Applications
Applications
Barrier Function of Cell Layers
Measurement of barrier function of cell layers in tissue culture via
ECIS has proven to be a significant and rapidly growing use of this
technology. The utilization of ECIS to monitor barrier function was
first demonstrated in 1992 using bovine pulmonary endothelial cells
exposed to thrombin [ "Electrical method for detection of endothelial
cell shape change in real time: assessment of endothelial barrier
function" Proc. Natl. Acad. Sci. USA. 89: 7919-7923, 1992
In the decade following that publication, endothelial cell biologist have increasingly found barrier function measurements via ECIS to be an attractive alternative to techniques involving the use of membrane inserts and Ussing chambers. In addition to its convenience in gathering transcellular resistance (TER) data with a minimum of labor, the measurement requires no tagged compounds and associated sampling/measuring techniques. The relatively low values of the transcellular resistance for most endothelial layers (for example 10 ohm cm^2 ) make precise determinations of this value difficult with other technologies such as resistance measurements of cell layers upon filter substrates.
True barrier function measurements
Cell layers with very high TER values such as endothelial cells from the brain and epithelial cells exhibiting tight junctions can be measured electrically using porated membrane supports as well as with ECIS, but there is an important distinction between the capabilities of these methods. In both cases, the true barrier function (the resistance of the paracellular pathway between the cells) is complicated by the constricted current flow in the space between the basal cell membrane and the cell support. This complication is equally true for measurements using tagged compound. This is shown in the figure below where we see a cartoon version of cells upon a porated filter. Note that for molecules to travel from the lower chamber to the space above the cell layer, they must also travel in the constricted space beneath the cells to reach the intercellular junctions. Because of this, one does not measure the true barrier function but rather a combination of the constricted paths beneath the cells and the transcellular pathway.
The ECIS method is unique in being able to distinguish between these two pathways, since they each affect the measured resistance and capacitance monitored by ECIS in different ways. Using a published model, the true barrier function due to intercellular junctions can be extracted from ECIS data.
Garcia, et al., have used this model to address the changes in impedance when bovine pulmonary arterial endothelial cell layers are exposed to VEGF. Data analyzed by the model show a drop in the true barrier function in response to the growth factor.
Lo, C.M., Keese, C.R., Giaever, I., "Cell-substrate contact: Another factor may influence transepithelial electrical resistance of cell layers cultured on permeable filters", Experimental Cell Research, 250 (2): 576-580 (1999)] For more details regarding true barrier function measurements and the complication associated with porated filters.
Barrier Function: Related ECIS Publications
The PI3K p110α isoform regulates endothelial adherens junctions via Pyk2 and Rac1. RJ Cain, B Vanhaesebroeck, and AJ Ridley. J Cell Biol. 2010; vol:188 iss:6 pg:863 -876
EGFR signaling contributes to house dust mite-induced epithelial barrier dysfunction. I.H. Heijink, A. van Oosterhout, and A. Kapus. Eur. Respir. J. published 29 March 2010, 10.1183/09031936.00125809.
Desmoglein 2-mediated adhesion is required for intestinal epithelial barrier integrity. Nicolas Schlegel, Michael Meir, Wolfgang-Moritz Heupel, Bastian Holthöfer, Rudolf E. Leube, and Jens Waschke. Am J Physiol Gastrointest Liver Physiol. 2010; 298:G774-G783.
Calcium/calmodulin-dependent protein kinase II delta6 (CaMKIIdelta6) and RhoA involvement in thrombin-induced endothelial barrier dysfunction. Zhen Wang, Roman Ginnan, Iskandar F. Abdullaev, Mohamed Trebak, Peter A. Vincent, and Harold A. Singer. J. Biol. Chem. published 4 May 2010, 10.1074/jbc.M110.120790.
