Increase In Net Cl Research Articles

Signature current of SO2-induced bronchitis in rabbit.

To investigate abnormalities of airway epithelial ion transport underlying chronic inflammatory airway diseases, we performed electrophysiological, histological, and molecular biological experiments using rabbits exposed to SO2 as a model of bronchitis. By comparison with control, the SO2-exposed trachea exhibited decreased short circuit current (Isc) and conductance associated with increased potential difference. In normal trachea, apical ATP induced a transient Isc activation followed by a suppression, whereas the bronchitis model exhibited a prolonged activation without suppression. This pathological ATP response was abolished by diphenylamine 2-carboxylate or Cl--free bath solution. A significant increase in net Cl- flux toward the lumen was observed after ATP in our bronchitis model. Isoproterenol or adenosine evoked a sustained Isc increase in SO2-exposed, but not in normal, tracheas. The Northern blot analysis showed a strong expression of cystic fibrosis transmembrane conductance regulator (CFTR) mRNA in SO2-exposed epithelium. The immunohistochemical study revealed a positive label of CFTR on cells located luminally only in SO2-exposed rabbits. We concluded that the prolonged ATP response in our bronchitis model was of a superimposed normal and adenosine-activated current. The latter current was also activated by isoproterenol and appeared as a signature current for the bronchitis model airway. This was likely mediated by CFTR expressed in the course of chronic inflammation.

Protein kinase C regulates prairie dog gallbladder ion transport.

Gallstone formation is characterized by increased biliary calcium (Ca2+) level and altered gallbladder absorption. Recent studies suggest that luminal Ca2+ regulates gallbladder ion transport via intracellular calcium ([Ca2+]ic). Ca2+-calmodulin and protein kinase C (PKC) are two major systems through which [Ca2+]ic carries out second-messenger functions in many cell types. We have previously shown that Ca2+-calmodulin regulates basal gallbladder ion transport in prairie dog. The present study tests the hypothesis that PKC is also essential in regulation of gallbladder ion transport in this model. The role of PKC in regulation of gallbladder ion transport was determined by studying the effects of phorbol esters, synthetic analogues of diacylglycerol, which directly activates PKC. Gallbladders were mounted in Ussing chambers, and standard electrophysiologic parameters were recorded after exposing tissues to either 10(-5) mol/L of 4-alpha-phorbol 12,13-didecanoate (PDD), 4-beta-phorbol 12-myristate 13-acetate, 4-beta-phorbol 12,13-dibutyrate (PDB), or 10(-4) mol/L serotonin. Unidirectional Na+, Cl-, and H2O fluxes were measured before and after treatment with only inactive PDD and most active PDB. Mucosal and serosal exposure of tissues to either 4-beta-phorbol 12-myristate 13-acetate or PDB resulted in a decrease in short-circuit current and transepithelial potential difference without any change in tissue resistance. Serotonin induced similar changes in gallbladder electrical properties. PDB caused an inhibition of mucosal to serosal fluxes of Na+, Cl-, and H2O, with a decrease in net Na+ absorption, an increase in net Cl- secretion, and a conversion of net H2O absorption to net H2O secretion. Serosal-to-mucosal fluxes of Na+, Cl-, and H2O did not change. Inactive PDD had no effect on either electrophysiologic parameters or ion and water fluxes. Pretreatment of tissues with PKC antagonist 1-(5-isoquinolinylsulfonyl)-2-methylpiperazine blocked the phorbol ester-induced inhibition of ion transport. PKC regulates gallbladder ion transport in the prairie dog by inhibiting Na+ absorption and stimulating Cl- secretion.

