Luminal CO2 Research Articles

Duodenal Luminal Chemosensing; Acid, ATP, and Nutrients

Intestinal chemosensing of endogenous and exogenous luminal compounds, including acid, CO2, bile acids and nutrients is an emerging area of gastrointestinal research, since gut hormones, particularly including incretins and glucagon-like peptide-2 (GLP-2) are released in response to luminal nutrients. Identification of luminal chemosensors such as nutrient-ligand G-protein coupled receptors (GPCRs) in enteroendocrine cells has linked luminal compounds to the corresponding gut hormone release. Mucosal chemical sensors are necessary to exert physiological responses such as secretion, digestion, absorption, and motility. We have been studying the mechanisms by which luminal compounds are sensed via mucosal acid sensors and GPCRs, which trigger mucosal defense mechanisms. In addition to luminal acid/CO2 sensing in the duodenum, recent studies also show that compounds present post-prandially such as amino acids, bile acids and fatty acids, enhance duodenal mucosal defenses, with digestion following the initial gastric processing. These studies may form the basis for therapies in which luminal nutrients release gut hormones that affect the mucosal protection, appetite, satiety, and systemic metabolisms.

The distinct roles of anion transporters Slc26a3 (DRA) and Slc26a6 (PAT-1) in fluid and electrolyte absorption in the murine small intestine

The mixing of gastric and pancreatic juice subjects the jejunum to unique ionic conditions with high luminal CO2 tension and HCO3 − concentration. We investigated the role of the small intestinal apical anion exchangers PAT-1 (Slc26a6) and DRA (Slc26a3) in basal and CO2/HCO3 −-stimulated jejunal fluid absorption. Single pass perfusion of jejunal segments was performed in anaesthetised wild type (WT) as well as in mice deficient in DRA, PAT-1, Na+/H+ exchanger 3 (NHE3) or NHE2, and in carbonic anhydrase II (CAII). Unbuffered saline (pH 7.4) perfusion of WT jejunum resulted in fluid absorption and acidification of the effluent. DRA-deficient jejunum absorbed less fluid than WT, and acidified the effluent more strongly, consistent with its action as a Cl−/HCO3 − exchanger. PAT-1-deficient jejunum also absorbed less fluid but resulted in less effluent acidification. Switching the luminal solution to a 5 % CO2/HCO3 − buffered solution (pH 7.4), resulted in a decrease in jejunal enterocyte pHi in all genotypes, an increase in luminal surface pH and a strong increase in fluid absorption in a PAT-1- and NHE3- but not DRA-, CAII, or NHE2-dependent fashion. Even in the absence of luminal Cl−, luminal CO2/HCO3 − augmented fluid absorption in WT, CAII, NHE2- or DRA-deficient, but not in PAT-1- or NHE3-deficient mice, indicating the likelihood that PAT-1 serves to import HCO3 − and NHE3 serves to import Na+ under these circumstances. The results suggest that PAT-1 plays an important role in jejunal Na+HCO3 – reabsorption, while DRA absorbs Cl− and exports HCO3 − in a partly CAII-dependent fashion. Both PAT-1 and DRA significantly contribute to intestinal fluid absorption and enterocyte acid/base balance but are activated by different ion gradients.Electronic supplementary materialThe online version of this article (doi:10.1007/s00424-013-1381-2) contains supplementary material, which is available to authorized users.

Tonometry as a predictor of inadequate splanchnic perfusion in an intra-abdominal hypertension animal model

BackgroundThe gastrointestinal system is the most sensitive to the presence of intra-abdominal hypertension. We aimed to assess the early prognostic value of gastric air tonometry as a predictor of inadequate splanchnic perfusion and determine its relation with abdominal perfusion pressure (APP). MethodsTwenty-five Large White swine were used for this study. A control group and two study groups were included, in which intra-abdominal pressure (IAP) was elevated with Co2 to 20 and 30 mmHg during 5 h. We measured the intramucosal gastric pH (pHim) and determined gastric luminal PCO2 (PgCO2) and PgCO2gap (gastric luminal CO2–arterial CO2) to evaluate gastric acidity. APP was indirectly obtained through IAP and mean arterial pressure. Additionally, histopathologic samples of small intestine were obtained and analyzed. ResultspHim showed a decrease in IAP groups, with statistical significance in the 30 mmHg group, 90 min after stabilization period (P < 0.01). Serum lactate showed delayed alteration when compared with pHim, with significant increase, 180 min after stabilization (P < 0.05). The values of PgCO2 and PCO2gap were increased in IAP groups, being statistically significant in the 30 mmHg group, 120 and 150 min, respectively, after stabilization. In increased IAP groups, there was a time progressive decrease of APP, with statistically significant differences observed between groups at 20 min (P < 0.001). The histopathology study revealed parenchymal injury of the intestine at 30 mmHg. ConclusionsTonometry is sensitive to the increase in IAP and relates to the reduction of APP generated by splanchnic hypoperfusion.

