THE EFFECT OF VALINOMYCIN ON THE TOAD BLADDER: ANTAGONISM TO VASOPRESSIN AND ALDOSTERONE

1969 ◽  
Vol 45 (2) ◽  
pp. 287-295 ◽  
Author(s):  
P. J. BENTLEY
Keyword(s):  
Macrolide Antibiotic ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Water Transfer ◽  
Toad Bladder ◽  
Na Transport ◽  
Bathing Medium ◽  
Sodium Transfer ◽  
Sites Of Action ◽  
Subsequent Addition

SUMMARY The macrolide antibiotic valinomycin decreased short-circuit current (SCC, Na transport) across the isolated bladder of the toad. This effect was not overcome by increasing the K+ levels in the bathing medium or by the action of amphotericin B. The effects of vasopressin on both sodium and water transfer across the toad bladder were inhibited by valinomycin and the latter inhibition is non-competitive. The action of theophylline in increasing water transfer across the bladder was also inhibited. Cyclic AMP also increased water and Na+ transfer across the bladder but its action was not reduced by the macrolide. These results suggest that valinomycin inhibits adenyl cyclase. Aldosterone increases sodium transport across the toad bladder and this action was abolished by previous incubation of the tissue with the macrolide. Once the steroid-induced effect had been established subsequent addition of valinomycin did not alter the sodium transfer. Valinomycin thus appears to have several sites of action on the toad bladder.

Selective effects of bumetanide on chloride transport in bullfrog cornea

1978 ◽  
Vol 234 (4) ◽  
pp. F297-F301
Author(s):  
O. A. Candia ◽  
H. F. Schoen
Keyword(s):  
Electrical Resistance ◽  
Loop Diuretic ◽  
Chloride Transport ◽  
Short Circuit ◽  
Ussing Chamber ◽  
Short Circuit Current ◽  
Selective Inhibitor ◽  
O2 Consumption ◽  
Na Transport ◽  
Bathing Medium

Frog corneas were mounted in a modified Ussing chamber and short-circuit current (SCC) and unidirectional Cl fluxes were measured. Bumetanide, a loop diuretic, at concentrations as low as 10(-7) M, reduced the SCC 29%. At 10(-5) M, bumetanide reduced the SCC 96% and increased transcorneal electrical resistance 20-51%. The forward Cl flux declined from 0.71 +/- 0.04 to 0.20 +/- 0.03 mueq/h.cm2 (n, 7), while, in separate experiments, the backward Cl flux did not change significantly (from 0.22 +/- 0.03 to 0.23 +/- 0.04; n, 7). When corneas were mounted in Cl-free Ringer and the net Na transport was stimulated with amphotericin B, 10(-5) M bumetanide had no effect on the SCC. In separate experiments the effect of 10(-5) M bumetanide on the O2 consumption was measured in a stirrer bath assembly. Bumetanide decreased the O2 consumption from 352 +/- 14 to 297 +/- 19 microliter/h.cm2 (significantly different from sham-treated controls). This decrease was similar to that obtained with furosemide or when Cl was removed from the bathing medium. We infer from these results that bumetanide is a selective inhibitor of active Cl transport in the bullfrog cornea.

scholarly journals THE EFFECT OF CALCIUM WITHDRAWAL ON THE STRUCTURE AND FUNCTION OF THE TOAD BLADDER

1965 ◽  
Vol 25 (3) ◽  
pp. 195-209 ◽  
Author(s):  
Richard M. Hays ◽  
Bayla Singer ◽  
Sasha Malamed
Keyword(s):  
Epithelial Cells ◽  
Sodium Transport ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Structure And Function ◽  
Toad Bladder ◽  
Active Sodium ◽  
Bathing Medium ◽  
Active Sodium Transport ◽  
And Function

Previous reports have indicated that calcium is necessary to support active sodium transport by the toad bladder, and may be required as well in the action of vasopressin on both toad bladder and frog skin. The structure and function of the toad bladder has been studied in the absence of calcium, and a reinterpretation of the previous findings now appears possible. When calcium is withdrawn from the bathing medium, epithelial cells detach from one another and eventually from their supporting tissue. The short-circuit current (the conventional means of determining active sodium transport) falls to zero, and vasopressin fails to exert its usual effect on short-circuit current and water permeability. However, employing an indirect method for the estimation of sodium transport (oxygen consumption), it is possible to show that vasopressin exerts its usual effect on Qoo2 when sodium is present in the bathing medium. Hence, it appears that the epithelial cells maintain active sodium transport when calcium is rigorously excluded from the bathing medium, and continue to respond to vasopressin. The failure of conventional techniques to show this can be attributed to the structural alterations in the epithelial layer in the absence of calcium. These findings may provide a model for the physiologic action of calcium in epithelia such as the renal tubule.

