Norepinephrine induces Na+-H+ and Cl -HCO3 exchange in Amphiuma intestine: locus and response to amiloride

1988 ◽  
Vol 255 (1) ◽  
pp. G18-G26
Author(s):  
J. F. White ◽  
C. F. Hinton

Catecholamines stimulate Na+-dependent acid secretion by Amphiuma small intestine. Studies were undertaken to localize the response within the mucosa and characterize the effect on Na+ and Cl- transport. Stripped segments of jejunum were mounted in tissue chambers that permitted isolation of villus or intervillus epithelium. In Cl-free medium, norepinephrine (NE) stimulated the transepithelial voltage (Vms) in both villus and intervillus epithelium, whereas galactose and valine elevated Vms predominately in the villus. Paired segments of whole mucosa were maintained under short circuit while the rate of acid secretion (JH) was measured by titration of the unbuffered serosal medium and unidirectional fluxes of Na+ were measured by 22Na. NE significantly stimulated net Na+ absorption (JNanet), short circuit current (Isc), and JH. Amiloride reduced JH and Isc in NE-stimulated tissues and blocked the stimulation of JNanet by NE. The NE-induced current was nearly completely and reversibly inhibited by replacement of luminal medium HCO3- or CO2. NE significantly stimulated net Cl- absorption without changing Isc or JH. It is concluded that cells throughout the mucosa respond to catecholamines with enhanced Na+ and Cl- absorption, possibly through induction or stimulation of Na+-H+ and Cl- -HCO3- exchange.

1980 ◽  
Vol 239 (6) ◽  
pp. G532-G535 ◽  
Author(s):  
A. Ayalon ◽  
A. Corcia ◽  
G. Klemperer ◽  
S. R. Caplan

The effect of furosemide on acid secretion and Cl- transport was studied in isolated fundic mucosa of the guinea pig. Furosemide (10(-3) M), applied to the serosal side produced an immediate effect on the short-circuit current (Isc), lowering it by 47 +/- 2%. Potential difference decreased by 29 +/- 3%, electrical conductance by 18 +/- 4%, acid secretion by 38 +/- 1%, and net flux of Cl- from serosal-to-mucosal side by 37%. Application of the drug to the mucosal side produced similar effects on acid secretion and on the electrical parameters. It is suggested that furosemide blocks the entrance of Cl-, by the Na+--Cl- cotransport mechanism, through the basolateral membrane of the secreting cell. The consequent reduction in electrogenic Cl- transport would cause Isc and acid secretion to decrease. A reduction of Cl- conductance of the apical membrane, upon mucosal application of the drug, would cause similar effects on acid secretion and Cl- transport.


1991 ◽  
Vol 155 (1) ◽  
pp. 455-467
Author(s):  
R. BRENT THOMSON ◽  
N. AUDSLEY ◽  
JOHN E. PHILLIPS

The commonly used method of passing short-circuit current (Isc) across insect epithelia through Ag-AgCl electrodes, without the use of salt bridges, leads to significant OH− production at the cathode (lumen side) when high currents are applied. The alkalization of the lumen previously reported when cyclic AMP was added to short-circuited locust hindgut is a result of this phenomenon rather than cyclic-AMP-mediated stimulation of acid-base transport in the hindgut. When salt bridges are used to pass short-circuit current across locust hindgut, acid secretion (JH) into the lumen equals alkaline movement (JOH) to the haemocoel side, and JH is similar under both open- and short-circuit conditions. JH is similar (1.5 μequiv cm−2 h−1) in recta and ilea. Addition of cyclic AMP inhibits JH across the rectum by 42–66%, but has no effect on the ileum when salt bridges are used. Electrical parameters (Isc, Vt, Rt) reflecting hindgut Cl− transport (JCL) before and after stimulation with cyclic AMP are the same whether or not salt bridges are used. We found no evidence of any coupling between JCl and JH/JOH.


1992 ◽  
Vol 262 (2) ◽  
pp. G312-G318 ◽  
Author(s):  
S. K. Nath ◽  
P. Dechelotte ◽  
D. Darmaun ◽  
M. Gotteland ◽  
M. Rongier ◽  
...  

L-Glutamine (Gln) fluxes and the effects of Gln on Na and Cl transport were studied across the ileum of healthy and rabbit diarrheagenic Escherichia coli (RDEC-1)-infected weanling rabbits. Stable ([alpha-15N]Gln) and radioisotopic ([U-14C]Gln) tracers provided identical estimates of Gln transport both in healthy (H) and infected (I) rabbits. RDEC-1 infection, however, decreased net Gln flux [Jnet[14C]Gln = 682 +/- 147 (H) vs. 278 +/- 63 (I); Jnet[15N]Gln = 739 +/- 160 vs. 225 +/- 110 nmol.h-1.cm-2] due to a reduction in mucosal-to-serosal flux. After addition of Gln, increases in net Na absorption [delta Jnet[15N]Gln = 1.87 +/- 0.45 (H) vs. 0.70 +/- 0.27 (I) microeq.h-1.cm-2] and short-circuit current (delta Isc) [1.80 +/- 0.40 (H) vs. 0.74 +/- 0.14 (I) microeq.h-1.cm-2] were also reduced in infected rabbits. Addition of glucose after Gln, however, stimulated Na absorption further. These results indicate that 1) Gln is actively absorbed as intact Gln molecule across rabbit ileum; 2) Gln stimulates an electrogenic Na absorption in a 1:2 ratio that may be further stimulated by glucose; and 3) in RDEC-1 infection electroneutral NaCl absorption, intact Gln absorption, and electrogenic stimulation of Na absorption by glutamine are reduced.


