title: Microtubule stabilization opposes the (TNF-alpha)-induced loss in the barrier integrity of corneal endothelium.
abstract: Microtubule disassembly breaks down the barrier integrity in a number of epithelial and endothelial monolayers. This study has investigated effects of TNF-alpha, which is implicated in corneal allograft rejection, on microtubules and barrier integrity in cultured bovine corneal endothelial cells. Exposure to TNF-alpha led to disassembly of the microtubules, and also caused disruption of the perijunctional actomyosin ring (PAMR). As a measure of barrier integrity, trans-endothelial electrical resistance (TER) was determined based on electrical cell-substrate impedance sensing in realtime. Exposure to TNF-alpha caused a slow decline in TER for > 20 h, and a similar exposure to cells grown on porous culture inserts led to a significant increase in permeability to FITC dextran. These changes, indicating a loss of barrier integrity, were also reflected by dislocation of ZO-1 at the cell border and disassembly of cadherins. These effects of TNF-alpha were inhibited upon stabilization of microtubules by pre-treatment with paclitaxel or epothilone B. Microtubule stabilization may be a useful strategy to overcome (TNF-alpha)-induced loss of the barrier integrity of corneal endothelium during inflammation associated with transplant rejection and uveitis.
authors.affiliation: School of Optometry, 800 E. Atwater Avenue, Indiana University, Bloomington, IN 47405, USA.
authors.forename: Mahesh
authors.initials: M
authors.lastname: Shivanna
authors.forename: Sangly P
authors.initials: SP
authors.lastname: Srinivas
mesh_terms: D015820:Cadherins / Q000378:metabolism; D002478:Cells, Cultured; D003911:Dextrans / Q000493:pharmacokinetics; D017097:Electric Impedance; D004728:Endothelium, Corneal / Q000187:drug effects* / Q000502:physiology / Q000648:ultrastructure; D034261:Epothilones / Q000494:pharmacology; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives / Q000493:pharmacokinetics; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism; D008870:Microtubules / Q000187:drug effects* / Q000502:physiology; D017239:Paclitaxel / Q000494:pharmacology; D010539:Permeability / Q000187:drug effects; D010750:Phosphoproteins / Q000378:metabolism; D050257:Tubulin Modulators / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*; D062826:Zonula Occludens-1 Protein
chemical_list: D015820:Cadherins; D003911:Dextrans; D034261:Epothilones; D008565:Membrane Proteins; D010750:Phosphoproteins; C569042:TJP1 protein, human; D050257:Tubulin Modulators; D014409:Tumor Necrosis Factor-alpha; D062826:Zonula Occludens-1 Protein; C015219:fluorescein isothiocyanate dextran; D016650:Fluorescein-5-isothiocyanate; D017239:Paclitaxel; C093788:epothilone B
references.citation: Traffic. 2004 Jan;5(1):1-9
references.pmid: 14675420
references.citation: Invest Ophthalmol Vis Sci. 2008 Feb;49(2):565-71
references.pmid: 18235000
references.citation: Exp Eye Res. 2005 Aug;81(2):183-92
references.pmid: 16080912
references.citation: Invest Ophthalmol Vis Sci. 2007 Apr;48(4):1518-27
references.pmid: 17389480
references.citation: Invest Ophthalmol Vis Sci. 2003 Aug;44(8):3326-31
references.pmid: 12882777
references.citation: Biochim Biophys Acta. 2000 May 17;1470(3):M79-91
references.pmid: 10799747
references.citation: J Cell Physiol. 2005 Sep;204(3):934-47
references.pmid: 15828024
references.citation: Circ Res. 2002 May 3;90(8):874-81
references.pmid: 11988488
references.citation: J Pharmacol Exp Ther. 2003 May;305(2):482-94
references.pmid: 12606598
references.citation: Curr Eye Res. 2007 Dec;32(12):1005-16
references.pmid: 18085464
references.citation: Invest Ophthalmol Vis Sci. 1996 Aug;37(9):1924-9
references.pmid: 8759364
references.citation: J Immunol. 2003 Dec 1;171(11):6164-72
references.pmid: 14634132
references.citation: Physiol Genomics. 2007 Sep 19;31(1):104-13
references.pmid: 17566077
references.citation: Exp Eye Res. 2009 Sep;89(3):333-43
references.pmid: 19345211
references.citation: J Cell Physiol. 2007 Oct;213(1):221-8
references.pmid: 17476691
references.citation: Am J Physiol Gastrointest Liver Physiol. 2004 Mar;286(3):G367-76
references.pmid: 14766535
references.citation: J Cell Physiol. 2004 Oct;201(1):55-70
references.pmid: 15281089
references.citation: Prog Retin Eye Res. 2003 May;22(3):359-89
references.pmid: 12852491
references.citation: FEBS Lett. 1997 Jun 2;409(1):57-62
references.pmid: 9199504
references.citation: J Biol Chem. 2001 Feb 16;276(7):4948-56
references.pmid: 11058585
references.citation: Am J Physiol Gastrointest Liver Physiol. 2005 Mar;288(3):G422-30
references.pmid: 15701621
references.citation: Environ Health Perspect. 1998 Oct;106 Suppl 5:1205-12
references.pmid: 9788899
references.citation: Am J Physiol. 1993 Apr;264(4 Pt 1):C894-905
references.pmid: 8476021
references.citation: Invest Ophthalmol Vis Sci. 2007 Jan;48(1):120-33
references.pmid: 17197525
references.citation: J Biol Chem. 1997 Jan 24;272(4):2534-41
references.pmid: 8999970
references.citation: Antioxid Redox Signal. 2003 Dec;5(6):723-30
references.pmid: 14588145
references.citation: Invest Ophthalmol Vis Sci. 2008 Nov;49(11):4816-27
references.pmid: 18614806
references.citation: Arterioscler Thromb Vasc Biol. 2008 May;28(5):899-905
references.pmid: 18292388
references.citation: Exp Eye Res. 2004 Oct;79(4):543-51
references.pmid: 15381038
references.citation: Exp Eye Res. 2004 Oct;79(4):477-86
references.pmid: 15381032
references.citation: Am J Physiol Gastrointest Liver Physiol. 2006 Mar;290(3):G496-504
references.pmid: 16474009
references.citation: J Biol Chem. 2009 Apr 24;284(17):11454-66
references.pmid: 19261619
references.citation: Drugs R D. 2008;9(1):1-10
references.pmid: 18095749
references.citation: Chem Biol. 1999 Mar;6(3):R65-9
references.pmid: 10074470
references.citation: Am J Transplant. 2004 May;4(5):678-85
references.pmid: 15084160
references.citation: Invest Ophthalmol Vis Sci. 2005 Jun;46(6):1927-33
references.pmid: 15914605
references.citation: Shock. 2002 Jul;18(1):82-5
references.pmid: 12095140
references.citation: Invest Ophthalmol Vis Sci. 2006 May;47(5):1754-67
references.pmid: 16638979
references.citation: Am J Physiol Lung Cell Mol Physiol. 2001 Jun;280(6):L1168-78
references.pmid: 11350795
references.citation: Am J Physiol Lung Cell Mol Physiol. 2006 Mar;290(3):L540-8
references.pmid: 16257999
references.citation: Am J Respir Cell Mol Biol. 2003 May;28(5):574-81
references.pmid: 12707013
references.citation: Invest Ophthalmol Vis Sci. 2006 Sep;47(9):4011-8
references.pmid: 16936117