Role of transglutaminase 1 in stabilisation of intercellular junctions of the vascular endothelium.
Microvascular endothelial monolayers from mouse myocardium (MyEnd) cultured for up to 5 days postconfluency became increasingly resistant to various barrier-compromising stimuli such as low extracellular Ca(2+) and treatment with the Ca(2+) ionophore A23187 and with the actin depolymerising compound cytochalasin D. In contrast, microvascular endothelial monolayers from mouse lung microvessels (PulmEnd) remained sensitive to these conditions during the entire culture period which corresponds to the well-known in vivo sensitivity of the lung microvasculature to Ca(2+) depletion and cytochalasin D treatment. One molecular difference between pulmonary and myocardial endothelial cells was found to be transglutaminase 1 (TGase1) which is strongly expressed in myocardial endothelial cells but is absent from pulmonary endothelial cells. Resistance of MyEnd cells to barrier-breaking conditions correlated strongly with translocation of TGase1 to intercellular junctions. Simultaneous inhibition of intracellular and extracellular TGase activity by monodansylcadaverine (MDC) strongly weakened barrier properties of MyEnd monolayers, whereas inhibition of extracellular TGases by the membrane-impermeable active site-directed TGase inhibitor R281 did not reduce barrier properties. Weakening of barrier properties could be also induced in MyEnd cells by downregulation of TGase1 expression using RNAi-based gene silencing. These findings suggest that crosslinking activity of intracellular TGase1 at intercellular junctions may play a role in controlling barrier properties of endothelial monolayers.
Baumgartner
Werner
W
Institute of Anatomy and Cell Biology, University of Würzburg, Koellikerstrasse 6, 97070 Würzburg, Germany. werner.baumgartner@mail.uni-wuerzburg.de
Golenhofen
Nikola
N
Weth
Agnes
A
Hiiragi
Takashi
T
Saint
Rob
R
Griffin
Martin
M
Drenckhahn
Detlev
D
D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D002463:Cell Membrane Permeability / Q000502:physiology; D004730:Endothelium, Vascular / Q000201:enzymology*; D020868:Gene Silencing; D007365:Intercellular Junctions / Q000502:physiology*; D008168:Lung / Q000098:blood supply; D051379:Mice; D008833:Microcirculation; D008856:Microscopy, Fluorescence; D009206:Myocardium; D034741:RNA, Small Interfering / Q000494:pharmacology; D011503:Transglutaminases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*
D034741:RNA, Small Interfering; D011503:Transglutaminases; C105804:transglutaminase 1