Duodenal epithelial sensing of luminal acid: role of carbonic anhydrases

Sensing the luminal contents is a prerequisite to activate appropriate gastrointestinal functions. A major task of the duodenal epithelium is to resist the repeated challenges of hydrochloric acid expelled from the stomach. Although extensive research in this field, the complete mechanisms providing this defence remain to be revealed. The duodenal epithelium exports bicarbonate into a submillimetre-thick mucus gel on top of the mucosal surface. Despite the very low pH of the luminal contents, the duodenal mucus-bicarbonate barrier provides a means of maintaining a virtually neutral pH at the epithelial surface. Instead of pH, CO₂ generated by the mixing of acid and bicarbonate at levels not found elsewhere in the body serves as the mediator for sensing the luminal acid. Carbonic anhydrases (CAs) catalyse the reversible hydration of CO₂ and are heavily expressed in the duodenal segment. Accumulating data support the key function of CAs in sensing luminal acid and CO₂. Recent advances demonstrate that the presence of CA II in upper villus plays a crucial role in enterocyte intracellular acidification preceding the secretory increase in response to luminal acid. However, CAs only have a minor role in the bicarbonate supply destined for duodenal bicarbonate secretion into the lumen. The purpose of this review is to summarize the current knowledge of how intraluminal acid is sensed by the duodenal mucosa, with a focus on the role of CAs.

Lafutidine, a Protective H2 Receptor Antagonist, Enhances Mucosal Defense in Rat Esophagus

BackgroundLuminal acid or CO2 induces a hyperemic response in the esophagus, via activation of acid sensors on capsaicin-sensitive afferent nerves (CSAN). Since disruption of the hyperemic response to luminal CO2 acidifies the interstitium of the esophageal mucosa, the hyperemic response may maintain interstitial pH (pHint). We hypothesized that acid-related hyperemia maintains pHint, preventing acid-induced injury in the esophageal mucosa.MethodsWe examined the effects of capsaicin (Cap) or lafutidine (Laf), a mucosal protective H2 antagonist, on the regulation of pHint and blood flow in rat esophagus using ratiometric microimaging and laser-Doppler measurements of the lower esophageal mucosa of living rats. The esophagus was topically superfused with pH 7.0 buffer, or a pH 1.0 or pH 1.0 + pepsin (1 mg/ml) solution with or without Laf.ResultsCap (30 or 100 µM) or Laf (0.1 or 1 mM) dose-dependently increased blood flow, accompanied by increased pHint. The pH 1.0 solution increased blood flow without pHint change, whereas Laf (1 mM) increased blood flow and pHint during acid exposure. The effects of Laf were abolished by ablation of CSAN. Perfusion of the acidified pepsin solution gradually decreased pHint, inhibited by Laf perfusion.ConclusionsActivation of CSAN by Laf with or without acid, accompanied by hyperemia, increased pHint, preventing acidified pepsin-induced interstitial acidification. Stimulation of the capsaicin pathway with compounds such as Laf enhances mucosal protection from acid-related injury in the upper gastrointestinal tract.

Duodenal acidity “sensing” but not epithelial HCO 3 − supply is critically dependent on carbonic anhydrase II expression

