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First published online 12 October 2004
doi: 10.1242/jcs.01416


Journal of Cell Science 117, 5521-5534 (2004)
Published by The Company of Biologists 2004
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Research Article

Spatial mapping of integrin interactions and dynamics during cell migration by Image Correlation Microscopy

Paul W. Wiseman1,*, Claire M. Brown2,*,{ddagger}, Donna J. Webb2, Benedict Hebert1, Natalie L. Johnson2, Jeff A. Squier3, Mark H. Ellisman4 and A. F. Horwitz2

1 Departments of Chemistry and Physics, McGill University, 801 Sherbrooke St. W. Montreal, Quebec H3A 2K6, Canada
2 Department of Cell Biology, University of Virginia, PO Box 800732, Charlottesville, VA 22908, USA
3 Department of Physics, Colorado School of Mines, 1523 Illinois Street, Golden, CO 80401, USA
4 Departments of Neurosciences and Bioengineering, and National Center for Microscopy and Imaging Research, University of California, San Diego, CA 92093, USA

{ddagger} Author for correspondence (e-mail: cmb8t{at}virginia.edu)

Accepted 21 July 2004

Image correlation microscopy methodology was extended and used to determine retrospectively the density, dynamics and interactions of {alpha}5-integrin in migrating cells. {alpha}5-integrin is present in submicroscopic clusters containing 3-4 integrins before it is discernibly organized. The integrin in nascent adhesions, as identified by the presence of paxillin, is ~1.4 times more concentrated, ~4.5 times more clustered and much less mobile than in surrounding regions. Thus, while integrins are clustered throughout the cell, they differ in nascent adhesions and appear to initiate adhesion formation, despite their lack of visible organization. In more mature adhesions where the integrin is visibly organized there are ~900 integrins µm–2 (about fivefold higher than surrounding regions). Interestingly, {alpha}5-integrin and {alpha}-actinin, but not paxillin, reside in a complex throughout the cell, where they diffuse and flow together, even in regions where they are not organized. During adhesion disassembly some integrins diffuse away slowly, {alpha}-actinin undergoes a directed movement at speeds similar to actin retrograde flow (0.29 µm min–1), while all of the paxillin diffuses away rapidly.

Key words: {alpha}5-integrin, Cell migration, Adhesions, {alpha}-actinin, Paxillin, Correlation microscopy, Cytoskeleton


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