Passive solute and fluid transport in brook trout (Salvelinus fontinalis) urinary bladder epithelium

1988 ◽  
Vol 66 (4) ◽  
pp. 912-918 ◽  
Author(s):  
William S. Marshall
Keyword(s):  
Hydraulic Conductivity ◽  
Urinary Bladder ◽  
Brook Trout ◽  
Fluid Transport ◽  
Salvelinus Fontinalis ◽  
Bladder Epithelium ◽  
Solute Permeability ◽  
Tight Epithelia

The passive transport of solutes across the brook trout (Salvelinus fontinalis) urinary bladder epithelium was examined in vitro in Ussing-style membrane chambers. The low transepithelial conductance (average 0.14–0.20 mS∙cm−2) and low mannitol permeability (6.9 ± 1.4 × 10−11 cm∙s−1, mean ± SE) indicate that both the transcellular and paracellular pathways have limited solute permeability. Fluid transport measurements in in vitro bag preparations indicate low hydraulic conductivity (1.6 ± 0.4 × 10−7 cm∙s−1∙atm−1; 1 atm = 101.325 kPa) and suggest that the absorbate is hyperosmotic, 5-fold more concentrated than the bathing solutions. Voltage clamping experiments with unidirectional 22Na+ and 36Cl− fluxes indicated that Na+ passive diffusion occurs primarily via a transcellular pathway, whereas the epithelium behaves as a simple resistive barrier to Cl−; thus, a diffusional portion of the Cl− flux may be paracellular. The balance of the Cl− serosa-to-mucosa flux is nonconductive and apparently represents anion exchange. Current–voltage relations were nonlinear as is typical of some tight epithelia. Bladder urine is highly hypotonic, with sodium, potassium and chloride contents of 2.00 ± 0.36, 0.76 ± 0.19 and 1.31 ± 0.20 mM, respectively. In addition to the previously demonstrated absorptive neutral NaCl active transport, present results indicate a barrier function of the urinary bladder epithelium in which hydraulic conductivity and ion and uncharged solute permeabilities are low. These characterisitics are in turn consistent with the production in vivo of a very dilute urine.

Independent Na+ and Cl- active transport by urinary bladder epithelium of brook trout

1986 ◽  
Vol 250 (2) ◽  
pp. R227-R234 ◽  
Author(s):  
W. S. Marshall
Keyword(s):  
Urinary Bladder ◽  
Brook Trout ◽  
Titratable Acidity ◽  
Bladder Epithelium ◽  
Nacl Transport ◽  
Cl Transport ◽  
Luminal Membrane ◽  
Full Independence ◽  
Free Media

Brook trout (Salvelinus fontinalis) urinary bladder in vitro had a low serosa-positive transepithelial potential (6.7 +/- 1.2 mV), low transmural conductance (0.23 +/- 0.03 mS. cm-2), and net absorptive transport of Na+ and Cl-. The net flux of Na+ was equivalent to that of Cl- and much larger than the membrane current, indicating neutral NaCl uptake. Na+ uptake was not coupled to that of Cl-, since absorptive Na+ transport continued in (Cl-)-free media. In the absence of Cl-, the net absorption of Na+ was accompanied by net secretion of titratable acidity, suggestive of Na+-H+ exchange on the luminal surface. Active Cl- transport persisted in dilute (2.0 mM) tetraethylammonium chloride with zero K+ and zero Na+, indicating full independence of the Cl- transport from cation coupling and suggesting the presence of Cl(-)-HCO-3 exchange. The kinetics of Cl- uptake (K1/2 = 35-37 mM, maximal transport rate = 3.0-3.4 mu eq . cm-2 . h-1) were not significantly affected by removal of mucosal Na+ . Cl- uptake was inhibited partially by 10(-4) M amiloride but not by 10(-4) M bumetanide. The results strongly support a model for active NaCl transport involving paired ion exchangers, likely located at the luminal membrane.

Investigation of Iodothyronine Binding to Plasma Proteins in Brook Trout, Salvelinus fontinalis, Using Precipitation, Dialysis, and Electrophoretic Methods

1973 ◽  
Vol 30 (8) ◽  
pp. 1131-1140 ◽  
Author(s):  
N. W. Falkner ◽  
J. G. Eales
Keyword(s):  
Brook Trout ◽  
Plasma Proteins ◽  
Trichloroacetic Acid ◽  
Salvelinus Fontinalis ◽  
In Vivo Studies ◽  
Electrophoretic Separation ◽  
Hormone Levels ◽  
Separation Of Proteins

