Inhibitory effect of phorbol ester on sodium transport in frog urinary bladder

1990 ◽  
Vol 259 (3) ◽  
pp. F425-F431
Author(s):  
T. Satoh ◽  
H. Endou
Keyword(s):  
Urinary Bladder ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Inhibitory Effect ◽  
Inhibitory Influence ◽  
Dependent Manner ◽  
Camp Accumulation ◽  
Frog Urinary Bladder ◽  
Na Transport

To confirm the role of protein kinase C (PKC) on epithelial Na transport, we studied the effects of phorbol 12-myristate 13-acetate (PMA) and dioctanoylglycerol (DiC8), activators of PKC, on short-circuit current (Isc) in frog urinary bladder and further examined the influence of sphingosine, an inhibitor of PKC, on PMA- or DiC8-modulated Isc. PMA reduced basal Isc in a dose-dependent manner, and sphingosine (10 and 100 microM) partially restored PMA-reduced Isc. On the other hand, DiC8 (5 x 10(-5) M) also reduced basal Isc, and this action was completely prevented by 100 microM sphingosine. Both PMA (4 x 10(-5) M) and DiC8 inhibited vasopressin (50 mU/ml)- and forskolin (5 x 10(-5) M)-stimulated increases in Isc. PMA (4 x 10(-5) M) also inhibited 8-bromoadenosine 3',5'-cyclic monophosphate (8-BrcAMP)-stimulated increase in Isc. Furthermore, PMA (4 x 10(-5) M) and DiC8 (5 x 10(-5) M) inhibited vasopressin (50 mU/ml)-stimulated cAMP accumulation. DiC8 also inhibited forskolin-stimulated cAMP accumulation. These results indicate that PMA exerts inhibitory influence on Na transport mainly by its own potency of PKC activation. In addition, it is suggested that there is a cross talk in epithelial Na transport between PKC and cAMP-dependent pathway in frog urinary bladder.

Effects of 3'-deoxycytidine on rRNA synthesis in toad bladder: analysis of response to aldosterone

1977 ◽  
Vol 232 (5) ◽  
pp. C174-C179 ◽  
Author(s):  
B. C. Rossier ◽  
P. A. Wilce ◽  
J. F. Inciardi ◽  
F. K. Yoshimura ◽  
I. S. Edelman
Keyword(s):  
Urinary Bladder ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Early Response ◽  
Rrna Synthesis ◽  
Mrna Synthesis ◽  
Na Transport ◽  
Polyadenylated Rna ◽  
Toad Bufo

Previous studies showed that aldosterone augments transepithelial active Na+ transport and the incorporation of [3H]uridine into polyadenylated RNA (poly(A)(+)-RNA) (putatively mRNA) early in the latent period. Soon thereafter, incorporation of [methyl-14C] groups, as well as [3H]uridine into rRNA is also increased. To evaluate the role of rRNA in mineralocorticoid action, the inhibitor 3'-deoxycytidine was used in studies on the urinary bladder of the toad Bufo marinus. 3'-deoxycytidine suppressed the incorporation of [methyl-14C] and [3H]uridine into nuclear precursors of rRNA and subunits of cytoplasmic rRNA. In contrast, 3'-deoxycytidine inhibited incorporation of ]3H]uridine into cytoplasmic poly(A)(+)-RNA minimally. In control experiments, 3'-deoxycytidine had no significant effect on Na+ transport, measured as the short-circuit current (scc), when given alone. 3'-Deoxycytidine also had no significant effect on the aldosterone-dependent increase in scc. In the presence of 3'-deoxycytidine, aldosterone enhanced both the scc and the incorporation of [3H]uridine into poly(A)(+)-RNA significantly. We conclude that during the first 3 h, the mineralocorticoid action of aldosterone is not sensitive to inhibition of rRNA synthesis. Previous studies, however, implicate mRNA synthesis in this early response.

