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Ursodeoxycholic acid action on the transport function of the small intestine in normal and cystic fibrosis mice.

Ursodeoxycholic acid possesses choleretic and cytoprotective properties and in cystic fibrosis (CF) it is used to treat the hepatobiliary symptoms of the disease. This study investigated the effects of this bile acid on the transport function of the small intestine in normal and CF mice. The effects of ursodeoxycholic acid were monitored as changes in short-circuit current (SCC) in stripped sheets of small intestine from normal (Swiss MF1) and transgenic CF (Cftr(tm2Cam)) mice. In ileal sheets from Swiss MF1 mice, mucosal ursodeoxycholic acid caused a biphasic increase in SCC. The first phase was reduced by lowering the mucosal Na+ concentration, while the second phase was inhibited by (Cl-)-free conditions, serosal furosemide or mucosal diphenylamine-2-carboxylic acid (DPC), suggesting an initial Na+-dependent bile acid absorption followed by a stimulation of electrogenic Cl- secretion. Serosal application of ursodeoxycholic acid to the ileum and mucosal or serosal application to the mid-intestine and jejunum elicited a secretory response only. Secretion was Ca2+-dependent, but did not involve neural mechanisms. Mucosal mast cells, histamine and serotonin (5-HT) were implicated in the secretory response. Responses in tissues from transgenic wild-type mice were similar to those obtained with Swiss MF1 mice, but the secretory response to mucosal or serosal application of the bile acid was impaired in CF tissues. In ilea from CF mice the initial absorptive phase of the response to mucosal ursodeoxycholic acid was still observed. It is concluded that ursodeoxycholic acid induces secretion throughout the murine small intestine by a mechanism that involves degranulation of mucosal mast cells. In the ileum Na+-dependent absorption can also be detected. The secretory response is defective in CF intestine, but the absorptive effect is still present.

Animals↗

Calcium transport by isolated skin of rainbow trout.

The skin overlying the cleithrum bone of freshwater-acclimated rainbow trout contains numerous mitochondria-rich (MR) cells, as detected by DASPEI fluorescence. This tissue was mounted in vitro in an Ussing-style chamber with fresh water on the mucosal surface and saline supplemented with bovine serum albumin on the serosal surface. The preparation developed a high transepithelial resistance and a small transepithelial potential (Vt), positive on the serosal side. Radioisotopic flux measurements indicated that the preparation actively transported Ca2+ from the mucosal to the serosal surface, as assessed by the Ussing flux ratio criterion. Ca2+ transport was positively correlated with MR cell density. Cortisol pretreatment in vivo reduced MR cell density and increased Vt but did not significantly alter Ca2+ fluxes. Ca2+ transport was unaffected by adrenergic agonists (10(-5) mol l-1 adrenaline, clonidine, isoprenaline) or cyclic AMP stimulants (10(-3) mol l-1 dibutyryl cyclic adenosine monophosphate, db-cAMP, plus 10(-4) mol l-1 isobutylmethylxanthine, IBMX) applied to the serosal surface. The Ca2+ ionophore ionomycin (1 x 10(-6)-3.2 x 10(-6) mol l-1 on the mucosal surface) increased both unidirectional Ca2+ fluxes and caused Ca2+ to accumulate within the epithelium. Lanthanum (10(-4) mol l-1) did not inhibit unidirectional Ca2+ fluxes, but apparently displaced Ca2+ from binding sites on the mucosal surface. Unlike Ca2+, movements of Na+ and Cl- across the epithelium were passive, as assessed by the flux ratio criterion, and neither adrenaline nor db-cAMP plus IBMX had any effect on Na+ or Cl- fluxes or electrical properties. These results indicate that ion transport across the skin mediated by MR cells ('chloride cells') contributes to Ca2+ but not to NaCl balance in freshwater trout.

Acclimatization↗

Mechanisms of gastrointestinal copper absorption in the African walking catfish: copper dose-effects and a novel anion-dependent pathway in the intestine.

