Apical acidification induces paracellular injury in canine gastric mucosal monolayers

1994 ◽  
Vol 267 (6) ◽  
pp. G1012-G1020 ◽  
Author(s):  
M. C. Chen ◽  
A. Chang ◽  
T. Buhl ◽  
M. Tanner ◽  
A. H. Soll
Keyword(s):  
Short Circuit ◽  
Phase 1 ◽  
Short Circuit Current ◽  
Paracellular Permeability ◽  
Ussing Chambers ◽  
Low Concentrations ◽  
Three Phase ◽  
Mucosal Cells ◽  
Primary Monolayer ◽  
Primary Monolayer Cultures

We used primary monolayer cultures of enzyme-dispersed canine oxyntic mucosal cells mounted in Ussing chambers to characterize the apical barrier to H+. [3H]mannitol flux (MF) and [14C]inulin flux (IF) were used as size probes for tight junctions. Apical H+ produced a three-phase effect. In phase 1, as the apical pH was decreased from 7 to about 2.5, resistance (R) increased, but short-circuit current (Isc) did not change. In phase 2, an increased paracellular permeability developed at pH below 2.5-1.7, evidenced by decreased R and increased MF but not IF. Size sieving and monolayer integrity were preserved, and this paracellular leak was either fully reversed or stabilized by apical neutralization, depending on the duration of the paracellular leak. In phase 3, after sustained exposure to an apical pH below approximately 2, transepithelial integrity was lost; R decreased to fluid R, and both MF and IF increased. Basolateral acidification below pH 5.5 produced rapid monolayer disruption. Low concentrations of cytochalasin D (CD) decreased R and increased MF but not IF; apical acidification to pH 4 after CD increased R and decreased the MF, indicating reduced paracellular permeability by apical H+. Apical amiloride did not alter Isc; however, after 48 h of treatment with hydrocortisone and insulin, an amiloride-sensitive Isc component became evident. Our data indicate that the increase in R observed with apical acidification reflects decreased paracellular permeability and that the earliest injury with apical acidification is a selective paracellular leak.

Neural control of intestinal ion transport and paracellular permeability is altered by nutritional status

2000 ◽  
Vol 278 (6) ◽  
pp. R1589-R1594 ◽  
Author(s):  
Ursula L. Hayden ◽  
Hannah V. Carey
Keyword(s):  
Ion Transport ◽  
Nicotinic Receptors ◽  
Neural Control ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Paracellular Permeability ◽  
Cholinergic Stimulation ◽  
Fed State ◽  
Ussing Chambers ◽  
Intestinal Ion Transport

This study examined the effect of fasting on the neural control of ion transport and paracellular permeability in piglet jejunum. Muscle-stripped tissues from fed or 48-h fasted piglets were mounted in Ussing chambers. Neural blockade with tetrodotoxin (TTX) or antagonists of muscarinic or nicotinic receptors caused reductions in basal short-circuit current that were approximately threefold greater in fasted piglets. The TTX-induced reduction in short-circuit current in fasted piglets was due to a decrease in residual ion flux and was abolished in the absence of[Formula: see text]. Intestinal paracellular permeability, as indicated by tissue conductance (Gt) and fluxes of inulin and mannitol, was significantly increased by fasting. TTX increased inulin flux and Gt in fed but not fasted piglets. In fasted piglets, carbachol reduced Gt by 29% and mannitol flux by 27% but had no effect on these parameters in the fed state. We conclude that fasting enhances enteric neural control of basal ion transport and increases paracellular permeability in piglet jejunum. Tonic release of enteric neurotransmitters regulates paracellular permeability in the fed state, and cholinergic stimulation restores fasting-induced elevations in paracellular permeability to fed levels.