Thyroxine and Na+ transport in toad: role in transition from poikilo- to homeothermy

1979 ◽  
Vol 236 (3) ◽  
pp. C117-C124 ◽  
Author(s):  
B. C. Rossier ◽  
M. Rossier ◽  
C. S. Lo
Keyword(s):  
Urinary Bladder ◽  
Atpase Activity ◽  
Short Circuit ◽  
Stage Iv ◽  
Short Circuit Current ◽  
Inhibitory Effect ◽  
Base Line ◽  
Na Transport ◽  
Tissue Resistance

The effects of thyroxine (T4) on Na+ transport, oxygen consumption (QO2), and Na+-K+-ATPase activity were studied in the urinary bladder and liver of the toad Bufo marinus. In the bladder, T4 in vitro (10(-8) to 10(-6) M) had no significant effect on these parameters during 15 h of incubation. When injected intraperitoneally (approximately 20 microgram/(kg body wt.day) for 6 days), T4 lowered base-line, short-circuit current by 62% (P less than 0.0025) and potential difference by 37% (P less than 0.001), increasing tissue resistance by 40% (P less than 0.02). T4 depressed QO2/DNA (-25%, P less than 0.05) with no significant effect on Na+-K+-ATPase activity. In liver, T4 increased the recovery per cell DNA of mitochondrial proteins by 32% (P less than 0.025), corresponding to an increased QO2 (stage IV) of isolated mitochondria per cell DNA (+54%, P less than 0.01). There was no significant effect on Na+-K+-ATPase activity. These results suggest that, unlike its function in the rat, T4 in the toad does not regulate cellular thermogenesis by inducing Na+-K+-ATPase. This major difference could account at least in part for the transition from poikilothermy to homeothermy. In addition, T4 has a distinct inhibitory effect on Na+ transport in the urinary bladder, which suggests an antagonism to the action of aldosterone.

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.

Na+ transport in normal and CF human bronchial epithelial cells is inhibited by BAY 39-9437

2001 ◽  
Vol 281 (1) ◽  
pp. L16-L23 ◽  
Author(s):  
Robert J. Bridges ◽  
Ben B. Newton ◽  
Joseph M. Pilewski ◽  
Daniel C. Devor ◽  
Christopher T. Poll ◽  
...  
Keyword(s):  
Serine Protease ◽  
Half Life ◽  
Inhibitory Concentration ◽  
Therapeutic Potential ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Inhibitory Effect ◽  
Human Bronchial Epithelial Cells ◽  
Na Transport ◽  
Bronchial Epithelial

To test the hypothesis that Na+ transport in human bronchial epithelial (HBE) cells is regulated by a protease-mediated mechanism, we investigated the effects of BAY 39-9437, a recombinant Kunitz-type serine protease inhibitor, on amiloride-sensitive short-circuit current of normal [non-cystic fibrosis (CF) cells] and CF HBE cells. Mucosal treatment of non-CF and CF HBE cells with BAY 39-9437 decreased the short-circuit current, with a half-life of ∼45 min. At 90 min, BAY 39-9437 (470 nM) reduced Na+ transport by ∼70%. The inhibitory effect of BAY 39-9437 was concentration dependent, with a half-maximal inhibitory concentration of ∼25 nM. Na+ transport was restored to control levels, with a half-life of ∼15 min, on washout of BAY 39-9437. In addition, trypsin (1 μM) rapidly reversed the inhibitory effect of BAY 39-9437. These data indicate that Na+transport in HBE cells is activated by a BAY 39-9437-inhibitable, endogenously expressed serine protease. BAY 39-9437 inhibition of this serine protease maybe of therapeutic potential for the treatment of Na+ hyperabsorption in CF.

HCO3-Cl exchange transport in the adaptive response to alkalosis by turtle bladder

1980 ◽  
Vol 239 (2) ◽  
pp. F167-F174
Author(s):  
L. Cohen
Keyword(s):  
Urinary Bladder ◽  
Adaptive Response ◽  
Intact Animal ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Na Transport ◽  
Acid Base ◽  
Turtle Bladder ◽  
Acid Base Status ◽  
Ph Stat

The isolated turtle urinary bladder acidifies its mucosal (M) solution, and the rate of acidification (JH) is equivalent to the short-circuit current after Na+ transport is abolished by ouabain. When HCO3(-) is present in the serosal solution it is secreted into M in an electroneutral exchange for absorbed Cl-. The rate of HCO3(-) secretion (JHCO3(-)) can be measured by pH stat titration after JH is nullified by an opposing pH gradient. With use of these methods JH and JHCO3 were measured sequentially in bladdes from control animals and animals fed NaHCO3 (alkalosis) or NH4Cl (acidosis). JH in alkalosis (57 +/- 6 micro A) was ot different from control values (53 +/- 7 micro A). JHCO3, however, was nearly 40% higher in alkalosis (1.63 +/- 0.11 vs. 1.17 +/- 0.14 mu mol x h-1 x 8 cm-2). In contrast, JHCO3 in acidosis was similar to control values (0.89 +/- 0.15 mu mol x h-1 x 8 cm-2) but JH was increased. As judged from Cl- fluxes, neither alkalosis nor acidosis altered the electroneutral coupling between HCO3(-) secretion and Cl- absorption. JH and JHCO3 appear to be independent processes in the turtle bladder that are capable of responding independently to physiologic changes in the acid-base status of the intact animal.