In mammals, copper (Cu) absorption occurs mostly in the small intestine, and some of the Cu transporters involved in its uptake have been characterised. In fish, however, the regions of the gut involved in Cu absorption and the membrane transport mechanisms responsible for gastrointestinal Cu uptake are unknown. Everted gut sacs and isolated perfused intestine of Clarias gariepinus were used to explore Cu absorption (at 22 degrees C). Gut sacs exposed to 100 micromol l(-1) mucosal solution Cu ([Cu](m)) showed that Cu was mostly (70 %) absorbed in the middle and hind intestine. Most of the accumulated Cu was located in the mucosa. In perfused intestines, cumulative Cu absorption from the mucosal solution to the serosal perfusate was greatest at 10 micromol l(-1) [Cu](m) and decreased at higher values of [Cu](m), while tissue accumulation of Cu showed a dose-dependent elevation. Absorption efficiency therefore declined with increasing Cu dose, and basolateral transport was the limiting factor in Cu uptake. Serosal applications of the P-type ATPase inhibitor vanadate (100 micromol l(-1)) or the anion transport inhibitor DIDS (100 micromol l(-1)) caused threefold increases in net Cu uptake (at [Cu](m)=10 micromol l(-1)). The vanadate effect was explained by a reduction in transepithelial potential rather than inhibition of Cu-ATPase, but the DIDS effect was not. Transepithelial potential, water transport and tissue [Cu] were not affected by DIDS, but tissue [K(+)] was elevated. Removal of Cl(-) simultaneously from both the mucosal and serosal solutions caused a 10-fold reduction in the rate of Cu uptake, while removal of Cl(-) from the mucosal solution only completely abolished Cu absorption to the serosal perfusate. Transepithelial potential effects are discussed. We conclude that Cu absorption occurs mostly in the intestine and is normally driven by a basolateral Cu/anion symport that prefers Cl(-).

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗

The mechanism of intestinal transport of sulfamethoxazole and the effect of chlorpromazine in rat everted intestine.

The mechanism of the intestinal transport of sulfonamides and the effect of chlorpromazine (CPZ) on it were studied using rat everted intestine in vitro. Sulfamethoxazole (SMZ) was accumulated in the serosal solution in the everted sac obeying the pH partition theory, while sulfisoxazole (SIX) was not accumulated despite the presence of the pH difference between the serosal and mucosal solutions. The reason was suggested that the microclimate pH on the mucosal surface of the intestine so decreased the amount of the unionized molecules of SIX that the transport rate of SIX was decreased to show no accumulation in the serosal solution within the sampling period. CPZ as well as metabolic inhibitors blocked the SMZ accumulation by inhibiting the growth of the pH difference. This effect was caused by the inhibition of the serosal alkalinization.

Animals↗

Contribution of an electrogenic sodium pump to the membrane potential in the intestinal epithelial cell.

The transmural potential difference of isolated rat duodenum and the membrane potential of its epithelial cell were examined during changes of the ionic medium at both warm (31--35 degrees C) and cold temperatures (2--5 degrees C). In the range of low external K+ concentrations, cooling produced an immediate and reversible depolarization of the membrane potential (around -10 mV) greater than that estimated from a simple thermodynamic temperature effect (RT/F). Such a temperature sensitive component of the membrane potential at 31 degrees C was completely abolished by anoxia and serosal ouabain. Readmission of K+ to the bathing fluid of duodenum previously deprived of K+ and loaded by Na+ caused a remarkable transient hyperpolarization of the epithelial membrane, which was abolished by serosal ouabain. Decreases in the value of transmural potential difference were also produced by cooling, anoxia and serosal ouabain, but these were so small that concurrent changes in the membrane potential could be regarded as chiefly reflecting changes in the effective emf of the cell. Thus, an electrogenic Na+-K+ exchange pump located on the serosal membrane of the duodenal epithelial cells was shown to contribute significantly to the membrane potential under normal conditions (up to around -7 mV). The coupling ratio of Na+-efflux and K+-influx associated with this pump was estimated by the Mullins-Noda equation (1963) to be approximately 4:3 under the physiological steady-state conditions.

Animals↗

Restoration by VIP of the carbachol-stimulated Cl- secretion in TTX-treated guinea pig distal colon.

