UMR168 – Laboratoire Physico-Chimie Curie

Publications de l’UMR 168

Année de publication : 2015

Sham Tlili, Cyprien Gay, François Graner, Philippe Marcq, François Molino, Pierre Saramito (2015 Nov 6)

Erratum to: Colloquium: Mechanical formalisms for tissue dynamics.

The European physical journal. E, Soft matter : 115 : DOI : 10.1140/epje/i2015-15115-3 En savoir plus
Résumé

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Mijo Simunovic, Gregory A Voth, Andrew Callan-Jones, Patricia Bassereau (2015 Nov 2)

When Physics Takes Over: BAR Proteins and Membrane Curvature.

Trends in cell biology : 780-92 : DOI : 10.1016/j.tcb.2015.09.005 En savoir plus
Résumé

Cell membranes Become highly curved During membrane trafficking, cytokinesis, infection, immune response, or cell motion. Bin / Amphiphysin / Rvs (BAR) domain proteins Intrinsically With Their curved shape anisotropy and are Involved in Many of These processes, aim with a wide spectrum of modes of action. In vitro experiments and computer simulations multiscale-have Contributed in Identifying a minimal set of physical parameters derived derived, namely protein density on the membrane, membrane voltage and membrane shape, That control how bound BAR domain proteins behave on the membrane. In this review, we summarize the multifaceted BAR coupling of proteins to membrane mechanics and offers a single-phase diagram That Recapitulates the effects of These parameters.

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Ahmed WW, Fodor É, Betz T (2015 Nov 1)

Active cell mechanics: measurement and theory

Biochimica et Biophysica Acta : 1853 : 3083-94 : DOI : 10.1016/j.bbamcr.2015.05.022 En savoir plus
Résumé

Living cells are active mechanical systems that are able to generate forces. Their structure and shape are primarily determined by biopolymer filaments and molecular motors that form the cytoskeleton. Active force generation requires constant consumption of energy to maintain the nonequilibrium activity to drive organization and transport processes necessary for their function. To understand this activity it is necessary to develop new approaches to probe the underlying physical processes. Active cell mechanics incorporates active molecular-scale force generation into the traditional framework of mechanics of materials. This review highlights recent experimental and theoretical developments towards understanding active cell mechanics. We focus primarily on intracellular mechanical measurements and theoretical advances utilizing the Langevin framework. These developing approaches allow a quantitative understanding of nonequilibrium mechanical activity in living cells. This article is part of a Special Issue entitled: Mechanobiology.

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Cáceres R, Abou-Ghali M, Plastino J (2015 Nov 1)

Reconstituting the actin cytoskeleton at or near surfaces in vitro

Biochimica et Biophysica Acta : 1853 : 3006-14 : DOI : 10.1016/j.bbamcr.2015.07.021 En savoir plus
Résumé

Actin filament dynamics have been studied for decades in pure protein solutions or in cell extracts, but a breakthrough in the field occurred at the turn of the century when it became possible to reconstitute networks of actin filaments, growing in a controlled but physiological manner on surfaces, mimicking the actin assembly that occurs at the plasma membrane during cell protrusion and cell shape changes. The story begins with the bacteria Listeria monocytogenes, the study of which led to the reconstitution of cellular actin polymerization on a variety of supports including plastic beads. These studies made possible the development of liposome-type substrates for filament assembly and micropatterning of actin polymerization nucleation. Based on the accumulated expertise of the last 15 years, many exciting approaches are being developed, including the addition of myosin to biomimetic actin networks to study the interplay between actin structure and contractility. The field is now poised to make artificial cells with a physiological and dynamic actin cytoskeleton, and subsequently to put these cells together to make in vitro tissues. This article is part of a Special Issue entitled: Mechanobiology.

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Grégory Beaune, Françoise M Winnik, Françoise Brochard-Wyart (2015 Oct 29)

Formation of Tethers from Spreading Cellular Aggregates.

