AVP reduces transepithelial resistance across IMCD cell monolayers

Mishler, Delta R.; Kraut, Jeffrey A.; Nagami, Glenn T.

doi:10.1152/ajprenal.1990.258.6.f1561

Cited by 5 publications

(6 citation statements)

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“…The most widely used system for measuring TEER in vitro is the commercially available TransWell culture environment and the “chopstick-style” recording electrode 35,36. For more reproducible and stable TEER measures of leaky cell barriers with lower TEER values, improved recording chambers with embedded electrodes have been constructed.…”

Section: Resultsmentioning

confidence: 99%

Fabrication of Two-Layered Channel System with Embedded Electrodes to Measure Resistance Across Epithelial and Endothelial Barriers

et al. 2010

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This manuscript describes a straightforward fabrication process for embedding Ag/AgCl electrodes within a two-layer PDMS microfluidic chip where an upper and a lower channel are separated by a semi-porous membrane. This system allows for the reliable real-time measurement of transendothelial and trans-epithelial electrical resistance (TEER), an accepted quantification of cell monolayer integrity, across cells cultured on membranes inside the microchannels using impedance spectroscopy. The technique eliminates the need for costly or specialized microelectrode fabrication, enabling commercially available wire electrodes to easily be incorporated into PDMS microsystems for measuring TEER under microfluidic environments. The capability of measuring impedance across a confluent cell monolayer is confirmed using (i) brain-derived endothelial cells (bEND.3), (ii) Madin Darby Canine Kidney Cells (MDCK-2), and mouse myoblast (C2C12) (all from ATCC, Manassas, VA). TEER values as a function of cell type and cell culture time were measured and both agree with previously published values from macro-scale culture techniques. This system opens new opportunities for conveniently resolving both trans-endothelial and trans-epithelial electrical resistance to monitor cell function in real-time in microfluidic cell cultures. KeywordsTrans-endothelial electrical resistance; Trans-epithelial electrical resistance; TEER; Blood Brain Barrier; bEND.3; Impedance Spectroscopy; Impedance Spectra; Barrier Integrity; Microfluidic Endothelium; Microfluidic Epithelium Trans-membrane electrical resistance offers a quantitative technique to measure the integrity of the tight junctions that govern solute transport across the paracellular space of endothelial SUPPORTING INFORMATION AVAILABLE: Matlab Programs for resolving TEER from Impedance Spectra (circuit_fit.m and circuit_fun.m)

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Section: Resultsmentioning

confidence: 99%

Fabrication of Two-Layered Channel System with Embedded Electrodes to Measure Resistance Across Epithelial and Endothelial Barriers

et al. 2010

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“…Detailed studies over several decades have identified a number of cellular level processes that are regulated in response to vasopressin in collecting duct cells (Table 1) (Ganote et al, 1968;Kirk et al, 1984;Star et al, 1988;Mishler et al, 1990;Champigneulle et al, 1993;Nielsen et al, , 1995Simon et al, 1993;Naruse et al, 1995;Sabolić et al, 1995;Chou et al, 2000Chou et al, , 2004Chou et al, , 2008Tamma et al, 2001;Brown, 2003;Yamaguchi et al, 2003;Nunes et al, 2008;Hasler et al, 2009;Nedvetsky et al, 2010; This article has not been copyedited and formatted. The final version may differ from this version.…”

Section: Cellular Physiology Of Aqp2-expressing Renal Collecting mentioning

confidence: 99%

Phosphoproteomic Identification of Vasopressin/cAMP/Protein Kinase A–Dependent Signaling in Kidney

et al. 2020

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Water excretion by the kidney is regulated by the neurohypophyseal peptide hormone vasopressin through actions in renal collecting duct cells to regulate the water channel protein, aquaporin-2. Vasopressin signalling is initiated by binding to a G-protein coupled receptor V2R, which signals through Gsα, adenylyl cyclase 6, and activation of the cAMP-regulated protein kinase (PKA). Signaling events coupling PKA activation and aquaporin-2 regulation were largely unknown until the advent of modern protein mass spectrometry techniques that allow proteomewide quantification of protein phosphorylation changes (phosphoproteomics). This short review documents phosphoproteomic findings in collecting duct cells describing the response to V2selective vasopressin agonists and antagonists, the response to CRISPR-mediated deletion of PKA, results from in vitro phosphorylation studies using recombinant PKA, the response to the broad spectrum kinase inhibitor H89, and the responses underlying lithium-induced nephrogenic diabetes insipidus. These phosphoproteomic datasets have been made available online for modelling vasopressin signalling and signalling downstream from other Gsα-coupled receptors.

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“…When the barrier is established, the cell layer constitutes the electrical resistance between the area beyond the cells and the cell-populated area [24, 25, 26]. This technique has a very high sensitivity for detecting damage of the barrier function, and changes in barrier function can be studied over time [27, 28, 29, 30]. Using this sensitive method, we reported effects of isopropyl unoprostone ophthalmic solution on cultured rabbit corneal epithelial cells [20]and Kawazu et al [19]reported permeation of a β-adrenergic antagonist crossing cultured rabbit corneal epithelial cells.…”

Section: Discussionmentioning

confidence: 99%

Safety of Ozonated Solution as an Antiseptic of the Ocular Surface prior to Ophthalmic Surgery

et al. 2001

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Purpose: To evaluate the safety of an ozonated solution as an antiseptic of the ocular surface prior to ophthalmic surgery. Methods: In experiment 1, a primary culture of rabbit corneal epithelium was established. Then, 0, 4 and 10 ppm ozonated solution and 1.25% povidone-iodine, respectively, were applied to confluent cells on collagen-coated filter inserts (Millicell-CM^®) for 10 min followed by replacement with fresh medium. The transepithelial electrical resistance (TER), which is a good indicator of cell barrier function, was sequentially measured for 30 min. In experiment 2, adult pigmented rabbit eyes were washed with 20 ml of 4 ppm ozonated solution, 1.25% povidone-iodine solution or saline. Slitlamp examinations were performed before and after washing. Results: In experiment 1, 4 ppm ozonated solution did not change the TER as compared with the control. 10 ppm ozonated solution and 1.25% povidone-iodine similarly reduced the TER values significantly as compared with those of the control and 4 ppm ozonated solution. In experiment 2, 4 ppm ozonated solution and saline showed mild superficial punctate keratitis (SPK) in 8.3% of eyes. However, 1.25% povidone-iodine resulted in mild SPK in 17% of eyes and moderate SPK in 25% of eyes. The prevalence of SPK between two groups was significantly different (p = 0.03). Conclusion: Ozonated solution may be safe and a useful antiseptic of the ocular surface prior to ophthalmic surgery.

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AVP reduces transepithelial resistance across IMCD cell monolayers

Cited by 5 publications

References 0 publications

Fabrication of Two-Layered Channel System with Embedded Electrodes to Measure Resistance Across Epithelial and Endothelial Barriers

Fabrication of Two-Layered Channel System with Embedded Electrodes to Measure Resistance Across Epithelial and Endothelial Barriers

Phosphoproteomic Identification of Vasopressin/cAMP/Protein Kinase A–Dependent Signaling in Kidney

Safety of Ozonated Solution as an Antiseptic of the Ocular Surface prior to Ophthalmic Surgery

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