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Sample records for actin flap dynamics

  1. Dynamics of active actin networks

    Koehler, Simone

    2014-03-01

    Local mechanical and structural properties of a eukaryotic cell are determined by its cytoskeleton. To adapt to their environment, cells rely on constant self-organized rearrangement processes of their actin cytoskeleton. To shed light on the principles underlying these dynamic self-organization processes we investigate a minimal reconstituted active system consisting of actin filaments, crosslinking molecules and molecular motor filaments. Using quantitative fluorescence microscopy and image analysis, we show, that these minimal model systems exhibit a generic structure formation mechanism. The competition between force generation by molecular motors and the stabilization of the network by crosslinking proteins results in a highly dynamic reorganization process which is characterized by anomalous transport dynamics with a superdiffusive behavior also found in intracellular dynamics. In vitro, these dynamics are governed by chemical and physical parameters that alter the balance of motor and crosslinking proteins, such as pH. These findings can be expected to have broad implications in our understanding of cytoskeletal regulation in vivo.

  2. Dynamics of an actin spring

    Riera, Christophe; Mahadevan, L.; Shin, Jennifer; Matsudaira, Paul

    2003-03-01

    The acrosome of the sperm of the horseshoe crab (Limulus Polyphemus) is an unusual actin based system that shows a spectacular dynamical transition in the presence of Ca++ that is present in abundance in the neighborhood of the egg. During this process, the bundle, which is initially bent and twisted uncoils and becomes straight in a matter of a few seconds. Based on microstructural data, we propose a model for the dynamics of uncoiling that is best represented by a triple-well potential corresponding to the different structural arrangements of the supertwisted filaments. Each of the false, true and coiled states corresponds to a local minimum of the energy, with the true state being the one with the lowest energy. Using an evolution equation derived by balancing torques, we investigate the nucleation and propagation of the phase transition and compare the results with those of experiments. Our model quantifies the hypothesis that the acrosomal bundle behaves like a mechano-chemical spring.

  3. A dynamical system for interacting flapping swimmers

    Oza, Anand; Ramananarivo, Sophie; Ristroph, Leif; Shelley, Michael

    2015-11-01

    We present the results of a theoretical investigation into the dynamics of interacting flapping swimmers. Our study is motivated by the recent experiments of Becker et al., who studied a one-dimensional array of self-propelled flapping wings that swim within each other's wakes in a water tank. They discovered that the system adopts certain ``schooling modes'' characterized by specific spatial phase relationships between swimmers. To rationalize these phenomena, we develop a discrete dynamical system in which the swimmers are modeled as heaving airfoils that shed point vortices during each flapping cycle. We then apply our model to recent experiments in the Applied Math Lab, in which two tandem flapping airfoils are free to choose both their speed and relative positions. We expect that our model may be used to understand how schooling behavior is influenced by hydrodynamics in more general contexts. Thanks to the NSF for its support.

  4. Reactive oxygen species (ROS)-induced actin glutathionylation controls actin dynamics in neutrophils

    Sakai, Jiro; Li, Jingyu; Subramanian, Kulandayan K.; Mondal, Subhanjan; Bajrami, Besnik; Hattori, Hidenori; Jia, Yonghui; Dickinson, Bryan C.; Zhong, Jia; Ye, Keqiang; Chang, Christopher J; Ho, Ye-Shih; Zhou, Jun; Luo, Hongbo R.