Duramycin enhances chloride secretion in airway epithelium

The effect of duramycin, a polypeptide antibiotic, on Cl- transport in canine tracheal epithelium mounted in Ussing chambers was studied. Over a narrow concentration range, duramycin increased short-circuit current (Isc) and net Cl- secretion and had no effect on mannitol flux when added to the mucosal bathing solution. The maximum increase in Isc was observed at a duramycin concentration of 2 X 10(-6) M and was associated with an increase in both unidirectional Cl- fluxes. Higher duramycin concentrations produced a decrease in Isc. Submucosal addition of duramycin had no effect on Isc except at high concentrations. Pretreatment of tissues with mucosal amiloride (10(-4) M) to reduce basal Na+ transport had no effect on the subsequent response to duamycin. In other tissues pretreated with 10(-3) M dibutyryl adenosine 3',5'-cyclic monophosphate (cAMP), duramycin produced a further increase in Isc and net Cl- secretion similar to its effect in nonpretreated tissues. In all instances the increase in Isc was entirely accounted for by an increase in net Cl- secretion. We conclude that duramycin increases Isc and Cl- secretion in airway epithelium. Although the mechanism of activation is not known, these data demonstrate that duramycin increases Cl- secretion by a pathway other than cAMP. An understanding of the mechanism of action of duramycin may further our understanding of Cl- secretion regulation in airway epithelium.

The secretory actions of histamine in rat small intestine.

1. Histamine caused a dose-dependent rise in the transintestinal potential difference in vivo that was competitively blocked by H1, but not by H2 antagonists. This effect of histamine was also reduced by the cyclo-oxygenase inhibitor indomethacin. 2. Histamine induced a net secretion of fluid by the small intestine and this effect was reduced by indomethacin. 3. In intestinal sheets histamine increased the potential difference, short-circuit current and tissue resistance. This response was decreased in the absence of Cl- and in the presence of furosemide, suggesting that Cl- secretion was responsible for the observed electrical changes. This was confirmed by direct measurement of ion fluxes which showed an increase in net Cl- secretion together with an inhibition of net Na+ absorption. 4. The response to histamine was Ca2+-dependent since it was inhibited by removal of serosal Ca2+ and by verapamil. 5. Indomethacin caused a dose-dependent reduction in the response of intestinal sheets to histamine, without affecting the rise in short-circuit current induced by mucosal glucose or serosal prostaglandin E2. Mepacrine also inhibited the response to histamine, but not that to prostaglandin E2. 6. It is concluded that histamine induces intestinal secretion by stimulating the production of prostaglandins which then activate the secretory process.

Sodium and chloride transport by the tracheal epithelium of fetal, new‐born and adult sheep.

In vitro measurements were made of Na+ and Cl- isotopic fluxes across the tracheal epithelium of mature fetal lambs (130-143 days gestation), new-born lambs (up to 41 days of age) and adult sheep under conditions of continuous short circuiting. The effects of a variety of drugs were examined, but only in the case of amiloride and isoprenaline were observations made in all three groups. Experiments designed to elucidate the mechanism of basal Cl- secretion were performed in adult trachea only. Under resting conditions the net flux of Na+ from lumen to submucosa exceeds that of Cl- in the reverse direction in fetal and adult trachea. In the new-born the two fluxes are more or less equivalent in magnitude. In none of the three groups is the sum of ion fluxes significantly different from the short-circuit current (Isc). Removal of Na+ from, or addition of furosemide (10(-3) M) to, the solution bathing the submucosal surface of adult trachea has the effect of reducing Isc by an amount which approximates to the Cl- current (29%). At a concentration of 10(-4) M on the submucosal side of adult trachea, ouabain causes potential difference and Isc to fall to zero within 70 min of addition to the bathing solution. Nevertheless, there remains a significant net Na+ flux from submucosa to lumen. The addition of isoprenaline (10(-4) M) to the medium bathing the submucosal surface of both fetal and adult trachea causes an increase in the one-way flux of Cl- from submucosa to lumen with consequent increase in net Cl- flux towards the lumen. (The Na+ fluxes are unchanged.) However, in the adult the Cl- secretory response to isoprenaline is very much less and is not accompanied by an increase in electrical conductance. As judged by the change in Isc, all the post-natal fall in beta-agonist responsiveness takes place within the 3 week period following birth. Whereas, in the fetus, the effect of luminally applied amiloride on the Na+ fluxes is negligible, in the adult the one-way flux of Na+ from lumen to submucosa is reduced by 35% with a consequent 60% fall in net Na+ flux towards the submucosa.(ABSTRACT TRUNCATED AT 400 WORDS)