Langmuir : the ACS journal of surfaces and colloids : 12984-92 : DOI : 10.1021/acs.langmuir.5b02785 En savoir plus
Résumé

Membrane tubes are commonly extruded from cells and vesicles when a point-like force is applied on the membrane. We report here the unexpected formation of membrane tubes from lymph node cancer prostate (LNCaP) cell aggregates in the absence of external applied forces. The spreading of LNCaP aggregates deposited on adhesive glass substrates coated with fibronectin is very limited because cell-cell adhesion is stronger than cell-substrate adhesion. Some cells on the aggregate periphery are very motile and try to escape from the aggregate, leading to the formation of membrane tubes. Tethered networks and exchange of cargos between cells were observed as well. Growth of the tubes is followed by either tube retraction or tube rupture. Hence, even very cohesive cells are successful in escaping aggregates, which may lead to epithelial mesenchymal transition and tumor metastasis. We interpret the dynamics of formation and retraction of tubes in the framework of membrane mechanics.

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Reza M Mohamadi, Zuzana Svobodova, Zuzana Bilkova, Markus Otto, Myriam Taverna, Stephanie Descroix, Jean-Louis Viovy (2015 Oct 22)

An integrated microfluidic chip for immunocapture, preconcentration and separation of β-amyloid peptides.

Biomicrofluidics : 054117 : DOI : 10.1063/1.4931394 En savoir plus
Résumé

We present an integrated microfluidic chip for detection of β-amyloid (Aβ) peptides. Aβ peptides are major biomarkers for the diagnosis of Alzheimer’s disease (AD) in its early stages. This microfluidic device consists of three main parts: (1) An immunocapture microcolumn based on self-assembled magnetic beads coated with antibodies specific to Aβ peptides, (2) a nano-porous membrane made of photopolymerized hydrogel for preconcentration, and (3) a microchip electrophoresis (MCE) channel with fluorescent detection. Sub-milliliter sample volume is either mixed off-chip with antibody coated magnetic beads and injected into the device or is injected into an already self-assembled column of magnetic beads in the microchannel. The captured peptides on the beads are then electrokinetically eluted and re-concentrated onto the nano-membrane in a few nano-liters. By integrating the nano-membrane, total assay time was reduced and also off-chip re-concentration or buffer exchange steps were not needed. Finally, the concentrated peptides in the chip are separated by electrophoresis in a polymer-based matrix. The device was applied to the capture and MCE analysis of differently truncated peptides Aβ (1-37, 1-39, 1-40, and 1-42) and was able to detect as low as 25 ng of synthetic Aβ peptides spiked in undiluted cerebrospinal fluid (CSF). The device was also tested with CSF samples from healthy donors. CSF samples were fluorescently labelled and pre-mixed with the magnetic beads and injected into the device. The results indicated that Aβ1-40, an important biomarker for distinguishing patients with frontotemporal lobe dementia from controls and AD patients, was detectable. Although the sensitivity of this device is not yet enough to detect all Aβ subtypes in CSF, this is the first report on an integrated or semi-integrated device for capturing and analyzing of differently truncated Aβ peptides. The method is less demanding and faster than the conventional Western blotting method currently used for research.

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A Y Grosberg, J-F Joanny (2015 Oct 15)

Nonequilibrium statistical mechanics of mixtures of particles in contact with different thermostats.

