Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 17 de 17
Filtrar
Mais filtros










Base de dados
Intervalo de ano de publicação
1.
J Cell Sci ; 135(16)2022 08 15.
Artigo em Inglês | MEDLINE | ID: mdl-35972759

RESUMO

Spectrins are large, evolutionarily well-conserved proteins that form highly organized scaffolds on the inner surface of eukaryotic cells. Their organization in different cell types or cellular compartments helps cells withstand mechanical challenges with unique strategies depending on the cell type. This Review discusses our understanding of the mechanical properties of spectrins, their very distinct organization in red blood cells and neurons as two examples, and the contribution of the scaffolds they form to the mechanical properties of these cells.


Assuntos
Citoesqueleto de Actina , Espectrina , Citoesqueleto de Actina/metabolismo , Axônios/metabolismo , Eritrócitos/metabolismo , Neurônios/metabolismo , Espectrina/metabolismo
2.
J Phys Condens Matter ; 33(8): 084003, 2021 Feb 24.
Artigo em Inglês | MEDLINE | ID: mdl-33171443

RESUMO

Rheological properties of a material often require to be probed under extensional deformation. Examples include fibrous materials such as spider-silk, high-molecular weight polymer melts, and the contractile response of living cells. Such materials have strong molecular-level anisotropies which are either inherent or are induced by an imposed extension. However, unlike shear rheology, which is well-established, techniques to perform extensional rheology are currently under development and setups are often custom-designed for the problem under study. In this article, we present a versatile device that can be used to conduct extensional deformation studies of samples at microscopic scales with simultaneous imaging. We discuss the operational features of this device and present a number of applications.

3.
Soft Matter ; 16(2): 487-493, 2020 Jan 02.
Artigo em Inglês | MEDLINE | ID: mdl-31803881

RESUMO

Spider silk possesses unique mechanical properties like large extensibility, high tensile strength, super-contractility, etc. Understanding these mechanical responses requires characterization of the rheological properties of silk beyond the simple force-extension relations which are widely reported. Here we study the linear and non-linear viscoelastic properties of dragline silk obtained from social spider Stegodyphus sarasinorum using a Micro-Extension Rheometer that we have developed. Unlike continuous extension data, our technique allows for the probing of the viscoelastic response by applying small perturbations about sequentially increasing steady-state strain values. In addition, we extend our analysis to obtain the characteristic stress relaxation times and the frequency responses of the viscous and elastic moduli. Using these methods, we show that in a small strain regime (0-4%) dragline silk of social spiders shows a strain softening response followed by a strain stiffening response at higher strains (>4%). The stress relaxation time, on the other hand, increases monotonically with increasing strain for the entire range. We also show that the silk stiffens while ageing within the typical lifetime of a web. Our results demand the inclusion of the kinetics of domain unfolding and refolding in the existing models to account for the relaxation time behavior.


Assuntos
Seda/química , Animais , Módulo de Elasticidade , Cinética , Reologia , Aranhas , Resistência à Tração , Viscosidade
4.
Biophys J ; 117(5): 880-891, 2019 09 03.
Artigo em Inglês | MEDLINE | ID: mdl-31427070

RESUMO

Axonal beading, or the formation of a series of swellings along the axon, and retraction are commonly observed shape transformations that precede axonal atrophy in Alzheimer's disease, Parkinson's disease, and other neurodegenerative conditions. The mechanisms driving these morphological transformations are poorly understood. Here, we report controlled experiments that can induce either beading or retraction and follow the time evolution of these responses. By making quantitative analysis of the shape modes under different conditions, measurement of membrane tension, and using theoretical considerations, we argue that membrane tension is the main driving force that pushes cytosol out of the axon when microtubules are degraded, causing axonal thinning. Under pharmacological perturbation, atrophy is always retrograde, and this is set by a gradient in the microtubule stability. The nature of microtubule depolymerization dictates the type of shape transformation, vis-à-vis beading or retraction. Elucidating the mechanisms of these shape transformations may facilitate development of strategies to prevent or arrest axonal atrophy due to neurodegenerative conditions.