    2012-01-01

    Summary The regulation of actin dynamics is pivotal for cellular processes such as cell adhesion, migration, and phagocytosis, and thus is crucial for neutrophils to fulfill their roles in innate immunity. Many factors have been implicated in signal-induced actin polymerization, however the essential nature of the potential negative modulators are still poorly understood. Here we report that NADPH oxidase-dependent physiologically generated reactive oxygen species (ROS) negatively regulate actin polymerization in stimulated neutrophils via driving reversible actin glutathionylation. Disruption of glutaredoxin 1 (Grx1), an enzyme that catalyzes actin deglutathionylation, increased actin glutathionylation, attenuated actin polymerization, and consequently impaired neutrophil polarization, chemotaxis, adhesion, and phagocytosis. Consistently, Grx1-deficient murine neutrophils showed impaired in vivo recruitment to sites of inflammation and reduced bactericidal capability. Together, these results present a physiological role for glutaredoxin and ROS- induced reversible actin glutathionylation in regulation of actin dynamics in neutrophils. PMID:23159440

  5. Actin dynamics and the elasticity of cytoskeletal networks

    2009-09-01

    Full Text Available The structural integrity of a cell depends on its cytoskeleton, which includes an actin network. This network is transient and depends upon the continual polymerization and depolymerization of actin. The degradation of an actin network, and a corresponding reduction in cell stiffness, can indicate the presence of disease. Numerical simulations will be invaluable for understanding the physics of these systems and the correlation between actin dynamics and elasticity. Here we develop a model that is capable of generating actin network structures. In particular, we develop a model of actin dynamics which considers the polymerization, depolymerization, nucleation, severing, and capping of actin filaments. The structures obtained are then fed directly into a mechanical model. This allows us to qualitatively assess the effects of changing various parameters associated with actin dynamics on the elasticity of the material.

  6. Dynamics of Flapping Flag in Axial Flow

    Abderrahmane, Hamid Ait; Fayed, Mohamed; Gunter, Amy-Lee; Paidoussis, Michael P.; Ng, Hoi Dick

    2010-11-01

    We investigate experimentally the phenomenon of the flapping of a flag, placed within a low turbulent axial flow inside a small scale wind tunnel test section. Flags of different sizes and flexural rigidities were used. Image processing technique was used and the time series of a given point on the edge of the flag was analyzed. The stability condition of the flag was obtained and compared to the recent theoretical models and numerical simulations. Afterwards, the nonlinear dynamics of the flapping was investigated using nonlinear time series method. The nonlinear dynamics is depicted in phase space and the correlation dimension of the attractors is determined. On the basis of observations made in this study, some conclusions on the existing models were drawn.

  7. Reactive oxygen species (ROS)-induced actin glutathionylation controls actin dynamics in neutrophils

    Sakai, Jiro; Li, Jingyu; Subramanian, Kulandayan K.; Mondal, Subhanjan; Bajrami, Besnik; Hattori, Hidenori; Jia, Yonghui; Dickinson, Bryan C; Zhong, Jia; Ye, Keqiang; Chang, Christopher J.; Ho, Ye-Shih; Zhou, Jun; Luo, Hongbo R.

    2012-01-01

    The regulation of actin dynamics is pivotal for cellular processes such as cell adhesion, migration, and phagocytosis, and thus is crucial for neutrophils to fulfill their roles in innate immunity. Many factors have been implicated in signal-induced actin polymerization, however the essential nature of the potential negative modulators are still poorly understood. Here we report that NADPH oxidase-dependent physiologically generated reactive oxygen species (ROS) negatively regulate actin poly...

  8. Bacterial Subversion of Host Actin Dynamics at the Plasma Membrane

    Carabeo, Rey

    2011-01-01

    Invasion of non-phagocytic cells by a number of bacterial pathogens involves the subversion of the actin cytoskeletal remodelling machinery to produce actin-rich cell surface projections designed to engulf the bacteria. The signalling that occurs to induce these actin-rich structures has considerable overlap amongst a diverse group of bacteria. The molecular organisation within these structures act in concert to internalise the invading pathogen. This dynamic process could be subdivided into ...

  9. Effects of primary rotor parameters on flapping dynamics

    Chen, R. T. N.