Cysteine effect on toad bladder response to vasopressin, cyclic AMP, and theophylline

1964 ◽  
Vol 206 (3) ◽  
pp. 505-509 ◽  
Author(s):  
Joseph S. Handler ◽  
Jack Orloff
Keyword(s):  
Urinary Bladder ◽  
Chemical Reduction ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Inhibitory Effect ◽  
Bathing Medium ◽  
Low Concentrations ◽  
All Solutions ◽  
Neurohypophyseal Hormone ◽  
Appreciable Reduction

The increase in permeability to water and in short-circuit current of the toad's urinary bladder in response to 1 mU/ml of neurohypophyseal hormone was inhibited by 1 mm cysteine. The response to 10 mm theophylline was inhibited by as little as 0.1 mm cysteine. Cysteine or thioglycollate had no effect on the response to 1–4 mm cyclic-3',5'-AMP. The inhibitory effect of cysteine was evident under circumstances that would not be expected to cause appreciable reduction of hormone or theophylline; namely, low concentrations of cysteine were used, all solutions were well oxygenated, and there was no preincubation of hormone or theophylline with cysteine. The results are interpreted as indicating that cysteine acts at least in part by directly affecting intracellular processes rather than solely by chemical reduction of the hormone or theophylline in the bathing medium.

Interaction of heptanol and pressure on sodium and chloride transport by toad skin

1990 ◽  
Vol 69 (5) ◽  
pp. 1883-1892 ◽  
Author(s):  
M. Li ◽  
S. K. Hong ◽  
J. M. Goldinger ◽  
M. E. Duffey
Keyword(s):  
Channel Blocker ◽  
Chloride Transport ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Inhibitory Effect ◽  
Inhibitory Effects ◽  
Na Transport ◽  
Toad Skin ◽  
Shunt Resistance ◽  
Cl Conductance

We examined the interaction of heptanol and hydrostatic pressure on Na+ and Cl- transport in isolated toad skin. In the presence of Cl-, heptanol decreased short-circuit current (Isc) and total transepithelial resistance (Rt). However, in the absence of Cl- in the mucosal bath, heptanol increased Rt, although it retained the same inhibitory effect on Isc. When transepithelial active Na+ transport was blocked by amiloride, heptanol had no effect on Isc whether or not Cl- was present, whereas it decreased the shunt resistance (Rs) only in the presence of Cl- in the mucosal bath. Moreover, this effect of heptanol on Rs was significantly smaller in the presence of diphenylamine-2-carboxylate (DPC), a known Cl- channel blocker. Pressure also decreased Isc through inhibition of active Na+ transport, but it increased Rs. When heptanol and pressure were applied together, their inhibitory effects on Isc were additive, but their effects on Rs were antagonistic. Furthermore, when a transepithelial Cl- current was produced by reducing the Cl- concentration of the serosal bath, heptanol stimulated this current, which was reversibly inhibited by pressure or DPC addition to the mucosal bath. When the heptanol-stimulated Cl- current was first inhibited by pressure, subsequent DPC addition had less or no effect. These results suggest that one site of an antagonistic interaction of heptanol and pressure in toad skin is an apical membrane Cl- conductance.

Tetracycline-induced inhibition of Na+ transport in the toad urinary bladder

1978 ◽  
Vol 235 (4) ◽  
pp. F359-F366 ◽  
Author(s):  
J. Guzzo ◽  
M. Cox ◽  
A. B. Kelley ◽  
I. Singer
Keyword(s):  
Urinary Bladder ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Lipid Solubility ◽  
Toad Urinary Bladder ◽  
Na Transport ◽  
Ussing Chambers ◽  
Incubation Periods ◽  
Active Conductance ◽  
Drug Protein Binding