Carbonic anhydrase (CA) is strongly expressed in the duodenum and has been implicated in a variety of physiological functions including enterocyte HCO(3)(-) supply for secretion and the "sensing" of luminal acid and CO(2). Here, we report the physiological role of the intracellular CAII isoform involvement in acid-, PGE(2,) and forskolin-induced murine duodenal bicarbonate secretion (DBS) in vivo. CAII-deficient and WT littermates were studied in vivo during isoflurane anesthesia. An approximate 10-mm segment of the proximal duodenum with intact blood supply was perfused under different experimental conditions and DBS was titrated by pH immediately. Two-photon confocal microscopy using the pH-sensitive dye SNARF-1F was used to assess duodenocyte pH(i) in vivo. After correction of systemic acidosis by infusion of isotonic Na(2)CO(3), basal DBS was not significantly different in CAII-deficient mice and WT littermates. The duodenal bicarbonate secretory response to acid was almost abolished in CAII-deficient mice, but normal to forskolin- or 16,16-dimethyl PGE(2) stimulation. The complete inhibition of tissue CAs by luminal methazolamide and i.v. acetazolamide completely blocked the response to acid, but did not significantly alter the response to forskolin. While duodenocytes acidified upon luminal perfusion with acid, no significant pH(i) change occurred in CAII-deficient duodenum in vivo. The results suggest that CA II is important for duodenocyte acidification by low luminal pH and for eliciting the acid-mediated HCO(3)(-) secretory response, but is not important in the generation of the secreted HCO(3)(-) ions.

CO2 chemosensing in rat oesophagus

Background:Acid in the oesophageal lumen is often sensed as heartburn. It was hypothesised that luminal CO2, a permeant gas, rather than H+, permeates through the epithelium, and is converted to...

CFTR inhibition augments NHE3 activity during luminal high CO2 exposure in rat duodenal mucosa.

We hypothesized that the function of duodenocyte apical membrane acid-base transporters are essential for H(+) absorption from the lumen. We thus examined the effect of inhibition of Na(+)/H(+) exchanger-3 (NHE3), cystic fibrosis transmembrane regulator (CFTR), or apical anion exchangers on transmucosal CO(2) diffusion and HCO(3)(-) secretion in rat duodenum. Duodena were perfused with a pH 6.4 high CO(2) solution or pH 2.2 low CO(2) solution with the NHE3 inhibitor, S3226, the anion transport inhibitor, DIDS, or pretreatment with the potent CFTR inhibitor, CFTR(inh)-172, with simultaneous measurements of luminal and portal venous (PV) pH and carbon dioxide concentration ([CO(2)]). Luminal high CO(2) solution increased CO(2) absorption and HCO(3)(-) secretion, accompanied by PV acidification and PV Pco(2) increase. During CO(2) challenge, CFTR(inh)-172 induced HCO(3)(-) absorption, while inhibiting PV acidification. S3226 reversed CFTR(inh)-associated HCO(3)(-) absorption. Luminal pH 2.2 challenge increased H(+) and CO(2) absorption and acidified the PV, inhibited by CFTR(inh)-172 and DIDS, but not by S3226. CFTR inhibition and DIDS reversed HCO(3)(-) secretion to absorption and inhibited PV acidification during CO(2) challenge, suggesting that HCO(3)(-) secretion helps facilitate CO(2)/H(+) absorption. Furthermore, CFTR inhibition prevented CO(2)-induced cellular acidification reversed by S3226. Reversal of increased HCO(3)(-) loss by NHE3 inhibition and reduced intracellular acidification during CFTR inhibition is consistent with activation or unmasking of NHE3 activity by CFTR inhibition, increasing cell surface H(+) available to neutralize luminal HCO(3)(-) with consequent CO(2) absorption. NHE3, by secreting H(+) into the luminal microclimate, facilitates net transmucosal HCO(3)(-) absorption with a mechanism similar to proximal tubular HCO(3)(-) absorption.

Assessment of a new prototype hydrogel CO 2 sensor; comparison with air tonometry

Gastrointestinal ischemia is always accompanied by an increased luminal CO(2). Currently, air tonometry is used to measure luminal CO(2). To improve the response time a new sensor was developed, enabling continuous CO(2) measurement. It consists of a pH-sensitive hydrogel which swells and shrinks in response to luminal CO(2), which is measured by the pressure sensor. We evaluated the potential clinical value of the sensor during an in vitro and in vivo study. The response time to immediate, and stepwise change in pCO(2) was determined between 5 and 15 kPa, as well as temperature sensitivity between 25 and 40 degrees C at two pCO(2) levels. Three sensors were compared to air tonometry (Tonocap) in healthy volunteers using a stepwise incremental exercise test, followed by a period of hyperventilation and an artificial CO(2)-peak. The in vitro response time to CO(2) increase and decrease was mean 5.9 and 6.6 min. The bias, precision and reproducibility were +5%, 3% and 2%, resp. Increase of 1 degrees C at constant pCO(2) decreased sensor signal by 8%. In vivo tests: The relation with the Tonocap was poor during the exercise test. The response time of the sensor was 3 min during hyperventilation and the CO(2) peak. The hydrogel carbon dioxide sensor enabled fast and accurate pCO(2) measurement in a controlled environment but is very temperature dependent. The current prototype hydrogel sensor is still too unstable for clinical use, and should therefore be improved.