    1980-01-01

    The effects of flapping dynamics of four main rotor design features that influence the agility, stability, and operational safety of helicopters are studied. The parameters include flapping hinge offset, flapping hinge restraint, pitch-flap coupling, and blade lock number. First, the flapping equations of motion are derived that explicitly contain the design parameters. The dynamic equations are then developed for the tip-path plane, and the influence of individual and combined variations in the design parameters determined. The steady state flapping response is examined with respect to control input and aircraft angular rate which leads to a feedforward control law for control decoupling through cross feed, and a feedback control law to decouple the steady state flapping response. The condition for achieving perfect decoupling of the flapping response due to aircraft pitch and roll rates without using feedback control is also found for the hover case. It is indicated that the frequency of the regressing flapping mode of the rotor system can become low enough to require consideration in the assessment of handling characteristics.

  10. Spontaneous actin dynamics in contractile rings

    Kruse, Karsten; Wollrab, Viktoria; Thiagarajan, Raghavan; Wald, Anne; Riveline, Daniel

    Networks of polymerizing actin filaments are known to be capable to self-organize into a variety of structures. For example, spontaneous actin polymerization waves have been observed in living cells in a number of circumstances, notably, in crawling neutrophils and slime molds. During later stages of cell division, they can also spontaneously form a contractile ring that will eventually cleave the cell into two daughter cells. We present a framework for describing networks of polymerizing actin filaments, where assembly is regulated by various proteins. It can also include the effects of molecular motors. We show that the molecular processes driven by these proteins can generate various structures that have been observed in contractile rings of fission yeast and mammalian cells. We discuss a possible functional role of each of these patterns. The work was supported by Agence Nationale de la Recherche, France, (ANR-10-LABX-0030-INRT) and by Deutsche Forschungsgemeinschaft through SFB1027.

  11. Dynamic response of a piezoelectric flapping wing

    Kumar, Alok; Khandwekar, Gaurang; Venkatesh, S.; Mahapatra, D. R.; Dutta, S.

    2015-03-01

    Piezo-composite membranes have advantages over motorized flapping where frequencies are high and certain coupling between bending and twisting is useful to generate lift and forward flight. We draw examples of fruit fly and bumble bee. Wings with Piezo ceramic PZT coating are realized. The passive mechanical response of the wing is characterized experimentally and validated using finite element simulation. Piezoelectric actuation with uniform electrode coating is characterized and optimal frequencies for flapping are identified. The experimental data are used in an empirical model and advanced ratio for a flapping insect like condition for various angular orientations is estimated.

  12. Wnt Signalling Promotes Actin Dynamics during Axon Remodelling through the Actin-Binding Protein Eps8.

    Eleanna Stamatakou

    Full Text Available Upon arrival at their synaptic targets, axons slow down their growth and extensively remodel before the assembly of presynaptic boutons. Wnt proteins are target-derived secreted factors that promote axonal remodelling and synaptic assembly. In the developing spinal cord, Wnts secreted by motor neurons promote axonal remodelling of NT-3 responsive dorsal root ganglia neurons. Axon remodelling induced by Wnts is characterised by growth cone pausing and enlargement, processes that depend on the re-organisation of microtubules. However, the contribution of the actin cytoskeleton has remained unexplored. Here, we demonstrate that Wnt3a regulates the actin cytoskeleton by rapidly inducing F-actin accumulation in growth cones from rodent DRG neurons through the scaffold protein Dishevelled-1 (Dvl1 and the serine-threonine kinase Gsk3β. Importantly, these changes in actin cytoskeleton occurs before enlargement of the growth cones is evident. Time-lapse imaging shows that Wnt3a increases lamellar protrusion and filopodia velocity. In addition, pharmacological inhibition of actin assembly demonstrates that Wnt3a increases actin dynamics. Through a yeast-two hybrid screen, we identified the actin-binding protein Eps8 as a direct interactor of Dvl1, a scaffold protein crucial for the Wnt signalling pathway. Gain of function of Eps8 mimics Wnt-mediated axon remodelling, whereas Eps8 silencing blocks the axon remodelling activity of Wnt3a. Importantly, blockade of the Dvl1-Eps8 interaction completely abolishes Wnt3a-mediated axonal remodelling. These findings demonstrate a novel role for Wnt-Dvl1 signalling through Eps8 in the regulation of axonal remodeling.

