Structural and cytochemical differentiation of membrane elements of the apical membrane of amphibian urinary bladder epithelial cells. A label fracture study

1985 ◽  
Vol 55 (3) ◽  
pp. 181-190 ◽  
Author(s):  
J. Chevalier ◽  
P. Pinto Silva ◽  
P. Ripoche ◽  
R. Gobin ◽  
X. Y. Wang ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Urinary Bladder ◽  
Apical Membrane ◽  
Amphibian Urinary Bladder ◽  
Fracture Study

Similarities of membrane structure in freeze-fractured Xenopus laevis kidney collecting tubule and urinary bladder

1980 ◽  
Vol 44 (1) ◽  
pp. 353-363
Author(s):  
D. Brown
Keyword(s):  
Epithelial Cells ◽  
Urinary Bladder ◽  
Xenopus Laevis ◽  
Apical Membrane ◽  
Collecting Duct ◽  
Large Diameter ◽  
Cell Plasma ◽  
Amphibian Urinary Bladder ◽  
Collecting Tubule ◽  
Apical Membranes

The collecting tubule of Xenopus laevis kidney is formed of two main types of cell: the socalled flask cells (or mitochondria-rich cells) and the remaining, more cuboidal epithelial cells. It has previously been shown that the flask-cell plasma membrane contains a population of elongated intramembrane particles similar to those found in mitochondria-rich cells of the toad bladder. It is now clear that the structure of the apical membrane of the remaining epithelial cells of the collecting duct is similar to the apical membrane of the amphibian urinary bladder granular cells. The P-face of the apical membrane has relatively few particles, whereas the E-face has many more. The E-face particles are of large diameter (16 nm), and many of them have an apical dense spot, which may represent a pit or depression in the particle. Such particles are not found on the lateral E-face below the level of the tight junctions. At the present time, the functional significance of these particles is unknown, but since vasopressin fails to elicit a hydrosmotic response in Xenopus laevis they are probably not involved in transepithelial water permeability. The fact that the different membrane specializations which characterize these mitochondria-rich and non-mitochondria-rich cells are found both in the bladder and the collecting tubule suggests that, at least in Xenopus, the 2 epithelia may share some common functions at the level of the apical membranes of their constituent cells.

ADH-induced water permeability and particle aggregates: alteration by a synthetic estrogen

1991 ◽  
Vol 261 (1) ◽  
pp. F144-F152 ◽  
Author(s):  
G. Calamita ◽  
Y. Le Guevel ◽  
J. Bourguet
Keyword(s):  
Urinary Bladder ◽  
Water Flow ◽  
Water Permeability ◽  
Glucose Transporter ◽  
Apical Membrane ◽  
Selective Inhibition ◽  
Synthetic Estrogen ◽  
Permeability Changes ◽  
Particle Aggregates ◽  
Amphibian Urinary Bladder

In the amphibian urinary bladder, the increase in water permeability induced by antidiuretic hormone (ADH) is accompanied by the appearance of apical intramembrane particle (IMP) aggregates that are believed to contain specific channels for water. In a previous work, we have shown that 3,3'-diallyldiethylstilbestrol (DADES), a synthetic estrogen which is a blocker of the glucose transporter, also inhibits the hydrosmotic response to ADH in the bladder. Our aim in the present study was to analyze the alterations of the membrane fine structure further and to correlate them with the water permeability changes. The results point to a selective inhibition of the ADH-induced net water flow, probably due to an interference with one of the last steps of the response to the hormone. This inhibition is associated with an increase in the density of the apical IMP aggregates, which are thus probably not operational. The resting net water flow is not inhibited and, surprisingly, typical IMP aggregates are frequently observed in the apical membrane after DADES treatment. The compound also induces the appearance of unusual loose IMP clusters that can only be seen on the apical membrane of the granular cells and that share several ultrastructural similarities with the ADH-induced aggregates. These results suggest that 1) apical DADES treatment stimulates the insertion of IMP aggregates in the apical membrane of the urinary bladder and 2) DADES inhibits the ADH-induced water flow by interfering with the aggregates and thus probably by blocking the specific water channels.

