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Differential role of Rho GTPases in endothelial barrier regulation dependent on endothelial cell origin

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Abstract

From studies using macrovascular endothelium, it was concluded that Rho A activation generally leads to endothelial barrier breakdown. Here, we characterized the role of Rho GTPases in endothelial barrier regulation in four different cell lines, both microvascular and macrovascular. Rho A activation by cytotoxic necrotizing factor y (CNFy) induced stress fiber formation in all cell lines. This was paralleled by gap formation and barrier breakdown in microvascular mesenteric endothelial cells (MesEnd), human dermal microvascular endothelial cells (HDMEC) as well as in macrovascular pulmonary artery endothelial cells (PAEC) but not in microvascular myocardial endothelial cells (MyEnd). In MyEnd cells, activation of Rac 1 and Cdc42 by CNF-1 strengthened barrier properties whereas in MesEnd, HDMEC and PAEC all three GTPases were activated which increased permeability in PAEC but not in MesEnd and HDMEC. In PAEC, CNF-1-induced decrease of barrier properties was blocked by the Rho kinase inhibitor Y27632 indicating that co-activation of Rho A dominated the barrier response. Inactivation of Rac 1 by toxin B or by lethal toxin (LT) compromised barrier properties in all cell lines. Taken together, Rac 1 requirement for endothelial barrier maintenance but not the destabilizing role of Rho A seems to be ubiquitous.

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Acknowledgments

We are grateful to Stefanie Imhof and Nadja Niedermeier for skilful technical assistance. We thank D. Vestweber, University of Münster, for supplying the VE-cadherin antibody and G. Schmidt and Torsten Giesemann (Department of Pharmacology and Toxicology, University of Freiburg) as well as Holger Barth (University of Ulm) for generously supplying bacterial toxins. These studies were supported in part by grants from the Deutsche Forschungsgemeinschaft (SFB 688, TP A4).

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Authors and Affiliations

  1. Institute of Anatomy and Cell Biology, University of Würzburg, Koellikerstr. 6, 97070, Würzburg, Germany

    Y. Baumer, S. Burger, N. Golenhofen, D. Drenckhahn & J. Waschke

  2. Department of Human Physiology and Membrane Biology, School of Medicine, University of California, Davis, CA, 95616, USA

    F. E. Curry

  3. Department of Anatomy and Cell Biology, University of Ulm, Albert-Einstein Allee 11, 89081, Ulm, Germany

    N. Golenhofen

  4. Institute of Anatomy and Cell Biology, Julius-Maximilians-University, Koellikerstr. 6, 97070, Würzburg, Germany

    J. Waschke

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  1. Y. Baumer
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  2. S. Burger
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  3. F. E. Curry
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  4. N. Golenhofen
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  5. D. Drenckhahn
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  6. J. Waschke
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Correspondence to J. Waschke.

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Y. Baumer and S. Burger contributed equally.

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Baumer, Y., Burger, S., Curry, F.E. et al. Differential role of Rho GTPases in endothelial barrier regulation dependent on endothelial cell origin. Histochem Cell Biol 129, 179–191 (2008). https://doi.org/10.1007/s00418-007-0358-7

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  • DOI: https://doi.org/10.1007/s00418-007-0358-7

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