To determine if vasoactive intestinal peptide (VIP) restores neural activity from tetrodotoxin (TTX) blockade, we studied the effects of VIP and related agents on carbachol (Cch)-induced Cl(-) secretion in control-isolated guinea pig distal colon and in that treated with TTX. The short circuit current (I(sc)) increased dose-dependently after serosal applications of Cch (10(-6) - 2 x 10(-5) M) and VIP (5 x 10(-9) - 10(-7) M). But no additive or synergistic increase in I(sc) was observed. Cch- and VIP-induced I(sc) was completely abolished by a serosal application of TTX (10(-6) M). However, a serosal application, not mucosal, of VIP (10(-7) M) and 8-bromo-cAMP (10(-3) M) restored the Cch-stimulated, TTX-inhibited I(sc) by 113% and 75.8%, respectively. Furthermore, mucosal and serosal applications of forskolin (aden late cyclase activator) restored the I(sc) by 43.9% and 65.3%, respectively. The restored I(sc) was completely abolished by atropine (muscarinic receptor antagonist). These results suggest that VIP may restore the cholinergic activity by increasing the level of intracellular cAMP, and that cholinergic neuron is very likely to be responsible for the regulation of Cl(-) secretion at neuroepithelial junctions. The exact mechanism of VIP's effect on the TTX-inhibited epithelial Cl(-) secretion, and its possible usefulness in the treatment of TTX-induced pathophysiological conditions, remain to be determined.

8-Bromo Cyclic Adenosine Monophosphate↗

Luminal leptin induces rapid inhibition of active intestinal absorption of glucose mediated by sodium-glucose cotransporter 1.

The effect of leptin on glucose transport was studied in rat jejunal mucosa in Ussing chambers. Leptin was added in the luminal or the serosal compartment before the tissues were challenged with 1, 10, or 50 mmol/l glucose. In response to 10 mmol/l glucose, the increase in short-circuit current (DeltaIsc) reached 26.8 +/- 2.1 microA/cm(2). Luminal addition of leptin dramatically decreased glucose-induced Isc (90.5% for 10 nmol/l leptin). Inhibition was maximal after 5 min and dose dependent (IC(50) = 0.13 nM). Western blot analysis showed that rapid inhibition of glucose-induced Isc by leptin was associated with a parallel decrease in the abundance of sodium-glucose transporter-1 in brush border membranes. Inhibition by luminal leptin of DeltaIsc was prevented by inhibitor of conventional protein kinase C isoforms. Serosal addition of leptin did not decrease glucose-induced Isc within 5 min and reached maximum after 10 min. The effect of leptin from serosal side was blocked by cholecystokinin (CCK) receptor-2 receptor antagonist YM022. Altogether, these data demonstrate that luminal leptin induces rapid inhibition of glucose entry into enterocyte. The slower action of leptin on the serosal side of mucosa seems indirect and is likely mediated by endogenous CCK. They demonstrate that gut leptin is a major regulator of rapid intestinal glucose transport.

Acetophenones↗

Effect of carboxymethylcellulose and a hyaluronate-carboxymethylcellulose membrane on healing of intestinal anastomoses in horses.

OBJECTIVE: To evaluate the effect of sodium carboxymethylcellulose (SCMC) or a hyaluronate-carboxymethylcellulose membrane (HA membrane) on healing of the small intestine in horses. ANIMALS: 18 healthy adult horses. PROCEDURE: Midline celiotomy and 2 jejunal resection-and-anastomosis surgeries were performed. In treated horses, SCMC (n = 6) or a HA membrane (6) was applied to the jejunum to cover the anastomosis. There were 6 untreated control horses. Horses were euthanatized 10 days after surgery. For each horse, 1 anastomosis was used for histologic examination, and the second was used to determine intestinal bursting strength. Intestinal bursting tension, serosal granulation tissue, serosal fibrin deposition, and width of the fibrous seal at the anastomosis were compared among groups. RESULTS: 3 control horses had adhesions associated with the anastomosis, but none of the treated horses had adhesions associated with the anastomosis. Mean thickness of fibrin deposited on the serosal surfaces for the SCMC and HA-membrane groups was significantly less than that for control horses. Mean thickness of serosal granulation tissue, width of fibrous seal between inverted musculature, inflammatory cell infiltrate scores, and bursting tension did not differ significantly among groups. CONCLUSIONS AND CLINICAL RELEVANCE: Use of SCMC or application of a HA membrane to small intestinal anastomoses in horses resulted in fewer adhesions and decreased fibrin deposition, and it did not adversely affect anastomotic healing. In horses at increased risk for intra-abdominal adhesions, SCMC or application of HA membranes may decrease the frequency of adhesions without adversely affecting healing of small intestinal anastomoses.