Isolated epithelial cells from amphibian urinary bladder express functional gap junctional hemichannels

1999 ◽  
Vol 276 (1) ◽  
pp. C279-C284 ◽  
Author(s):  
Carlos G. Vanoye ◽  
Leoncio A. Vergara ◽  
Luis Reuss
Keyword(s):  
Epithelial Cells ◽  
Urinary Bladder ◽  
Cell Damage ◽  
Divalent Cations ◽  
Membrane Conductance ◽  
Bladder Epithelium ◽  
Necturus Maculosus ◽  
Pathological Conditions ◽  
Amphibian Urinary Bladder ◽  
Gap Junctional

Exposure of the urinary bladder epithelium of Necturus maculosus (NUB) to protease and collagenase yields ∼50% isolated polarized cells. These cells express a membrane current slowly activated by depolarization or by removal of external divalent cations. The biophysical and pharmacological properties of the current are largely consistent with those of gap junctional hemichannels. After removal of divalent cations, the cells can also be loaded with 5(6)-carboxyfluorescein, a hydrophilic fluorescent anionic dye, and exposure to dye reduces the current in a manner dependent on membrane voltage and side of application. In contrast, Necturus gallbladder (NGB) cells exhibit no membrane conductance attributable to gap junctional hemichannels, although previous studies reveal the persistence of gap junction plaques on the plasma membrane. We conclude that functional gap junctional hemichannels can be expressed on the surface of certain isolated epithelial cells and that this is not a necessary consequence of the isolation procedure. These structures may contribute to cell damage under pathological conditions involving cell detachment.

Role of PKC isozyme III (γ) in water transport in amphibian urinary bladder

1992 ◽  
Vol 50 (1) ◽  
pp. 796-797
Author(s):  
A.J Mia ◽  
L.X. Oakford ◽  
P.D. Thompson ◽  
Z.H. Ning ◽  
T. Yorio
Keyword(s):  
Urinary Bladder ◽  
Water Flow ◽  
Phorbol Esters ◽  
Apical Membrane ◽  
Protein A ◽  
Water Channels ◽  
Renal Epithelia ◽  
Amphibian Urinary Bladder ◽  
Pkc Isozymes

Vasopressin stimulated water flow across renal epithelia is thought to occur through a V2 receptor coupled to adenylcyclase. The increase in water flow occurs as a result of a fusion of water channels with the apical membrane and is indicative of an increase in membrane capacitance following hormone addition.What controls the cycling of water channels and their insertion into the membrane is uncertain. Our laboratory has demonstrated that renal epithelia as well as amphibian urinary bladder membranes, contain a vasopressin V1 receptor which upon activation results in the breakdown of phosphoinositide and the formation of inositol triphosphate and diacylglycerol, the latter an activator of protein kinase C (PKC). The initiation of transepithelial water flow also appears to involve V1 receptors and possibly activation of PKC. To test this hypothesis, we have been using activators of PKC, such as phorbol esters and mezerein, as pharmacological tools to determine if PKC activation results in similar physiological responses as the hormone. Several PKC isozymes, upon activation, are known to be translocated to the apical membrane as visualized by FITC immunofluorescence. Previously, we reported co-localization of PKC subtypes I (γ) and II (β) in toad urinary bladders using monoclonal antibodies and protein A-gold probes. This report includes the localization of PKC subtype III (α) and its distribution pattern using immunogold labeling.

Selected polyclonal antibodies and ADH challenge in frog urinary bladder: a label-fracture study

1992 ◽  
Vol 262 (2) ◽  
pp. F267-F274
Author(s):  
G. Calamita ◽  
G. Valenti ◽  
M. Svelto ◽  
J. Bourguet
Keyword(s):  
Urinary Bladder ◽  
Binding Sites ◽  
Apical Membrane ◽  
Polyclonal Antibodies ◽  
Initial Study ◽  
Frog Urinary Bladder ◽  
Fusion Images ◽  
Fracture Study ◽  
Induced Changes ◽  
Triton X

It is clearly established that the changes induced by antidiuretic hormone (ADH) in its target epithelial cells result from the insertion in the apical membrane of new components that contain channels for water. We have already undertaken an initial study of these channels by raising polyclonal antibodies against Triton X-100 apical extracts from ADH-treated bladders and approached their purification by different adsorption steps. In the present study, we used the label-fracture technique to investigate the localization of the binding sites of the obtained polyclonal antibodies on the apical membrane of ADH-stimulated frog urinary bladder. The results obtained clearly demonstrated a preferential labeling by the selected antibodies of morphological structures such as the groove arrays and the fusion images that are generally accepted as being involved in ADH-induced changes in water permeability of the apical membrane.