Over 95% of 3,5,3′-triiodo-L-thyronine (T3) or L-thyroxine (T4) up to added hormone levels of at least 5 μg/ml, were precipitated by trichloroacetic acid with plasma proteins of brook trout using a semimicro method. Hormone recovery in the precipitate was higher than with precipitation methods previously used on fish plasma.Equilibrium dialysis showed over 99% of T4 or T3 bound to plasma proteins of trout up to added hormone levels of at least 5 μg/ml.Acrylamide gel was superior to paper and particularly cellulose polyacetate as a medium for electrophoretic separation of proteins responsible for binding T3 or T4. In vitro studies at high hormone levels showed that T3 and T4 bound mainly to prealbumin-like proteins. In vivo studies at more physiological levels showed T4 bound to fast prealbumin-like, albuminlike, and β-globulin-like proteins, while T3 bound to slow prealbumin-like and probably the same albumin-like and β-globulin-like proteins as T4.

scholarly journals Erythrocyte-derived sphingosine-1-phosphate stabilizes basal hydraulic conductivity and solute permeability in rat microvessels

2012 ◽  
Vol 303 (7) ◽  
pp. H825-H834 ◽  
Author(s):  
F. E. Curry ◽  
J. F. Clark ◽  
R. H. Adamson
Keyword(s):  
Hydraulic Conductivity ◽  
Lipid Mediator ◽  
Rat Erythrocytes ◽  
Apparent Permeability ◽  
Solute Permeability ◽  
Permeability Coefficients ◽  
Rat Mesentery ◽  
Sphingosine 1 Phosphate

Exogenous sphingosine-1-phosphate (S1P), a lipid mediator in blood, attenuates acute microvascular permeability increases via receptor S1P1 to stabilize the endothelium. To evaluate the contribution of erythrocytes as an endogenous source of S1P to the regulation of basal permeability, we measured permeability coefficients in intact individually perfused venular microvessels of rat mesentery. This strategy also enabled the contributions of other endogenous S1P sources to be evaluated. Apparent permeability coefficients ( PS) to albumin and α-lactalbumin and the hydraulic conductivity of mesenteric microvessels were measured in the presence or absence of rat erythrocytes or rat erythrocyte-conditioned perfusate. Rat erythrocytes added to the perfusate were the principal source of S1P in these microvessels. Basal PS to albumin was stable and typical of blood-perfused microvessels (mean 0.5 × 10−6 cm/s) when erythrocytes or erythrocyte-conditioned perfusates were present. When they were absent, PS to albumin or α-lactalbumin increased up to 40-fold (over 10 min). When exogenous S1P was added to perfusates, permeability returned to levels comparable with those seen in the presence of erythrocytes. Addition of SEW 2871, an agonist specific for S1P1, in the absence of red blood cells reduced PSBSA (40-fold reduction) toward basal. The specific S1P1 receptor antagonist (W-146) reversed the stabilizing action of erythrocytes and increased permeability (27-fold increase) in a manner similar to that seen in the absence of erythrocytes. Erythrocytes are a primary source of S1P that maintains normal venular microvessel permeability. Absence of erythrocytes or conditioned perfusate in in vivo and in vitro models of endothelial barriers elevates basal permeability.

Surface Labeling with Adhesion Protein FimH Improves Binding of Immunotherapeutic Agent Salmonella Ty21a to the Bladder Epithelium

2020 ◽  
pp. 1-12
Author(s):  
Maroeska J. Burggraaf ◽  
Lisette Waanders ◽  
Mariska Verlaan ◽  
Janneke Maaskant ◽  
Diane Houben ◽  
...  
Keyword(s):  
Bladder Cancer ◽  
Antitumor Activity ◽  
Bladder Wall ◽  
Bladder Epithelium ◽  
Modest Improvement ◽  
Adhesion Protein ◽  
Survival Experiment ◽  
Muscle Invasive

BACKGROUND: Bladder cancer is the ninth most common cancer in men. 70% of these tumors are classified as non-muscle invasive bladder cancer and those patients receive 6 intravesical instillations with Mycobacterium bovis BCG after transurethral resection. However, 30% of patients show recurrences after treatment and experience severe side effects that often lead to therapy discontinuation. Recently, another vaccine strain, Salmonella enterica typhi Ty21a, demonstrated promising antitumor activity in vivo. Here we focus on increasing bacterial retention in the bladder in order to reduce the number of instillations required and improve antitumor activity. OBJECTIVE: To increase the binding of Ty21a to the bladder wall by surface labeling of the bacteria with adhesion protein FimH and to study its effect in a bladder cancer mouse model. METHODS: Binding of Ty21a with surface-labeled FimH to the bladder wall was analyzed in vitro and in vivo. The antitumor effect of a single instillation of Ty21a+FimH in treatment was determined in a survival experiment. RESULTS: FimH-labeled Ty21a showed significant (p <  0.0001) improved binding to mouse and human cell lines in vitro. Furthermore, FimH labeled bacteria showed ∼5x more binding to the bladder than controls in vivo. Enhanced binding to the bladder via FimH labeling induced a modest improvement in median but not in overall mice survival. CONCLUSIONS: FimH labeling of Ty21a significantly improved binding to bladder tumor cells in vitro and the bladder wall in vivo. The improved binding leads to a modest increase in median survival in a single bladder cancer mouse study.

