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Published online 19 May 2003. doi:10.1083/jcb.200212037
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© The Rockefeller University Press, 0021-9525/2003/5/697 $5.00
The Journal of Cell Biology, Volume 161, Number 4, 697-705

Article

 Macrophage podosomes assemble at the leading lamella by growth and fragmentation

James G. Evans1,2, Ivan Correia2, Olga Krasavina2, Nicki Watson2 and Paul Matsudaira1,2,3

1 BioImaging Center, Cambridge, MA 02142
2 Whitehead Institute for Biomedical Research, Cambridge, MA 02142
3 Department of Biology and Division of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139

Address correspondence to James G. Evans, Whitehead Institute, 9 Cambridge Center, Cambridge, MA 02142. Tel.: (617) 324-0300. Fax: (617) 258-7226. E-mail: jgevans{at}wi.mit.edu

Podosomes are actin- and fimbrin-containing adhesions at the leading edge of macrophages. In cells transfected with ß-actin–ECFP and L-fimbrin–EYFP, quantitative four-dimensional microscopy of podosome assembly shows that new adhesions arise at the cell periphery by one of two mechanisms; de novo podosome assembly, or fission of a precursor podosome into daughter podosomes. The large podosome cluster precursor also appears to be an adhesion structure; it contains actin, fimbrin, integrin, and is in close apposition to the substratum. Microtubule inhibitors paclitaxel and demecolcine inhibit the turnover and polarized formation of podosomes, but not the turnover rate of actin in these structures. Because daughter podosomes and podosome cluster precursors are preferentially located at the leading edge, they may play a critical role in continually generating new sites of cell adhesion.

Key Words: fluorescence microscopy; adhesion; microtubules; actin; kymography

The online version of this article includes supplemental material.

* Abbreviations used in this paper: 2-, 3-, and 4-D, two, three, and four dimensional; IRM, interference reflection microscopy; PCP, podosome cluster precursor.


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