Calpain- and talin-dependent control of microvascular pericyte contractility and cellular stiffness
Pericytes surround capillary endothelial cells and exert contractile forces modulating microvascular tone and endothelial growth. We previously described pericyte contractile phenotype to be Rho GTPase- and α-smooth muscle actin (αSMA)-dependent. However, mechanisms mediating adhesion-dependent shap...
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| Format: | Article |
| Language: | en_US |
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Elsevier
2015
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| Online Access: | http://hdl.handle.net/1721.1/99426 https://orcid.org/0000-0001-5735-0560 |
| _version_ | 1811094946973745152 |
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| author | Kotecki, Maciej Zeiger, Adam S. Van Vliet, Krystyn J. Herman, Ira M. |
| author2 | Massachusetts Institute of Technology. Department of Biological Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Biological Engineering Kotecki, Maciej Zeiger, Adam S. Van Vliet, Krystyn J. Herman, Ira M. |
| author_sort | Kotecki, Maciej |
| collection | MIT |
| description | Pericytes surround capillary endothelial cells and exert contractile forces modulating microvascular tone and endothelial growth. We previously described pericyte contractile phenotype to be Rho GTPase- and α-smooth muscle actin (αSMA)-dependent. However, mechanisms mediating adhesion-dependent shape changes and contractile force transduction remain largely equivocal. We now report that the neutral cysteine protease, calpain, modulates pericyte contractility and cellular stiffness via talin, an integrin-binding and F-actin associating protein. Digital imaging and quantitative analyses of living cells reveal significant perturbations in contractile force transduction detected via deformation of silicone substrata, as well as perturbations of mechanical stiffness in cellular contractile subdomains quantified via atomic force microscope (AFM)-enabled nanoindentation. Pericytes overexpressing GFP-tagged talin show significantly enhanced contractility (~ two-fold), which is mitigated when either the calpain-cleavage resistant mutant talin L432G or vinculin are expressed. Moreover, the cell-penetrating, calpain-specific inhibitor termed CALPASTAT reverses talin-enhanced, but not Rho GTP-dependent, contractility. Interestingly, our analysis revealed that CALPASTAT, but not its inactive mutant, alters contractile cell-driven substrata deformations while increasing mechanical stiffness of subcellular contractile regions of these pericytes. Altogether, our results reveal that calpain-dependent cleavage of talin modulates cell contractile dynamics, which in pericytes may prove instrumental in controlling normal capillary function or microvascular pathophysiology. |
| first_indexed | 2024-09-23T16:08:03Z |
| format | Article |
| id | mit-1721.1/99426 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T16:08:03Z |
| publishDate | 2015 |
| publisher | Elsevier |
| record_format | dspace |
| spelling | mit-1721.1/994262022-10-02T06:33:18Z Calpain- and talin-dependent control of microvascular pericyte contractility and cellular stiffness Kotecki, Maciej Zeiger, Adam S. Van Vliet, Krystyn J. Herman, Ira M. Massachusetts Institute of Technology. Department of Biological Engineering Massachusetts Institute of Technology. Department of Materials Science and Engineering Zeiger, Adam S. Van Vliet, Krystyn J. Pericytes surround capillary endothelial cells and exert contractile forces modulating microvascular tone and endothelial growth. We previously described pericyte contractile phenotype to be Rho GTPase- and α-smooth muscle actin (αSMA)-dependent. However, mechanisms mediating adhesion-dependent shape changes and contractile force transduction remain largely equivocal. We now report that the neutral cysteine protease, calpain, modulates pericyte contractility and cellular stiffness via talin, an integrin-binding and F-actin associating protein. Digital imaging and quantitative analyses of living cells reveal significant perturbations in contractile force transduction detected via deformation of silicone substrata, as well as perturbations of mechanical stiffness in cellular contractile subdomains quantified via atomic force microscope (AFM)-enabled nanoindentation. Pericytes overexpressing GFP-tagged talin show significantly enhanced contractility (~ two-fold), which is mitigated when either the calpain-cleavage resistant mutant talin L432G or vinculin are expressed. Moreover, the cell-penetrating, calpain-specific inhibitor termed CALPASTAT reverses talin-enhanced, but not Rho GTP-dependent, contractility. Interestingly, our analysis revealed that CALPASTAT, but not its inactive mutant, alters contractile cell-driven substrata deformations while increasing mechanical stiffness of subcellular contractile regions of these pericytes. Altogether, our results reveal that calpain-dependent cleavage of talin modulates cell contractile dynamics, which in pericytes may prove instrumental in controlling normal capillary function or microvascular pathophysiology. National Science Foundation (U.S.) (CAREER Award) American Society for Engineering Education. National Defense Science and Engineering Graduate Fellowship 2015-10-23T13:49:18Z 2015-10-23T13:49:18Z 2010-08 Article http://purl.org/eprint/type/JournalArticle 00262862 1095-9319 http://hdl.handle.net/1721.1/99426 Kotecki, Maciej, Adam S. Zeiger, Krystyn J. Van Vliet, and Ira M. Herman. “Calpain- and Talin-Dependent Control of Microvascular Pericyte Contractility and Cellular Stiffness.” Microvascular Research 80, no. 3 (December 2010): 339–348. https://orcid.org/0000-0001-5735-0560 en_US http://dx.doi.org/10.1016/j.mvr.2010.07.012 Microvascular Research Creative Commons Attribution-Noncommercial-NoDerivatives http://creativecommons.org/licenses/by-nc-nd/4.0/ application/pdf Elsevier PMC |
| spellingShingle | Kotecki, Maciej Zeiger, Adam S. Van Vliet, Krystyn J. Herman, Ira M. Calpain- and talin-dependent control of microvascular pericyte contractility and cellular stiffness |
| title | Calpain- and talin-dependent control of microvascular pericyte contractility and cellular stiffness |
| title_full | Calpain- and talin-dependent control of microvascular pericyte contractility and cellular stiffness |
| title_fullStr | Calpain- and talin-dependent control of microvascular pericyte contractility and cellular stiffness |
| title_full_unstemmed | Calpain- and talin-dependent control of microvascular pericyte contractility and cellular stiffness |
| title_short | Calpain- and talin-dependent control of microvascular pericyte contractility and cellular stiffness |
| title_sort | calpain and talin dependent control of microvascular pericyte contractility and cellular stiffness |
| url | http://hdl.handle.net/1721.1/99426 https://orcid.org/0000-0001-5735-0560 |
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