In vitro and in vivo relaxation of urinary bladder smooth muscle by the selective myosin II inhibitor, blebbistatin

2011 ◽  
Vol 107 (2) ◽  
pp. 310-317 ◽  
Author(s):  
Xinhua Zhang ◽  
Dwaraka Srinivasa R. Kuppam ◽  
Arnold Melman ◽  
Michael E. DiSanto
Keyword(s):  
Smooth Muscle ◽  
Urinary Bladder ◽  
Myosin Ii ◽  
Bladder Smooth Muscle ◽  
Urinary Bladder Smooth Muscle

Evaluation of the bioactivity about anti-sca-1/basic fibroblast growth factor-urinary bladder matrix scaffold for pelvic reconstruction

2018 ◽  
Vol 33 (6) ◽  
pp. 808-818 ◽  
Author(s):  
Jiankui Li ◽  
Xi Chen ◽  
Kaijian Ling ◽  
Zhiqing Liang ◽  
Huicheng Xu
Keyword(s):  
Growth Factor ◽  
Urinary Bladder ◽  
Fibroblast Growth Factor ◽  
Basic Fibroblast Growth Factor ◽  
Pelvic Organ ◽  
Fibroblast Growth ◽  
Pelvic Reconstruction ◽  
Urinary Bladder Matrix

Introduction and hypothesis: Pelvic support structure injury is the major cause of pelvic organ prolapse. At present, polypropylene-based filler material has been suggested as a common method to treat pelvic organ prolapse. However, it cannot functionally rehabilitate the pelvic support structure. In addition to its poor long-term efficiency, the urinary bladder matrix was the most suitable biological scaffold material for pelvic floor repair. Here, we hypothesize that anti-sca-1 monoclonal antibody and basic fibroblast growth factor were cross-linked to urinary bladder matrix to construct a two-factor bioscaffold for pelvic reconstruction. Methods Through a bispecific cross-linking reagent, sulfosuccinimidyl 4-[N-maleimidomethyl] cyclohexane-1-carboxylate (sulfo-smcc) immobilized anti-sca-1 and basic fibroblast growth factor to urinary bladder matrix. Then scanning electron microscope and plate reader were used to detect whether the anti-sca-1/basic fibroblast growth factor-urinary bladder matrix scaffold was built successfully. After that, the capacity of enriching sca-1 positive cells was measured both in vitro and in vivo. In addition, we evaluated the differentiation capacity and biocompatibility of the scaffold. Finally, western blotting was used to detect the level of fibulin-5 protein. Results The scanning electron microscope and plate reader revealed that the double-factor biological scaffold was built successfully. The scaffold could significantly enrich a large number of sca-1 positive cells both in vitro and in vivo, and obviously accelerate cells and differentiate functional tissue with good biocompatibility. Moreover, the western blotting showed that the scaffold could improve the expression of fibulin-5 protein. Conclusion The anti-sca-1/basic fibroblast growth factor-urinary bladder matrix scaffold revealed good biological properties and might serve as an ideal scaffold for pelvic reconstruction.

Murine colonic mucosa hyperproliferation. I. Elevated CFTR expression and enhanced cAMP-dependent Cl−secretion

2000 ◽  
Vol 278 (5) ◽  
pp. G753-G764 ◽  
Author(s):  
Shahid Umar ◽  
Jason Scott ◽  
Joseph H. Sellin ◽  
William P. Dubinsky ◽  
Andrew P. Morris
Keyword(s):  
Cystic Fibrosis ◽  
Fluid Transport ◽  
Cellular Proliferation ◽  
Neck Region ◽  
Anion Channel ◽  
Cellular Level ◽  
Cystic Fibrosis Gene ◽  
Mrna And Protein Expression

