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Dynamics of cell-cell junction reinforcement probed with high resolution microscopy and microstructured surfaces
We present a new experimental approach to study the dynamics of formation of adherens junctions. Cells are individually added to 3D microwells to form duplexes or triplexes. The initial contact time is well defined and the 3D microstructure allows us to control the physical and chemical environment surrounding the cells. Using an appropriate geometry, we form cell-cell contacts parallel to the focal plane of the microscope objective. The dynamics of the contact area is then imaged at high spatial and temporal resolution by spinning disc microscopy. Few minutes after cell contact is established, the cortical actin is disrupted within the contact zone and rapidly forms an actin ring at the growing rim of the contact area. The E-cadherins cluster into five to ten foci per micron2 that subsequently assemble into a punctuated ring colocalized with the actin ring. We then study the dynamics of patch formation depending on the preexisting engagement of the cells in integrin based adhesion with the surrounding substrate. Lastly we show the existence of a load-dependent recruitement of E-cadherins at the junction.
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Contact : mathilde.reyssat@espci.fr