Source and distribution of factors in locust nervous system which stimulate rectal Cl− transport

1985 ◽  
Vol 63 (1) ◽  
pp. 37-41 ◽  
Author(s):  
J. Proux ◽  
B. Proux ◽  
J. Phillips
Keyword(s):  
Chloride Transport ◽  
Short Circuit ◽  
Active Form ◽  
Short Circuit Current ◽  
Neurosecretory Cells ◽  
Neuroendocrine Cells ◽  
Response Curves ◽  
Ussing Chambers ◽  
Cl Transport ◽  
Low Concentrations

Corpus cardiacum (CC) of locusts contains a potent stimulant of rectal Cl transport, i.e., chloride transport stimulating hormone (CTSH). Extracts of other locust tissues which contain neuroendocrine cells were assayed for their ability to stimulate Cl-dependent, short-circuit current (Isc) across recta mounted in Ussing chambers. Dose–response curves indicated that only brain (particularly the pars intercerebralis, PI) contained substantial stimulatory activity, but this was still at very low concentrations compared with CTSH activity in CC. Destruction of either the PI or the nerve tract from PI to CC (i.e., NCC I) reduced CTSH levels in the CC. In contrast, destruction of lateral neurosecretory cells (NSCs) of the brain had no effect. The results suggest that CTSH is synthesized in NSCs of the PI and is transported down NCC I to the CC where it may mature to a more active form.

Protein kinase C potentiates cAMP-stimulated mouse duodenal mucosal bicarbonate secretion in vitro

2004 ◽  
Vol 286 (5) ◽  
pp. G814-G821 ◽  
Author(s):  
Bi-Guang Tuo ◽  
Jimmy Y. C. Chow ◽  
Kim E. Barrett ◽  
Jon I. Isenberg
Keyword(s):  
Short Circuit ◽  
Short Circuit Current ◽  
Duodenal Mucosa ◽  
Bicarbonate Secretion ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Ussing Chambers ◽  
Duodenal Bicarbonate Secretion

PKC has been shown to regulate epithelial Cl- secretion in a variety of models. However, the role of PKC in duodenal mucosal bicarbonate secretion is less clear. We aimed to investigate the role of PKC in regulation of duodenal mucosal bicarbonate secretion. Bicarbonate secretion by murine duodenal mucosa was examined in vitro in Ussing chambers using a pH-stat technique. PKC isoform expression and activity were assessed by Western blotting and in vitro kinase assays, respectively. PMA (an activator of PKC) alone had no effect on duodenal bicarbonate secretion or short-circuit current ( Isc). When PMA and dibutyryl-cAMP (db-cAMP) were added simultaneously, PMA failed to alter db-cAMP-stimulated duodenal bicarbonate secretion or Isc ( P > 0.05). However, a 1-h preincubation with PMA potentiated db-cAMP-stimulated duodenal bicarbonate secretion and Isc in a concentration-dependent manner (from 10-8 to 10-5M) ( P < 0.05). PMA preincubation had no effects on carbachol- or heat-stable toxin-stimulated bicarbonate secretion. Western blot analysis revealed that PKCα, -γ, -ϵ, -θ, -μ, and -ι/λ were expressed in murine duodenal mucosa. Ro 31–8220 (an inhibitor active against PKCϵ, -α, -β, and -γ), but not Gö 6983 (an inhibitor active against PKCα, -γ, -β, and -δ), reversed the potentiating effect of PMA on db-cAMP-stimulated bicarbonate secretion. PMA also time- and concentration-dependently increased the activity of PKCϵ, an effect that was prevented by Ro 31–8220 but not Gö 6983. These results demonstrate that activation of PKC potentiates cAMP-stimulated duodenal bicarbonate secretion, whereas it does not modify basal secretion. The effect of PKC on cAMP-stimulated bicarbonate secretion is mediated by the PKCϵ isoform.