  13. Lift Enhancement by Dynamically Changing Wingspan in Forward Flapping Flight

    Wang, Shizhao; He, Guowei; Liu, Tianshu

    2013-01-01

    Stretching and retracting wingspan has been widely observed in the flight of birds and bats, and its effects on the aerodynamic performance particularly lift generation are intriguing. The rectangular flat-plate flapping wing with a sinusoidally stretching and retracting wingspan is proposed as a simple model of biologically-inspired dynamic morphing wings. Direct numerical simulations of the low-Reynolds-number flows around the flapping morphing wing in a parametric space are conducted by using immersed boundary method. It is found that the instantaneous and time-averaged lift coefficients of the wing can be significantly enhanced by dynamically changing wingspan in a flapping cycle. The lift enhancement is caused not only by changing the lifting surface area, but also manipulating the flow structures that are responsible to the generation of the vortex lift. The physical mechanisms behind the lift enhancement are explored by examining the three-dimensional flow structures around the flapping wing.

  14. Flap motion of helicopter rotors with novel, dynamic stall model

    Han Wei

    2016-07-01

    Full Text Available In this paper, a nonlinear flapping equation for large inflow angles and flap angles is established by analyzing the aerodynamics of helicopter blade elements. In order to obtain a generalized flap equation, the Snel stall model was first applied to determine the lift coefficient of the helicopter rotor. A simulation experiment for specific airfoils was then conducted to verify the effectiveness of the Snel stall model as it applies to helicopters. Results show that the model requires no extraneous parameters compared to the traditional stall model and is highly accurate and practically applicable. Based on the model, the relationship between the flapping angle and the angle of attack was analyzed, as well as the advance ratio under the dynamic stall state.

  15. Dynamic buckling of actin within filopodia

    Leijnse, Natascha; Oddershede, Lene B; Bendix, Pól Martin

    2015-01-01

    Filopodia are active tubular structures protruding from the cell surface which allow the cell to sense and interact with the surrounding environment through repetitive elongation-retraction cycles. The mechanical behavior of filopodia has been studied by measuring the traction forces exerted on...... external substrates.(1) These studies have revealed that internal actin flow can transduce a force across the cell surface through transmembrane linkers like integrins. In addition to the elongation-retraction behavior filopodia also exhibit a buckling and rotational behavior. Filopodial buckling in...

  16. Flapping dynamics of a flexible flag in a uniform flow

    Hwa Lee, Jae; Jin Sung, Hyung [Department of Mechanical Engineering, KAIST 291 Daehak-ro, Yuseong-gu, Daejeon 305-701 (Korea, Republic of); Huang, Wei-Xi, E-mail: hjsung@kaist.ac.kr, E-mail: hwx@tsinghua.edu.cn [Department of Engineering Mechanics, Tsinghua University, Beijing 100084 (China)

    2014-10-01

    Numerical simulations using the immersed boundary method were performed to investigate the flapping dynamics of a flexible flag in a uniform flow. The relevant parameters related to the problem are the mass ratio (ρ=ρ{sub 1}/ρ{sub f}L), the bending rigidity of the flag (K{sub B}=EI/ρ{sub f}U{sub ∞}{sup 2}L{sup 3}) and the Reynolds number (Re=U{sub ∞}L/ν), where ρ{sub 1} denotes the line density difference between the flag and the surrounding fluid. By varying the parameters over the ranges 0 not <= ρ not <= 10 and 10{sup -4} not < = K{sub B} not <= 10{sup -1} for Re = 200, we identified three dynamical states: the stretched–straight state (I), the regular flapping state (II), and the irregular flapping state (III) with a region in which violent snapping events occurred, characterized by rapid changes in velocity, tension and drag. The description of the flapping dynamics at each dynamic state was provided through a sequence of time-evolving instantaneous fields, and was examined in detail with respect to the frequency and the energy rank based on the dynamic mode decomposition technique. Furthermore, the hysteresis phenomena with varying the mass ratio and the Reynolds number was examined, and the results showed that the hysteresis loop width was varied for approximately 20–65%, depending on the values of the relevant parameters employed. (paper)

