PGE2-mediated podosome loss in dendritic cells is dependent on actomyosin contraction downstream of the RhoA–Rho-kinase axis

Helden, Suzanne F. G. van; Oud, Machteld M.; Joosten, Ben; Peterse, Niels; Figdor, Carl G.; Leeuwen, Frank N. van

doi:10.1242/jcs.020289

Cited by 77 publications

(84 citation statements)

References 59 publications

(72 reference statements)

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“…Myosin IIA localizes into F-actin radial network at the vicinity of podosomes (Fig. S5) and has been shown to regulate podosome dynamics (3,21); therefore, we assessed the role of myosin II activity in the regulation of podosome biophysical properties. For this purpose, macrophages were treated with 50 μM of blebbistatin (Blebb) and analyzed by fluorescent microscopy and AFM.…”

Section: Biophysical Properties Of Podosomes Reflect F-actin Core Promentioning

confidence: 99%

Dynamics of podosome stiffness revealed by atomic force microscopy

Labernadie

Thibault

Vieu

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

147

163

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Podosomes are unique cellular entities specifically found in macrophages and involved in cell-matrix interactions, matrix degradation, and 3D migration. They correspond to a core of F-actin surrounded at its base by matrix receptors. To investigate the structure/function relationships of podosomes, soft lithography, atomic force microscopy (AFM), and correlative fluorescence microscopy were used to characterize podosome physical properties in macrophages differentiated from human blood monocytes. Podosome formation was restricted to delineated areas with micropatterned fibrinogen to facilitate AFM analyses. Podosome height and stiffness were measured with great accuracy in living macrophages (578 ± 209 nm and 43.8 ± 9.3 kPa) and these physical properties were independent of the nature of the underlying matrix. In addition, time-lapse AFM revealed that podosomes harbor two types of overlapping periodic stiffness variations throughout their lifespan, which depend on F-actin and myosin II activity. This report shows that podosome biophysical properties are amenable to AFM, allowing the study of podosomes in living macrophages at nanoscale resolution and the analysis of their intimate dynamics. Such an approach opens up perspectives to better understand the mechanical functionality of podosomes under physiological and pathological contexts.cytoskeleton | Young's modulus | microcontact printing

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Section: Biophysical Properties Of Podosomes Reflect F-actin Core Promentioning

confidence: 99%

Dynamics of podosome stiffness revealed by atomic force microscopy

Labernadie

Thibault

Vieu

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

147

163

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“…Less clear is the role of PGE 2 in focal adhesion formation. Different reports suggest that PGE 2 can either promote or inhibit focal adhesion formation, depending on the cell type (37,49). To better understand the role of PGE 2 in the organization of actin cytoskeleton in macrophages, we examined cells, using confocal microscopy, that were treated or not with PGE 2 (2 mM) for 18 h. Visualization of actin filaments (stained with phalloidin) and vinculin showed a significant reduction in the number of cells containing podosomes after treatment with PGE 2 (Fig.…”

Section: /2mentioning

confidence: 99%

“…PGE 2 signals through the EP2/EP4 receptors and switches Cdc42 on and Rac-1 off. This switch promotes podosome dissociation and focal adhesion formation (37). Cdc42, Rac, and Rho are direct targets of PI3K (38,39).…”

mentioning

confidence: 99%

Cyclooxygenase-2 Deficiency in Macrophages Leads to Defective p110γ PI3K Signaling and Impairs Cell Adhesion and Migration

Díaz-Muñoz

Osma-García

Íñiguez

et al. 2013

The Journal of Immunology

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Cyclooxygenase (Cox)-2 dependent PGs modulate several functions in many pathophysiological processes, including migration of immune cells. In this study, we addressed the role of Cox-2 in macrophage migration by using in vivo and in vitro models. Upon thioglycolate challenge, CD11b+ F4/80+ macrophages showed a diminished ability to migrate to the peritoneal cavity in cox-2−/− mice. In vivo migration of cox-2−/− macrophages from the peritoneal cavity to lymph nodes, as well as cell adhesion to the mesothelium, was reduced in response to LPS. In vitro migration of cox-2−/− macrophages toward MCP-1, RANTES, MIP-1α, or MIP-1β, as well as cell adhesion to ICAM-1 or fibronectin, was impaired. Defects in cell migration were not due to changes in chemokine receptor expression. Remarkably, cox-2−/− macrophages showed a deficiency in focal adhesion formation, with reduced phosphorylation of paxillin (Tyr188). Interestingly, expression of the p110γ catalytic subunit of PI3K was severely reduced in the absence of Cox-2, leading to defective Akt phosphorylation, as well as cdc42 and Rac-1 activation. Our results indicate that the paxillin/p110γ-PI3K/Cdc42/Rac1 axis is defective in cox-2−/− macrophages, which results in impaired cell adhesion and migration.

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“…Non-muscle myosin-II has been shown to localise to podosome regions (Chabadel et al, 2007;van Helden et al, 2008). MyosinIIa was localised using confocal microscopy to determine whether it was associated with DC podosomes.…”

Section: The Myosin-ii Inhibitor Blebbistatin Decreases the Length Ofmentioning

confidence: 99%

“…Surrounding the core is a ring region that contains scaffolding, signalling and integrin-binding proteins, including the typical focal adhesion proteins FAK, vinculin and paxillin (Block et al, 2008). Non muscle myosins are also associated with podosomes (Kopp et al, 2006;van Helden et al, 2008). Together, the core and the ring contain an extraordinary array of molecular components some of which are essential for podosome assembly (Gimona et al, 2008).…”

Section: Introductionmentioning

confidence: 99%

Dendritic cell podosomes are protrusive and invade the extracellular matrix using metalloproteinase MMP-14

Gawden-Bone

Zhou

King

et al. 2010

Journal of Cell Science

126

123

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SummaryPodosomes are spot-like actin-rich structures formed at the ventral surface of monocytic and haematopoietic cells. Podosomes degrade extracellular matrix and are proposed to be involved in cell migration. A key question is whether podosomes form protrusions similar to the invadopodia of cancer cells. We characterised podosomes of immature dendritic cells using electron microscopy combined with both conventional and novel high-resolution structured illumination light microscopy. Dendritic cell podosomes are composed of actin foci surrounded by a specialised ring region that is rich in material containing paxillin. We found that podosomes were preferential sites for protrusion into polycarbonate filters impregnated with crosslinked gelatin, degrading up to 2 mm of matrix in 24 hours. Podosomeassociated uptake of colloidal gold-labelled gelatin matrix appeared to occur via large phagosome-like structures or narrow tubular invaginations. The motor protein myosin-II was excluded from ring or core regions but was concentrated around them and the myosin-II inhibitor Blebbistatin reduced the length of podosome protrusions. Finally, we found that degradation, protrusion and endocytosis in this system are dependent on the matrix metalloproteinase MMP-14. We propose that podosomes mediate migration of dendritic cells through tissues by means of myosin-II-dependent protrusion coupled to MMP-14-dependent degradation and endocytosis.

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PGE2-mediated podosome loss in dendritic cells is dependent on actomyosin contraction downstream of the RhoA–Rho-kinase axis

Cited by 77 publications

References 59 publications

Dynamics of podosome stiffness revealed by atomic force microscopy

Dynamics of podosome stiffness revealed by atomic force microscopy

Cyclooxygenase-2 Deficiency in Macrophages Leads to Defective p110γ PI3K Signaling and Impairs Cell Adhesion and Migration

Dendritic cell podosomes are protrusive and invade the extracellular matrix using metalloproteinase MMP-14

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