Effect of pH on chloride absorption in the flounder intestine

1988 ◽  
Vol 255 (2) ◽  
pp. G247-G252 ◽  
Author(s):  
A. N. Charney ◽  
J. I. Scheide ◽  
P. M. Ingrassia ◽  
J. A. Zadunaisky
Keyword(s):  
Small Intestine ◽  
Short Circuit ◽  
Ph Effect ◽  
Short Circuit Current ◽  
Bathing Solution ◽  
Solution Ph ◽  
Transport Pathway ◽  
Ussing Chambers ◽  
Chloride Absorption ◽  
In Series

Chloride absorption in the small intestine of the winter flounder, Pseudopleuronectes americanus, is reported to be sensitive to ambient pH. We studied this sensitivity in isolated stripped intestinal mucosa mounted in modified Ussing chambers. Unidirectional 36Cl fluxes (JClm----s, JCls----m) were measured under short-circuited conditions in bathing solutions containing various combinations of HCO3- (0-20 mM), partial pressure of CO2 (0-36 mmHg), and pH (6.77-7.85). We found that JClm----s, net 36Cl flux (JClnet), and short-circuit current (Isc) increased and JCls----m decreased predominately in response to increases in bathing solution pH. There was a linear relationship between pH and both JClnet (r = 0.92, P less than 0.01) and Isc (r = 0.96, P less than 0.005) between pH 6.77 and 7.74. The pH effect was completely reversible, did not require either CO2 or HCO3-, and was not affected by the presence of mucosal barium at 1 mM. Mucosal bumetanide (0.1 mM) completely inhibited the pH effect. These data suggest that the process by which Cl- is absorbed in the flounder intestine is sensitive to pH. The data do not indicate whether pH affects Na+-K+-2Cl- cotransport or a Cl- transport pathway in series with this process. The direction of Cl- absorption in response to pH contrasts with inverse relation of pH and Cl- absorption in mammalian small intestine.

Active electrolyte transport in mammalian buccal mucosa

1988 ◽  
Vol 255 (3) ◽  
pp. G286-G291 ◽  
Author(s):  
R. C. Orlando ◽  
N. A. Tobey ◽  
V. J. Schreiner ◽  
R. D. Readling
Keyword(s):  
Buccal Mucosa ◽  
Short Circuit ◽  
Basolateral Membrane ◽  
Electrical Potential ◽  
Short Circuit Current ◽  
Electrolyte Transport ◽  
Electrical Potential Difference ◽  
Ussing Chambers ◽  
Net Fluxes

The transmural electrical potential difference (PD) was measured in vivo across the buccal mucosa of humans and experimental animals. Mean PD was -31 +/- 2 mV in humans, -34 +/- 2 mV in dogs, -39 +/- 2 mV in rabbits, and -18 +/- 1 mV in hamsters. The mechanisms responsible for this PD were explored in Ussing chambers using dog buccal mucosa. After equilibration, mean PD was -16 +/- 2 mV, short-circuit current (Isc) was 15 +/- 1 microA/cm2, and resistance was 1,090 +/- 100 omega.cm2, the latter indicating an electrically "tight" tissue. Fluxes of [14C]mannitol, a marker of paracellular permeability, varied directly with tissue conductance. The net fluxes of 22Na and 36Cl were +0.21 +/- 0.05 and -0.04 +/- 0.02 mueq/h.cm2, respectively, but only the Na+ flux differed significantly from zero. Isc was reduced by luminal amiloride, serosal ouabain, or by reducing luminal Na+ below 20 mM. This indicated that the Isc was determined primarily by active Na+ absorption and that Na+ traverses the apical membrane at least partly through amiloride-sensitive channels and exits across the basolateral membrane through Na+-K+-ATPase activity. We conclude that buccal mucosa is capable of active electrolyte transport and that this capacity contributes to generation of the buccal PD in vivo.

