Force mapping in epithelial cell migration.

Olivia du Roure; Alexandre Saez; Axel Buguin; Robert H Austin; Philippe Chavrier; Pascal Silberzan; Pascal Siberzan; Benoit Ladoux

doi:10.1073/pnas.0408482102

PubMed · 15695588

Force mapping in epithelial cell migration.

Olivia du Roure·Alexandre Saez·Axel Buguin·Robert H Austin·Philippe Chavrier·Pascal Silberzan·Pascal Siberzan·Benoit Ladoux

View original source ↗Export metadata ↓

Abstract

We measure dynamic traction forces exerted by epithelial cells on a substrate. The force sensor is a high-density array of elastomeric microfabricated pillars that support the cells. Traction forces induced by cell migration are deduced from the measurement of the bending of these pillars and are correlated with actin localization by fluorescence microscopy. We use a multiple-particle tracking method to estimate the mechanical activity of cells in real time with a high-spatial resolution (down to 2 microm) imposed by the periodicity of the post array. For these experiments, we use differentiated Madin-Darby canine kidney (MDCK) epithelial cells. Our data provide definite information on mechanical forces exerted by a cellular assembly. The maximum intensity of the forces is localized on the edge of the epithelia. Hepatocyte growth factor promotes cell motility and induces strong scattering activity of MDCK cells. Thus, we compare forces generated by MDCK cells in subconfluent epithelia versus isolated cells after hepatocyte growth factor treatment. Maximal-traction stresses at the edge of a monolayer correspond to higher values than those measured for a single cell and may be due to a collective behavior.

Explore related subjects

Actins Animals Biomechanical Phenomena Biophysical Phenomena Biophysics Cell Adhesion Cell Line Cell Movement Dogs Epithelial Cells Hepatocyte Growth Factor Microscopy, Electron, Scanning Microscopy, Fluorescence Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

Keep this discovery

Explore connections, maps & timelines

BibTeX RIS

Olivia du Roure, Alexandre Saez, Axel Buguin, Robert H Austin, Philippe Chavrier, Pascal Silberzan, Pascal Siberzan, Benoit Ladoux. 2005-02-04. Force mapping in epithelial cell migration.. https://doi.org/10.1073/pnas.0408482102

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related citations

PubMed2009

Nap1-mediated actin remodeling is essential for mammalian myoblast fusion.

Myoblast fusion is crucial for the formation, growth, maintenance and regeneration of healthy skeletal muscle. Unfortunately, the molecular machinery, cell behaviors, and membrane and cytoskeletal remodeling events that govern fusion and myofiber formation remain poorly understood. Using time-lapse imaging approaches on mouse C2C12 myoblasts, we identify discrete and specific molecular events at myoblast membranes during fusion and myotube formation. These events include rearrangement of cell shape from fibroblast to spindle-like morphologies, changes in lamellipodial and filopodial extensions during different periods of differentiation, and changes in membrane alignment and organization during fusion. We find that actin-cytoskeleton remodeling is crucial for these events: pharmacological inhibition of F-actin polymerization leads to decreased lamellipodial and filopodial extensions and to reduced myoblast fusion. Additionally, shRNA-mediated inhibition of Nap1, a member of the WAVE actin-remodeling complex, results in accumulations of F-actin structures at the plasma membrane that are concomitant with a decrease in myoblast fusion. Our data highlight distinct and essential roles for actin cytoskeleton remodeling during mammalian myoblast fusion, provide a platform for cellular and molecular dissection of the fusion process, and suggest a functional conservation of Nap1-regulated actin-cytoskeleton remodeling during myoblast fusion between mammals and Drosophila.

Actins↗

PubMed2006

Curcumin blocks activation of pancreatic stellate cells.

Activated pancreatic stellate cells (PSCs) play a pivotal role in the pathogenesis of pancreatic fibrosis and inflammation. Inhibition of activation and cell functions of PSCs is a potential target for the treatment of pancreatic fibrosis and inflammation. The polyphenol compound curcumin is the yellow pigment in curry, and has anti-inflammatory and anti-fibrotic properties. We here evaluated the effects of curcumin on the activation and cell functions of PSCs. PSCs were isolated from rat pancreas tissue and used in their culture-activated, myofibroblast-like phenotype unless otherwise stated. The effects of curcumin on proliferation, alpha-smooth muscle actin gene expression, monocyte chemoattractant protein (MCP)-1 production, and collagen expression were examined. The effect of curcumin on the activation of freshly isolated cells in culture was also assessed. Curcumin inhibited platelet-derived growth factor (PDGF)-induced proliferation, alpha-smooth muscle actin gene expression, interleukin-1beta- and tumor necrosis factor (TNF)-alpha-induced MCP-1 production, type I collagen production, and expression of type I and type III collagen genes. Curcumin inhibited PDGF-BB-induced cyclin D1 expression and activation of extracellular signal-regulated kinase (ERK). Curcumin inhibited interleukin-1beta- and TNF-alpha-induced activation of activator protein-1 (AP-1) and mitogen-activated protein (MAP) kinases (ERK, c-Jun N-terminal kinase (JNK), and p38 MAP kinase), but not of nuclear factor-kappaB (NF-kappaB). In addition, curcumin inhibited transformation of freshly isolated cells to myofibroblast-like phenotype. In conclusion, curcumin inhibited key cell functions and activation of PSCs.

Actins↗

PubMed2006

The effect of hydroxyapatite nanocrystals on microvascular endothelial cell viability and functions.

To favor bone reconstitution with biomaterials endothelial cells should maintain proper functions to drive angiogenesis. To this aim nanocrystals of hydroxyapatite (HA) have been synthesized and characterized on endothelial cells. Microvascular endothelial cells have been exposed to stoichiometric HA nanocrystals. Cell morphology and organization of cytoskeletal proteins have been monitored by SEM analysis and immunofluorescence. Biochemical markers of physiological and pathological responses of endothelial cells, endothelial constitutive nitric oxide synthase, and cycloxygenase-2 (ecNOS and COX-2, respectively) have been measured by immunofluorescence. Crystallized HA sustained endothelial survival without any cytotoxic effect. At the observation with SEM, endothelial cell morphology was maintained in the presence of HA. The localization and organization of beta-actin documented the formation of stress fibers, indicating an activation of endothelial cells induced by HA nanocrystals. Immunohistochemistry for biochemical key signaling pathways in endothelium demonstrated that nanocrystals of HA maintained the expression of ecNOS and did not increase COX-2 expression. In conclusion, the present findings indicate that HA nanocrystals exhibit high biocompatibility for microvascular endothelium. In the presence of HA nanocrystals endothelial cells maintain biochemical markers of healthy endothelium. They do not acquire a proinflammatory or thrombogenic phenotype, but express markers of functioning endothelium that might contribute to angiogenesis.

Actins↗