Biology subjects

Berthoz, R.

Publications and source records attributed to Berthoz, R..

4 recordsLinked to original sources

bioRxiv2025

Interplay between Rac1/RhoA and actin waves in giant epithelial cells : experiment and theory

The acto-myosin cytoskeleton is a key driver of cellular shape changes in vivo and in vitro. Acto-myosin organization results from actin assembly and interactions between actin and myosin, which are both regulated by small Rho GTPases like Rac1 and RhoA. To uncover principles governing cytoskeletal organization, we analyzed actin patterns using live microscopy and theory. In giant Madin-Darby Canine Kidney (MDCK) epithelial cells and REF52 fibroblasts, we observed acto-myosin stress fibres and propagating waves. Stress fibres were stationary and correlated with homogeneous distributions of Rac and RhoA activity. Waves propagated at {approx} 1m/min and were associated with density variations of actin, Rac and active RhoA. Some waves transported cellular components or generated protrusions at the cell edge. Essential features of wave propagation are captured by a polar reaction-diffusion system for actin and Rac. Notably, two colliding waves annihilate each other. In cells, myosin activity was not required for the emergence of waves, but tended to suppress them. Consistently, local activation of RhoA slowed down or stopped and broke waves. These results highlight how the coupling between acto-myosin and Rho GTPase generates a variety of cytoskeletal structures and dynamics. O_TEXTBOXSIGNIFICANCEThe cytoskeleton shapes cells in vitro and in vivo. Its molecular mechanisms have been extensively documented, but the mesoscopic structures resulting from the interplay between the activator and the motor activity have been poorly characterized. Here we show that two structures are conserved in two systems, stress fibres with RhoA activity and diffusion actin waves regulated by Rac1. The annihilation of waves and the alteration of the wave dynamics are reproduced by a minimal model based on a polar reaction-diffusion model. This approach could pave the way to a generic description of regulated active gels in cells with potential biological functions, such as stress generation for fibres and probing of space and homogenization of the cortex by actin waves. C_TEXTBOX

biophysics↗

bioRxiv2025

Teach your microscope how to print: Low-cost and rapid-iteration microfabrication for biology

The application of traditional microfabrication techniques to biological research is hindered by their reliance on clean rooms, expensive or toxic materials, and slow iteration cycles. We present an accessible microfabrication workflow that addresses these challenges by integrating consumer 3D printing techniques and repurposing standard fluorescence microscopes equipped with DMDs for maskless photolithography. Our method achieves micrometer-scale precision across centimeter-sized areas without clean room infrastructure, using affordable and readily available consumables. We demonstrate the versatility of this approach through four biological applications: inducing cytoskeletal protrusions via 1 m-resolution surface topographies; micropatterning to standardize cell and tissue morphology; fabricating multilayer microfluidic devices for confined cell migration studies; imprinting agar chambers for long-time tracking of C. elegans. Our protocol drastically reduces material costs compared to conventional methods and enables design-to-device turnaround within a day. By leveraging open-source microscope control software and existing lab equipment, our workflow lowers the entry barrier to micro-fabrication, enabling labs to prototype custom solutions for diverse experimental needs while maintaining compatibility with soft lithography and downstream biological assays.

cell biology↗

bioRxiv2024

Myosin cluster dynamics determines epithelial wound ring constriction

Collection of myosin motors and actin filaments can self-assemble into submicrometric clusters under the regulation of RhoA. Emergent dynamics of these clusters have been reported in a variety of morphogenetic systems, ranging from Drosophila to acto-myosin assays in vitro. In single cell cytokinetic rings, acto-myosin clusters are associated with stress generation when radial and transport when tangential with respect to the ring closure. Here, we show that these phenomena hold true for acto-myosin multi-cellular rings during wound closure in epithelial monolayers. We assessed the activity of RhoA using FRET sensors, and we report that cluster dynamics does not correlate with RhoA activity. Nevertheless, we show that bursts of RhoA activation precede recruitment of myosin. Altogether myosin clusters dynamics is conserved between single and multi-cellular systems and this suggests that they could be used as generic read-outs for mapping and predicting stress generation and shape changes in morphogenesis.

biophysics↗

bioRxiv2022

Polarity-driven three-dimensional spontaneous rotation of a cell doublet

Cell mechanical interactions play a fundamental role in the self-organisation of organisms. How these interactions drive coordinated cell movement in three-dimensions remains unclear. Here we report that cell doublets embedded in a 3D extracellular matrix undergo spontaneous rotations and we investigate the rotation mechanism using live cell imaging, quantitative measurements, mechanical perturbations, and theory. We find that rotation is driven by a polarized distribution of myosin within cell cortices. The mismatched orientation of this polarized distribution breaks the doublet mirror symmetry. In addition, cells adhere at their interface through adherens junctions and with the extracellular matrix through focal contacts near myosin clusters. Using a physical theory describing the doublet as two interacting active surfaces, we find that rotation is driven by myosin-generated gradients of active tension, whose profiles are dictated by interacting cell polarity axes. We show that interface three-dimensional shapes can be understood from the Curie principle: shapes symmetries are related to broken symmetries of myosin distribution in cortices. To test for the rotation mechanism, we suppress myosin clusters using laser ablation and we generate new myosin clusters by optogenetics. Our work clarifies how polarity-oriented active mechanical forces drive collective cell motion in three dimensions.

biophysics↗