© The Rockefeller University Press,
0021-9525/1999//109 $5.00
The Journal of Cell Biology, Volume 145, Number 1,
, 1999 109-122
Direct Observations of the Mechanical Behaviors of the Cytoskeleton in Living Fibroblasts
Steven R. Heidemann
,
Stefanie Kaech*,
Robert E. Buxbaum
, and
Andrew Matus*
* Friedrich Miescher Institute, Basel CH-4002, Switzerland; and
Department of Physiology, Michigan State University, East Lansing, Michigan 48824-1101
Cytoskeletal proteins tagged with green fluorescent protein were used to directly visualize the mechanical role of the cytoskeleton in determining cell shape. Rat embryo (REF 52) fibroblasts were deformed using glass needles either uncoated for purely physical manipulations, or coated with laminin to induce attachment to the cell surface. Cells responded to uncoated probes in accordance with a three-layer model in which a highly elastic nucleus is surrounded by cytoplasmic microtubules that behave as a jelly-like viscoelastic fluid. The third, outermost cortical layer is an elastic shell under sustained tension. Adhesive, laminin-coated needles caused focal recruitment of actin filaments to the contacted surface region and increased the cortical layer stiffness. This direct visualization of actin recruitment confirms a widely postulated model for mechanical connections between extracellular matrix proteins and the actin cytoskeleton. Cells tethered to laminin-treated needles strongly resisted elongation by actively contracting. Whether using uncoated probes to apply simple deformations or laminin-coated probes to induce surface-to-cytoskeleton interaction we observed that experimentally applied forces produced exclusively local responses by both the actin and microtubule cytoskeleton. This local accomodation and dissipation of force is inconsistent with the proposal that cellular tensegrity determines cell shape.
Key Words: cytoskeleton cytomechanics biorheology integrins cell shape
Abbreviations used in this paper: GFP, green fluorescent protein; MT, microtubule.
Address correspondence to Dr. Heidemann, Department of Physiology, Michigan State University, East Lansing, MI 48824-1101. Tel.: (517) 355-6475, ext. 1236. Fax: (517) 355-5125. E-mail: heidemann{at}psl.msu.edu
The first two authors contributed equally to this paper.

CiteULike
Complore
Connotea
Del.icio.us
Digg
Facebook
Reddit
Technorati
Twitter What's this?
This article has been cited by other articles:
-
Stolarska, M. A., Kim, Y., Othmer, H. G.
(2009). Multi-scale models of cell and tissue dynamics. Phil Trans R Soc A
367: 3525-3553
[Abstract]
[Full Text]
-
Whipple, R. A., Balzer, E. M., Cho, E. H., Matrone, M. A., Yoon, J. R., Martin, S. S.
(2008). Vimentin Filaments Support Extension of Tubulin-Based Microtentacles in Detached Breast Tumor Cells. Cancer Res.
68: 5678-5688
[Abstract]
[Full Text]
-
Gill, K. S., Beier, F., Goldberg, H. A.
(2008). Rho-ROCK signaling differentially regulates chondrocyte spreading on fibronectin and bone sialoprotein. Am. J. Physiol. Cell Physiol.
295: C38-C49
[Abstract]
[Full Text]
-
Sun, Z., Martinez-Lemus, L. A., Hill, M. A., Meininger, G. A.
(2008). Extracellular matrix-specific focal adhesions in vascular smooth muscle produce mechanically active adhesion sites. Am. J. Physiol. Cell Physiol.
295: C268-C278
[Abstract]
[Full Text]
-
Dixon, R. D. S., Arneman, D. K., Rachlin, A. S., Sundaresan, N. R., Costello, M. J., Campbell, S. L., Otey, C. A.
(2008). Palladin Is an Actin Cross-linking Protein That Uses Immunoglobulin-like Domains to Bind Filamentous Actin. J. Biol. Chem.
283: 6222-6231
[Abstract]
[Full Text]
-
Brangwynne, C. P., MacKintosh, F. C., Kumar, S., Geisse, N. A., Talbot, J., Mahadevan, L., Parker, K. K., Ingber, D. E., Weitz, D. A.
(2006). Microtubules can bear enhanced compressive loads in living cells because of lateral reinforcement. JCB
173: 733-741
[Abstract]
[Full Text]
-
Helmke, B. P.
(2005). Molecular Control of Cytoskeletal Mechanics by Hemodynamic Forces. Physiology
20: 43-53
[Abstract]
[Full Text]
-
Huang, H., Sylvan, J., Jonas, M., Barresi, R., So, P. T. C., Campbell, K. P., Lee, R. T.