Carbonic Anhydrases and Mucosal Vanilloid Receptors Help Mediate the Hyperemic Response to Luminal CO2 in Rat Duodenum

The duodenal mucosa is exposed to PCO(2) >200 mm Hg due to the luminal mixture of gastric acid with secreted bicarbonate, which augments mucosal protective mechanisms. We examined the hyperemic response to elevated luminal PCO(2) in the duodenum of anesthetized rats luminally exposed to high CO(2) saline to help elucidate luminal acid-sensing mechanisms. Blood flow was measured by laser Doppler, and intracellular pH of epithelial cells by measured by ratio microimaging. The permeant carbonic anhydrase (CA) inhibitor methazolamide, relatively impermeant CA inhibitor benzolamide, vanilloid receptor antagonist capsazepine, or sodium-hydrogen exchanger 1 (NHE-1) inhibitor dimethyl amiloride were perfused with or without the high CO(2) solution. The high CO(2) solution increased duodenal blood flow, which was abolished by pretreatment with methazolamide or capsazepine or by dimethyl amiloride coperfusion. Sensory denervation with capsaicin also abolished the CO(2) effects. Benzolamide dose-dependently inhibited CO(2)-induced hyperemia and at 100 nmol/L inhibited CO(2)-induced intracellular acidification. The membrane-bound CA isoforms IV, IX, XII, and XIV and cytosolic CA II and the vanilloid receptor 1 (TRPV1) were expressed in duodenum and stomach. Dorsal root ganglion and nodose ganglion expressed all isoforms except for CA IX. The duodenal hyperemic response to luminal CO(2) is dependent on cytosolic and membrane-bound CA isoforms, NHE-1, and TRPV1. CO(2)-induced intracellular acidification was inhibited by selective extracellular CA inhibition, suggesting that CO(2) diffusion across the epithelial apical membrane is mediated by extracellular CA. NHE-1 activation preceding TRPV1 stimulation suggests that luminal CO(2) is sensed as H(+) in the subepithelium.

Blood in the gastrointestinal tract delays and blunts the PCO2 response to transient mucosal ischaemia.

To determine whether the presence of blood in the gastrointestinal tract impairs the detection of splanchnic mucosal ischaemia by reducing the rate and magnitude of rise in gut luminal PCO2. Prospective observational study. University Research Laboratory. Four adult male Wistar rats with four controls from a concurrent study. Four adult male Wistar rats were anaesthetised with sodium pentobarbitone and ventilated with oxygen and isoflurane to an initial PaCO2 of 30-40 torr. Electrochemical-fiberoptic gas sensors inserted into silastic tubing placed in the ileal lumen measured luminal PCO2 at 1-s intervals. Distal aortic pressure was monitored continuously. Six 2-min episodes of aortic hypotension were induced in each rat to a mean pressure of 30 mmHg by intermittent elevation of a silk sling placed around the proximal aorta. Before the last three episodes in each rat 0.75-1.0 ml blood was injected into the ileal lumen via a 25-gauge needle. Four control rats from a concurrent experiment were treated in an identical fashion except that the rats were subjected to five ischaemic episodes, and there was no intraluminal injection of blood, although a 20-gauge cannula was placed in the proximal ileal lumen. The presence of blood in the lumen significantly decreased the ischaemic deltaPCO2 response from 33 +/- 8 to 15 +/- 4 torr (P < 0.001) and also significantly increased the peak response time from 188 +/- 12 to 227 +/- 24 s (P < 0.001). The corresponding measurements in the concurrent controls differed only from the values after blood injection in the experimental group. In this animal model the presence of intraluminal blood significantly delayed the rate and the amplitude of luminal PCO2 increases in response to transient ischaemia. This raises questions about the validity of luminal CO2 measurements as an indicator of splanchnic ischaemia in the presence of gastrointestinal haemorrhage.

