Action of adenosine on chloride active transport of isolated frog cornea

1979 ◽  
Vol 237 (2) ◽  
pp. F121-F127
Author(s):  
B. S. Spinowitz ◽  
J. A. Zadunaisky

Addition of adenosine (10–7 to 10–4 M) to the tear side of isolated corneas (Rana catesbeiana) produced a rapid, sustained increase in short-circuit current, potential difference, and radioisotopic chloride net flux. The increased net chloride flux accounted for the increased short-circuit current. Adenosine, a known activator of adenyl cyclase in other tissues, exerted its effects on chloride transport through a receptor different from the one described for epinephrine and prostaglandins in the corneal epithelium. Propranolol inhibited the epinephrine response but not the adenosine effect. Dipolyphloretin phosphate inhibited prostaglandin responses but did not affect the adenosine stimulation of chloride transport. Adenine and/or ribose, parts of the adenosine molecule, had no stimulatory effect, but 5'-AMP had a partial effect.The activation of the chloride pump with DBcAMP blocked the response to adenosine. Adenosine interacted with the effects of theophylline. Adenosine, a naturally occurring molecule, stimulated chloride transport by activation of adenyl cyclase through a separate membrane receptor in the corneal eqithelium.

The interior of the rumen in cattle and sheep is normally maintained at a potential of about — 40 mV relative to the blood. This potential depends primarily on the occurrence of an active transport of sodium from rumen to blood, since the potential, short-circuit current and the net sodium flux are simultaneously abolished by anoxia, ouabain and removal of sodium from the bathing solutions. There is an appreciable net flux of potassium from blood to rumen. There is also a substantial active transport of chloride in the same direction as sodium and it can be reduced by treatment with acetazolamide without affecting the potential or the sodium system. Nevertheless, sodium transport is reduced by the removal of chloride ions. Omasum epithelium is similar to rumen epithelium. However, the chloride pump appears to work in both directions in this tissue. Short-circuited omasum epithelium can also transport magnesium from omasum to blood.


1978 ◽  
Vol 56 (8) ◽  
pp. 1879-1882 ◽  
Author(s):  
J. Spring ◽  
J. Hanrahan ◽  
J. Phillips

Rates of ion transport across locust recta were monitored in vitro by following fluxes of 22Na+ and 36Cl−, short-circuit current (Isc), and open-circuit electropotential difference (PD) across this epithelium for several hours. Corpora cardiaca (CC) homogenates, cAMP, theophylline, and hemolymph of recently fed locusts all stimulate electrogenic transport of Cl− across locust rectum, as indicated by a two- to three-fold increase in 36Cl− net flux, Isc, and PD. Cyclic AMP caused a Cl-dependent increase in PD across the lumen-facing but not the hemocoel-facing plasma membrane of the epithelial cells. We propose that a blood-borne factor, possibly from the CC, causes an elevation in cAMP levels in rectal tissue and that this second messenger acts by increasing Cl− entry into the cell from the rectal lumen. Additional fluid absorption accompanies the resulting increase in transport of NaCl, leading to an increase in the hemolymph volume of previously dehydrated locusts.


1963 ◽  
Vol 205 (4) ◽  
pp. 631-637 ◽  
Author(s):  
John G. Forte ◽  
Alan H. Nauss

Removal of calcium from isolated bullfrog gastric mucosae by serial washings in calcium-free solutions or by the addition of EDTA or citrate caused a fall in transmucosal potential difference, a rise in the mucosal conductance, and an apparent decrease in the rate of acid secretion as measured by the pH stat method. Unless the potential difference fell to values very nearly zero the short-circuit current was not significantly decreased. Unidirectional fluxes of chloride in both directions across the mucosa were increased by EDTA, but the net flux, or actively transported chloride, was not significantly altered. This suggests that the removal of calcium by these methods does not interfere with active chloride transport. The increased movement of sodium and sucrose across the mucosa after EDTA suggests that a large part of the permeability change is occurring at the spaces between cells. Because of the large permeability changes accompanying removal of calcium it is postulated that the process of H+ production is not directly inhibited.


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.