(2005). Cell stiffness and receptors: evidence for cytoskeletal subnetworks. Am. J. Physiol. Cell Physiol.
288: C72-C80
[Abstract]
[Full Text]
-
Hu, S., Eberhard, L., Chen, J., Love, J. C., Butler, J. P., Fredberg, J. J., Whitesides, G. M., Wang, N.
(2004). Mechanical anisotropy of adherent cells probed by a three-dimensional magnetic twisting device. Am. J. Physiol. Cell Physiol.
287: C1184-C1191
[Abstract]
[Full Text]
-
Trepat, X., Grabulosa, M., Puig, F., Maksym, G. N., Navajas, D., Farre, R.
(2004). Viscoelasticity of human alveolar epithelial cells subjected to stretch. Am. J. Physiol. Lung Cell. Mol. Physiol.
287: L1025-L1034
[Abstract]
[Full Text]
-
Hu, S., Chen, J., Fabry, B., Numaguchi, Y., Gouldstone, A., Ingber, D. E., Fredberg, J. J., Butler, J. P., Wang, N.
(2003). Intracellular stress tomography reveals stress focusing and structural anisotropy in cytoskeleton of living cells. Am. J. Physiol. Cell Physiol.
285: C1082-C1090
[Abstract]
[Full Text]
-
Lammerding, J., Kazarov, A. R., Huang, H., Lee, R. T., Hemler, M. E.
(2003). Tetraspanin CD151 regulates {alpha}6{beta}1 integrin adhesion strengthening. Proc. Natl. Acad. Sci. USA
100: 7616-7621
[Abstract]
[Full Text]
-
Ingber, D. E.
(2003). Tensegrity I. Cell structure and hierarchical systems biology. J. Cell Sci.
116: 1157-1173
[Abstract]
[Full Text]
-
Davies, P. F., Zilberberg, J., Helmke, B. P.
(2003). Spatial Microstimuli in Endothelial Mechanosignaling. Circ. Res.
92: 359-370
[Abstract]
[Full Text]
-
Humphrey, J. D.
(2002). On Mechanical Modeling of Dynamic Changes in the Structure and Properties of Adherent Cells. Mathematics and Mechanics of Solids
7: 521-539
[Abstract]
-
Collinsworth, A. M., Zhang, S., Kraus, W. E., Truskey, G. A.
(2002). Apparent elastic modulus and hysteresis of skeletal muscle cells throughout differentiation. Am. J. Physiol. Cell Physiol.
283: C1219-C1227
[Abstract]
[Full Text]
-
Delhaas, T., van Engeland, S., Broers, J., Bouten, C., Kuijpers, N., Ramaekers, F., Snoeckx, L. H. E. H.
(2002). Quantification of cytoskeletal deformation in living cells based on hierarchical feature vector matching. Am. J. Physiol. Cell Physiol.
283: C639-C645
[Abstract]
[Full Text]
-
Beningo, K. A., Wang, Y.-l.
(2002). Fc-receptor-mediated phagocytosis is regulated by mechanical properties of the target. J. Cell Sci.
115: 849-856
[Abstract]
[Full Text]
-
LANGEVIN, H. M., CHURCHILL, D. L., CIPOLLA, M. J.
(2001). Mechanical signaling through connective tissue: a mechanism for the therapeutic effect of acupuncture. FASEB J.
15: 2275-2282
[Abstract]
[Full Text]
-
Witke, W., Li, W., Kwiatkowski, D. J., Southwick, F. S.
(2001). Comparisons of CapG and gelsolin-null macrophages: demonstration of a unique role for CapG in receptor-mediated ruffling, phagocytosis, and vesicle rocketing. JCB
154: 775-784
[Abstract]
[Full Text]
-
Wang, N., Naruse, K., Stamenovic', D., Fredberg, J. J., Mijailovich, S. M., Tolic'-Norrelykke, I. M., Polte, T., Mannix, R., Ingber, D. E.
(2001). Mechanical behavior in living cells consistent with the tensegrity model. Proc. Natl. Acad. Sci. USA
98: 7765-7770
[Abstract]
[Full Text]
-
Berrios, J. C., Schroeder, M. A., Hubmayr, R. D.
(2001). Mechanical properties of alveolar epithelial cells in culture. J. Appl. Physiol.
91: 65-73
[Abstract]
[Full Text]
-
Yvon, A.-M. C., Gross, D. J., Wadsworth, P.
