transepithelial potential
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Membranes ◽  
2020 ◽  
Vol 10 (3) ◽  
pp. 43 ◽  
Author(s):  
Miroslaw Zajac ◽  
Andrzej Lewenstam ◽  
Piotr Bednarczyk ◽  
Krzysztof Dolowy

We measured concentration changes of sodium, potassium, chloride ions, pH and the transepithelial potential difference by means of ion-selective electrodes, which were placed on both sides of a human bronchial epithelial 16HBE14σ cell line grown on a porous support in the presence of ion channel blockers. We found that, in the isosmotic transepithelial concentration gradient of either sodium or chloride ions, there is an electroneutral transport of the isosmotic solution of sodium chloride in both directions across the cell monolayer. The transepithelial potential difference is below 3 mV. Potassium and pH change plays a minor role in ion transport. Based on our measurements, we hypothesize that in a healthy bronchial epithelium, there is a dynamic balance between water absorption and secretion. Water absorption is caused by the action of two exchangers, Na/H and Cl/HCO3, secreting weakly dissociated carbonic acid in exchange for well dissociated NaCl and water. The water secretion phase is triggered by an apical low volume-dependent factor opening the Cystic Fibrosis Transmembrane Regulator CFTR channel and secreting anions that are accompanied by paracellular sodium and water transport.



2020 ◽  
Vol 318 (3) ◽  
pp. C675-C694 ◽  
Author(s):  
Sima Jonusaite ◽  
Klaus W. Beyenbach ◽  
Heiko Meyer ◽  
Achim Paululat ◽  
Yasushi Izumi ◽  
...  

Septate junctions (SJs) are occluding cell-cell junctions that have roles in paracellular permeability and barrier function in the epithelia of invertebrates. Arthropods have two types of SJs, pleated SJs and smooth SJs (sSJs). In Drosophila melanogaster, sSJs are found in the midgut and Malpighian tubules, but the functions of sSJs and their protein components in the tubule epithelium are unknown. Here we examined the role of the previously identified integral sSJ component, Mesh, in the Malpighian tubule. We genetically manipulated mesh specifically in the principal cells of the tubule at different life stages. Tubules of flies with developmental mesh knockdown revealed defects in epithelial architecture, sSJ molecular and structural organization, and lack of urine production in basal and kinin-stimulated conditions, resulting in edema and early adult lethality. Knockdown of mesh during adulthood did not disrupt tubule epithelial and sSJ integrity but decreased the transepithelial potential, diminished transepithelial fluid and ion transport, and decreased paracellular permeability to 4-kDa dextran. Drosophila kinin decreased transepithelial potential and increased chloride permeability, and it stimulated fluid secretion in both control and adult mesh knockdown tubules but had no effect on 4-kDa dextran flux. Together, these data indicate roles for Mesh in the developmental maturation of the Drosophila Malpighian tubule and in ion and macromolecular transport in the adult tubule.



Sensors ◽  
2019 ◽  
Vol 19 (8) ◽  
pp. 1881 ◽  
Author(s):  
Mirosław Zając ◽  
Andrzej Lewenstam ◽  
Magdalena Stobiecka ◽  
Krzysztof Dołowy

Cystic Fibrosis (CF) is the most common fatal human genetic disease, which is caused by a defect in an anion channel protein (CFTR) that affects ion and water transport across the epithelium. We devised an apparatus to enable the measurement of concentration changes of sodium, potassium, chloride, pH, and transepithelial potential difference by means of ion-selective electrodes that were placed on both sides of a 16HBE14σ human bronchial epithelial cell line that was grown on a porous support. Using flat miniaturized ISE electrodes allows for reducing the medium volume adjacent to cells to approximately 20 μL and detecting changes in ion concentrations that are caused by transport through the cell layer. In contrast to classic electrochemical measurements, in our experiments neither the calibration of electrodes nor the interpretation of results is simple. The calibration solutions might affect cell physiology, the medium composition might change the direction of actions of the membrane channels and transporters, and water flow that might trigger or cut off the transport pathways accompanies the transport of ions. We found that there is an electroneutral transport of sodium chloride in both directions of the cell monolayer in the isosmotic transepithelial concentration gradient of sodium or chloride ions. The ions and water are transported as an isosmotic solution of 145 mM of NaCl.