Directly measured tissue pH is an earlier indicator of splanchnic acidosis than tonometric parameters during hemorrhagic shock in swine

To compare tissue pH in the stomach, bowel, and abdominal wall muscle during hemorrhagic shock and recovery using tissue electrodes; also, to compare tissue electrode pH measurements to gastric intramucosal pH (pHi), gastric luminal PCO2, and PCO2 gap (gastric luminal CO2--arterial CO2) measured with an air-equilibrated tonometer. Prospective animal study. University animal research laboratory. Eight anesthetized, mechanically ventilated Yorkshire swine. Hemorrhagic shock was initiated by withdrawing blood over a 15-min period to lower systolic blood pressure to 45 mm Hg. Shock was maintained for 45 mins and was followed by a 5-min resuscitation to normal blood pressure with a blood/lactated Ringer's (1:2) mixture. Recovery was monitored for 60 mins. pH was measured with electrodes in the submucosa of the stomach, the submucosa of the small bowel, and the abdominal wall muscle. Gastric luminal PCO2 was measured with an air-equilibrated tonometer and pHi and PCO2 gap were calculated. Each organ showed a different sensitivity to shock and resuscitation. The bowel pH responded most rapidly to the onset of hemorrhagic shock and had the largest change in tissue pH. The bowel also showed the most rapid recovery during resuscitation. The submucosal pH of the stomach responded more slowly than the bowel, but faster than the abdominal wall muscle pH, gastric PCO2 gap, or pHi. The smallest changes in organ pH as a result of hemorrhagic shock were seen in the abdominal wall muscle and the stomach as assessed by gastric tonometry. Direct measurement of tissue pH indicates that intra-abdominal organ pH varies during hemorrhagic shock. The small bowel pH changes the most in magnitude and rapidity compared with stomach pH or abdominal wall muscle pH. Tonometrically derived parameters were not as sensitive in the detection of tissue acidosis during shock and resuscitation as pH measured directly in the submucosa of the stomach or small bowel.

Subcutaneous oxygen tensions provide similar information to ileal luminal CO2 tensions in an animal model of haemorrhagic shock.

The cutaneous and splanchnic circulations undergo early vasoconstriction in shock. Methodological problems and insufficient information on subcutaneous carbon dioxide partial pressures limit the usefulness of previous studies on splanchnic and subcutaneous gas tensions in shock. Little comparative data exist on the responses of these tissues to shock and resuscitation. We therefore compared continuous subcutaneous PO2 (PO2sc) and PCO2 (PCO2sc) with simultaneous continuous gut luminal PCO2 (PCO2gi) in an animal model of haemorrhagic shock and resuscitation. Prospective observational study. Intensive care laboratory in a teaching hospital. Five anaesthetised rats. Electrochemical-fiberoptic gas sensors inserted into Silastic tubing placed in the subcutaneous tissue and in the ileal lumen measured PCO2sc, PO2sc and PCO2gi continuously in five anaesthetised rats. After steady state conditions, hypotension [mean arterial blood pressure (MAP) 40 mmHg] was induced by controlled haemorrhage. The rats were allowed to remain hypotensive for 15 min and then resuscitated with shed blood and crystalloids. Arterial plasma lactate concentrations were measured at defined periods during the study. Hypovolaemia resulted in a significant decrease in PO2sc (P < 0.01) and a significant increase in PCO2gi and PCO2sc (P < 0.05). These values returned to baseline with resuscitation. PO2sc appeared to respond to haemorrhage earlier than PCO2gi and PCO2sc (P = 0.02). PO2sc was inversely correlated with PCO2gi (r2 0.7, P < 0.001). There were no significant changes in arterial plasma lactate concentrations. In our rat model, subcutaneous oxygen tension provided similar information to ileal luminal PCO2 and was more rapidly responsive than subcutaneous carbon dioxide tensions and arterial lactate during evolving haemorrhagic shock and resuscitation.

The effects of airway CO2 and cooling on ventilation and upper airway resistance in anaesthetized rats.

Upper airway (UA) luminal CO2 modulates the activity of superior laryngeal nerve afferents (Boushey et al, 1974, Bradford et al, 1990) and affects breathing and UA muscle activity through a superior laryngeal nerve-mediated reflex (Nolan et al, 1990) Therefore, airway CO2 may influence the patency of the UA, especially in obstructive apnoea and other circumstances of altered UA CO2 concentration However, the effects of UA luminal CO2 on airway patency have not been studied

