Effect of cAMP derivates on assembly and maintenance of tight junctions in human umbilical vein endothelial cells

<p>Abstract</p> <p>Background</p> <p>Endothelial tight and adherens junctions control a variety of physiological processes like adhesion, paracellular transport of solutes or trafficking of activated leukocytes. Formation and maintenance of endothelial junctions largely...

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Main Authors: Beese Michaela, Wyss Kristin, Haubitz Marion, Kirsch Torsten
Format: Article
Language:English
Published: BMC 2010-09-01
Series:BMC Cell Biology
Online Access:http://www.biomedcentral.com/1471-2121/11/68
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author Beese Michaela
Wyss Kristin
Haubitz Marion
Kirsch Torsten
author_facet Beese Michaela
Wyss Kristin
Haubitz Marion
Kirsch Torsten
author_sort Beese Michaela
collection DOAJ
description <p>Abstract</p> <p>Background</p> <p>Endothelial tight and adherens junctions control a variety of physiological processes like adhesion, paracellular transport of solutes or trafficking of activated leukocytes. Formation and maintenance of endothelial junctions largely depend on the microenvironment of the specific vascular bed and on interactions of the endothelium with adjacent cell types. Consequently, primary cultures of endothelial cells often lose their specific junctional pattern and fail to establish tight monolayer <it>in vitro</it>. This is also true for endothelial cells isolated from the vein of human umbilical cords (HUVEC) which are widely used as model for endothelial cell-related studies.</p> <p>Results</p> <p>We here compared the effect of cyclic 3'-5'-adenosine monophosphate (cAMP) and its derivates on formation and stabilization of tight junctions and on alterations in paracellular permeability in HUVEC. We demonstrated by light and confocal laser microscopy that for shorter time periods the sodium salt of 8-bromoadenosine-cAMP (8-Br-cAMP/Na) and for longer incubation periods 8-(4-chlorophenylthio)-cAMP (pCPT-cAMP) exerted the greatest effects of all compounds tested here on formation of continuous tight junction strands in HUVEC. We further demonstrated that although all compounds induced protein kinase A-dependent expression of the tight junction proteins claudin-5 and occludin only pCPT-cAMP slightly enhanced paracellular barrier functions. Moreover, we showed that pCPT-cAMP and 8-Br-cAMP/Na induced expression and membrane translocation of tricellulin.</p> <p>Conclusions</p> <p>pCPT-cAMP and, to a lesser extend, 8-Br-cAMP/Na improved formation of continuous tight junction strands and decreased paracellular permeability in primary HUVEC. We concluded that under these conditions HUVEC represent a feasible <it>in vitro </it>model to study formation and disassembly of endothelial tight junctions and to characterize tight junction-associated proteins</p>
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spelling doaj.art-8b916a7a963c4ae4bcd041a1833d72012022-12-21T22:10:07ZengBMCBMC Cell Biology1471-21212010-09-011116810.1186/1471-2121-11-68Effect of cAMP derivates on assembly and maintenance of tight junctions in human umbilical vein endothelial cellsBeese MichaelaWyss KristinHaubitz MarionKirsch Torsten<p>Abstract</p> <p>Background</p> <p>Endothelial tight and adherens junctions control a variety of physiological processes like adhesion, paracellular transport of solutes or trafficking of activated leukocytes. Formation and maintenance of endothelial junctions largely depend on the microenvironment of the specific vascular bed and on interactions of the endothelium with adjacent cell types. Consequently, primary cultures of endothelial cells often lose their specific junctional pattern and fail to establish tight monolayer <it>in vitro</it>. This is also true for endothelial cells isolated from the vein of human umbilical cords (HUVEC) which are widely used as model for endothelial cell-related studies.</p> <p>Results</p> <p>We here compared the effect of cyclic 3'-5'-adenosine monophosphate (cAMP) and its derivates on formation and stabilization of tight junctions and on alterations in paracellular permeability in HUVEC. We demonstrated by light and confocal laser microscopy that for shorter time periods the sodium salt of 8-bromoadenosine-cAMP (8-Br-cAMP/Na) and for longer incubation periods 8-(4-chlorophenylthio)-cAMP (pCPT-cAMP) exerted the greatest effects of all compounds tested here on formation of continuous tight junction strands in HUVEC. We further demonstrated that although all compounds induced protein kinase A-dependent expression of the tight junction proteins claudin-5 and occludin only pCPT-cAMP slightly enhanced paracellular barrier functions. Moreover, we showed that pCPT-cAMP and 8-Br-cAMP/Na induced expression and membrane translocation of tricellulin.</p> <p>Conclusions</p> <p>pCPT-cAMP and, to a lesser extend, 8-Br-cAMP/Na improved formation of continuous tight junction strands and decreased paracellular permeability in primary HUVEC. We concluded that under these conditions HUVEC represent a feasible <it>in vitro </it>model to study formation and disassembly of endothelial tight junctions and to characterize tight junction-associated proteins</p>http://www.biomedcentral.com/1471-2121/11/68
spellingShingle Beese Michaela
Wyss Kristin
Haubitz Marion
Kirsch Torsten
Effect of cAMP derivates on assembly and maintenance of tight junctions in human umbilical vein endothelial cells
BMC Cell Biology
title Effect of cAMP derivates on assembly and maintenance of tight junctions in human umbilical vein endothelial cells
title_full Effect of cAMP derivates on assembly and maintenance of tight junctions in human umbilical vein endothelial cells
title_fullStr Effect of cAMP derivates on assembly and maintenance of tight junctions in human umbilical vein endothelial cells
title_full_unstemmed Effect of cAMP derivates on assembly and maintenance of tight junctions in human umbilical vein endothelial cells
title_short Effect of cAMP derivates on assembly and maintenance of tight junctions in human umbilical vein endothelial cells
title_sort effect of camp derivates on assembly and maintenance of tight junctions in human umbilical vein endothelial cells
url http://www.biomedcentral.com/1471-2121/11/68
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AT haubitzmarion effectofcampderivatesonassemblyandmaintenanceoftightjunctionsinhumanumbilicalveinendothelialcells
AT kirschtorsten effectofcampderivatesonassemblyandmaintenanceoftightjunctionsinhumanumbilicalveinendothelialcells