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Mechanisms of angiotensin II signaling on cytoskeleton of podocytes

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Abstract

Podocytes are significant in establishing the glomerular filtration barrier. Sustained rennin–angiotensin system (RAS) activation is crucial in the pathogenesis of podocyte injury and causes proteinuria. This study demonstrates that angiotensin II (Ang II) caused a reactive oxygen species (ROS)-dependent rearrangement of cortical F-actin and a migratory phenotype switch in cultured mouse podocytes with stable Ang II type 1 receptor (AT1R) expression. Activated small GTPase Rac-1 and phosphorylated ezrin/radixin/moesin (ERM) proteins provoked Ang II-induced F-actin cytoskeletal remodeling. This work also shows increased expression of Rac-1 and phosphorylated ERM proteins in cultured podocytes, and in glomeruli of podocyte-specific AT1R transgenic rats (Neph-hAT1 TGRs). The free radical scavenger DMTU eliminated Ang II-induced cell migration, ERM protein phosphorylation and cortical F-actin remodeling, indicating that ROS mediates the influence of Rac-1 on podocyte AT1R signaling. Heparin, a potent G-coupled protein kinase 2 inhibitor, was found to abolish ERM protein phosphorylation and cortical F-actin ring formation in Ang II-treated podocytes, indicating that phosphorylated ERM proteins are the cytoskeletal effector in AT1R signaling. Moreover, Ang II stimulation triggered down-regulation of α actinin-4 and reduced focal adhesion expression in podocytes. Signaling inhibitor assay of Ang II-treated podocytes reveals that Rac-1, RhoA, and F-actin reorganization were involved in expressional regulation of α actinin-4 in AT1R signaling. With persistent RAS activation, the Ang II-induced phenotype shifts from being dynamically stable to adaptively migratory, which may eventually exhaust podocytes with a high actin cytoskeletal turnover, causing podocyte depletion and focal segmental glomerulosclerosis.

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Acknowledgements

This work was partly funded by Chang Gung medical research project grant (CMRPG33082). Furthermore, Dr. H. H. Hsu is a recipient of the Interdepartmental Graduate-Program for Experimental Life Sciences (iGEL) fellowship from the University of Muenster, Germany, while Dr. A. Schwab was supported by a grant from IZKF Münster (Schw2/030/08) and Pa1/011/08.

This work was partially presented at the 6th International Podocyte Congress; June 8 through 11, 2006, Helsinki, Finland; the German Congress for Nephrology; September 23 through 26, 2006, Essen, Germany; and the German Congress for Nephrology; September 22 through 25, 2007, Munich, Germany.

The authors wish to thank Dr. G. Ciarimboli and U. Neugebauer (Department of Medicine D, Experimental Nephrology, University Hospital Muenster, Germany) for assisting with the intracellular calcium measurements, and S. Mally (Institute of Physiology II, University of Muenster, Germany) for assistance with life cell imaging, and Dr. C. Reinhardt (Center for Cancer Research, Department of Biology, Massachusetts Institute of Technology, MA, USA) for helping with the retroviral gene delivery method. Ted Knoy is appreciated for his editorial assistance.

Disclosures

We declare that no financial conflict of interest exists in relation to the publication of this work.

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Authors and Affiliations

  1. Department of Medicine D, Division of General Internal Medicine and Nephrology, University Hospital Muenster, Albert-Schweitzer-Str. 33, 48149, Muenster, Germany

    Hsiang-Hao Hsu, Thomas Weide, Eberhard Schlatter & Hermann Pavenstädt

  2. Kidney Institute and Department of Nephrology, Chang Gung Memorial Hospital, Chang Gung University, Taipei, Taiwan

    Hsiang-Hao Hsu

  3. Medical Research Center, University of Heidelberg, Mannheim, Germany

    Sigrid Hoffmann

  4. Department of Anatomy, University of Greifswald, Greifswald, Germany

    Nicole Endlich

  5. Institute of Physiology and Centre for Integrative Human Physiology, University of Zurich, Zurich, Switzerland

    Ana Velic

  6. Institute of Physiology II, University of Muenster, Muenster, Germany

    Albrecht Schwab

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  1. Hsiang-Hao Hsu
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Correspondence to Hermann Pavenstädt.

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Hsu, HH., Hoffmann, S., Endlich, N. et al. Mechanisms of angiotensin II signaling on cytoskeleton of podocytes. J Mol Med 86, 1379–1394 (2008). https://doi.org/10.1007/s00109-008-0399-y

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