacing perfusion fluid NA+ or Cl- by Tris+ or SO42- respectively. Readmission of Na+ or Cl- caused a transient increase above the normal control level of both fluid and amylase output. Exposure to K+-free solution severely reduced fluid output and K+ readmission resulted in a transient increase in secretory rate. 3. Maximal stimulation with ACh (10(-7) M), CCK-PZ (1-5 X 10(-10) M) or caerulein (10(-10) M) caused marked sustained fluid and amylase secretion. Maximal secretin stimulation (5-7 X 10(-9) M) caused marked sustained fluid but only a small sustained amylase secretion following an initial transient. 4. Under continuous secretin stimulation, replacement of the CO2/HCO3-buffered control fluid by a CO2/HCO3-free Tris buffered solution caused a sharp decrease in pancreatic juice flow. In the absence of extracellular CO2/HCO3-secretin did not evoke fluid or enzyme secretion. In contrast the effects of ACh, CCK-PZ or caerulein were independent on CO2/HCO3-. Monobutyryl cyclic AMP (10(-3) M) caused marked sustained fluid secretion and transient enzyme secretion. The effect was entirely dependent on the presence of CO2/HCO3-in the perfusion fluid. 5. Ouabain (10(-4)-10(-3) M) markedly inhibited both secretin- and caerulein-evoked fluid secretion while caerulein-evoked amylase secretion was hardly affected. Similar findings were made with K+-free solution. 6. The effect of maximal secretin stimulation on amylase secretion was greatly augmented in the presence of a maximally stimulating concentration of caerulein. The effects on fluid secretion of secretin and caerulein were simply additive. The effects of secretin on both amylase and fluid secretion, in the presence of caerulein, were entirely dependent on the presence of CO2/HCO3- in the perfusion fluid. 7. We conclude that two different fluid secretion processes occur in the rat exocrine pancreas. One stimulated by ACh and CCK-PZ, that is independent of extracellular CO2/HCO3- and another stimulated by secretin involving H+ or HCO3-transport. Only the effects of secretin seem to be mediated by intracellular cyclic AMP.     

Response of Amphiuma small intestine to theophylline: effect on bicarbonate transport.

The electrolyte transport properties of isolated proximal segments of Amphiuma small intestine and their response to theophylline were observed under various conditions. In the absence of theophylline the intestine generates a transepithelial potential (psi ms) serosa negative to mucosa when Cl- and HCO3- are present in the bath. Replacement of Cl- or HCO3- reduced the magnitude and usually reversed the sign of psi ms. Acetazolamide (10(-4) M) nearly abolished the serosa negative psi ms. Theophylline (10 mM) drove psi ms serosa positive, the magnitude depending on the bath Na+ and HCO3- concentrations. Simultaneously it increased the short-circuit current (Isc) and tissue resistance (Rt). The increase in Isc was not due to increase net Na+ transport in Cl-free buffer and was attributed to a residual ion flux. Acetazolamide reduced the Isc, Rt, and the net residual flux observed in theophylline-treated intestine. The magnitude of the acetazolamine effect on Isc was proportional to the Na+ and HCO3- concentrations of the bath. The results suggest that in the absence of theophylline, HCO3-, and Cl- transport are related. Furthermore, acetazolamide inhibits the movement of an ion, possibly HCO3-, secreted in response to theophylline.     

Coupling between chloride absorption and base excretion in isolated skin of Rana esculenta.

The net excretory fluxes of base (HCO3- or OH-) and the unidirectional fluxes of chloride were measured and their relationship examined in isolated frog skin maintained in open- or short-circuit (OC and SC) conditions. When the mucosal solution was a 2 mM choline chloride solution and the serosal solution a Ringer solution buffered with a HCO3-/CO2 mixture, the rate of base excretion was -105 +/- 10 in OC and -60 +/- 7 neq h-1 cm-2 in SC. A highly significant correlation was observed between the influx of chloride and the excretion of base. As a function of external chloride both these parameters followed saturation kinetics, Vmax being obtained for a chloride concentration below 2 mM. The removal of chloride in the external solution was followed by a 70 or 100% inhibition of base excretion in OC and SC conditions, respectively. Chloride transport is dependent on the presence of a HCO3-/CO2 mixture in the internal or the external medium. This transport, as well as base excretion, is inhibited to a considerable extent by removal of HCO3-/CO2 or by acetazolamide (10(-3) M). This investigation characterizes a saturable transport system in which chloride absorption and base excretion are coupled.     