Frog urinary bladder epithelial cells express TLR4 and respond to bacterial LPS by increase of iNOS expression andl-arginine uptake

2012 ◽  
Vol 303 (10) ◽  
pp. R1042-R1052 ◽  
Author(s):  
Svetlana Nikolaeva ◽  
Vera Bachteeva ◽  
Ekaterina Fock ◽  
Sabine Herterich ◽  
Elena Lavrova ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Epithelial Cells ◽  
Urinary Bladder ◽  
Inos Expression ◽  
Arginase Activity ◽  
No Production ◽  
Arginine Uptake ◽  
Amphibian Urinary Bladder ◽  
Arginase Ii ◽  
Stimulation Of

As in mammals, epithelium of the amphibian urinary bladder forms a barrier to pathogen entry and is a first line of defense against penetrating microorganisms. We investigated the effect of Escherichia coli LPS on generation of nitric oxide (NO), a critically important mediator during infectious processes, by primary cultured frog ( Rana temporaria) urinary bladder epithelial cells (FUBEC). It was found that FUBEC constitutively express Toll-like receptor 4 (TLR4), a receptor of LPS, and respond to LPS (10 μg/ml) by stimulation of inducible nitric oxide synthase (iNOS) mRNA/protein expression and NOS activity measured by nitrite produced in the culture medium and by citrulline assay. We characterized uptake of l-arginine, a precursor in NO synthesis, by FUBEC and showed that it is mediated mainly by the y+ cationic amino acid transport system. LPS stimulated l-arginine uptake, and this effect was blocked by the iNOS inhibitor 1400W. Arginase II was found to be expressed in FUBEC. Inhibition of arginase activity by (S)-(boronoethyl)-l-cysteine increased generation of NO, suggesting contribution of arginase to NO production via competing with NOS for the substrate. LPS altered neither total arginase activity nor arginase II expression. Among epithelial cells, phagocytic macrophage-like cells were observed, but they did not contribute to LPS-induced NO production. These data demonstrate that amphibian urinary bladder epithelial cells recognize LPS and respond to it by increased generation of NO via stimulation of iNOS expression and l-arginine uptake, which appears to be essential for the regulation of the innate immune response and the inflammation in bladder epithelium.

Effect of verapamil on cytoplasmic distribution of granules and microfilaments in amphibian urinary bladder

1988 ◽  
Vol 46 ◽  
pp. 324-325
Author(s):  
A.J. Mia ◽  
L.X. Oakford ◽  
T. Yorio
Keyword(s):  
Urinary Bladder ◽  
Water Flow ◽  
Morphological Changes ◽  
Granular Cell ◽  
Cytosolic Calcium ◽  
Calcium Storage ◽  
Amphibian Urinary Bladder ◽  
Vesicular Membrane ◽  
High Concentrations ◽  
Cytoskeletal System

The amphibian urinary bladder has been used as a ‘model’ system for studies of the mechanism of action of antidiuretic hormone (ADH) in stimulating transepithelial water flow. The increase in water permeability is accompanied by morphological changes that include the stimulation of apical microvilli, mobilization of microtubules and microfilaments and vesicular membrane fusion events . It has been shown that alterations in the cytosolic calcium concentrations can inhibit ADH transmembrane water flow and induce alterations in the epithelial cell cytomorphology, including the cytoskeletal system . Recently, the subapical granules of the granular cell in the amphibian urinary bladder have been shown to contain high concentrations of calcium, and it was suggested that these cytoplasmic constituents may act as calcium storage sites for intracellular calcium homeostasis. The present study utilizes the calcium antagonist, verapamil, to examine the effect of calcium deprivation on the cytomorphological features of epithelial cells from amphibian urinary bladder, with particular emphasis on subapical granule and microfilament distribution.

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