1975 ◽  
Vol 63 (2) ◽  
pp. 313-320
Author(s):  
J. L. Wood ◽  
A. M. Jungreis ◽  
W. R. Harvey

1. The 28Mg-measured net flux of magnesium from lumen-side to haemolymph-side of the isolated and short-circuited midgut was 1.97 +/− 0.28 mu-equiv cm(−2) /(−1) in 8 mM-Mg2+. 2. The magnesium-influx shows a delay before the tracer steady-state is attained, indicating the existence of a magnesium-transport pool equivalent to 6.7 mu-equiv/g wet weight of midgut tissue. 3. Magnesium depresses the short-circuit current produced the midgut but not the potassium transport, the depression being equal to the rate of magnesium transport. 4. Magnesium transport yields a linear Lineweaver-Burk plot with an apparent Km of 34 mM-Mg2+ and an apparent Vmax of 14.9 mu-equiv cm(−1) /(−1). 5. Magnesium is actively transported across the midgut and contributes to the regulation of the haemolymph magnesium concentration in vivo.


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.


1985 ◽  
Vol 116 (1) ◽  
pp. 153-167
Author(s):  
J. W. HANRAHAN ◽  
J. E. PHILLIPS

1. Electrophysiological and tracer flux techniques were used to studyregulation of KC1 reabsorption across locust recta. Physiologically high K+levels (100 mmolI−1) on the lumen side stimulated net 36Cl flux and reduced the theoretical energy cost of anion transport under open-circuit conductions. 2. The stimulation of short-circuit current (Ibc i.e. active C− absorption) by crude corpora cardiaca extracts (CC) was not dependent on exogenous Ca2+. Stimulations of Ibc were greatly enhanced in the presence of theophylline, indicating that the rate of synthesis of cAMP is increased by CC extracts. High CC levels lowered transepithelial resistance (Rt), suggesting that chloride transport stimulating hormone (CTSH) regulates both active Cl− absorption and counter-ion (K+) permeability. 3. High mucosal osmolarity or K+ concentration decreased Ibc and caused a disproportionately large increase in Rt, consistent with a decrease in theshunt (K+) conductance. Measurements of relative mucosal-to-serosal membrane resistance confirmed that high mucosal K+ levels reduced apical membrane conductance. Lowering mucosal pH to values observed in vivo atthe end of resorptive cycles also inhibited Ibc, apparently without affecting K+ permeability.


1990 ◽  
Vol 259 (6) ◽  
pp. L459-L467 ◽  
Author(s):  
G. J. Tessier ◽  
T. R. Traynor ◽  
M. S. Kannan ◽  
S. M. O3'Grady

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.


1964 ◽  
Vol 207 (5) ◽  
pp. 1010-1014 ◽  
Author(s):  
José A. Zadunaisky ◽  
Felisa W. De Fisch

Several aspects of chloride passage through isolated amphibian skin were studied. The chloride transport performed by the skin of the frog Leptodactylus ocellatus or the passive chloride fluxes observed in the skin of the toad Bufo arenarum Hensel are not affected by antidiuretic hormone. The chloride transport produces a negative potential and a short-circuit current in sodium-free solutions, though the unidirectional fluxes of chloride are greatly reduced under these conditions. The short-circuit current due to the chloride transport is smaller than the net chloride flux. It was found that this disagreement could be ascribed to a loss of sodium toward the inside from the sodium pool of the skin. Antidiuretic hormone did not affect the chloride current, nor the sodium loss from the skin. The isolated skin of the toad Bufo arenarum Hensel does not transport chloride ions. Thus the active transport of chloride observed in isolated skins of the frog Leptodactylus ocellatus does not depend on environmental conditions, since both animals live in the same surroundings.


1984 ◽  
Vol 247 (3) ◽  
pp. G253-G260 ◽  
Author(s):  
K. A. Hubel

Electrical field stimulation (EFS) depolarizes nerves and causes chloride secretion by mucosa of rabbit ileum mounted in a flux chamber. To test the hypothesis that the transmitter is a peptide, we determined whether the EFS response was prevented by the endopeptidase chymotrypsin (CT). Serosal, but not mucosal, addition of CT (200 micrograms/ml) reduced the short-circuit current (Isc) response to EFS by 90% or more. CT also reduced Cl absorption by decreasing the mucosal-to-serosal flux, but it did not affect net Na absorption. CT prevented the response to vasoactive intestinal polypeptides, but the response returned when CT activity was eliminated. The response to EFS did not return, however, implying that CT damaged cells that released transmitter or epithelial target cells. CT reduced the Isc response to serotonin by 69% and to A23187 by 10% and did not affect the theophylline response. We conclude that 1) the effects of CT on cell function limit its usefulness in identifying peptide neurotransmitters in epithelium, 2) CT irreversibly inhibits ion transport responses to EFS and to serotonin, and 3) CT reduces absorption of Cl probably by affecting a calcium pathway that modifies Cl transport.


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