A minimal model for spontaneous cell polarization and edge activity in oscillating, rotating and migrating cells

How the cells break symmetry and organize their edge activity to move directionally is a fun- damental question in cell biology. Physical models of cell motility commonly rely on gradients of regulatory factors and/or feedback from the motion itself to describe polarization of edge activity. Theses approaches, however, fail to explain cell behavior prior to the onset of polarization. Our analysis using the model system of polarizing and moving fish epidermal keratocytes suggests a novel and simple principle of self-organization of cell activity in which local cell-edge dynamics depends on the distance from the cell center, but not on the orientation with respect to the front-back axis. We validate this principle with a stochastic model that faithfully reproduces a range of cell-migration behaviors. Our findings indicate that spontaneous polarization, persistent motion, and cell shape are emergent properties of the local cell-edge dynamics controlled by the distance from the cell center.Comment: 8 pages, 5 figure

Weak Force Stalls Protrusion at the Leading Edge of the Lamellipodium

AbstractProtrusion, the first step of cell migration, is driven by actin polymerization coupled to adhesion at the cell’s leading edge. Polymerization and adhesive forces have been estimated, but the net protrusion force has not been measured accurately. We arrest the leading edge of a moving fish keratocyte with a hydrodynamic load generated by a fluid flow from a micropipette. The flow arrests protrusion locally as the cell approaches the pipette, causing an arc-shaped indentation and upward folding of the leading edge. The effect of the flow is reversible upon pipette removal and dependent on the flow direction, suggesting that it is a direct effect of the external force rather than a regulated cellular response. Modeling of the fluid flow gives a surprisingly low value for the arresting force of just a few piconewtons per micrometer. Enhanced phase contrast, fluorescence, and interference reflection microscopy suggest that the flow does not abolish actin polymerization and does not disrupt the adhesions formed before the arrest but rather interferes with weak nascent adhesions at the very front of the cell. We conclude that a weak external force is sufficient to reorient the growing actin network at the leading edge and to stall the protrusion

Dynamic measurement of the height and volume of migrating cells by a novel fluorescence microscopy technique

We propose a new technique to measure the volume of adherent migrating cells. The method is based on a negative staining where a fluorescent, non cell-permeant dye is added to the extracellular medium. The specimen is observed with a conventional fluorescence microscope in a chamber of uniform height. Given that the fluorescence signal depends on the thickness of the emitting layer, the objects excluding the fluorescent dye (i.e., cells) appear dark, and the decrease of the fluorescent signal with respect to the background is expected to give information about the height and the volume of the object. Using a glass microfabricated pattern with steps of defined heights, we show that the drop in fluorescence intensity is indeed proportional to the height of the step and obtain calibration curves relating fluorescence intensity to height. The technique, termed fluorescence displacement method, is further validated by comparing our measurements with the ones obtained by atomic force microscopy (AFM). We apply our method to measure the real-time volume dynamics of migrating fish epidermal keratocytes subjected to osmotic stress. The fluorescence displacement technique allows fast and precise monitoring of cell height and volume, thus providing a valuable tool for characterizing the three-dimensional behaviour of migrating cells

Force transmission in migrating cells

Analysis of the relationship between actin network velocity and traction forces at the substrate shows that force transmission mechanisms vary with distinct regions of the cell

Comparative Dynamics of Retrograde Actin Flow and Focal Adhesions: Formation of Nascent Adhesions Triggers Transition from Fast to Slow Flow

Dynamic actin network at the leading edge of the cell is linked to the extracellular matrix through focal adhesions (FAs), and at the same time it undergoes retrograde flow with different dynamics in two distinct zones: the lamellipodium (peripheral zone of fast flow), and the lamellum (zone of slow flow located between the lamellipodium and the cell body). Cell migration involves expansion of both the lamellipodium and the lamellum, as well as formation of new FAs, but it is largely unknown how the position of the boundary between the two flow zones is defined, and how FAs and actin flow mutually influence each other. We investigated dynamic relationship between focal adhesions and the boundary between the two flow zones in spreading cells. Nascent FAs first appeared in the lamellipodium. Within seconds after the formation of new FAs, the rate of actin flow decreased locally, and the lamellipodium/lamellum boundary advanced towards the new FAs. Blocking fast actin flow with cytochalasin D resulted in rapid dissolution of nascent FAs. In the absence of FAs (spreading on poly-L-lysine-coated surfaces) retrograde flow was uniform and the velocity transition was not observed. We conclude that formation of FAs depends on actin dynamics, and in its turn, affects the dynamics of actin flow by triggering transition from fast to slow flow. Extension of the cell edge thus proceeds through a cycle of lamellipodium protrusion, formation of new FAs, advance of the lamellum, and protrusion of the lamellipodium from the new base

The mechanisms of spatial and temporal patterning of cell-edge dynamics

Adherent cells migrate and change their shape by means of protrusion and retraction at their edges. When and where these activities occur defines the shape of the cell and the way it moves. Despite a great deal of knowledge about the structural organization, components, and biochemical reactions involved in protrusion and retraction, the origins of their spatial and temporal patterns are still poorly understood. Chemical signaling circuitry is believed to be an important source of patterning, but recent studies highlighted mechanisms based on physical forces, motion, and mechanical feedback

Cell Polarization: Mechanical Switch for a Chemical Reaction

SummaryAnterior–posterior polarity in the Caenorhabditis elegans zygote depends on two groups of PAR proteins, as well as on cortical flow. Recent work now demonstrates that this polarization results from a transition in a bistable reaction–diffusion system of PAR proteins that is triggered by cortical flow

Cell–Matrix Adhesion: Slip and Immobilization under Force

SummaryWhen force is applied to cell–matrix adhesion complexes, they respond by growing larger and stronger. It emerges that strengthening involves transient motion of the transmembrane integrin receptors and their eventual immobilization to the extracellular matrix