Group V Phospholipase A2 Mediates Barrier Disruption of Human Pulmonary Endothelial Cells Caused by LPS in Vitro. Steven M. Dudek, Nilda M. Muñoz, Anjali Desai, Christopher M. Osan, Angelo Y. Meliton, and Alan R. Leff. Am. J. Respir. Cell Mol. Biol. published 6 May 2010, 10.1165/rcmb.2009-0446OC.
Adenosine protected against pulmonary edema through transporter- and receptor A2-mediated endothelial barrier enhancement. Qing Lu, Elizabeth O. Harrington, Julie Newton, Brian Casserly, Gregory Radin, Rod Warburton, Yang Zhou, Michael R. Blackburn, and Sharon Rounds. Am J Physiol Lung Cell Mol Physiol. 2010; 298:L755-L767.
Claudin-2, a component of the tight junction, forms a paracellular water channel. Rita Rosenthal, Susanne Milatz, Susanne M. Krug, Beibei Oelrich, Jörg-Dieter Schulzke, Salah Amasheh, Dorothee Günzel, and Michael Fromm. J. Cell Sci. 2010; 123:1913-1921.
Regulation of CovR expression in Group B streptococcus impacts blood-brain barrier penetration. A Lembo, MA Gurney, K Burnside, A Banerjee, M de Los Reyes, JE Connelly, WJ Lin, KA Jewell, A Vo, CW Renken, KS Doran, and L Rajagopal. Mol Microbiol. 2010.
Cellular mechanisms of IL-17-induced blood-brain barrier disruption. Jula Huppert, Dorothea Closhen, Andrew Croxford, Robin White, Paulina Kulig, Eweline Pietrowski, Ingo Bechmann, Burkhard Becher, Heiko J. Luhmann, Ari Waisman, and Christoph R. W. Kuhlmann. FASEB J. 2010 Apr;24(4):1023-34. Epub 2009 Nov 25.
Kei Sarai, Kenichi Shikata, Yasushi Shikata, Kazuyoshi Omori, Naomi Watanabe, Motofumi Sasaki, Shingo Nishishita, Jun Wada, Noriko Goda, Noriyuki Kataoka, and Hirofumi Makino. Endothelial barrier protection by FTY720 under hyperglycemic condition: involvement of focal adhesion kinase, small GTPases, and adherens junction proteins. Am J Physiol Cell Physiol. 2009; 297:C945-C954.
Jula Huppert, Dorothea Closhen, Andrew Croxford, Robin White, Paulina Kulig, Eweline Pietrowski, Ingo Bechmann, Burkhard Becher, Heiko J. Luhmann, Ari Waisman, and Christoph R. W. Kuhlmann. Cellular mechanisms of IL-17-induced blood-brain barrier disruption. FASEB J. published 25 November 2009, 10.1096/fj.09-141978.
Donghong He, Yanlin Su, Peter V. Usatyuk, Ernst Wm. Spannhake, Paul Kogut, Julian Solway, Viswanathan Natarajan, and Yutong Zhao, “Lysophosphatidic Acid Enhances Pulmonary Epithelial Barrier Integrity and Protects Endotoxin-induced Epithelial Barrier Disruption and Lung Injury.” The Journal of Biological Chemistry, Vol. 284, NO. 36, pp. 24123–24132, September 4, 2009.
Tripathi, A. K., Sullivan, D.J., Stins, M.F., "Plasmodium falciparum-Infected Erthyrocytes Decrease the Integrity of Human Blood-Brain Barrier Endothelial Cell Monolayers" J. of Infectious Diseases: 942-950 (2007).
Litkouhi, B., Kwong, J., Lo, C.M., Smedley, J.G., McClane, B.A, Aponte, M., Gao, Z., Sarno, J.L., Hinners, J., Welch, W.R., Berkowitz, R.S., Mok, S.C., Garner, E.I.O., "Claudin-4 Overexpression in Epithelial Ovarian Cancer Is Associated with Hypomethylation and Is a Potential Target for Modulation of Tight Junction Barrier Function Using a C-Terminal Fragment of Clostridium perfringens Enterotoxin." Neoplasia: 304-314 (2007).