Ba2+ inhibition of VIP- and A23187-stimulated Cl- secretion by T84 cell monolayers

Addition of either 10(-8) M vasoactive intestinal polypeptide (VIP) or 10(-6) M A23187 to T84 cell monolayers, grown on permeable supports and mounted in Ussing chambers, stimulated net Cl- secretion. The effect of 10(-6) M A23187 on Cl- flux was consistently smaller than that observed with 10(-8) M VIP. In both cases the increase in net Cl- secretion accounted for the entire change in the observed short-circuit current (Isc). Since Cl- enters the cells through a basolaterally localized Na+-K+-Cl(-)-cotransport system (J. Clin. Invest. 75: 462, 1985), the fate of K+, which is cotransported with Cl- during VIP, and A23187-mediated Cl- secretion was explored. Unidirectional and net transepithelial 42K+ flux rates were negligible compared with 36Cl- flux rates (less than 4% of Cl- flux), indicating that little K+ was secreted along with Cl-. K+ recycling across the basolateral membrane was suggested from experiments in which 86Rb+ efflux (as a tracer for K+) was measured across the apical and basolateral membranes of 86Rb+ -preloaded monolayers under voltage-clamped conditions. In the absence of secretagogues, 86Rb+ efflux was 10-fold higher across the basolateral membrane than across the apical membrane. 86Rb+ efflux across the basolateral membrane was accelerated two- to threefold by addition of either VIP or A23187. In each case accelerated efflux was inhibited by 5 mM Ba2+. Cl- secretion induced by VIP or A23187 was also inhibited by serosal addition of Ba2+.(ABSTRACT TRUNCATED AT 250 WORDS)

The effect of barium chloride on intestinal secretion in the rat.

The addition of BaCl2 to the serosal solution, at concentrations from 0 X 25 mM to 10 mM, caused increases in potential difference, short-circuit current and resistance across stripped sheets of rat mid-intestine, although mucosal application had little effect. The response to BaCl2 was significantly greater than that obtained with the same concentration of CaCl2. An increase in net Cl- secretion entirely accounted for the rise in short-circuit current induced by BaCl2. BaCl2 inhibited net fluid uptake by everted sacs. It also enhanced the accumulation of fluid by intestinal loops in vivo and this was associated with an increased potential difference. The response to BaCl2 in vitro was not reduced in the absence of serosal Ca2+ ions. The effect of BaCl2 was abolished by trifluoperazine and also by TMB-8 (8-(N,N-diethylamino)-octyl-3,4,5-trimethoxybenzoate hydrochloride). BaCl2 did not alter cyclic AMP production by isolated enterocytes. It is concluded that BaCl2 induces intestinal secretion by releasing Ca2+ from intracellular stores which then combines with calmodulin to stimulate the secretory process.

THE ACTION OF LOPERAMIDE IN INHIBITING PROSTAGLANDIN‐INDUCED INTESTINAL SECRETION IN THE RAT

1 The mechanisms by which loperamide inhibits the intestinal secretion induced by prostaglandin E2 were investigated in rat jejunum. 2 In vivo loperamide prevented prostaglandin-induced fluid secretion but did not reduce the associated rise in the transintestinal potential difference. 3 In intestinal sheets the electrical response to prostaglandin E2 was enhanced in the presence of loperamide. 4 The ionic basis of these changes was determined by measuring Na+ and Cl- fluxes across intestinal sheets. Loperamide did not reduce the prostaglandin-induced increase in net Cl- secretion, although it prevented the inhibition of mucosal-to-serosal Na+ movement. 5 Loperamide does not alter cyclic adenosine 3',5'-monophosphate (cyclic AMP) levels by a direct action at the enterocyte, since in isolated enterocytes neither basal nor prostaglandin-stimulated cyclic AMP levels were affected by the drug.