(2001). Antagonistic forces generated by myosin II and cytoplasmic dynein regulate microtubule turnover, movement, and organization in interphase cells. Proc. Natl. Acad. Sci. USA
10.1073/pnas.141224198v1
[Abstract]
[Full Text]
-
Riveline, D., Zamir, E., Balaban, N. Q., Schwarz, U. S., Ishizaki, T., Narumiya, S., Kam, Z., Geiger, B., Bershadsky, A. D.
(2001). Focal Contacts as Mechanosensors: Externally Applied Local Mechanical Force Induces Growth of Focal Contacts by an Mdia1-Dependent and Rock-Independent Mechanism. JCB
153: 1175-1186
[Abstract]
[Full Text]
-
Hamill, O. P., Martinac, B.
(2001). Molecular Basis of Mechanotransduction in Living Cells. Physiol. Rev.
81: 685-740
[Abstract]
[Full Text]
-
Rusan, N. M., Fagerstrom, C. J., Yvon, A.-M. C., Wadsworth, P.
(2001). Cell Cycle-Dependent Changes in Microtubule Dynamics in Living Cells Expressing Green Fluorescent Protein-{alpha} Tubulin. Mol. Biol. Cell
12: 971-980
[Abstract]
[Full Text]
-
Putnam, A. J., Schultz, K., Mooney, D. J.
(2001). Control of microtubule assembly by extracellular matrix and externally applied strain. Am. J. Physiol. Cell Physiol.
280: C556-C564
[Abstract]
[Full Text]
-
Ko, K., Arora, P., Bhide, V, Chen, A, McCulloch, C.
(2001). Cell-cell adhesion in human fibroblasts requires calcium signaling. J. Cell Sci.
114: 1155-1167
[Abstract]
-
Yvon, A.-M. C., Wadsworth, P.
(2000). Region-Specific Microtubule Transport in Motile Cells. JCB
151: 1003-1012
[Abstract]
[Full Text]
-
Stamenovic, D., Wang, N.
(2000). Cellular Responses to Mechanical Stress: Invited Review: Engineering approaches to cytoskeletal mechanics. J. Appl. Physiol.
89: 2085-2090
[Abstract]
[Full Text]
-
Maksym, G. N., Fabry, B., Butler, J. P., Navajas, D., Tschumperlin, D. J., Laporte, J. D., Fredberg, J. J.
(2000). Mechanical properties of cultured human airway smooth muscle cells from 0.05 to 0.4 Hz. J. Appl. Physiol.
89: 1619-1632
[Abstract]
[Full Text]
-
Ingber, D. E., Heidemann, S. R., Lamoureux, P., Buxbaum, R. E.
(2000). Opposing views on tensegrity as a structural framework for understanding cell mechanics. J. Appl. Physiol.
89: 1663-1678
[Full Text]
-
Galbraith, C. G., Sheetz, M. P.
(1999). Keratocytes Pull with Similar Forces on Their Dorsal and Ventral Surfaces. JCB
147: 1313-1324
[Abstract]
[Full Text]
-
Xu, J., Tseng, Y., Wirtz, D.
(2000). Strain Hardening of Actin Filament Networks. REGULATION BY THE DYNAMIC CROSS-LINKING PROTEIN alpha -ACTININ. J. Biol. Chem.
275: 35886-35892
[Abstract]
[Full Text]
-
Yvon, A.-M. C., Gross, D. J., Wadsworth, P.
(2001). Antagonistic forces generated by myosin II and cytoplasmic dynein regulate microtubule turnover, movement, and organization in interphase cells. Proc. Natl. Acad. Sci. USA
98: 8656-8661
[Abstract]
[Full Text]
-
Wang, N., Tolic-Norrelykke, I. M., Chen, J., Mijailovich, S. M., Butler, J. P., Fredberg, J. J., Stamenovic, D.
(2002). Cell prestress. I. Stiffness and prestress are closely associated in adherent contractile cells. Am. J. Physiol. Cell Physiol.
282: C606-C616
[Abstract]
[Full Text]
-
Stamenovic, D., Mijailovich, S. M., Tolic-Norrelykke, I. M., Chen, J., Wang, N.
(2002). Cell prestress. II. Contribution of microtubules. Am. J. Physiol. Cell Physiol.
282: C617-C624
[Abstract]
[Full Text]
-
Gokina, N. I., Osol, G.
(2002). Actin cytoskeletal modulation of pressure-induced depolarization and Ca2+ influx in cerebral arteries. Am. J. Physiol. Heart Circ. Physiol.
282: H1410-H1420
[Abstract]
[Full Text]