Hartmann, C., Zozulya, A., Wegener, J., Galla, H.J., "The impact of glia-derived extracellular matrices on the barrier function of cerebral endothelial cells: An in vitro study" Experimental Cell Research (2007).
Chiang, E.T., Persaud-Sawin, D.A., Kulkarni, S., Garcia, J.G., Imani, F., "Bluetongue Virus and Double-Stranded RNA Increase Human Vascular Permeability: Role of p38 MAPK" J Clin Immunol. [Epub ahead of print] (2006).
Yin F, Watsky MA. "LPA and S1P Increase Corneal Epithelial and Endothelial Cell Transcellular Resistance" Invest Ophthalmol Vis Sci. 46(6):1927-33 (2005).
Weidenfeller, C., Schrot, S., Zozulya, A., Galla, H.J., "Murine brain
capillary endothelial cells exhibit improved barrier properties under
the influence of hydrocortisone" Brain Res. 1053(1-2):162-74 (2005).
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McLaughlin, J.N., Shen, L., Holinstat, M., Brooks, J.D., DiBenedetto, E., Hamm, H.E., "Functional Selectivity of G Protein Signaling by Agonist Peptides and Thrombin for the Protease-activated Receptor-1." J. Biol. Chem. 280(26): 25048-25059 (2005).
McLaughlin, J.N., Mazzoni, M.R., Cleator, J.H., Earls, L., Perdigoto, A.L., Brooks, J.D., Muldowney, J.A.S., Vaughan, D.E., Hamm, H.E. , "Thrombin Modulates the Expression of a Set of Genes Including Thrombospondin-1 in Human Microvascular Endothelial Cells" J. Biol. Chem. 280(23): 22172-22180 (2005).
Wu, M.H., "Endothelial focal adhesions and barrier function" J. Physiology. 569(2): 359 (2005).
Treeratanapiboon, L., Psathaki, K., Wegener, J., Looareesuwan, S., Galla, H.J., Udomsangpetch, R. "In vitro study of malaria parasite induced disruption of blood-brain barrier" Biochem Biophys Res Commun. 335(3):810-8 (2005).
Aigul Moldobaeva, Laura E Welsh-Servinsky, Larissa A Shimoda, R. Scott Stephens, Alexander D Verin, Rubin M Tuder, and David B Pearse "Role of Protein Kinase G in Barrier Protective Effects of cGMP in Human Pulmonary Artery Endothelial Cells" Am J Physiol Lung Cell Mol Physiol, 0: 4342005 (2005).
Anna A Birukova, Djanybek Adyshev, Boris Gorshkov, Gary M Bokoch, Konstantin G Birukov, and Alexander A Verin "GED-H1 is Involved in Agonist-induced Human Pulmonary Entotherlial Barrier Dysfunction" Am J Physiol Lung Cell Mol Physiol, 0: 2592005 (2005).
Grab, D.J., Nikolskaia, O., Kim, Y.V., Lonsdale-Eccles, J.D., Ito, S., Hara, T., Fukuma, T., Nyarko, E., Kim, K.J., Stins, M.F., Delannoy, M.J., Rodgers, J., Kim, K.S., "African Trypanosome Interactions With an In Vitro Model of the Human Blood-Brain Barrier" J Parasitology, 90(5): 970-979 (2004).
Birukova, A.A., Birukov, K.G., Smurova, K., Adyshev, D., Kaibuchi, K., Alieva, I., Garcia, J.G.N., Verin, A.D., "Novel role of microtubules in thrombin-induced endothelial barrier dysfunction" The FASEB Journal. 18:1879-1890, (2004).
Jacobson, J.R., Dudek, S.M., Birukov, K.G., Ye, S.Q., Grigoryev,D.N., Girgis, R.E., Garcia, J.G.N. "Cytoskeletal Activation and Altered Gene Expression in Endothelial Barrier Regulation by Simvastatin" American Journal of Respiratory Cell and Molecular Biology. Vol. 30, pp. 662-670, (2004).
