|
|
|
|
|
|
|
|
|||||
| Home Help Feedback Subscriptions Archive Search Table of Contents | |||||
Journal of Cell Science, Vol 108, Issue 6 2311-2320, Copyright © 1995 by Company of Biologists
JOURNAL ARTICLES |
DJ Mooney, R Langer and DE Ingber
Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, USA.
This study was undertaken to analyze how cell binding to extracellular matrix produces changes in cell shape. We focused on the initial process of cell spreading that follows cell attachment to matrix and, thus, cell 'shape' changes are defined here in terms of alterations in projected cell areas, as determined by computerized image analysis. Cell spreading kinetics and changes in microtubule and actin microfilament mass were simultaneously quantitated in hepatocytes plated on different extracellular matrix substrata. The initial rate of cell spreading was highly dependent on the matrix coating density and decreased from 740 microns 2/h to 50 microns 2/h as the coating density was lowered from 1000 to 1 ng/cm2. At approximately 4 to 6 hours after plating, this initial rapid spreading rate slowed and became independent of the matrix density regardless of whether laminin, fibronectin, type I collagen or type IV collagen was used for cell attachment. Analysis of F-actin mass revealed that cell adhesion to extracellular matrix resulted in a 20-fold increase in polymerized actin within 30 minutes after plating, before any significant change in cell shape was observed. This was followed by a phase of actin microfilament disassembly which correlated with the most rapid phase of cell extension and ended at about 6 hours; F-actin mass remained relatively constant during the slow matrix-independent spreading phase. Microtubule mass increased more slowly in spreading cells, peaking at 4 hours, the time at which the transition between rapid and slow spreading rates was observed. However, inhibition of this early rise in microtubule mass using either nocodazole or cycloheximide did not prevent this transition. Use of cytochalasin D revealed that microfilament integrity was absolutely required for hepatocyte spreading whereas interference with microtubule assembly (using nocodazole or taxol) or protein synthesis (using cycloheximide) only partially suppressed cell extension. In contrast, cell spreading could be completely inhibited by combining suboptimal doses of cytochalasin D and nocodazole, suggesting that intact microtubules can stabilize cell form when the microfilament lattice is partially compromised. The physiological relevance of the cytoskeleton and cell shape in hepatocyte physiology was highlighted by the finding that a short exposure (6 hour) of cells to nocodazole resulted in production of smaller cells 42 hours later that exhibited enhanced production of a liver-specific product (albumin). These data demonstrate that spreading and flattening of the entire cell body is not driven directly by net polymerization of either microfilaments or microtubules.(ABSTRACT TRUNCATED AT 400 WORDS)
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati 
Twitter What's this?
This article has been cited by other articles:
|
|
 |
|
  Y. Huang, P. Arora, C. A. McCulloch, and W. F. Vogel The collagen receptor DDR1 regulates cell spreading and motility by associating with myosin IIA J. Cell Sci., May 15, 2009; 122(10): 1637 - 1646. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y. Pan, R. Jing, A. Pitre, B. J. Williams, and O. Skalli Intermediate filament protein synemin contributes to the migratory properties of astrocytoma cells by influencing the dynamics of the actin cytoskeleton FASEB J, September 1, 2008; 22(9): 3196 - 3206. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. A. Whipple, E. M. Balzer, E. H. Cho, M. A. Matrone, J. R. Yoon, and S. S. Martin Vimentin Filaments Support Extension of Tubulin-Based Microtentacles in Detached Breast Tumor Cells Cancer Res., July 15, 2008; 68(14): 5678 - 5688. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. Xia, C. K. Thodeti, T. P. Hunt, Q. Xu, M. Ho, G. M. Whitesides, R. Westervelt, and D. E. Ingber Directional control of cell motility through focal adhesion positioning and spatial control of Rac activation FASEB J, June 1, 2008; 22(6): 1649 - 1659. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