scholarly journals The Coupling of the Short-Circuit Current to Metabolism in the Urinary Bladder of the Toad

1963 ◽  
Vol 46 (4) ◽  
pp. 733-754 ◽  
Author(s):  
Roy H. Maffly ◽  
I. S. Edelman
Keyword(s):  
Tricarboxylic Acid Cycle ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Metabolic Effects ◽  
Metabolic Inhibitors ◽  
Na Transport ◽  
Bathing Medium ◽  
Free Media ◽  
Tricarboxylic Acid Cycle Intermediates

The relationship of the short-circuit current to metabolism was studied in the toad bladder in vitro. Substrates and inhibitors were added to the bathing medium and the effect on the short-circuit current was determined. The spontaneous decline in the short-circuit current that occurred in substrate-free media was prevented or reversed by the addition of glucose, pyruvate, lactate, or ß-hydroxybutyrate, whereas acetate and tricarboxylic acid cycle intermediates had no effect. A variety of metabolic inhibitors depressed the short-circuit current; depression by iodoacetate and by malonate was delayed by prior addition of pyruvate or lactate but not by glucose. The ability of a substrate to stimulate the current did not correlate with its rate of oxidation to CO2. On the basis of earlier studies, the metabolic effects on the short-circuit current were assumed to reflect equivalent effects on the rate of active Na transport. It is suggested that the energy for Na transport is provided not by a general cellular metabolic pool but by a specific metabolic pathway or pathways spatially linked to the transport mechanism.

Aldosterone and insulin effects on driving force of Na+ pump in toad bladder

1976 ◽  
Vol 230 (6) ◽  
pp. 1603-1608 ◽  
Author(s):  
B Siegel ◽  
MM Civan
Keyword(s):  
Urinary Bladder ◽  
Driving Force ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Toad Bladder ◽  
Na Transport ◽  
Na Pump ◽  
Before And After

Both aldosterone and insulin increase active Na+ transport across the urinary bladder of the toad. Recent data have provided further support to the concept that aldosterone acts primarily to increase Na+ entry from the mucosal medium into the transporting cells, whereas insulin acts to increase active Na+ extrusion into the serosal medium. To examine this concept further, the driving force (E(Na)) of the Na+ pump was measured, by the technique described by Yonath and Civan (48), before and after hormonal administration. Both hormones increased short-circuit current, but only insulin increased E(Na). The validity of the technique was further explored by imposing periods of hypoxia upon a series of experimental hemibladders; as expected, hypoxia reversibly decreased E(Na). The data indicate that insulin stimulates Na+ transport, in part by directly stimulating the Na+ pump. The results are also consistent with the concept that aldosterone stimulates net Na+ movement solely by enhancing Na+ entry into the transporting cells, but are subject to alternative interpretations.

Effects of high potassium concentration on theophylline responses of toad bladder

1977 ◽  
Vol 232 (2) ◽  
pp. F173-F177
Author(s):  
S. A. Mendoza ◽  
K. K. Nakamoto
Keyword(s):  
Cyclic Amp ◽  
Water Permeability ◽  
Potassium Concentration ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Toad Bladder ◽  
Osmotic Water Permeability ◽  
High Potassium ◽  
Bathing Medium ◽  
Osmotic Water

It has been demonstrated previously that a high concentration of potassium in the serosal bathing medium (5–21.5 mM) potentiates the increase in short-circuit current caused by vasopressin or exogenous cyclic AMP. The same concentration of potassium in the bathing medium inhibited the increase in short-circuit current caused by theophylline. The increases in osmotic water permeability caused by vasopressin or cyclic AMP were unaffected by a serosal potassium concentration of 21.5 mM. The increase in osmotic water permeability caused by theophylline was inhibited by 21.5 mM potassium. The concentration of cyclic AMP in either intact total bladder or isolated toad bladder cells was increased two- or three-fold by theophylline. Increasing the concentration of potassium to 21.5 mM did not alter cyclic AMP concentration in either the absence of presence of theophylline. One interpretation of these results is that theophylline increases osmotic water flow and short-circuit current by a mechanism other than by inhibition of cyclic nucleotide phosphodiesterase.