Zeng, W., Matter, W.F., Yan, S.B., Um, S.L., Vlahos, C.J., Liu, L., "Effect of drotrecogin alfa (activated) on human endothelial cell permeability and Rho kinase signaling" Critical Care Medicine. 32(5): S302-S308 (2004).
Regard, J.B., Scheek, S., Borbiev, T., Lanahan, A.A., Schneider, A., Demetriades, A.M., Hiemisch, H., Barnes, C.A., Verin, A.D., Worley, P.F. , "Verge: A Novel Vascular Early Reponse Gene" The Journal of Neuroscience. 24(16):4092-4103 (2004)
Kilani, M.M., Mohammed, K.A., Nasreen, N., Hardwick, J.A., Kaplan, M.H., Tepper, R.S., Antony, V.B. "Respiratory syncytial virus causes increased bronchial epithelial permeability" Chest 126(1):186-91 (2004)
Tsikitis, V., Morin, N., Harrington, E., Albina, J., Reichner, J. "The Lectin-Like Domain of Comple,emt Receptor 3 Protects Endotherlial Barrier Function from Activated Neutrophils" The Journal of Immunology 173: 1284-1291 (2004).
Iyer, S., Ferreri, D.M., DeCocco, N.C., Minnear, F.L., Vincent, P.A. "VE-cadherin-p120 interaction is required for maintenance of endothelial barrier function" Am J Physiol Lung Cell Mol Physiol. 286(6):L1143-53. (2004) Dowload pdf file
Pearse, P.M., Shimoda, L.A., Verin, A.D., Bogatcheva, N., Moon, C., Ronnett., G.V., Welsh, L.A., Becker, P.M., "Effect of cGMP on hydrogen peroxide- induced barrier dysfunction in lung microvascular endothelium" Endothelium, 10:309-317, (2003).
Tar, K., Birukova, A.A., Csortos, C., Bakó, E., Garcia, J.G.N., Verin, A.D., "Phosphatase 2A is involved in endothelial cell microtubule remodeling and barrier regulation" Journal of Cellular Biochemistry Vol 92, Issue 3 , 534 - 546, (2003).
Schaphorst, K.L., Chiang, E., Jacobs, K.N., Zaiman, A., Natarajan, V., Wigley, F., Garcia, J.G.N., "Role of sphingosine-1 phosphate in the enhancement of endothelial barrier integrity by platelet-released products" Am J Physiol Lung Cell Mol Physiol 285: L258-L267, (2003).
Usatyuk, P.V., Fomin, V.P., Shi, S., Garcia, J.G.N., Schaphorst, K., Natarajan, V., "Role of Ca2+ in diperoxovanadate-induced cytoskeletal remodeling and endothelial cell barrier function" Am J Physiol Lung Cell Mol Physiol 285: L1006-L1017, (2003).
Lum, Hazel, Qiao, Jing, Walter, Robert J., Huang, Fei, Subbaiah, Papasani V., Kim, Kwang S., and Holian, Oksana . "Inflammatory Stress Increases Receptor for Lysophosphatidylcholine in Human Microvasucal Endothelial Cells", Amer. J. Physiol., in press, (2003).
Birukov, K.G., Jacobson, J.R., Flores, A.A., Ye, S.Q., Birukova, A.A., Verin, A.D., Garcia, J.G.N. , "Magnitude-dependent regulation of pulmonary endothelial call barrier function by cyclic stretch" Am J Physiol Lung Cell Mol Physiol. 285: L785-L797 (2003).
van Wetering, S., van den Berk, N., van Buul, J.D., Mul, F.P.J., Lommerse, I., Mous, R., ten Klooster, J.P., Zwaginga, J.J,. Hordijk, P.L. , "VCAM-1-mediated Rac signaling controls endothelial cell-cell contacts and leukocyte transmigration", American Journal of Physiology-Cell Physiology, 285 (2): C343-C352 (2003).