1981 ◽  
Vol 241 (5) ◽  
pp. G389-G396 ◽  
Author(s):  
J. F. White ◽  
M. A. Imon

Isolated segments of jejunum from Amphiuma bathed in Cl--free (SO42(-)) media usually generated serosa-negative electrical potentials when HCO3(-) was present in the media. Bidirectional isotope fluxes under short circuit revealed a negligible absorption of Na+ and a residual flux consistent with anion absorption. Acetazolamide (10(-4) M) eliminated the short-circuit current and the residual flux. Segments of jejunum exposed on the mucosal surface to HCO3(-) (pH 7.4) alkalinized the unbuffered serosal fluid at a rate of about 1.1 mueq . h-1 . cm-2, as measured by the pH-stat while clamped at zero transepithelial potential. Acetazolamide, anoxia, and 2,4-dinitrophenol lowered the rate of alkalinization and simultaneously reduced the short-circuit current by an equal amount. Absorption was constant above a [HCO3(-)] of 35 meq/l and uninfluenced by applied transepithelial voltage gradients. HCO3(-) absorption was not reduced after replacement of media Na+ or Cl- but was reduced on addition of ouabain or removal of K+. It is concluded that the jejunum actively absorbs HCO3(-) by an electrogenic process that does not involve Na+-H+ exchange.


1998 ◽  
Vol 201 (11) ◽  
pp. 1753-1762 ◽  
Author(s):  
T M Clark ◽  
T K Hayes ◽  
G M Holman ◽  
K W Beyenbach

The mechanism of action of synthetic CCRF-DP, the corticotropin-releasing factor (CRF)-related diuretic peptide of the salt marsh mosquito Culex salinarius, was investigated in isolated Malpighian tubules of the yellow fever mosquito Aedes aegypti. A low concentration of CCRF-DP (10(-9)mol l-1) caused a small but insignificant increase in transepithelial secretion of NaCl and fluid, but significantly reduced transepithelial voltage and resistance without a change in short-circuit current, pointing to the stimulation of passive Cl- transport through the paracellular pathway as the principal mechanism of a mild diuresis. Significant changes in voltage and resistance but not in short-circuit current were duplicated by the ionophore A23187 (0.4 micromol l-1), suggesting Ca2+ as a second messenger at 10(-9)mol l-1 CCRF-DP. A high concentration of CCRF-DP (10(-7)mol l-1) significantly increased transepithelial secretion of NaCl and fluid and significantly increased short-circuit current, pointing to the stimulation of active Na+ transport through the transcellular pathway as the mechanism of a strong diuresis. This effect was mimicked by dibutyryl-cAMP, suggesting cAMP as a second messenger at 10(-7)mol l-1 CCRF-DP. Dibutyryl-cGMP had no effects. These results suggest dose-dependent, receptor-mediated effects of CCRF-DP that target discrete transport pathways via discrete second messengers: low concentrations of CCRF-DP cause a mild diuresis, apparently via Ca2+-mediated effects on paracellular Cl- transport, and high concentrations cause a strong diuresis via cAMP-mediated effects on active transcellular Na+ transport in addition to the effects on the paracellular pathway.


1981 ◽  
Vol 90 (1) ◽  
pp. 123-142
Author(s):  
M. M. P. RAMOS ◽  
J. C. ELLORY

1. The tissue was found to have a serosa negative potential, and short-circuit currents equivalent to the net Cl transport. 2. A significant part of the Cl uptake was Na dependent and a similar fraction of the Na uptake was Cl dependent. 3. Short-circuit current and uptake of both ions were inhibited by loop diuretics and analogues. 4. I80 and P.D. were abolished by ouabain. 5. The observations are consistent with the idea of a coupled NaCl entry into the cell, using the energy inherent in the Na gradient; Na being pumped out of the cells by the Na pump and followed electrically by Cl−. Net chloride transport and the serosa negative potential would be a consequence of the permselective properties of the junctions allowing Na but not Cl to recycle back to the mucosal solution.


1982 ◽  
Vol 99 (1) ◽  
pp. 349-362
Author(s):  
M. CHAMBERLIN ◽  
J. E. PHILLIPS

1. Recta of desert locusts were short-circuited and depleted of endogenous substrates by exposing them to saline containing cyclic AMP but no metabolites. Individual substrates were then added to substrate-depleted recta and the change in short-circuit current (Isc) monitored. 2. Proline or glucose (50 mM) caused by far the largest increase in Isc of all substrates tested. Stimulation of the Isc by proline was not dependent upon external sodium, but did require external chloride. 3. Physiological levels of proline also caused a large increase in Isc, while physiological levels of glucose produced a much smaller stimulation. Over 90% of the proline-dependent Isc stimulation can be produced by adding 15 mM proline solely to the lumen side of the tissue. 4. These results are discussed with regard to rectal oxidative metabolism and availability of metabolic substrates in vivo. High levels of proline in Malpighian tubule fluid are probably the major substrate source for rectal Cl−transport. Note:


1988 ◽  
Vol 255 (2) ◽  
pp. G247-G252 ◽  
Author(s):  
A. N. Charney ◽  
J. I. Scheide ◽  
P. M. Ingrassia ◽  
J. A. Zadunaisky

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.


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