Anastomosis, Surgical↗

Absorption of 2-hydroxy-4-(methylthio)butanoic acid by isolated sheep ruminal and omasal epithelia.

Alimet (Novus Inter., Inc., St. Louis, MO) feed supplement (an 88% aqueous solution of 2-hydroxy-4-(methylthio) butanoic acid; HMB) is a source of L-Met commonly used in nonruminants and ruminants. The absorption of HMB across ovine omasal and ruminal epithelia was evaluated in this study. Ruminal and omasal epithelia were collected from eight lambs (BW = 67.6 kg +/- 9.1) and mounted in parabiotic chambers that were repeatedly sampled throughout a 60-min incubation. The appearance of HMB (using DL-[5-14C]-HMB as a radiolabeled marker) in serosal buffers increased quadratically (P < .004) with time in both tissues. More (P < .001) HMB appeared in the serosal buffers with omasal than with ruminal epithelia. Both tissues responded similarly, and, after 60 min of incubation, the accumulation of HMB within the tissues increased linearly (P < .001) as substrate concentration (.375, .75, 1.5, 3.0, 6.0, and 12.0 mM) increased in mucosal buffers. As the concentration of HMB in the mucosal buffers increased, there was a quadratic (P < .001) increase in the appearance of HMB in the serosal buffer of the omasal epithelium, indicating some saturation of the system. The increase in serosal appearance of HMB was linear (P < .001) with ruminal tissue. The results indicate that there are probably multiple mechanisms involved in the absorption of HMB. Because saturation was observed in the omasum, it is likely that mediated transport accounts for at least a portion of the absorption of HMB in the omasum. Other mechanisms (e.g., diffusion and(or) paracellular absorption) are responsible for the balance of the absorption. Omasal epithelium appears to have a greater capacity for HMB absorption than ruminal epithelium. The enzymes involved in the conversion of HMB to 2-keto-4-(methylthio)butanoic acid were found in ruminal and omasal epithelia, liver and kidney. These results indicate that HMB can be absorbed across ruminal and omasal epithelium and that HMB can be used as a source of L-methionine.

Animals↗

Assessment of blood flow in the small intestine with laser Doppler flowmetry.

Blood flow in the small intestine was assessed in 48 patients by laser Doppler flowmetry. Mucosal and serosal flowmeter signals were compared during 'resting' conditions, vascular occlusion, and reactive hyperemia. Serosal flowmeter recordings were compared with the total blood flow of a bowel segment as measured by venous collection. The magnitudes of the mucosal (n = 49) and serosal (n = 49) flowmeter signals were comparable throughout the whole range of flowmeter signals (r = 0.97; p less than 0.001). A correlation coefficient of 0.95 (n = 51; p less than 0.001) was obtained between serosal flowmeter signals and total blood flow during 'resting' and reduced blood flows. During vasodilatation after a vascular occlusion, blood flow was underestimated by the flowmeter. A calibration curve could be constructed for approximate interpretation of the flowmeter signals in absolute flow units. The present study underlines the potential of laser Doppler flowmetry in the assessment of blood flow in the human small intestine.

Adolescent↗

Measurement of colonic blood flow with laser Doppler flowmetry.

Colonic blood flow was measured with laser Doppler flowmetry during operation in 62 patients and during coloscopy in 15 patients. In 18 subjects mucosal and serosal laser Doppler signals were compared during 'resting' conditions, vascular occlusion, and reactive hyperemia. The mucosal (n = 36) and serosal (n = 36) flowmeter signals were of the same order of magnitude throughout the whole range of flowmeter signals (r = 0.96; p less than 0.001). In eight subjects total venous outflow from a colonic segment was measured simultaneously with flowmeter recordings from the serosal side. A correlation coefficient of 0.95 (n = 46; p less than 0.001) was obtained between total intestinal blood flow and serosal flowmeter signal during 'resting' and reduced blood flows. In this flow range a calibration curve was constructed for interpretation of the flowmeter signals in absolute flow units. The wall thickness of the bowel determined the quantitative relationship between flowmeter signal and total intestinal blood flow. It is concluded that laser Doppler flowmetry represents a potentially very interesting non-invasive, continuous method for the quantitative study of human intestinal blood flow.