Fluid transport in the large intestine is mediated by the cystic fibrosis gene product and cAMP-dependent anion channel cystic fibrosis transmembrane conductance regulator (CFTR). cAMP-mediated Cl−secretion by gastrointestinal cell lines in vitro has been positively correlated with the insertion of CFTR into the apical membrane of differentiated senescent colonocytes and negatively correlated with the failure of CFTR to insert into the plasma membrane of their undifferentiated proliferating counterparts. In native tissues, this relationship remains unresolved. We demonstrate, in a transmissible murine colonic hyperplasia (TMCH) model, that (8-fold) colonocyte proliferation was accompanied by increased cellular CFTR mRNA and protein expression (8.3- and 2.4-fold, respectively) and enhanced mucosal cAMP-dependent Cl−secretion (2.3-fold). By immunofluorescence microscopy, cellular CFTR expression was restricted to the apical pole of cells at the base of the epithelial crypt. In contrast, increased cellular proliferation in vivo led to increases in both the cellular level and the total number of cells expressing this anion channel, with cellular CFTR staining extending into the crypt neck region. Hyperproliferating colonocytes accumulated large amounts of CFTR in apically oriented subcellular perinuclear compartments. This novel mode of CFTR regulation may explain why high endogenous levels of cellular CFTR mRNA and protein within the TMCH epithelium were not matched with larger increases in transmucosal CFTR Cl−current.

scholarly journals MiADMSA ameliorate arsenic induced urinary bladder carcinogenesis in vivo and in vitro

2020 ◽  
Vol 128 ◽  
pp. 110257 ◽  
Author(s):  
Kshirod Sathua ◽  
Sakshi Srivastava ◽  
S.J.S. Flora
Keyword(s):  
Urinary Bladder ◽  
Bladder Carcinogenesis

scholarly journals Signature-Tagged Mutagenesis of Klebsiella pneumoniae To Identify Genes That Influence Biofilm Formation on Extracellular Matrix Material

2006 ◽  
Vol 74 (8) ◽  
pp. 4590-4597 ◽  
Author(s):  
Jennifer D. Boddicker ◽  
Rebecca A. Anderson ◽  
Jennifer Jagnow ◽  
Steven Clegg
Keyword(s):  
Extracellular Matrix ◽  
Klebsiella Pneumoniae ◽  
Biofilm Formation ◽  
Matrix Material ◽  
Infection Process ◽  
Bladder Epithelium ◽  
Tract Infections ◽  
Type 3

ABSTRACT Klebsiella pneumoniae causes urinary tract infections, respiratory tract infections, and septicemia in susceptible individuals. Strains of Klebsiella frequently produce extended-spectrum beta-lactamases, and infections with these strains can lead to relatively high mortality rates (approximately 15%). Other virulence factors include production of an antiphagocytic capsule and outer membrane lipopolysaccharide (LPS), which mediates serum resistance, as well as fimbriae on the surface of the bacteria. Type 1 fimbriae mediate adherence to many types of epithelial cells and may facilitate adherence of the bacteria to the bladder epithelium. Type 3 fimbriae can bind in vitro to the extracellular matrix of urinary and respiratory tissues, suggesting that they mediate binding to damaged epithelial surfaces. In addition, type 3 fimbriae are required for biofilm formation by Klebsiella pneumoniae on plastics and human extracellular matrix; thus, they may facilitate the formation of treatment-resistant biofilm on indwelling plastic devices, such as catheters and endotracheal tubing. The presence of these devices may cause tissue damage, allowing Klebsiella to grow as a biofilm on exposed tissue basement membrane components. Though in vivo biofilm growth may be an important step in the infection process, little is known about the genetic factors required for biofilm formation by Klebsiella pneumoniae. Thus, we performed signature-tagged mutagenesis to identify factors produced by K. pneumoniae strain 43816 that are required for biofilm formation. We identified mutations in the cps capsule gene cluster, previously unidentified transcriptional regulators, fimbrial, and sugar phosphotransferase homologues, as well as genetic loci of unknown function, that affect biofilm formation.

A reinvestigation of the function of the mammalian urinary bladder

1977 ◽  
Vol 232 (3) ◽  
pp. F187-F195 ◽  
Author(s):  
S. A. Lewis
Keyword(s):  
Urinary Bladder ◽  
Membrane Surface ◽  
Apical Cell ◽  
Membrane Damage ◽  
In Vitro Experiments ◽  
Apical Cell Membrane ◽  
Cell Membrane Damage ◽  
Using Data

The function of adult mammalian urinary bladder is evaluated in light of recent in vitro experiments. The discrepancy between in vivo and in vitro experimental results is examined and a possible solution proposed. Techniques for eliminating edge damage and measuring apical membrane surface area are described. A new chamber design for microelectrode studies is illustrated. The possibility of apical cell membrane damage caused by microelectrodes is critically examined and tested using the polyene antibiotic Nystatin. Using data from transepithelial and microelectrode experiments, a model for net Na+ transport across the bladder is proposed and then critically analyzed. The possible clinical implications of the in vitro experiments are briefly discussed.

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