Physical review. E, Statistical, nonlinear, and soft matter physics : 032118 : DOI : 10.1103/PhysRevE.92.032118 En savoir plus
Résumé

We introduce a novel type of locally driven systems made of two types of particles (or a polymer with two types of monomers) subject to a chaotic drive with approximately white noise spectrum, but different intensity; in other words, particles of different types are in contact with thermostats at different temperatures. We present complete systematic statistical mechanics treatment starting from first principles. Although we consider only corrections to the dilute limit due to pairwise collisions between particles, meaning we study a nonequilibrium analog of the second virial approximation, we find that the system exhibits a surprisingly rich behavior. In particular, pair correlation function of particles has an unusual quasi-Boltzmann structure governed by an effective temperature distinct from that of any of the two thermostats. We also show that at sufficiently strong drive the uniformly mixed system becomes unstable with respect to steady states consisting of phases enriched with different types of particles. In the second virial approximation, we define nonequilibrium « chemical potentials » whose gradients govern diffusion fluxes and a nonequilibrium « osmotic pressure, » which governs the mechanical stability of the interface.

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Thibaut J Lagny, Patricia Bassereau (2015 Oct 15)

Bioinspired membrane-based systems for a physical approach of cell organization and dynamics: usefulness and limitations.

Interface focus : 20150038 : DOI : 10.1098/rsfs.2015.0038 En savoir plus
Résumé

Being at the Periphery of Each cell compartment and enclosing the Entire cell while interacting with a large part of cell components, cell membranes Participate in MOST of the cell’s vital functions. Biologists-have for a long time Worked on deciphering how membranes are Organized, How They contribuer to trafficking, motility, cytokinesis, cell-cell communication, transport information, etc., using top-down Approaches and always more advanced techniques. In contrast, physicists-have Developed bottom-up Approaches and minimal model membrane systems of growing complexity in order to build up general models That explain how cell membranes work and How They interact with proteins, eg the cytoskeleton. We review the different model membrane systems That ares currently available, and How They can help deciphering cell functioning, goal aussi Their list limitations. Model membrane systems are aussi used in synthetic biology and can-have potential applications beyond basic research. We can the Chat the synergy entre le development of complex membrane systems in vitro in a biological context and for technological applications. Questions That Could aussi be Discussed are: what can we still do with synthetic systems, where do we stop building up and qui are the alternative solutions?

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Nicolas Christophorou, Thomas Rubin, Isabelle Bonnet, Tristan Piolot, Marion Arnaud, Jean-René Huynh (2015 Oct 13)

Microtubule-driven nuclear rotations promote meiotic chromosome dynamics.

Nature cell biology : 1388-400 : DOI : 10.1038/ncb3249 En savoir plus
Résumé

At the onset of meiosis, each chromosome needs to find its homologue and pair to ensure proper segregation. In Drosophila, pairing occurs during the mitotic cycles preceding meiosis. Here we show that germ cell nuclei undergo marked movements during this developmental window. We demonstrate that microtubules and Dynein are driving nuclear rotations and are required for centromere pairing and clustering. We further found that Klaroid (SUN) and Klarsicht (KASH) co-localize with centromeres at the nuclear envelope and are required for proper chromosome motions and pairing. We identified Mud (NuMA in vertebrates) as co-localizing with centromeres, Klarsicht and Klaroid. Mud is also required to maintain the integrity of the nuclear envelope and for the correct assembly of the synaptonemal complex. Our findings reveal a mechanism for chromosome pairing in Drosophila, and indicate that microtubules, centrosomes and associated proteins play a crucial role in the dynamic organization of chromosomes inside the nucleus.

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D Ferraro, Y Lin, B Teste, D Talbot, L Malaquin, S Descroix, A Abou-Hassan (2015 Oct 6)

Continuous chemical operations and modifications on magnetic γ-Fe2O3 nanoparticles confined in nanoliter droplets for the assembly of fluorescent and magnetic SiO2@γ-Fe2O3.

Chemical communications (Cambridge, England) : 16904-7 : DOI : 10.1039/c5cc07044a En savoir plus
Résumé

We present a microfluidic platform that allows undergoing different chemical operations in a nanoliter droplet starting from the colloidal suspension of magnetic iron oxide (γ-Fe2O3) nanoparticles « NPs » (ferrofluid). These operations include: mixing, flocculation, magnetic decantation, colloidal redispersion, washing, surface functionalization, heating and colloidal assembly. To prove the platform capabilities, we produced fluorescent and magnetic nanoassemblies composed of fluorescent silica and magnetic NPs.