Control of pH of airway surface liquid of the ferret trachea in vitro

We measured the pH of airway surface liquid (ASL) secreted by the ferret trachea in vitro by using a catheter-tipped pH electrode implanted in a collecting cannula close to the airway epithelium. Mucus secretion was promoted by methacholine (0.02 mmol/l) in the organ bath. The pH of the ASL was 6.85 +/- 0.03 (SE) compared with a bath value of 7.39 +/- 0.01, when the bath was bubbled with 5.65% CO2. Changing the bath CO2 from 0 to 20.93% CO2 altered the bath pH from 8.06 to 6.96, but the ASL pH only varied from 6.92 to 6.85. This homeostasis of ASL pH was not the result of the buffering powers of the ASL, because ex situ buffer curves for secreted ASL were similar to those for Krebs-Henseleit solution. Changing the luminal CO2 content by blowing gases through the trachea changed ASL pH by values similar to that ex situ. However, when external organ bath CO2 was changed, the luminal CO2 changes were proportionately far smaller. Measurement of rates of diffusion of CO2 across the tracheal wall indicated that this was not a limiting factor in the results. Similarly, measurement of metabolic rate CO2 production in the tracheal lumen indicated that this did not significantly affect the results. We conclude that the pH of ASL is significantly on the acid side of the pH or interstitial fluid and plasma and that it is maintained relatively constant despite large changes in external pH.

Mechanism for high PCO2 in gastric juice: roles of bicarbonate secretion and CO2 diffusion.

The partial pressure of CO2 (PCO2) in gastric juice often exceeds the PCO2 of blood. CO2 in gastric juice may originate from blood and enter luminal fluid by diffusion, or CO2 may be produced in the lumen of the stomach from the reaction of HCO3- and H+. Because CO2 production from HCO3- is dependent on acid (low pH), we suppressed acid secretion with intravenous cimetidine to estimate to what extent appearance of CO2 in luminal fluid is due to production from HCO3-. When denervated fundic pouches of dogs were distended with saline, the PCO2 of the solution increased to the PCO2 of blood in approximately 20 min, with the initial rate of appearance of CO2 in the pouch solution only minimally affected by cimetidine. Thereafter, PCO2 of luminal fluid continued to increase to 50-60 mmHg in the absence of cimetidine, whereas PCO2 of luminal fluid remained approximately equal to that of blood when cimetidine was infused (P less than 0.001, cimetidine vs. control). The mean pH in the pouch solution remained between 6.3 and 6.9 during cimetidine infusion but decreased to 4.75 without cimetidine (P less than 0.001). In additional experiments, an acidic solution with high PCO2 (242 +/- 3 mmHg) was infused into the fundic pouches. PCO2 in luminal fluid decreased rather slowly toward plasma PCO2, requiring 240 min for luminal fluid PCO2 to decrease to 56 +/- 2 mmHg. Thus the permeability of the gastric mucosa to luminal CO2 was relatively low.(ABSTRACT TRUNCATED AT 250 WORDS)

Ion transport in rat proximal colon in vivo

Active Na+ absorption by the rat proximal colon in vivo is for the most part electrically silent. The rheogenic Na+ flux makes up only 8%. To elucidate the underlying transport pathways, the following experimental approaches were used: ion substitution experiments such as choline for Na+, cyclamate for Cl-, variation of luminal pH; administration of known inhibitors; and determination of changes in luminal CO2 tension and pH. The transcolonic ion fluxes as well as the electrical parameters potential difference, specific electrical resistance, and short-circuit current were monitored. Na+ transport was drastically reduced in the absence of luminal Cl-, and vice versa Cl- absorption was blocked at zero Na+. NaCl absorption was blocked by amiloride (10(-3) M) and 4-acetamido-4'-isothiocyanostilbene-2, 2'-disulfonic acid and was lowered by acetazolamide. Colonic NaCl absorption was not influenced by luminal furosemide. Na+ absorption increased with alkalinization of the luminal fluid. Tris instead of HCO-3 buffer at constant pH favored Cl- uptake. The results may easily be explained by the operation of a Na+-H+ antiport functionally coupled to a Cl(-)-HCO-3 antiport. These transport processes are supposed to be present in the columnar cells of the colonic epithelium. There is good evidence for the association of K+ secretion with rheogenic Cl- secretion by the crypt cells.