Effects of insulin, ADH, and cyclic AMP on sodium transport in the toad bladder

1977 ◽  
Vol 232 (4) ◽  
pp. F307-F314
Author(s):  
W. P. Wiesmann ◽  
S. Sinha ◽  
S. Klahr
Keyword(s):  
Protein Synthesis ◽  
Cyclic Amp ◽  
Synergistic Effects ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Toad Bladder ◽  
Na Transport ◽  
Delayed Effect ◽  
Rapid Phase ◽  
Delayed Effects

These studies further define the mechanisms by which insulin stimulates Na transport in the toad bladder. Serosal but not mucosal addition of insulin, 100-1,000 mU/ml, stimulated short-circuit current (SCC) by 25-50%. The initial rise in SCC occurred at 5 min and the peak response at 15-25 min. Doses of insulin greater than 250 mU/ml increased SCC values for up to 3 h. Actinomycin D did not block the early rise in SCC produced by insulin, but it blocked the delayed effects. Insulin increased SCC in substrate-depleted bladders, although the increase in SCC was less (P less than 0.01) than in nonsubstrate-depleted bladders. Pyruvate addition to substrate-depleted bladders restored to normal the rise in SCC observed after insulin. Simultaneous addition of ADH and insulin led to an increase in SCC that was greater than the sum of the responses observed when each hormone was added independently. Synergistic effects on SCC were also obtained with cyclic AMP and insulin. Insulin did not increase cyclic AMP levels in toad bladder epithelial cells. It is suggested that insulin stimulates active Na transport by two mechanisms: 1) a rapid phase, which may involve unmasking of pump sites within the membrane, and 2) a delayed effect which seems to require protein synthesis. The synergism of which seems to require protein synthesis. The synergism of insulin with ADH or cyclic AMP may reflect a facilitative effect of insulin on ADH or cyclic AMP-sensitive pump sites or, alternatively, the uncovering of latent pump sites that then may be available to stimulation by ADH or cyclic AMP.

Testosterone: a specific competitive antagonist of aldosterone in the toad bladder

1980 ◽  
Vol 239 (5) ◽  
pp. F433-F439 ◽  
Author(s):  
B. C. Rossier ◽  
K. Geering ◽  
H. P. Gaggeler ◽  
M. Claire ◽  
P. Corvol
Keyword(s):  
Binding Sites ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Toad Bladder ◽  
Molar Ratio ◽  
Base Line ◽  
Na Transport ◽  
Competitive Antagonist ◽  
Antagonist Action

Testosterone (100 nM to 40 microM) antagonized the effect of aldosterone (10 nM) on Na+ transport in the toad bladder measured in vitro as short-circuit current (SCC). Half-maximal inhibition occurred at an antagonist-agonist molar ratio of 150:1. The antagonist action of testosterone was reversed by addition of more aldosterone. The antagonism was specific in the sense that testosterone (20 microM) did not inhibit the response of the SCC to oxytocin (50 mU/ml). By itself, testosterone (up to 20 microM) had no agonist activity on base-line SCC. Finally, testosterone (500 nM to 20 microM) specifically displaced [3H]aldosterone (5 nm) from its cytoplasmic and nuclear binding sites in bladders incubated in vitro at 25 or 0 degrees C and labeled at steady state. There was a significant linear correlation between the effect of testosterone on the aldosterone-dependent SCC and its effect on [3H]aldosterone binding sites in the cytoplasm and in the nucleus. We conclude that 1) testosterone is a specific competitive antagonist of aldosterone, and 2) [3H]aldosterone nuclear and cytoplasmic binding sites could be mineralocorticoid receptors, mediating the action of aldosterone on Na+ transport.

ACTIONS OF VASOPRESSIN AND ALDOSTERONE ON THE TOAD BLADDER: INHIBITION BY ETHACRYNIC ACID

1969 ◽  
Vol 43 (3) ◽  
pp. 347-357 ◽  
Author(s):  
P. J. BENTLEY
Keyword(s):  
Sodium Transport ◽  
Ethacrynic Acid ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Binding Activity ◽  
Toad Bladder ◽  
Na Transport ◽  
Atpase Inhibitor ◽  
Structural Analogues ◽  
Osmotic Effects