Adolescent↗

Stimulation of Cl- secretion by AlF4- and vanadate in T84 cells.

We investigated the mechanism of Cl- secretion by fluoroaluminate(AlF4-) and sodium orthovanadate(vanadate) using the human colonic T84 cell line. T84 cell monolayers grown on collagen-coated filters were mounted in Ussing chambers to measure short circuit current(ISC). Serosal addition of AlF4- or vanadate to T84 monolayers produced a sustained increase in ISC. Removal of Ca2+ from the serosal bathing solution partially inhibited AlF4-(-)and vanadate-induced ISC, and readministration of Ca2+ restored AlF4-(-)and vanadate-induced ISC. Carbachol application in the presence of forskolin, AlF4- or vanadate induced a synergistic increase of ISC. Forskolin and vanadate significantly increased cellular cAMP level, while carbachol and AlF4- did not. Carbachol, AlF4- and vanadate significantly increased [Ca2+]i. After Na+ in mucosal bathing solution was replaced with K+, and the mucosal membrane of T84 cell was permeabilized with amphotericin B, AlF4-, vanadate, and carbachol increased K+ conductance, but forskolin did not. After sodium chloride in serosal bathing solution was replaced with sodium gluconate and the serosal membrane was permeabilized with nystatin, forskolin, AlF4-, and vanadate increased Cl- conductance, but carbachol did not. AlF4-(-)induced ISC was remarkably inhibited by the pretreatment of pertussis toxin(2 micrograms/ml) for 2 hours. These results indicate that AlF4- and vanadate can increase Cl- secretion via simultaneous stimulation of Cl- channel and K+ channel in T84 cells. However, the AlF4- action is mostly attributed to stimulation of pertussis toxin-sensitive G-proteins, whereas the vanadate action mostly results from G protein-independent mechanisms.

Aluminum↗

Nucleotide stimulation of Cl(-) secretion in the pigmented rabbit conjunctiva.

We evaluated the role of extracellular UTP and other nucleotides in the regulation of active ion transport across the pigmented rabbit conjunctiva. When added to the mucosal side of the conjunctiva, UTP (0.01-1000 microM), increased the short-circuit current by up to 14. 6 +/- 2.1 microA/cm(2). The half-maximal concentration was 11.4 +/- 2.3 microM. The serosal absence of Cl(-), serosal presence of 10 microM bumetanide, and mucosal presence of 0.3 mM N-phenylanthranilic acid significantly reduced the change in the short-circuit current (DeltaIsc) induced by 10 microM UTP by 78, 77, and 42%, respectively. Mucosal 10 microM UTP significantly increased (36)Cl flux in the serosal-to-mucosal direction by 0.17 microEq/cm(2)/h, while not affecting mucosal-to-serosal (36)Cl flux. By contrast, (22)Na transport in either direction was unaffected. The rank order of DeltaIsc elicited by adenosine and nucleotides was consistent with the predominant involvement of P2Y purinergic receptors in the UTP effect on conjunctival ion transport. Moreover, the DeltaIsc elicited by UTP was inhibited by 0.05 and 1 mM suramin (a P2-purinergic receptor antagonist), resulting in a rightward shift of the half-maximal concentration to 106.7 +/- 1.3 microM. In conclusion, the primary effect of UTP on ion transport in the pigmented rabbit conjunctiva is stimulation of Cl(-) secretion, possibly at the P2Y(2) and/or the P2Y(4) receptor on the mucosal side of the tissue. Because of the coupling of fluid flow with Cl(-) secretion, UTP or its analogs may be considered for stimulating transconjunctival fluid flow in the dry-eye state.

Adenosine↗

Involvement of an organic anion transporter (canalicular multispecific organic anion transporter/multidrug resistance-associated protein 2) in gastrointestinal secretion of glutathione conjugates in rats.