ENGINEERING ANALYSIS OF THE THREE-PHASE SHORT CIRCUIT MODE IN THE 10 KV ELECTRIC NETWORK TAKING INTO ACCOUNT THE INFLUENCE OF LOAD CURRENTS

2021 ◽  
pp. 74-83
Author(s):  
YURI D. VOLCHKOV ◽  
Keyword(s):  
Remote Monitoring ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Load Current ◽  
Electric Network ◽  
Operating Modes ◽  
Three Phase ◽  
Feed Network ◽  
Short Circuits ◽  
Switching Devices

Abstract. The load current aff ects the value of the short-circuit current in the electric network and, consequently, the voltage value. In some cases, this infl uence must be taken into account for the correct choice of switching devices, remote monitoring the operating modes of electric networks, and determining the modes. It is possible to disconnect loads connected through magnetic starters and contactors. Failure to consider the infl uence of the load current can lead to an incorrect interpretation of the identifi ed grid operating modes during remote monitoring and, as a result, incorrect dispatcher’s decisions. In addition, it is also insuffi cient to specify the choice of switching devices in the 10 kV feed network. The article describes a method for analyzing the three-phase short circuit mode in a 10 kV feed network, taking into account the infl uence of load currents. The method is exemplifi ed by the case of an actual electric network – the 10 kV ring feed network containing reclosers and receiving power from diff erent sections of lowvoltage buses of the “Kulikovskaya” 110/35/10 kV substation, belonging to the Branch of PJSC «DGC of Center”-“Orelenergo.” For this network, the values of the three-phase short-circuit currents at points with diff erent distances from the substation buses have been determined. The authors have fi guredout the values of the load currents and their shares in the total short-circuit current. The voltage values at different points of the network in the case of short circuits have also been determined. The research proves that the effect of the load current on the total short-circuit current should be taken into account for the case of remote short circuits.

Mechanisms of sodium and chloride transport across equine tracheal epithelium

1990 ◽  
Vol 259 (6) ◽  
pp. L459-L467 ◽  
Author(s):  
G. J. Tessier ◽  
T. R. Traynor ◽  
M. S. Kannan ◽  
S. M. O3'Grady
Keyword(s):  
Chloride Transport ◽  
Short Circuit ◽  
Basolateral Membrane ◽  
Short Circuit Current ◽  
Ringer Solution ◽  
Tracheal Epithelium ◽  
Ussing Chambers ◽  
Cl Secretion ◽  
Cotransport System ◽  
Ethanesulfonic Acid

Equine tracheal epithelium, stripped of serosal muscle, mounted in Ussing chambers, and bathed in plasmalike Ringer solution generates a serosa-positive transepithelial potential of 10–22 mV and a short-circuit current (Isc) of 70–200 microA/cm2. Mucosal amiloride (10 microM) causes a 40–60% decrease in Isc and inhibits the net transepithelial Na flux by 95%. Substitution of Cl with gluconate resulted in a 30% decrease in basal Isc. Bicarbonate substitution with 20 mM N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid decreased the Isc by 21%. The Cl-dependent Isc was inhibited by serosal addition of 1 mM amiloride. Bicarbonate replacement or serosal amiloride (1 mM) inhibits the net Cl flux by 72 and 69%, respectively. Bicarbonate replacement significantly reduces the effects of serosal amiloride (1 mM) on Isc, indicating its effect is HCO3 dependent. Addition of 8-bromoadenosine 3',5'-cyclic monophosphate (8-BrcAMP; 100 microM) causes a 40% increase in Isc. This effect is inhibited by subsequent addition of 10 microM serosal bumetanide. Bumetanide (10 microM) reduces net Cl secretion following stimulation with 8-BrcAMP (100 microM). Serosal addition of BaCl2 (1 mM) causes a reduction in Isc equal to that following Cl replacement in the presence or absence of 100 microM cAMP. These results suggest that 1) Na absorption depends on amiloride-inhibitable Na channels in the apical membrane, 2) Cl influx across the basolateral membrane occurs by both a Na-H/Cl-HCO3 parallel exchange mechanism under basal conditions and by a bumetanide-sensitive Na-(K?)-Cl cotransport system under cAMP-stimulated conditions, and 3) basal and cAMP-stimulated Cl secretion depends on Ba-sensitive K channels in the basolateral membrane.

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