The shape of a cell membrane is largely defined by the underlying actin cytoskeleton and membrane mechanics. The actin cytoskeleton asserts contractile forces on the membrane that can be divided... Show moreThe shape of a cell membrane is largely defined by the underlying actin cytoskeleton and membrane mechanics. The actin cytoskeleton asserts contractile forces on the membrane that can be divided in isotropic and directed forces. We present a theory which is an extension of the Young-Laplace equation. It models cell edges as parts of one uniform ellipse, which changes from cell to cell. The ellipse parameters are characterized by the ratio of isotropic to directed contractility of the cell. We demonstrate the capabilities of this model using fibroblasts seeded on an elastic micro-pillar array. In this way adhesion forces exerted by the cell at single adhesion sites are measured. We show that isotropic and directed forces balance the line tension in cortical actin. Furthermore, for cells with homogeneous contractile forces and a single orientation of stress-fibers any part of the cell edge follows a universal ellipse, enabling us to calculate the magnitude of isotropic and directed contractility in a single cell. We show that in 3T3 fibroblasts the directed contractility is about three times as strong as the isotropic contractility. If myosin motors are inhibited, however, directed contractility decreases, effectively disabling forces generated by stress-fibers, and the elliptical cell cortex turns into a circular shape predicted for an isotropic contractile cytoskeleton. Our analysis shows that a simple two-parameter model explains polarity, shape of the cell cortex and cellular forces as experimentally observed. Potentially this model can be used to predict stresses and forces on the extracellular matrix and tissue. Show less
Integrin adhesion receptors connect the extracellular matrix (ECM) to the cytoskeleton and serve as bidirectional mechanotransducers. During development, angiogenesis, wound healing and cancer... Show moreIntegrin adhesion receptors connect the extracellular matrix (ECM) to the cytoskeleton and serve as bidirectional mechanotransducers. During development, angiogenesis, wound healing and cancer progression, the relative abundance of fibronectin receptors, including integrins α5β1 and αvβ3, changes, thus altering the integrin composition of cell-matrix adhesions. Here, we show that enhanced αvβ3 expression can fully compensate for loss of α5β1 and other β1 integrins to support outside-in and inside-out force transmission. α5β1 and αvβ3 each mediate actin cytoskeletal remodeling in response to stiffening or cyclic stretching of the ECM. Likewise, α5β1 and αvβ3 support cellular traction forces of comparable magnitudes and similarly increase these forces in response to ECM stiffening. However, cells using αvβ3 respond to lower stiffness ranges, reorganize their actin cytoskeleton more substantially in response to stretch, and show more randomly oriented traction forces. Centripetal traction force orientation requires long stress fibers that are formed through the action of Rho kinase (ROCK) and myosin II, and that are supported by α5β1. Thus, altering the relative abundance of fibronectin-binding integrins in cell-matrix adhesions affects the spatiotemporal organization of force transmission. Show less
Life’s building block is a cell. Different cell types are differentiated by specific functional properties. A white blood cell, for instance, can get rid of bacteria and many muscle cells contract... Show moreLife’s building block is a cell. Different cell types are differentiated by specific functional properties. A white blood cell, for instance, can get rid of bacteria and many muscle cells contract together for proper muscle function. Deformation and force exertion play important roles in these processes. Bacteria have to be physically engulfed by the white blood cell, and the muscle cell has to contract in the right way. In this research we measured how much force cells exert and simultaneously visualized specific proteins. A newly developed technique enabled the visualization of the nanometer-structure of cellular adhesions. We also examined the relationship between cellular shape and orientation of an intracellular network of protein (actin). We discovered that the signal of yet another protein (p130Cas) alters the mechanical behavior of the cell when the stiffness outside the cell changes. Finally, we also examined the structure of other proteins (tubulin and H2B) during cell division. In all these processes we measured how much force a cell exerts on its environment. The results provide important insights in the mechanical component of cellular function and their role in life Show less
Ribeiro, M.C.; Hoorn, H. van; Monshouwer-Kloots, J.J.; Mummery, C.L.; Schmidt, T.; Passier, R. 2013