Assuntos
Axônios/metabolismo , Microtúbulos/metabolismo , Actinas/metabolismo , Animais , Atrofia , Axônios/efeitos dos fármacos , Fenômenos Biomecânicos , Compostos Bicíclicos Heterocíclicos com Pontes/farmacologia , Embrião de Galinha , Cones de Crescimento/efeitos dos fármacos , Cones de Crescimento/metabolismo , Imageamento Tridimensional , Membranas , Microtúbulos/efeitos dos fármacos , Nocodazol/farmacologia , Polimerização , Tiazolidinas/farmacologia
5.
Biophys J ; 115(4): 713-724, 2018 08 21.
Artigo em Inglês | MEDLINE | ID: mdl-30054033

RESUMO

Mechanotransduction is likely to be an important mechanism of signaling in thin, elongated cells such as neurons. Maintenance of prestress or rest tension may facilitate mechanotransduction in these cells. In recent years, functional roles for mechanical tension in neuronal development and physiology are beginning to emerge, but the cellular mechanisms regulating neurite tension remain poorly understood. Active contraction of neurites is a potential mechanism of tension regulation. In this study, we have explored cytoskeletal mechanisms mediating active contractility of neuronal axons. We have developed a simple assay in which we evaluate contraction of curved axons upon trypsin-mediated detachment. We show that curved axons undergo contraction and straighten upon deadhesion. Axonal straightening was found to be actively driven by actomyosin contractility, whereas microtubules may subserve a secondary role. We find that although axons show a monotonous decrease in length upon contraction, subcellularly, the cytoskeleton shows a heterogeneous contractile response. Further, using an assay for spontaneous development of tension without trypsin-induced deadhesion, we show that axons are intrinsically contractile. These experiments, using novel experimental approaches, implicate the axonal cytoskeleton in tension homeostasis. Our data suggest that although globally, the axon behaves as a mechanical continuum, locally, the cytoskeleton is remodeled heterogeneously.


Assuntos
Axônios/metabolismo , Citoesqueleto/metabolismo , Mecanotransdução Celular , Actomiosina/metabolismo , Animais , Adesão Celular , Galinhas , Microtúbulos/metabolismo , Tripsina/metabolismo
6.
Phys Biol ; 15(4): 046006, 2018 05 09.
Artigo em Inglês | MEDLINE | ID: mdl-29629709

RESUMO

Changes in cell-substrate adhesion are believed to signal the onset of cancer metastasis, but such changes must be quantified against background levels of intrinsic heterogeneity between cells. Variations in cell-substrate adhesion strengths can be probed through biophysical measurements of cell detachment from substrates upon the application of an external force. Here, we investigate, theoretically and experimentally, the detachment of cells adhered to substrates when these cells are subjected to fluid shear. We present a theoretical framework within which we calculate the fraction of detached cells as a function of shear stress for fast ramps as well as the decay in this fraction at fixed shear stress as a function of time. Using HEK and 3T3 fibroblast cells as experimental model systems, we extract characteristic force scales for cell adhesion as well as characteristic detachment times. We estimate force-scales of ∼500 pN associated to a single focal contact, and characteristic time-scales of [Formula: see text] s representing cell-spread-area dependent mean first passage times to the detached state at intermediate values of the shear stress. Variations in adhesion across cell types are especially prominent when cell detachment is probed by applying a time-varying shear stress. These methods can be applied to characterizing changes in cell adhesion in a variety of contexts, including metastasis.


Assuntos
Adesão Celular , Resistência ao Cisalhamento , Estresse Mecânico , Células 3T3 , Animais , Fenômenos Biomecânicos , Células HEK293 , Humanos , Camundongos , Modelos Biológicos
7.
Biophys J ; 108(3): 489-97, 2015 Feb 03.
Artigo em Inglês | MEDLINE | ID: mdl-25650917

RESUMO

Mechanical properties of cell membranes are known to be significantly influenced by the underlying cortical cytoskeleton. The technique of pulling membrane tethers from cells is one of the most effective ways of studying the membrane mechanics and the membrane-cortex interaction. In this article, we show that axon membranes make an interesting system to explore as they exhibit both free membrane-like behavior where the tether-membrane junction is movable on the surface of the axons (unlike many other cell membranes) as well as cell-like behavior where there are transient and spontaneous eruptions in the tether force that vanish when F-actin is depolymerized. We analyze the passive and spontaneous responses of axonal membrane tethers and propose theoretical models to explain the observed behavior.


Assuntos
Axônios/fisiologia , Membrana Celular/fisiologia , Citoesqueleto/fisiologia , Actinas/metabolismo , Animais , Fenômenos Biomecânicos , Galinhas , Fricção , Células HeLa , Humanos
8.
PLoS One ; 9(9): e107895, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25251154

RESUMO

It is well known that substrate properties like stiffness and adhesivity influence stem cell morphology and differentiation. Recent experiments show that cell morphology influences nuclear geometry and hence gene expression profile. The mechanism by which surface properties regulate cell and nuclear properties is only beginning to be understood. Direct transmission of forces as well as chemical signalling are involved in this process. Here, we investigate the formal aspect by studying the correlation between cell spreading and nuclear deformation using Mesenchymal stem cells under a wide variety of conditions. It is observed that a robust quantitative relation holds between the cell and nuclear projected areas, irrespective of how the cell area is modified or when various cytoskeletal or nuclear components are perturbed. By studying the role of actin stress fibers in compressing the nucleus we propose that nuclear compression by stress fibers can lead to enhanced cell spreading due to an interplay between elastic and adhesion factors. The significance of myosin-II in regulating this process is also explored. We demonstrate this effect using a simple technique to apply external compressive loads on the nucleus.