V. L. Tsang, A. A. Chen, L. M. Cho, K. D. Jadin, R. L. Sah, S. DeLong, J. L. West, and S. N. Bhatia Fabrication of 3D hepatic tissues by additive photopatterning of cellular hydrogels FASEB J, March 1, 2007; 21(3): 790 - 801. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y.-H. Wang, W.-T. Chiu, Y.-K. Wang, C.-C. Wu, T.-L. Chen, C.-F. Teng, W.-T. Chang, H.-C. Chang, and M.-J. Tang Deregulation of AP-1 Proteins in Collagen Gel-induced Epithelial Cell Apoptosis Mediated by Low Substratum Rigidity J. Biol. Chem., January 5, 2007; 282(1): 752 - 763. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. Gui, K. Wojciechowski, C. D. Gildner, H. Nedelkovska, and D. C. Hocking Identification of the Heparin-binding Determinants within Fibronectin Repeat III1: ROLE IN CELL SPREADING AND GROWTH J. Biol. Chem., November 17, 2006; 281(46): 34816 - 34825. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. Solon, P. Streicher, R. Richter, F. Brochard-Wyart, and P. Bassereau Vesicles surfing on a lipid bilayer: Self-induced haptotactic motion PNAS, August 15, 2006; 103(33): 12382 - 12387. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. Werner, A. P. Kowalczyk, and V. Faundez Anthrax Toxin Receptor 1/Tumor Endothelium Marker 8 Mediates Cell Spreading by Coupling Extracellular Ligands to the Actin Cytoskeleton J. Biol. Chem., August 11, 2006; 281(32): 23227 - 23236. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. J. Juszczak Comparative Vibrational Spectroscopy of Intracellular Tau and Extracellular Collagen I Reveals Parallels of Gelation and Fibrillar Structure J. Biol. Chem., February 27, 2004; 279(9): 7395 - 7404. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. F. Rotundo, T. M. Curtis, M. D. Shah, B. Gao, A. Mastrangelo, S. E. LaFlamme, and T. M. Saba TNF-alpha disruption of lung endothelial integrity: reduced integrin mediated adhesion to fibronectin Am J Physiol Lung Cell Mol Physiol, February 1, 2002; 282(2): L316 - L329. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. J. Putnam, K. Schultz, and D. J. Mooney Control of microtubule assembly by extracellular matrix and externally applied strain Am J Physiol Cell Physiol, March 1, 2001; 280(3): C556 - C564. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. E. Drees, K. M. Andrews, and M. C. Beckerle Molecular Dissection of Zyxin Function Reveals Its Involvement in Cell Motility J. Cell Biol., December 27, 1999; 147(7): 1549 - 1560. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. A. de la Houssaye, K. Mikule, D. Nikolic, and K. H. Pfenninger Thrombin-Induced Growth Cone Collapse: Involvement of Phospholipase A2 and Eicosanoid Generation J. Neurosci., December 15, 1999; 19(24): 10843 - 10855. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. INGBER How cells (might) sense microgravity FASEB J, May 1, 1999; 13(9001): 3 - 15. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. Castelo and D. G. Jay Radixin Is Involved in Lamellipodial Stability during Nerve Growth Cone Motility Mol. Biol. Cell, May 1, 1999; 10(5): 1511 - 1520. [Abstract] [Full Text]
|
|
|
|
|
|
P. Jones, F. Jones, B Zhou, and M Rabinovitch Induction of vascular smooth muscle cell tenascin-C gene expression by denatured type I collagen is dependent upon a beta3 integrin-mediated mitogen-activated protein kinase pathway and a 122-base pair promoter element J. Cell Sci., January 2, 1999; 112(4): 435 - 445. [Abstract] [PDF]
|
|
|
|
|
|
S. Huang, C. S. Chen, and D. E. Ingber Control of Cyclin D1, p27Kip1, and Cell Cycle Progression in Human Capillary Endothelial Cells by Cell Shape and Cytoskeletal Tension Mol. Biol. Cell, November 1, 1998; 9(11): 3179 - 3193. [Abstract] [Full Text]
|
|
|
|
|
|
T. M. Curtis, R. F. Rotundo, P. A. Vincent, P. J. McKeown-Longo, and T. M. Saba TNF-alpha -induced matrix Fn disruption and decreased endothelial integrity are independent of Fn proteolysis Am J Physiol Lung Cell Mol Physiol, July 1, 1998; 275(1): L126 - L138. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Putnam, J. Cunningham, R. Dennis, J. Linderman, and D. Mooney Microtubule assembly is regulated by externally applied strain in cultured smooth muscle cells J. Cell Sci., January 11, 1998; 111(22): 3379 - 3387. [Abstract] [PDF]
|
|
|
|
|
|
P. L. Jones, J. Crack, and M. Rabinovitch Regulation of Tenascin-C, a Vascular Smooth Muscle Cell Survival Factor that Interacts with the {alpha}v{beta}3 Integrin to Promote Epidermal Growth Factor Receptor Phosphorylation and Growth J. Cell Biol., October 6, 1997; 139(1): 279 - 293. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. Drees, E. Friederich, J. Fradelizi, D. Louvard, M. C. Beckerle, and R. M. Golsteyn Characterization of the Interaction between Zyxin and Members of the Ena/Vasodilator-stimulated Phosphoprotein Family of Proteins J. Biol. Chem., July 14, 2000; 275(29): 22503 - 22511. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. Chen, W. Xu, P. He, E. E. Medrano, and S. W. Whiteheart Gaf-1, a gamma -SNAP-binding Protein Associated with the Mitochondria J. Biol. Chem., April 13, 2001; 276(16): 13127 - 13135. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. Spizz and P. J. Blackshear Overexpression of the Myristoylated Alanine-rich C-kinase Substrate Inhibits Cell Adhesion to Extracellular Matrix Components J. Biol. Chem., August 17, 2001; 276(34): 32264 - 32273. [Abstract] [Full Text] [PDF]
|
|