Author(s):  
George M. Solomon ◽  
Inez Bronsveld ◽  
Kathryn Hayes ◽  
Michael Wilschanski ◽  
Paola Melotti ◽  
...  


2016 ◽  
Vol 1 (2) ◽  
pp. 48-58
Author(s):  
Aurelio Giancarlo Mauri ◽  
Lorenzo Sala ◽  
Paolo Airoldi ◽  
Giovanni Novielli ◽  
Riccardo Sacco ◽  
...  

Purpose: to theoretically investigate the role of bicarbonate ion (HCO−3 ) on the nonpigmented transepithelial potential dierence Vm, the sodium potassium pump (Na/K) and the active secretion of aqueous humor.Methods: a three-dimensional mathematical model is proposed to isolate the roles of HCO−3 and Na+, which are diicult to investigate experimentally. The model combines the velocity-extended Poisson-Nernst-Planck equations to describe ion electrodiusion and the Stokes equations to describe aqueous humor flow into the basolateral space adjacent to the nonpigmented ephitelial cells.Results: Computations showthat Vm is close to baseline experimental measurements (on monkeys) in the range [−2.7,−2.3]mVonly if HCO−3 is included in the simulation. The model is also capable of reproducing the flow of Na+ exiting the cell and the flow of K+ entering the cell, in accordance with the physiology of the Na/K pump. The simulated Na/K ratio is 1.53, which is in very good agreement with the theoretical value of 1.5.Conclusion: Model simulations suggest that HCO−3 inhibition may prevent physiologically correct baseline values of the nonpigmented transepithelial potential difference and Na/K ATPase function. This may provide useful indication in the design of medications that decrease the active secretion of aqueous humor, and supports the advantage of using mathematical models as a noninvasive complement of animal models.



2015 ◽  
Vol 309 (6) ◽  
pp. F523-F530 ◽  
Author(s):  
Donghai Wen ◽  
Yang Yuan ◽  
Ryan J. Cornelius ◽  
Huaqing Li ◽  
Paige C. Warner ◽  
...  

In many circumstances, the pathogenesis of distal renal tubular acidosis (dRTA) is not understood. In the present study, we report that a mouse model lacking the electrogenic Na+-HCO3− cotransporter [NBCe2/Slc4a5; NBCe2 knockout (KO) mice] developed dRTA after an oral acid challenge. NBCe2 expression was identified in the connecting tubule (CNT) of wild-type mice, and its expression was significantly increased after acid loading. NBCe2 KO mice did not have dRTA when on a standard mouse diet. However, after acid loading, NBCe2 KO mice exhibited complete features of dRTA, characterized by insufficient urinary acidification, hyperchloremic hypokalemic metabolic acidosis, and hypercalciuria. Additional experiments showed that NBCe2 KO mice had decreased luminal transepithelial potential in the CNT, as revealed by micropuncture. Further immunofluorescence and Western blot experiments found that NBCe2 KO mice had increased expression of H+-ATPase B1 in the plasma membrane. These results showed that NBCe2 KO mice with acid loading developed increased urinary K+ and Ca2+ wasting due to decreased luminal transepithelial potential in the CNT. NBCe2 KO mice compensated to maintain systemic pH by increasing H+-ATPase in the plasma membrane. Therefore, defects in NBCe2 can cause dRTA, and NBCe2 has an important role to regulate urinary acidification and the transport of K+ and Ca2+ in the distal nephron.



2012 ◽  
Vol 111 (3) ◽  
pp. 505-513 ◽  
Author(s):  
Bryony M. Dunning-Davies ◽  
Christopher H. Fry ◽  
Dina Mansour ◽  
Douglas R. Ferguson


2011 ◽  
Vol 182 (2) ◽  
pp. 247-258 ◽  
Author(s):  
Chris M. Wood ◽  
Harold L. Bergman ◽  
Adalto Bianchini ◽  
Pierre Laurent ◽  
John Maina ◽  
...  




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