Calpain‐2 regulation of VEGF‐mediated angiogenesis
- 1 July 2006
- journal article
- research article
- Published by Wiley in The FASEB Journal
- Vol. 20 (9), 1443-1451
- https://doi.org/10.1096/fj.05-5354com
Abstract
Angiogenesis is a complex process involving endothelial cell migration, proliferation, and differentiation as well as tube formation. These processes are stimulated by a variety of growth factors such as vascular endothelial growth factor (VEGF). VEGF-induced cytoskeletal reorganization plays a crucial role in the angiogenic processes. In the present study, we evaluated the role of calpain in VEGF-induced angiogenesis in vitro and in vivo. Human pulmonary microvascular endothelial cells (PMEC) were incubated with VEGF (10-60 ng/ml) for 2-24 h, after which we measured calpain activities, protein contents of the calpain subunits and of calpastatin, endothelial monolayer wound repair, tube formation, and actin cytoskeleton changes. Incubation of PMEC with VEGF resulted in dose- and time-dependent increases in calpain activity and protein content of calpain-2. VEGF did not change the protein contents of calpain-1 and the small subunit or of calpastatin. Incubation of PMEC with a VEGF receptor blocker prevented the VEGF-induced increase in calpain activity. Inhibition of calpain activity by siRNA directed against calpain-2 and by overexpression of calpastatin prevented VEGF-induced increases in actin stress fibers in endothelial cells and angiogenesis. Overexpression of calpastatin also inhibits vessel formation in subcutaneous (s.c.) matrigel plugs in mice. These results indicate that calpain mediates VEGF-induced angiogenic effects by modulating actin cytoskeletal organization.Keywords
Funding Information
- U.S. Department of Veterans Affairs (HL52136, HL67951)
- Flight Attendant Medical Research Institute (032040)
- American Heart Association (0555322B)
This publication has 42 references indexed in Scilit:
- Cytoskeletal Rearrangement and Caspase Activation in Sphingosine 1-Phosphate-Induced Lung Capillary Tube FormationStem Cells and Development, 2004
- Rho activity critically and selectively regulates endothelial cell organization during angiogenesisProceedings of the National Academy of Sciences of the United States of America, 2004
- The Calpain SystemPhysiological Reviews, 2003
- Involvement of RhoA/Rho Kinase Signaling in VEGF-Induced Endothelial Cell Migration and Angiogenesis In VitroArteriosclerosis, Thrombosis, and Vascular Biology, 2003
- CalpainThe International Journal of Biochemistry & Cell Biology, 2002
- Calpain Cleaves RhoA Generating a Dominant-negative Form That Inhibits Integrin-induced Actin Filament Assembly and Cell SpreadingOnline Journal of Public Health Informatics, 2002
- Rapid Transactivation of the Vascular Endothelial Growth Factor Receptor KDR/Flk-1 by the Bradykinin B2 Receptor Contributes to Endothelial Nitric-oxide Synthase Activation in Cardiac Capillary Endothelial CellsOnline Journal of Public Health Informatics, 2002
- VEGF Receptor Signaling and Endothelial FunctionIUBMB Life, 2001
- Strategies for Regulating Calpain Activities in Living CellsMethods in molecular biology (Clifton, N.J.), 1999
- ReviewBiological Chemistry Hoppe-Seyler, 1995