Qiao, Jing, Huang, Fei , and Lum, Hazel. "PKA Inhibits RhoA Activation: A Protection Mechanism Against Endothelial Barrier Dysfunation", Amer. J. Physiol, 284: L972-L980, (2003).
Tiruppathi, C., Freichel, M., Vogel, S.M., Paria, B.C., Mehta, D., Flockerzi, V., Malik, A.B. "Impairment of Store-Operated Ca ^ 2+ Entry in TRPC4 ^ -/- Mice Interferes With Increase in Lung Microvascular Permeability" Circulation Research 91: 70-76 (2002). Download pdf file
Goldberg PL, MacNaughton DE, Clements RT, Minnear FL, Vincent PA."p38 MAPK activation by TGF-beta1 increases MLC phosphorylation and endothelial monolayer permeability" Am J Physiol Lung Cell Mol Physiol. 282(1):L146-54. PMID,(2002). Download pdf file
Kataoka, N., Iwaki, K., Hashimoto, K., Mochizuki, S., Ogasawara, Y., Sato, M., Tsujioka, K., Kajiya, F., "Measurements of endothelial cell-to-cell and cell-to-substrate gaps and micromechanical properties of endothelial cells during monocyte adhesion", The National Academy of Sciences of the United Sates of America, 99 (24): 15638-15643 (2002).
Antony, A.B., Tepper, R.S., Mohammed, K.A., "Cockroach extract antigen increases bronchial airway epithelial permeability", Journal of Allergy and Clinical Immunology, 110 (4): 589-595 (2002).
Lum, Hazel, Hao, Zengping, Gayle, Dave, Kumar, Priyadarsini,Patterson, Carolyn E., and Uhler, Michael D. . "Vascular Endothelial Cells Express Isoforms of Protein Kinase A Inhibitor", Amer J. Physiol., 282: C59-C66, (2002).
Garcia, J.G.N., Liu, F., Verin, A.D., Birukova, A., Dechert, M.A., Gerthoffer, W.T., Bamberg, J.R., English, D., "Sphingosine 1-phosphate promotes endothelial cell barrier integrity by Edg-dependent cytoskeletal rearrangement" J Clin Invest. 108(5): 689?701, (2001).
Minnear FL, Patil S, Bell D, Gainor JP, Morton CA. "Platelet lipid(s) bound to albumin increases endothelial electrical resistance: mimicked by LPA" Am J Physiol Lung Cell Mol Physiol. 281(6):L1337-44. PMID, (2001). Download pdf file
Gainor JP, Morton CA, Roberts JT, Vincent PA, Minnear FL."Platelet-conditioned medium increases endothelial electrical resistance independently of cAMP/PKA and cGMP/PKG" Am J Physiol Heart Circ Physiol. Nov;281(5):H1992-2001. PMID (2001). Dowload pdf file
Becker,Patrice M. , Verin, Alexander D., Booth, Mary Ann, Liu,Feng, Birukova, Anna and Garcia, Joe G. N., "Differential regulation of diverse physiological responses to VEGF in pulmonary endothelial cells", Amer J. Physiol., 281: L1500-L1511, (2001).
Hillebrandt,H. Abdelghani,A. Abdelghani-Jacquin,C. Aepfelbacher,M. Sackmann, ., ">Electrical and optical characterization of thrombin-induced permeability of cultured endothelial cell monolayers on semiconductor electrode arrays", Applied Physics A-Material Science & Processing, 73 (5): 539-546 (2001).
Lum, Hazel, Podolski, J.L., Gurnack, K.L., Schulz, I.T., Huang, F. and Holian, O. "Protein Phosphatase 2B Inhibitor Potentiates PKC Activity and Endothelial Barrier Dysfunction", Amer. J. Physiol., 281: L546-555, (2001).
Sandoval, R., A. B. Malik., T. Naqvi, D. Metha, and C. Tiruppathi. "Requirement for Ca2+ signaling in the mechanism of thrombin-induced increase in endothelial permeability", Am. J. Physiol. 280:L239-L247, (2001).