We investigated the role of cMOAT/MRP2 (canalicular multispecific organic anion transporter/multidrug resistance-associated protein 2) in the intestinal secretion of organic anions by comparing the behavior in Sprague-Dawley (SD) rats and Eisai hyperbilirubinemic rat (EHBR) whose cMOAT/MRP2 is hereditarily defective. After i.v. administration of 1-chloro-2,4-dinitrobenzene (30 micromol/kg), the biliary and intestinal excretion of its glutathione conjugate 2, 4-dinitrophenyl-S-glutathione (DNP-SG), a substrate for cMOAT/MRP2, was significantly reduced in EHBR compared with SD rats. This result also was confirmed by Ussing chamber studies; DNP-SG showed 1.5-fold greater serosal-to-mucosal flux compared with the mucosal-to-serosal flux in SD rats, whereas a similar flux was observed in both directions in EHBR. In addition, metabolic inhibitors reduced the preferential serosal-to-mucosal flux of DNP-SG in SD rats. In everted sac studies, intestinal secretion clearance, defined as the efflux rate of DNP-SG into the mucosal side divided by the area under the curve on the serosal side, was significantly lower in the jejunum of EHBR than that in SD rats. Northern blot analyses demonstrated the highest mRNA level of cMOAT/MRP2 in the jejunum, which is in good agreement with the results of the everted sac studies. These results suggest that cMOAT/MRP2 is involved in the secretion of organic anions in the small intestine.

ATP-Binding Cassette Transporters↗

Functional characterization of organic cation drug transport in the pigmented rabbit conjunctiva.

PURPOSE: To characterize carrier-mediated organic cation drug transport in the rabbit conjunctiva. METHODS: The transport of [14C]guanidine, the model substrate, in the excised pigmented rabbit conjunctiva was evaluated in the modified Ussing chamber. Tetraethylammonium (TEA) transport also was investigated to determine substrate specificity. RESULTS: The apparent permeability coefficient for guanidine and TEA in the mucosal-to-serosal (ms) direction was 5.4 and 49.6 times greater than that in the serosal-to-mucosal (sm) direction, respectively. Guanidine transport in the ms (but not sm) direction revealed temperature and concentration dependency over 0.02 to 10 mM with an apparent Michaelis-Menten constant of 3.1 mM and a maximal flux of 11.4 nmol/(cm2 x h). Net guanidine transport measured at 0.1 mM across the conjunctiva was decreased by 71% or 82%, respectively, on the addition of 1 microM valinomycin (a K+ ionophore) in both bathing fluids or in a high K+ buffer in the mucosal fluid. Interestingly, net guanidine transport was reduced, rather than enhanced, by 63% upon acidifying the mucosal bathing fluid. By contrast, net guanidine transport was not affected by the serosal presence of 0.5 mM ouabain (a Na+, K+-ATPase inhibitor), by the mucosal and serosal presence of 0.1 microM monensin (a Na+ ionophore) or 0.3 microM carbonyl cyanide p-(trifluoromethoxy)phenyl-hydrazone (FCCP, a H+ ionophore). Guanidine transport in the ms direction was polyspecific, as indicated by the 48% to 82% inhibition by structurally diverse amines. In particular, guanidine ms transport was inhibited by the antiglaucoma drugs dipivefrine (72%), brimonidine (70%), and carbachol (78%). CONCLUSIONS: A carrier-mediated organic cation transport process appears to exist in the conjunctiva, mediating the absorption of organic amines, including certain amine-type ophthalmic drugs. This process may be driven by an inside-negative apical membrane potential difference.

Animals↗

[Functional role of V1- and V2-receptors of the apical and basolateral membranes of the frog bladder epithelial cells].

Arginine vasotocin, 0.02--1 nM, increases osmotic water permeability of frog urinary bladder, arginine vasotocin after a simultaneous addition to the mucosal and serosal Ringer solutions rises the water permeability to a lesser degree than on the hormone addition only to the serosal solution. 1 nM remestyp, an agonist of V1-receptors, from the apical membrane decreases the hydroosmotic effect of arginine vasotocin added to the serosal Ringer solution. When added to the mucosal solution, combination of the same concentrations of arginine vasotocin and SR 49059, an antagonist of V--receptors, or desmopressin, agonist of V2-receptor alone, increases the effect of the same concentration of arginine vasotocin added to the serosal solution. 1 nM arginine vasotocin at the luminal membrane increases secretion into the Ringer solution of prostaglandin E, and prostaglandin E1 but not of prostaglandin F2 alpha. The data obtained indicate the presence of the arginine vasotocin receptors responsible for the hydroosmotic effect only in the basolateral membranes, while arginine prostaglandin E, participation is shown in modulation of the arginine vasotocin effect.

