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Lamellipodia nucleation by filopodia depends on integrin occupancy and downstream Rac1 signaling

Abstract : Time-lapse video-microscopy unambiguously shows that fibroblast filopodia are the scaffold of lamellipodia nucleation that allows anisotropic cell spreading. This process was dissected into elementary stages by monitoring cell adhesion on micropatterned extracellular matrix arrays of various pitches. Adhesion structures are stabilized by contact with the adhesive plots and subsequently converted into lamellipodia-like extensions starting at the filopodia tips. This mechanism progressively leads to full cell spreading. Stable expression of the dominant-negative Rac1 N17 impairs this change in membrane extension mode and stops cell spreading on matrix arrays. Similar expression of the dominant-negative Cdc42 N17 impairs cell spreading on homogenous and structured substrate, suggesting that filopodia extension is a prerequisite for cell spreading in this model. The differential polarity of the nucleation of lamellipodial structures by filopodia on homogenous and structured surfaces starting from the cell body and of filopodia tip, respectively, suggested that this process is triggered by areas that are in contact with extracellular matrix proteins for longer times. Consistent with this view, wild-type cells cannot spread on microarrays made of function blocking or neutral anti-beta 1 integrin antibodies. However, stable expression of a constitutively active Rac1 mutant rescues the cell ability to spread on these integrin microarrays. Thereby, lamellipodia nucleation by filopodia requires integrin occupancy by matrix substrate and downstream Rac1 signaling.
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Submitted on : Wednesday, March 12, 2008 - 9:50:48 AM
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Hervé Guillou, Adeline Depraz-Depland, Emmanuelle Planus, Benoit Vianay, Jacques Chaussy, et al.. Lamellipodia nucleation by filopodia depends on integrin occupancy and downstream Rac1 signaling. Experimental Cell Research, Elsevier, 2008, 314 (3), pp.478-488. ⟨10.1016/j.yexcr.2007.10.026⟩. ⟨inserm-00263420⟩



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