Studies on locust rectum. III. Stimulation of electrogenic chloride transport by hemolymph

Hemolymph was collected from recently fed desert locusts either by adsorption onto filter paper, or by centrifugation and methanol extraction. Whole hemolymph caused both the short-circuit current (Isc) and open-circuit transepithelial electropotential difference (PD) across locust recta mounted in Ussing-type chambers to double during the steady-state period. Methanol extracts of hemolymph caused similar but smaller increases in Isc. The transepithelial resistance (R) did not change. Simultaneous measurements of 36Cl− fluxes indicated that all of the increase in Isc following stimulation could be accounted for by a parallel increase in net Cl− absorption from the lumen side. With the exception of an initial small biphasic fluctuation in Isc, stimulation by hemolymph exhibited identical characteristics to those produced by submaximal dosages of corpora cardiaca (CC). Cardiatectomy drastically reduced the stimulatory activity of hemolymph, suggesting that this neuroendocrine organ is the source of the active factor (chloride transport stimulating hormone (CTSH)) in hemolymph.

Bile salts and bioelectric properties of rat jejunum.

Addition of conjugated bile salts increased transmural- and transserosal-potential differences of sheets of rat jejunum. Removal of Cl- from buffer solutions minimized the bile-sale induced bioelectric changes. Bile-salt induced doubling of tissue resistance was not explained by an observed increase in net Cl- serosa leads to mucosa flux. Electrical effects were unrelated to concentration and were observed only when bile-salt solutions perfused the jejunal mucosa. The molecular events associated with bile-salt interactions with the plasma membrane affecting ion fluxes may relate to their unique effects on sterol absorption.

Ba++ on the resting frog stomach: effects on electrical and secretory parameters.

Ba++ added to the nutrient solution bathing the resting frog stomach increased resistance, decreased the PD, and stimulated acid secretion. Under short-circuit conditions, the increase in H+-secretory rate was accompanied by a decrease in short-circuit current (I-sc). These changes were reversed by NaSCN (10 mM), suggesting that Ba++ had not impaired the current-generating mechanism per se. Histamine-induced acid secretion was associated with an increase in net Cl- flux, particularly in the N yields S flux (JNS). Ba++ increased acid secretion with no increase in JNS and a decrease in net Cl- flux. The effects of Ba++ were amplified by low-Cl- solutions. Histamine, in the presence of Ba++ and low-Cl- solutions, increased acid secretion and transmucosal resistance, suggesting the operation of a neutral pump in the secretion of HCl. It is concluded that Ba++ limits Cl- entry and also acts as a secretagogue.

Interactions between lysine, Na+ and Cl− transport in rat jejunum

Abstract The effect of l -lysine (10 mM) on transmural electrical potential difference and short-circuit currents was studied in jejunal and ileal preparations from rat small intestine. It was seen that the jejunal response was much less than the ileal one but rose to the ileal level when Cl − was substituted for by SO 4 2− , whereas the ileal response was independent of the type of anion. In the short-circuited jejunum the lysine (10 mM) transport was found to be active and Na + dependent. It was further associated with an increase of the net flux of Na + from the mucosal to the serosal side, both in Cl − and in SO 4 2− media. With the short-circuited jejunal preparation it was also demonstrated that lysine (10 mM) increased the net flux of Cl − from the mucosal to the serosal side. With the short-circuited ileal preparation lysine did not influence the Cl − net flux. In the absence of lysine the short-circuited jejunum exhibited significant Cl − secretion. It is concluded that rat jejunal transport of lysine is active and Na + dependent and that the failure to observe lysine-induced increments in jejunal short-circuiting current in excess of what can be accounted for by the net flux of lysine results from a lysine-induced increase in net Cl − flux from the mucosal to the serosal side of the gut wall, that is by an inhibition of Cl − secretion. Based on current evidence it is concluded that measurements of changes in electrical transmural potential difference and short-circuiting current are at best poor and often plainly misleading substitutes for actual flux measurements.