Assuntos
Citoesqueleto de Actina/metabolismo , Forma do Núcleo Celular , Células-Tronco Mesenquimais/citologia , Citoesqueleto de Actina/ultraestrutura , Actinas/análise , Actinas/metabolismo , Actinas/ultraestrutura , Animais , Adesão Celular , Células Cultivadas , Células-Tronco Mesenquimais/metabolismo , Camundongos
9.
Rev Sci Instrum ; 84(10): 105107, 2013 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-24182163

RESUMO

We discuss the design, instrumentation, and calibration of a versatile force transducer with feedback control, called the Micro-Extensional Rheometer (MER). A force range of eight decades (1-10(8) pN) and a displacement range of four decades (10-10(5) nm) with a spatial resolution of the order of nanometers are accessible with the instrument. A feedback-loop algorithm is used to control the commanded force or the extensional strain on the sample and implement different rheometric protocols such as step-strain, step-force, exponential strain, among others. The device may also be used to measure the forces exerted by active suspensions, pulling neurons, etc.


Assuntos
Microtecnologia/instrumentação , Fibras Ópticas , Transdutores , Calibragem , Dimetilpolisiloxanos/química , Escherichia coli , Modelos Lineares , Reologia
10.
Biophys J ; 99(11): 3571-9, 2010 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-21112281

RESUMO

We present investigations on volume regulation and beading shape transitions in PC12 neurites, conducted using a flow-chamber technique. By disrupting the cell cytoskeleton with specific drugs, we investigate the role of its individual components in the volume regulation response. We find that microtubule disruption increases both swelling rate and maximum volume attained, but does not affect the ability of the neurite to recover its initial volume. In addition, investigation of axonal beading-also known as pearling instability-provides additional clues on the mechanical state of the neurite. We conclude that volume recovery is driven by passive diffusion of osmolites, and propose that the initial swelling phase is mechanically slowed down by microtubules. Our experiments provide a framework to investigate the role of cytoskeletal mechanics in volume homeostasis.


Assuntos
Forma Celular , Tamanho Celular , Citoesqueleto/metabolismo , Neuritos/metabolismo , Animais , Elasticidade , Modelos Biológicos , Dinâmica não Linear , Concentração Osmolar , Pressão Osmótica , Células PC12 , Pressão , Ratos , Temperatura , Viscosidade
11.
Biophys J ; 98(4): 515-23, 2010 Feb 17.
Artigo em Inglês | MEDLINE | ID: mdl-20159147

RESUMO

We investigate the mechanical response of PC12 neurites subjected to a drag force imposed by a laminar flow perpendicular to the neurite axis. The curvature of the catenary shape acquired by an initially straight neurite under the action of the drag force provides information on both elongation and tension of the neurite. This method allows us to measure the rest tension and viscoelastic parameters of PC12 neurites and active behavior of neurites. Measurement of oscillations in the strain rate of neurites at constant flow rate provides insight on the response of molecular motors and additional support for the presence of a negative strain-rate sensitivity region in the global mechanical response of PC12 neurites.


Assuntos
Neuritos/metabolismo , Animais , Fenômenos Biomecânicos , Compostos Bicíclicos Heterocíclicos com Pontes/farmacologia , Forma Celular/efeitos dos fármacos , Elasticidade/efeitos dos fármacos , Neuritos/efeitos dos fármacos , Nocodazol/farmacologia , Células PC12 , Ratos , Estresse Mecânico , Tiazolidinas/farmacologia , Fatores de Tempo , Viscosidade/efeitos dos fármacos
12.
Phys Rev E Stat Nonlin Soft Matter Phys ; 80(1 Pt 1): 011708, 2009 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-19658719

RESUMO

Phase-separation dynamics of an asymmetric mixture of an isotropic dopant in a nematogenic fluid is presented. We show that, on steady cooling, the nucleating nematic drops move down the dopant concentration gradient, with a velocity that is dependent on the cooling rate and concentration gradient. This propulsion of the drops leads to a mechanism of droplet coarsening, where radius of a drop scales with time as R(t) approximately t. Various mechanisms for droplet propulsion are discussed.