Sandoval, R., A. B. Malik., R. D. Minshall, P. Kouklis, C. A. Ellis, and C. Tiruppathi. "Ca 2+ signaling and PKC? activate increased endothelial permeability by disassembly of VE-cadherin junctions", J. Physiol. (London).533:433-445, (2001).
Sandoval, R., A. B. Malik., R. D. Minshall, P. Kouklis, C. A. Ellis, and C. Tiruppathi. "Ca 2+ signaling and PKC? activate increased endothelial permeability by disassembly of VE-cadherin junctions", J. Physiol. (London).533:433-445, (2001).
Minnear FL. Patil S, Bell D, Gainor JP, Morton CA. "Related Articles Platelet lipid(s) bound to albumin increases endothelial electrical resistance: mimicked by LPA", Am J Physiol Lung Cell Mol Physiol. 281(6):L1337-44, (2001).
Ellen, R.P., Ko, K.S.S., Lo, C.M., Grove, D.A., Ishihara, K. "Insertional Inactivation of the prtP Gene of Treponema denticola Confirms Dentilisin's Disruption of Epithelial Junctions" J. Mol. Microbiol. Biotechnol. 2(4): 581-586, (2000).
Tsukahara, H., Noiri, E., Jiang, M.Z., Hiraoka, M., Mayumi, M. "Role of nitric oxide in human pulmonary microvascular endothelial cell adhesion", Life Sci. 67(1):1-11 (2000).
Ellen, R.P., Ko, K.S., Lo, C.M., Grove, D.A., Ishihara, K. "Insertional inactivation of the prtP gene of Treponema denticola confirms dentilisin's disruption of epithelial junctions", J Mol Microbiol Biotechnol. 2(4):581-6 (2000).
Patterson, C.E., Lum, H. Schaphorst, K.L., Verin, A.D., and Garcia, J.G.N. "Regualtion of Endothelial Barrier Function by the cAMP-dependent Protein Kinase", Endotherlium, 7(4): 287-308, 2000
Waypa GB, Morton CA, Vincent PA, Mahoney JR Jr, Johnston WK III, and Minnear FL. "Oxidant-increases endothelial permeability: prevention with phosphodiesterase inhibition vs cAMP production", J Appl Physiol 88:835-842, 2000.
Lo, C.M., Keese, C.R., Giaever, I. , "Cell-substrate contact: Another factor may influence transepithelial electrical resistance of cell layers cultured on permeable filters", Experimental Cell Research, 250 (2): 576-580 (1999).
Ellis, C. A., C. Tiruppathi, R. Sandoval, W.D. Niles, and Malik. A. B. "Time course of recovery of endothelial cell surface thrombin receptor (PAR-1) expression", Am. J. Physiol. 276:C38-C45,1999.
Moy, A.B., Winter, M., Kamath, A., Blackwell, K., Reyes, G., Giaever, I., Keese, C., Shasby, D.M., "Histamine alters endothelial barrier function at cell-cell and cell-matrix sites", American Journal of Physiology-Lung Cellular and Molecular Physiology, 278 (5): L888-L898 (2000). Download pdf file
Patil S, Kaplan JE, Minnear FL. "Protein, not adenosine or adenine nucleotides, mediates platelet decrease in endothelial permeability" Am J Physiol. Nov;273(5 Pt 2):H2304-11. PMID (1997) Download pdf file
Tiruppathi, C., W. Song., M. Bergenfeldt. P. Sass and A.B. Malik. "Gp60 activation mediates albumin transcytosis in endothelial cells bytyrosine kinase-dependent pathway", J. Biol. Chem. 272:25968-25975,
Tiruppathi, C., Malik, A.B., Del Vecchio, P.J., Keese, C.R., and Giaever,I., "Electrical method for detection of endothelial cell shape change in real time", PNAS USA 89, 7919-7923 (1992).