Alprostadil↗

Studies on the mechanism of action of calciferol. VIII. The effects of dietary vitamin D and the polyene antibiotic, filipin, in vitro, on the intestinal cellular uptake of calcium.

Intestinal transport and cellular uptake of calcium were studied in vitro in ileal segments obtained from calciferol (vitamin D)-deficient (minus D3) chicks or in calciferol-treated chicks (+ D3). When calcium flux (J) was measured in both the mucosal yields serosal (J-ms) and serosal yields mucosal (J-sm) directions it was found that calcium transport in the +D3 system was an active process. Calcium was not actively transported in the minus D3 system or by either the +D3 or minus D3 serosal tissues which underlie the intestinal mucosa tissue. It was also found that the serosal tissue was not the rate-limiting step in calcium translocation. When calcium uptake was measured at the mucosal or brush border surface it was found to be a cation-oriented, saturable process in both the +D3 and minus D3 systems and was enhanced by calciferol supplementation. The rate of uptake was found to exhibit a two component transport phenomenon, one a saturable process at low Ca2+ concentrations and the other a linear of diffusion process at higher Ca2+ concentrations (greater than 5 mM). The polyene antibiotic, filipin, was used to study the transport and uptake of calcium in both the +D and minusD chick ileum in vitro. Filipin (10 mug/ml), when added in vitro to solutions bathing the mucosal surface of the ileum, stimulated the calcium flux, J-ms, of minus D ileal tissue by 150 to 200%, but had little or no stimulatory effect on J-ms of +D3 ileal tissue. However, in contrast it stimulated calcium uptake across the mucosal membrane in both the +D3 and minus D3 ileal tissue (50 to 100%). The effect of filipin is specific for calcium uptake as compared to Rb+, P-i, SO4(2-), glycerol, thiourea, and urea uptake. Isolated brush borders from +D3 chicks were found to bind calcium to a greater extent than the minus D3 brush borders, but both membrane fractions had the same cholesterol content. The sum of this and other evidence suggests that of uptake of calcium is a calciferol-mediated event which is not the rate-limiting step in the total transport of this ion across the intestinal epithelial cell. The filipin effect appears to affect a structural reorganization of the brush border membrane in both the +D3 and minus D3 membranes in a manner specific for calcium translocation. It now appears that filipin treatment somehow makes both transport systems more efficient in the uptake of calcium, but in a manner independent of the mode of action of calciferol. However, these data suggest that other components associated with calcium uptake are calciferol-dependent in an as yet unknown fashion.

Animals↗

[The effect of ATP and acetylcholine on the sodium transport in bullfrog large intestine (author's transl)].

By Ussing's flux chamber method the effect of ATP and acetylcholine (ACh) on the sodium transport was studied in bullfrog colon. The results obtained are as follows; 1. ATP added to the mucosal medium caused biphasic changes in the transmural potential difference (P.D.) and short-circuit current (S.C.C.), although serosal ATP was ineffective. After an initial rapid and transient rise, both the P.D. and S.C.C. increased in parallel to reach a peak in about 10 min suggesting that the tissue conductance is little affected by ATP. Addition of ouabain to the serosal fluid depressed both the P.D. and S.C.C. and abolished the electrical responses to ATP. The application of ouabain to the mucosal side did not cause any significant depression. These results can be explained in terms of stimulation of sodium pump by ATP added to the mucosal medium. 2. ACh added to either the mucosal or the serosal medium caused increased in the P.D. and the S.C.C. The serosal application was more effective than the mucosal application. The increase in S.C.C. was more remarkable than that in the P.D., indicating an increase in the tissue conductance. It is suggested that ACh stimulates ion transport systems by changing the membrane permeability of the colon.

Acetylcholine↗