Spatial Distribution of Cytoskeletal Mechanical Properties in Vascular Smooth Muscle Cells

Qifeng Jiang^1*, Ping Chen², Jinhe Li²

¹Department of Biochemistry and Molecular Biology, Chongqing Medical University, Chongqing, China.
²Department of Medical Information, Chongqing Medical University, Chongqing, China.

DOI: 10.4236/jbise.2015.85033                

Abstract

The mechanical properties on a whole basis of cell are extensively studied whereas regional mechanical properties of the cytoskeleton (CSK) have not been characterized. To fill this gap, here we provide detailed intracellular maps of regional CSK stiffness and traction forces, as well as the structural rearrangement in vascular smooth muscle cells (VSMCs). We used micropatterning to restrict VSMC to a single island. Regional cell stiffness was measured with optical magnetic twisting cytometry and traction force microscopy was used to measure regional traction forces. Finally, we used F-actin staining to measure the regional F-actin distribution. From traction and cell height data we computed the prestress within a cell using finite element methods. Compared with the cell center, cell edges were systematically stiffer, more fluid-like and supported higher traction forces, and at the same time had slower remodeling dynamics. These findings provide the most detailed characterization to date concerning regional mechanical properties of the intact cytoskeleton.

Keywords

Cell Stiffness, Traction Forces, Cytoskeleton, Prestress

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	Chen, C.S., et al. (1997) Geometric Control of Cell Life and Death. Science, 276, 1425-1428. http://dx.doi.org/10.1126/science.276.5317.1425
[2]	Small, J.V. and Resch, G.P. (2005) The Comings and Goings of Actin: Coupling Protrusion and Retraction in Cell Motility. Current Opinion in Cell Biology, 17, 517-523. http://dx.doi.org/10.1016/j.ceb.2005.08.004
[3]	Li, S., Guan, J.L. and Chien, S. (2005) Biochemistry and Biomechanics of Cell Motility. Annual Review of Biomedical Engineering, 7, 105-150. http://dx.doi.org/10.1146/annurev.bioeng.7.060804.100340
[4]	Yamazaki, D., Kurisu, S. and Takenawa, T. (2005) Regulation of Cancer Cell Motility through Actin Reorganization. Cancer Science, 96, 379-386. http://dx.doi.org/10.1111/j.1349-7006.2005.00062.x
[5]	Matzke, R., Jacobson, K. and Radmacher, M. (2001) Direct, High-Resolution Measurement of Furrow Stiffening during Division of Adherent Cells. Nature Cell Biology, 3, 607-610. http://dx.doi.org/10.1038/35078583
[6]	Ingber, D.E. (2006) Mechanical Control of Tissue Morphogenesis during Embryological Development. The International Journal of Developmental Biology, 50, 255-266. http://dx.doi.org/10.1387/ijdb.052044di
[7]	Fredberg, J.J. and Kamm, R.D. (2006) Stress Transmission in the Lung: Pathways from Organ to Molecule. Annual Review of Physiology, 68, 507-541. http://dx.doi.org/10.1146/annurev.physiol.68.072304.114110
[8]	Ingber, D.E. (2003) Mechanosensation through Integrins: Cells Act Locally But Think Globally. Proceedings of the National Academy of Sciences of the United States of America, 100, 1472-1474. http://dx.doi.org/10.1073/pnas.0530201100
[9]	Heidemann, S.R. and Wirtz, D. (2004) Towards a Regional Approach to Cell Mechanics. Trends in Cell Biology, 14, 160-166. http://dx.doi.org/10.1016/j.tcb.2004.02.003
[10]	Parker, K.K., et al. (2002) Directional Control of Lamellipodia Extension by Constraining Cell Shape and Orienting Cell Tractional Forces. The FASEB Journal, 16, 1195-1204. http://dx.doi.org/10.1096/fj.02-0038com
[11]	Brock, A., Chang, E., Ho, C.-C., LeDuc, P., Jiang, X.Y., Whitesides, G.M. and Ingber, D.E. (2003) Geometric Determinants of Directional Cell Motility Revealed Using Microcontact Printing. Langmuir, 19, 1611-1617. http://dx.doi.org/10.1021/la026394k
[12]	Kidoaki, S. and Matsuda, T. (2007) Shape-Engineered Vascular Endothelial Cells: Nitric Oxide Production, Cell Elasticity, and Actin Cytoskeletal Features. Journal of Biomedical Materials Research Part A, 81A, 728-735. http://dx.doi.org/10.1002/jbm.a.31112
[13]	Trepat, X., Deng, L.H., An, S.S., Navajas, D., Tschumperlin, D.J., Gerthoffer, W.T., et al. (2007) Universal Physical Responses to Stretch in the Living Cell. Nature, 447, 592-595. http://dx.doi.org/10.1038/nature05824
[14]	Fabry, B., Maksym, G.N., Shore, S.A., Moore, P.E., Panettieri Jr., R.A., Butler, J.P. and Fredberg, J.J. (2001) Selected Contribution: Time Course and Heterogeneity of Contractile Responses in Cultured Human Airway Smooth Muscle Cells. Journal of Applied Physiology, 91, 986-994.
[15]	Butler, J.P., Tolic-Norrelykke, I.M., Fabry, B. and Fredberg, J.J. (2002) Traction Fields, Moments, and Strain Energy That Cells Exert on Their Surroundings. AJP: Cell Physiology, 282, C595-C605. http://dx.doi.org/10.1152/ajpcell.00270.2001
[16]	Tee, S.Y., Fu, J., Chen, C.S. and Janmey, P.A. (2011) Cell Shape and Substrate Rigidity Both Regulate Cell Stiffness. Biophysical Journal, 100, L25-L27.
[17]	Solon, J., Levental, I., Sengupta, K., Georges, P.C. and Janmey, P.A. (2007) Fibroblast Adaptation and Stiffness Matching to Soft Elastic Substrates. Biophysical Journal, 93, 4453-4461. http://dx.doi.org/10.1529/biophysj.106.101386
[18]	Lu, Y.B., Iandiev, I., Hollborn, M., Kirber, N., Ulbricht, E., Hirrlinger, P.G., et al. (2011) Reactive Glial Cells: Increased Stiffness Correlates with Increased Intermediate Filament Expression. The FASEB Journal, 25, 624-631.
[19]	Dembo, M. and Wang, Y.L. (1999) Stresses at the Cell-to-Substrate Interface during Locomotion of Fibroblasts. Biophysical Journal, 76, 2307-2316. http://dx.doi.org/10.1016/S0006-3495(99)77386-8
[20]	Gardel, M.L., Shin, J.H., MacKintosh, F.C., Mahadevan, L., Matsudaira, P. and Weitz, D.A. (2004) Elastic Behavior of Cross-Linked and Bundled Actin Networks. Science, 304, 1301-1305. http://dx.doi.org/10.1126/science.1095087
[21]	Gardel, M.L., Nakamura, F., Hartwig, J.H., Crocker, J.C., Stossel, T.P. and Weitz, D.A. (2006) Prestressed F-Actin Networks Cross-Linked by Hinged Filamins Replicate Mechanical Properties of Cells. Proceedings of the National Academy of Sciences of the United States of America, 103, 1762-1767. http://dx.doi.org/10.1073/pnas.0504777103