Effect of side-chain shortening on the physiologic properties of bile acids: Hepatic transport and effect on biliary secretion of 23-nor-ursodeoxycholate in rodents

To define whether side-chain length influences the physiologic properties of bile acids, nor-ursodeoxycholate (nor-UDC), the C23-nor derivative of ursodeoxycholate (UDC), was synthesized in both nonradioactive and radioactive forms (23-14C). Its hepatic translocation, hepatic biotransformation, and effect on bile flow, biliary bicarbonate, and biliary lipid secretion were compared with that of UDC and those of their respective glycine and taurine conjugates in anesthetized biliary fistula hamsters, rats, and guinea pigs, as well as the isolated perfused hamster liver. Hepatic uptake and biliary output of nor-UDC was slower than that of UDC or cholyltaurine in the isolated perfused hamster liver. In biliary fistula animals, nor-UDC was secreted only in bile. Biliary recovery of nor-UDC as compared to that of UDC was prolonged in the rat and hamster, although not in the guinea pig. Hepatic biotransformation, assessed by chromatography of bile, showed that conjugation of nor-UDC was inefficient, as unconjugated nor-UDC was present in bile; there was little amidation with glycine or taurine in any species, but sulfates and glucuronides, as well as other metabolites, were formed, with the pattern of biotransformation varying among species. When infused over a dosage range of 0.2–30 μmol/kg · min, nor-UDC induced a striking choleresis of canalicular origin. The bile acid-dependent flow was increased threefold in hamsters, ninefold in rats, and nearly twofold in guinea pigs when compared to that induced by UDC. The choleresis was associated with a linear increase in bicarbonate output and concentration in bile, and little phospholipid or cholesterol secretion was induced. A competition experiment in the bile fistula hamster indicated that nor-UDC or its metabolites, or both, appeared to compete for canalicular transport of ursocholyltaurine (a cholyltaurine epimer) when the latter was secreted under its Vmax conditions. Conjugates of nor-UDC and UDC were promptly and almost completely recovered in bile without appreciable hepatic biotransformation; the conjugates did not induce a hypercholeresis or increase biliary bicarbonate concentration. It is proposed that a fraction of nor-UDC is secreted into canalicular bile in the unconjugated form and is protonated by a hydrogen ion derived from carbonic acid that was generated by the hydration of luminal CO2 by carbonic anhydrase present in biliary ductular cells.

Colonic absorption of inorganic ions and volatile fatty acids in the rabbit

1. 1. Net fluxes of water, Na +, K +, Cl −, HCO 3 −and volatile fatty acids (VFA) were investigated in three different segments of rabbit colon. 2. 2. Two opposite phenomena occurred: secretion of water and inorganic ions in the oral part of the colon and absorption in the remaining colon; VFA were always absorbed. 3. 3. The movement of cations was closely correlated with those of VFA and Cl −. Results are consistent with the presence of exchange: Na +/H +, K +/H +, in the colon brush border membrane. In fact net absorption of cations and VFA seems linked to the availability of protons. 4. 4. In the absence of net cation transport an additional source of protons may be provided by hydratation of luminal CO 2. So VFA could enter mucosa by passive diffusion as the undissociated acids.

On the mechanism of luminal CO2 generation during jejunal bicarbonate absorption

The mechanism of the increase in luminal CO2 tension (PCO2) that accompanies jejunal HCO-3 absorption is unknown. One possibility is that mucosal metabolism and the reaction of absorbed HCO-3 with blood buffers in mucosal capillaries govern luminal PCO2. To evaluate this possibility, jejunal segments of anesthetized adult male Sprague-Dawley rats were perfused in vivo with modified Ringer solutions with varying PCO2 levels (0-141 mmHg). Arterial PCO2 was varied by mechanical ventilation (20-70 mmHg). The net flux of CO2 gas into bulk luminal fluid varied directly with the transmucosal PCO2 gradient both in the absence (r = 0.98, P less than 0.001) and presence of net HCO-3 absorption (r = 0.97, P less than 0.001). The apparent permeability coefficient for CO2 gas across the jejunal mucosa was relatively low (2 X 10(-4) cm3 X s-1 X cm length -1) and was not affected by net HCO-3 absorption. Under conditions of equal arterial blood and perfusate PCO2 levels (40 mmHg), the PCO2 in mucosa and effluent was 51 and 44 mmHg, respectively, in the absence and 62 and 48 mmHg, respectively, in the presence of net HCO-3 absorption. These results suggest that CO2 diffuses from the mucosal region into bulk liminal fluid during net HCO-3 absorption in the rat jejunum. In addition, we found in vitro that the elevation of mucosal PCO2 above arterial PCO2 during net HCO-3 absorption can be quantitatively accounted for by metabolism and the admixture of absorbed HCO-3 with blood buffers.