The effects of three tetracyclines, demethylchlortetracycline (DMC), minocycline (MNC), and oxytetracycline (OTC), on Na+ transport (measured as short-circuit current) were examined in toad urinary bladders mounted in modified Ussing chambers. During a 1-h incubation period serosal DMC (but not MNC or OTC) inhibited basal Na+ transport, whereas MNC (but not DMC or OTC) inhibited ADH-stimulated Na+ transport. MNC also inhibited cyclic AMP-stimulated Na+ transport. During longer incubation periods all three drugs inhibited basal Na+ transport. The DMC-induced inhibition of basal Na+ transport and the MNC-induced inhibition of ADH-stimulated Na+ transport were paralleled by an inhibition of the active conductance of the bladders. Thus, although all three drugs inhibit basal Na+ transport, only MNC inhibits ADH-stimulated Na+ transport. This effect does not correlate with the known effects of the tetracyclines on ADH-stimulated water flow or with drug-protein binding, and may be related to the greater lipid solubility of MNC.

Modification of carboxyl of Na+ channel inhibits aldosterone action on Na+ transport

1983 ◽  
Vol 245 (6) ◽  
pp. F726-F734 ◽  
Author(s):  
J. Kipnowski ◽  
C. S. Park ◽  
D. D. Fanestil
Keyword(s):  
Amphotericin B ◽  
Short Circuit ◽  
Short Circuit Current ◽  
Na Channel ◽  
Dependent Manner ◽  
Na Channels ◽  
Na Transport ◽  
Maximal Increase ◽  
Osmotic Water ◽  
Stimulation Of

We investigated the effect of the carboxyl-selective reagent N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ) on aldosterone stimulation of Na+ transport in the urinary bladder of the toad. Na+ transport, measured as the short-circuit current (SCC), was irreversibly inhibited by EEDQ in a dose- and time-dependent manner prior to addition of aldosterone. The greater the percentage inhibition by EEDQ (X), the smaller was the maximal increase of SCC after aldosterone (Y). This relationship gave the regression equation Y = 128.41 - 1.73X, r = -0.99 (n = 35). Evidence that the inhibition of SCC produced by EEDQ was limited to effects at the mucosal membrane was attested by the following: 1) EEDQ did not alter the stimulation by aldosterone of the osmotic water flow response to antidiuretic hormone; 2) whereas inhibition of protein synthesis by cycloheximide prevented this effect of aldosterone; 3) amphotericin B fully restored SCC previously inhibited by EEDQ to the level produced in tissues not inhibited by EEDQ; 4) comparison of the effects of amiloride vs. EEDQ pretreatment on the SCC response to aldosterone and amphotericin B revealed nearly identical characteristics; 5) in contrast, amphotericin B stimulation of SCC was limited when Na+ transport was limited by antimycin A (an inhibitor of energy production) or by ouabain. The findings fail to provide positive evidence for the hypothesis that aldosterone induces the synthesis of new Na+ channels but are consistent with hormonal activation of previously existing but nonfunctioning Na+ channels.

Effect of brefeldin A on ADH-induced transport responses of toad bladder

1994 ◽  
Vol 266 (4) ◽  
pp. C1069-C1076 ◽  
Author(s):  
K. Weng ◽  
J. B. Wade
Keyword(s):  
Water Permeability ◽  
Short Circuit ◽  
Brefeldin A ◽  
Membrane Traffic ◽  
Short Circuit Current ◽  
Toad Urinary Bladder ◽  
Membrane Biogenesis ◽  
Na Transport ◽  
Stimulation Of

We have used brefeldin A (BFA) to examine the role of membrane traffic in the short-circuit current (ISC) and water permeability responses of the toad urinary bladder. BFA treatment of 1 or 5 micrograms/ml had a complex effect on the response of the ISC to antidiuretic hormone (ADH) or forskolin stimulation. Although the responses to initial challenges by ADH were not impaired by BFA, subsequent ISC responses were progressively reduced. Similarly, while the response to an initial challenge by forskolin was modestly reduced by BFA, subsequent responses were markedly reduced. Inhibition of protein synthesis with cycloheximide (CHM) affected ISC responses similarly. Neither BFA nor CHM had an effect on water permeability responses. These observations show that although the membrane traffic responsible for the water permeability response is insensitive to inhibition by BFA or CHM, the stimulation of Na+ transport becomes increasingly sensitive to these inhibitors with successive challenges by ADH or forskolin. Although initial increases in Na+ transport utilize preexisting components, subsequent responses appear to require an intact system for membrane biogenesis.

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