13.
Biophys J ; 95(2): 978-85, 2008 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-18375512

RESUMO

The freshwater polyp Hydra has considerable regeneration capabilities. A small fragment of tissue excised from an adult animal is sufficient to regenerate an entire Hydra in the course of a few days. During the initial stages of the regeneration process, the tissue forms a hollow sphere. Then the sphere exhibits shape oscillations in the form of repeated cycles of swelling and collapse. We propose a biophysical model for the swelling mechanism. Our model takes the osmotic pressure difference between Hydra's inner and outer media and the elastic forces of the Hydra shell into account. We validate the model by a comprehensive experimental study including variations in initial medium concentrations, Hydra sphere sizes and temperatures. Numerical simulations of the model provide values for the swelling rates that are in agreement with the ones measured experimentally. Based on our results we argue that the shape oscillations are a consequence of Hydra's osmoregulation.


Assuntos
Relógios Biológicos/fisiologia , Hydra/fisiologia , Mecanotransdução Celular/fisiologia , Modelos Biológicos , Oscilometria/métodos , Regeneração/fisiologia , Equilíbrio Hidroeletrolítico/fisiologia , Animais , Simulação por Computador
14.
Phys Rev Lett ; 99(1): 018301, 2007 Jul 06.
Artigo em Inglês | MEDLINE | ID: mdl-17678192

RESUMO

The mechanical response of PC12 neurites under tension is investigated using a microneedle technique. Elastic response, viscoelastic relaxation, and active contraction are observed. The mechanical model proposed by Dennerll et al. [J. Cell Biol. 109, 3073 (1989).10.1083/jcb.109.6.3073], which involves three mechanical devices--a stiff spring kappa coupled with a Voigt element that includes a less stiff spring k and a dashpot gamma--has been improved by adding a new element to describe the main features of the contraction of axons. This element, which represents the action of molecular motors, acts in parallel with viscous forces defining a global tension response of axons T against elongation rates delta(k). Under certain conditions, axons show a transition from a viscoelastic elongation to active contraction, suggesting the presence of a negative elongation rate sensitivity in the curve T vs delta(k).


Assuntos
Axônios/metabolismo , Animais , Elasticidade , Neuritos/metabolismo , Células PC12 , Ratos , Viscosidade
15.
Phys Rev Lett ; 97(11): 115701, 2006 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-17025900

RESUMO

We report a novel phase separation dynamics, mediated by self-propelled motion of the nucleated drops, in a mixture of a nematogen and an isotropic dopant. We show that surface flow, induced by the gradient in the concentration of the dopant expelled by the growing drops, provides the driving force for the propulsion of nematic droplets. While the liquid crystal-isotropic transition is used here to demonstrate the phenomenon, self-propulsion should be observable in many other systems in which the dynamics of a conserved order parameter is coupled to a nonconserved order parameter.

16.
Phys Rev Lett ; 96(4): 048104, 2006 Feb 03.
Artigo em Inglês | MEDLINE | ID: mdl-16486900

RESUMO

We report a cylindrical-peristaltic shape transformation in axons exposed to a controlled osmotic perturbation. The peristaltic shape relaxes and the axon recovers its original geometry within minutes. We show that the shape instability depends critically on the swelling rate and that volume and membrane area regulation are responsible for the shape relaxation. We propose that volume regulation occurs via leakage of ions driven by elastic pressure, and analyze the peristaltic shape dynamics taking into account the internal structure of the axon. The results obtained provide a framework for understanding peristaltic shape dynamics in nerve fibers occurring in vivo.


Assuntos
Axônios/fisiologia , Animais , Transporte Biológico Ativo , Forma Celular , Embrião de Galinha , Membranas Intracelulares/metabolismo , Íons/metabolismo , Modelos Neurológicos , Neuritos/fisiologia , Pressão Osmótica , Células PC12 , Ratos , Água/metabolismo
17.
Biophys J ; 90(10): 3796-805, 2006 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-16461394

RESUMO

Cell mechanical functions such as locomotion, contraction, and division are controlled by the cytoskeleton, a dynamic biopolymer network whose mechanical properties remain poorly understood. We perform single-cell uniaxial stretching experiments on 3T3 fibroblasts. By superimposing small amplitude oscillations on a mechanically prestressed cell, we find a transition from linear viscoelastic behavior to power law stress stiffening. Data from different cells over several stress decades can be uniquely scaled to obtain a master relation between the viscoelastic moduli and the average force. Remarkably, this relation holds independently of deformation history, adhesion biochemistry, and intensity of active contraction. In particular, it is irrelevant whether force is actively generated by the cell or externally imposed by stretching. We propose that the master relation reflects the mechanical behavior of the force-bearing actin cytoskeleton, in agreement with stress stiffening known from semiflexible filament networks.


Assuntos
Citoesqueleto/fisiologia , Modelos Biológicos , Fibras de Estresse/fisiologia , Células 3T3 , Animais , Simulação por Computador , Elasticidade , Camundongos , Estresse Mecânico , Viscosidade
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...