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Monica Rolando, Caroline Stefani, Anne Doye, Maria I Acosta, Orane Visvikis, Hannah G Yevick, Carmen Buchrieser, Amel Mettouchi, Patricia Bassereau, Emmanuel Lemichez (2015 Sep 26)

Contractile actin cables induced by Bacillus anthracis lethal toxin depend on the histone acetylation machinery.

Cytoskeleton (Hoboken, N.J.) : 542-56 : DOI : 10.1002/cm.21256 En savoir plus
Résumé

It remains a challenge to decode the molecular basis of the long-term actin cytoskeleton rearrangements That are-governed by the reprogramming of gene expression. Bacillus anthracis lethal toxin (LT) Inhibits mitogen-activated protein kinase (MAPK) signaling, thereby modulating gene expression, with major consequences for actin cytoskeleton organization and the loss of endothelial barrier function. Using a laser ablation approach, we caractérisé the contractile and tensile mechanical properties of LT-induced stress fibers. These actin cables resist pulling force That Transmitted are at cell-matrix interfaces and at cell-cell adherens junctions discontinuous. We carry That Treating the cells with trichostatin A (TSA), a broad inhibitor of histone deacetylases Range (HDACs), or with MS-275, qui targets HDAC1, 2 and 3, induces stress fibers. LT Decreased the cellular levels of HDAC1, 2 and 3 and the Reduced total HDAC activity in the nucleus. Both the LT and TSA treatments Rnd3 induced expression, qui est required for the LT-mediated induction of actin stress fibers. Furthermore, we reveal That Treating the cells intoxicated with LT-garcinol, an inhibitor of histone acetyl transferases (HATs) disrupts the stress fibers and limits the monolayer barrier dysfunctions. These data Demonstrate the importance of modulating the flow of protein acetylation in order to control actin cytoskeleton organization and the endothelial cell monolayer barrier.

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Maria-Elena Fernandez-Sanchez, Thibaut Brunet, Jens-Christian Röper, Emmanuel Farge (2015 Sep 24)

Mechanotransduction’s impact on animal development, evolution, and tumorigenesis.

Annual review of cell and developmental biology : 373-97 : DOI : 10.1146/annurev-cellbio-102314-112441 En savoir plus
Résumé

Mechanotransduction translates mechanical signals into biochemical signals. It is based on the soft-matter properties of biomolecules or membranes that deform in response to mechanical loads to trigger activation of biochemical reactions. The study of mechanotransductive processes in cell-structure organization has been initiated in vitro in many biological contexts, such as examining cells’ response to substrate rigidity increases associated with tumor fibrosis and to blood flow pressure. In vivo, the study of mechanotransduction in regulating physiological processes has focused primarily on the context of embryogenesis, with an increasing number of examples demonstrating its importance for both differentiation and morphogenesis. The conservation across species of mechanical induction in early embryonic patterning now suggests that major animal transitions, such as mesoderm emergence, may have been based on mechanotransduction pathways. In adult animal tissues, permanent stiffness and tissue growth pressure contribute to tumorigenesis and appear to reactivate such conserved embryonic mechanosensitive pathways.

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Delabre U, Feld K, Crespo E, Whyte G, Sykes C, Seifert U, Guck J (2015 Aug 14)