SUMMARY Ethacrynic acid inhibited short-circuit current (SCC, Na transport) across the toad bladder. Some structural analogues had similar effects and their potency varied with their ability to combine with SH groups. The Na-K ATPase inhibitor ouabain was more effective than ethacrynic acid and its action and that of ethacrynic acid was additive. The natriferic and hydro-osmotic effects of vasopressin were inhibited by ethacrynic acid (10−5 and 10−4m); the antagonism was competitive at 10− and 10−4 m (but not at 10−3 m). Certain analogues of ethacrynic acid also exhibited this effect but it was not clearly related to their SH-combining ability. The natriferic and hydro-osmotic effects of caffeine and cyclic AMP were not changed by 10−4 m-ethacrynic acid. Ethacrynic acid (10−4 m), which does not affect basal sodium transport in bladders pretreated for studying the action of aldosterone, prevented the initiation of the increase in sodium transport by aldosterone but was without effect when added to a toad bladder already under the influence of aldosterone. This suggests that ethacrynic acid interferes with metabolic processes underlying the action of aldosterone. Dihydroethacrynic acid which has no SH-binding activity was without effect.

OSMOTIC INHIBITION OF THE NATRIFERIC RESPONSE OF THE TOAD URINARY BLADDER TO VASOPRESSIN

1975 ◽  
Vol 67 (1) ◽  
pp. 119-125
Author(s):  
P. J. BENTLEY
Keyword(s):  
Urinary Bladder ◽  
Water Movement ◽  
Short Circuit ◽  
Electrical Potential ◽  
Short Circuit Current ◽  
Toad Bladder ◽  
Toad Urinary Bladder ◽  
Electrical Potential Difference ◽  
Osmotic Gradient

SUMMARY The electrical potential difference and short-circuit current (scc, reflecting active transmural sodium transport) across the toad urinary bladder in vitro was unaffected by the presence of hypo-osmotic solutions bathing the mucosal (urinary) surface, providing that the transmural flow of water was small. Vasopressin increased the scc across the toad bladder (the natriferic response), but this stimulation was considerably reduced in the presence of a hypo-osmotic solution on the mucosal side, conditions under which water transfer across the membrane was also increased. This inhibition of the natriferic response did not depend on the direction of the water movement, for if the osmotic gradient was the opposite way to that which normally occurs, the response to vasopressin was still reduced. The natriferic response to cyclic AMP was also inhibited in the presence of an osmotic gradient. Aldosterone increased the scc and Na+ transport across the toad bladder but this response was not changed when an osmotic gradient was present. The physiological implications of these observations and the possible mechanisms involved are discussed.

11-Dehydrocorticosterone, a glucocorticoid metabolite, inhibits aldosterone action in toad bladder

1991 ◽  
Vol 261 (5) ◽  
pp. F873-F879 ◽  
Author(s):  
A. S. Brem ◽  
K. L. Matheson ◽  
J. L. Barnes ◽  
D. J. Morris
Keyword(s):  
Sodium Transport ◽  
Cell Layer ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Hydroxysteroid Dehydrogenase ◽  
Toad Bladder ◽  
Compound A ◽  
Dose Dependent Fashion ◽  
Epithelial Cell Layer ◽  
Dose Dependent

The enzyme 11 beta-hydroxysteroid dehydrogenase (11 beta-OHSD) metabolizes glucocorticoid hormones and diminishes their ability to induce sodium transport. In these studies, we determined the location of this enzyme in toad bladder and assessed the biological role for its 11-dehydro end product. Employing a polyclonal antibody directed toward 11 beta-OHSD and immunofluorescence techniques, we located the enzyme in the epithelial cell layer of the toad bladder. Although corticosterone (10(-7) M) can partially suppress aldosterone (10(-7) M)-stimulated short-circuit current (SCC), a clear excess of corticosterone (10(-6) M) did not inhibit the aldosterone-induced induced (10(-8) M) rise in SCC (n = 6). The 11-dehydro product of corticosterone, 11-dehydrocorticosterone (compound A) added to the serosal bath suppressed aldosterone (10(-8) M) peak SCC (360 min) in a dose-dependent fashion reaching 46 +/- 5% of control values at 10(-5) M (n = 6; P less than 0.001). Compound A (10(-5) M) in the mucosal bath also was capable of partially inhibiting the peak aldosterone rise in SCC to 63 +/- 7% of control values with aldosterone at 10(-8) M (n = 6; P less than 0.01) and to 64 +/- 10% of control values with aldosterone at 10(-7) M (n = 9; P less than 0.01). Compound A alone at 10(-5) M did not have any effect on SCC. Isolated toad bladders were not able to transform compound A (at 10(-8) and 10(-5) M) back to corticosterone. Thus the 11-dehydro end product of 11 beta-OHSD (compound A) may play a biologic role by regulating a component of mineralocorticoid-induced sodium transport.

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