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Vol. 14, Issue 2, 407-416, February 2003
*Laboratoire de Biologie Moléculaire et Cellulaire,
Unité Mixte Recherche 5665, Centre National de la Recherche
Scientifique/ENS, INRA 913, Lyon, France; and
Podosomes, small actin-based adhesion structures, differ from focal
adhesions in two aspects: their core structure and their ability to
organize into large patterns in osteoclasts. To address the mechanisms
underlying these features, we imaged live preosteoclasts expressing
green fluorescent protein-actin during their differentiation. We
observe that podosomes always form inside or close to podosome groups,
which are surrounded by an actin cloud. Fluorescence recovery after
photobleaching shows that actin turns over in individual podosomes in
contrast to cortactin, suggesting a continuous actin polymerization in
the podosome core. The observation of podosome assemblies during
osteoclast differentiation reveals that they evolve from simple
clusters into rings that expand by the continuous formation of new
podosomes at their outer ridge and inhibition of podosome formation
inside the rings. This self-organization of podosomes into dynamic
rings is the mechanism that drives podosomes at the periphery of the
cell in large circular patterns. We also show that an additional step
of differentiation, requiring microtubule integrity, stabilizes the
podosome circles at the cell periphery to form the characteristic
podosome belt pattern of mature osteoclasts. These results therefore
provide a mechanism for the patterning of podosomes in osteoclasts and
reveal a turnover of actin inside the podosome.
Laboratoire de Physique, Unité Mixte
Recherche 5672, Centre National de la Recherche Scientifique-ENS Lyon
Ecole Normale Supérieure de Lyon 46, allée d'Italie, 69007 Lyon, France
These authors contributed equally to this work.
§
Present address: Cell and Developmental Biology
Department, University of California San Diego, La Jolla, CA
92093-0347.
Corresponding author. E-mail address:
fabard{at}biomail.ucsd.edu.
Online version of this article
contains video material for some figures. Online version available at
www.molbiolcell.org.
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