Deformation of phospholipid vesicles in an optical stretcher

Soft Matter : 11 : 6075-88 : DOI : 10.1039/c5sm00562k En savoir plus
Résumé

Phospholipid vesicles are common model systems for cell membranes. Important aspects of the membrane function relate to its mechanical properties. Here we have investigated the deformation behaviour of phospholipid vesicles in a dual-beam laser trap, also called an optical stretcher. This study explicitly makes use of the inherent heating present in such traps to investigate the dependence of vesicle deformation on temperature. By using lasers with different wavelengths, optically induced mechanical stresses and temperature increase can be tuned fairly independently with a single setup. The phase transition temperature of vesicles can be clearly identified by an increase in deformation. In the case of no heating effects, a minimal model for drop deformation in an optical stretcher and a more specific model for vesicle deformation that takes explicitly into account the angular dependence of the optical stress are presented to account for the experimental results. Elastic constants are extracted from the fitting procedures, which agree with literature data. This study demonstrates the utility of optical stretching, which is easily combined with microfluidic delivery, for the future serial, high-throughput study of the mechanical and thermodynamic properties of phospholipid vesicles.

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Byung-Kuk Yoo, Axel Buguin, Zoher Gueroui (2015 Jul 30)

Biochemical perturbations of the mitotic spindle in Xenopus extracts using a diffusion-based microfluidic assay.

Biomicrofluidics : 044101 : DOI : 10.1063/1.4926324 En savoir plus
Résumé

A microfluidic device is a powerful tool to manipulate in a controlled manner at spatiotemporal scales for biological systems. Here, we describe a simple diffusion-based assay to generate and measure the effect of biochemical perturbations within the cytoplasm of cell-free extracts from Xenopus eggs. Our approach comprises a microliter reservoir and a model cytoplasm that are separated by a synthetic membrane containing sub-micrometric pores through which small molecules and recombinant proteins can diffuse. We have used this system to examine the perturbation of elements of the mitotic spindle, which is a microtubule-based bipolar structure involved in the segregation of the replicated genome to daughter cells during cell division. First, we used the small molecule inhibitor monastrol to target kinesin-5, a molecular motor that maintains the microtubule spindle bipolarity. Next, we explored the dynamics of the mitotic spindle by monitoring the exchange between unpolymerized and polymerized tubulin within microtubule fibers. These results show that a simple diffusion-based system can generate biochemical perturbations directly within a cell-free cytoplasm based on Xenopus egg extracts at the time scale of minutes. Our assay is therefore suitable for monitoring the dynamics of supramolecular assemblies within cell-free extracts in response to perturbations. This strategy opens up broad perspectives including phenotype screening or mechanistic studies of biological assembly processes and could be applied to other cell-free extracts such as those derived from mammalian or bacterial cells.

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Thanh Duc Mai, Iago Pereiro, Mohamed Hiraoui, Jean-Louis Viovy, Stéphanie Descroix, Myriam Taverna, Claire Smadja (2015 Jul 25)

Magneto-immunocapture with on-bead fluorescent labeling of amyloid-β peptides: towards a microfluidized-bed-based operation.

The Analyst : 5891-900 : DOI : 10.1039/c5an01179e En savoir plus
Résumé

A new sample treatment approach for sensitive determination of three amyloid-β peptides (Aβ 1-42, Aβ 1-40 and Aβ 1-38) with capillary electrophoresis coupled with laser induced fluorescent detection is reported herein. These Aβ peptides are considered an important family of biomarkers in the cerebrospinal fluid (CSF) for early diagnosis of Alzheimer’s disease (AD). Due to their extremely low abundance in CSF (down to sub nM ranges), batch-wise preconcentration via magneto-immunocapture with enrichment factors up to 100 was implemented. The Aβ peptides were first captured onto magnetic micro-beads. Then, on-beads fluorescent labeling of the captured Aβ peptides were carried out to avoid the unwanted presence of extra fluorescent dye in the eluent as in the case of in-solution labeling. Finally thermal elution was performed and eluted labeled peptides were analyzed off line with CE-LIF. The Aβ-capturing efficiencies of different commercially available antibodies grafted onto magnetic beads were tested. Aβ peptides in CSF samples collected from AD’s patients and healthy persons (used as controls) were measured and evaluated. As a proof of concept, the developed strategy was adapted into a miniaturized fluidized bed configuration that has the potential for coupling with a microchip separation system.

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