The underlying cellular mechanism in the effect of tetramethylpyrazine on the anion secretion of colonic mucosa

The present study investigated the underlying cellular mechanism in the effect of ligustrazine (tetramethylpyrazine, TMP) on the anion secretion of colonic mucosa in rats using a short-circuit current (I(sc)) technique in conjunction with "tool drugs." (i) After a pretreatment of the tissues by bathing the bilateral surface with Cl(-)-free Krebs-Henseleit (K-H) solution for over an hour, a basolateral application of 1 mmol/l TMP produced an increase in I(sc), and the total charges transported for 30 min were about 8.7 +/- 1.4 mC/cm(2); an apical pretreatment of DPC and a basolateral addition of acetazolamide decreased the TMP-induced I(sc) by about 60% (P < 0.01) and 45% (P < 0.05), respectively; a basolateral application of 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS), the inhibitor of Na(+)-HCO(3)(-) cotransporter (NBC), did not alter the TMP-induced I(sc). (ii) After the bilateral surface of mucosa was bathed with HCO(3)(-)-free K-H solution for over an hour, a basolateral application of 1 mmol/l TMP produced an increase in I(sc), and the total charges transported in 30 min were about 8.3 +/- 1.9 mC/cm(2); an apical pretreatment of DPC (1 mmol/l), the inhibitor of Cl(-) channels, decreased the TMP-induced Isc by about 84% (P < 0.01). The basolateral presence of bumetanide (0.1 mmol/l), the inhibitor of Na(+)-K(+)-Cl(-) cotransporter (NKCC), significantly reduced the TMP-evoked I(sc) by about 86% (P < 0.01). In conclusion, (i) ligustrazine could promote colonic mucosa secretion Cl(-) via apical Cl(-) channels and basolateral NKCC; (ii) ligustrazine could promote colonic mucosa secretion HCO(3)(-) via apical Cl(-) channels and the basolateral diffusion of CO(2).     

Functional characterization of Cl-/HCO3- exchange in villous cells of the mouse ileum

At least three kinds of Cl(-)/HCO(3)(-) exchangers, SLC26A3, SLC26A6 and AE2, have been demonstrated to be expressed in the intestinal epithelial cell. To examine the functional expression of these exchangers in the native enterocyte, we studied the Cl(-)/HCO(3)(-)- exchange activity in isolated villi from the mouse ileum by microfluorometric intracellular pH (pH(i)) measurement. The pH(i) value increased upon Cl(-) removal when the villus was superfused with an HCO(3)(-)/CO(2)-buffered solution, while the response was blunted when superfused with an HCO(3)(-)/CO(2)-free, Hepes-buffered solution. The recovery of pH(i) value induced by Cl(-) re-addition (after initial Cl(-) removal) was totally or partially mimicked by the addition of Br(-), I(-), F(-), NO(3)(-), or SO(4)(2-) (in the absence of Cl(-)). The increase in pH(i) value induced by Cl(-) removal was partially inhibited in the presence of DIDS (30 muM), tenidap (10 muM), niflumic acid (30 muM) or NPPB (30 muM). Increasing the K(+) concentration from 5 mM to 60 mM in the superfusion solution induced a reversible increase in pH(i) value under the HCO(3)(-)/CO(2)-buffered condition, while it had hardly any effect on pH(i) under the Hepesbuffered condition. The K(+)-induced pH(i) changes were partially suppressed by removing Cl(-) from the superfusion solution. These results, together with the reported findings of mouse slc26a3, slc26a6 and AE2 in heterologously expressed systems, suggest the possibility that these three exchangers may all be functionally expressed in mouse ileal villous cells.     

In vitro potassium transport in the mouse small intestine

Ingested K+ is believed to be absorbed mainly in the small intestine by passive diffusion through the paracellular pathway. To further clarify K+ absorption in the small intestine, we determined the unidirectional flux values of Rb+ in vitro by atomic absorption spectroscopy in the mouse ileum mounted in Ussing chambers under short-circuit conditions. The mucosal-to-serosal Rb+ flux (J(ms)) was larger than the serosal-to-mucosal Rb+ flux (J(sm)), resulting in positive net Rb+ absorption (J(net)). The effect of changing the transmucosal potential (V(t)) showed that J(ms) was composed of both a V(t)-dependent diffusion component and a V(t)-independent non-diffusion component, while J(sm) was composed mainly of a V(t)-dependent component. A forskolin treatment eliminated J(net) mainly due to the increase in J(sm). When animals were fed a low-Na diet, J(net) was mainly eliminated as a result of the increase in J(sm). These findings suggest that K+ is absorbed not only by passive diffusion through the paracellular pathway, but also by an active transport mechanism operating through the cellular pathway. In addition, cAMP and aldosterone may be involved in regulating intestinal K+ transport.     

Studies of the urinary acidification defect induced by lithium

Experiments were carried out in rats and isolated turtle bladders to study the defect in H+ transport induced by LiCl. After 3-4 days of intraperitoneal LiCl, rats developed urinary findings of "distal" renal tubular acidosis. Proximal tubular fluid pH measured in situ by glass microelectrodes was higher in lithium-treated rats than in acidotic controls. Proximal fluid total CO2 [tCO2] was also higher, and the fraction of tCO2 leaving the proximal tubule was 14 vs. 7% (P less than 0.001). Impaired acidification was also apparent beyond distal convoluted tubules, as judged by normal distal tCO2 reabsorption but increased HCO3(-) in the urine. During NaHCO3 loading, the proximal defect was ameliorated but not the distal. Turtle bladder studies showed that mucosal lithium inhibits H+ secretion secondary to reducing transepithelial electrical potential, presumably by hyperpolarization of the luminal membrane. A similar mechanism may be responsible for lithium's effect on the distal nephron. Inhibition of proximal tubular HCO3(-) reabsorption is probably not attributable to electrical potential changes but might be due to interference of luminal membrane Na+ entry by Li+ and reduced (Na+ + Li+)-H+ exchange.