  17. Dynamic Actin Controls Polarity Induction de novo in Protoplasts

    Beatrix Zaban; Jan Maisch; Peter Nick

    2013-01-01

    Cell polarity and axes are central for plant morphogenesis.To study how polarity and axes are induced de novo,we investigated protoplasts of tobacco Nicotiana tabacum cv.BY-2 expressing fluorescentlytagged cytoskeletal markers.We standardized the system to such a degree that we were able to generate quantitative data on the temporal patterns of regeneration stages.The synthesis of a new cell wall marks the transition to the first stage of regeneration,and proceeds after a long preparatory phase within a few minutes.During this preparatory phase,the nucleus migrates actively,and cytoplasmic strands remodel vigorously.We probed this system for the effect of anti-cytoskeletal compounds,inducible bundling of actin,RGD-peptides,and temperature.Suppression of actin dynamics at an early stage leads to aberrant tripolar cells,whereas suppression of microtubule dynamics produces aberrant sausagelike cells with asymmetric cell walls.We integrated these data into a model,where the microtubular cytoskeleton conveys positional information between the nucleus and the membrane controlling the release or activation of components required for cell wall synthesis.Cell wall formation is followed by the induction of a new cell pole requiring dynamic actin filaments,and the new cell axis is manifested as elongation growth perpendicular to the orientation of the aligned cortical microtubules.

  18. Dynamic organization of actin cytoskeleton during the polarity formation and germination of pollen protoplasts

    XU Xia; Zl Huijun; SUN Yina; REN Haiyun

    2004-01-01

    The formation of the polarity of pollen protoplast and the dynamics of actin cytoskeleton were observed by non-fixation, Alexa-Phalloidin probing and confocal laser scanning microscopy. Our results showed that the protoplast obtained from stored pollen contained numerous crystalline fusiform bodies to constitute a storage form of actin. When dormant pollen was hydrated, the actin cytoskeleton forms a fine network spreading uniformly in the protoplast. In the process of polarity formation and germination of pollen protoplast, actin filaments marshaled slowly to the brim, and then formed multilayer continuous actin filament bundles surrounding the cortical of the protoplast. When the protoplast was exposed to actin filament-disrupting drugs, such as Latrunculin A and Cytochalasin D, continuously arranged actin bundles were disturbed and in this condition, the protoplast could not germinate. But when exposed to actin filament stabiling drug-phalliodin, the dynamics of actin filaments in the protoplasts behaved normally and the protoplasts could germinate normally. These results were also confirmed by the pharmacology experiments on pollen grains. And when Latrunculin A or Cytochalasin D was washed off, the ratio of pollen germination was resumed partly. All the results above show that the dynamic organization of the actin cytoskeleton are critical in the cell polarity formation and germination of pollen protoplast, and that the reorganization of actin cytoskeleton is mainly due to the rearrangement of actin filament arrays.

  19. Internal dynamics of F-actin and myosin subfragment-1 studied by quasielastic neutron scattering

    Various biological functions related to cell motility are driven by the interaction between the partner proteins, actin and myosin. To obtain insights into how this interaction occurs, the internal dynamics of F-actin and myosin subfragment-1 (S1) were characterized by the quasielastic neutron scattering measurements on the solution samples of F-actin and S1. Contributions of the internal motions of the proteins to the scattering spectra were separated from those of the global macromolecular diffusion. Analysis of the spectra arising from the internal dynamics showed that the correlation times of the atomic motions were about two times shorter for F-actin than for S1, suggesting that F-actin fluctuates more rapidly than S1. It was also shown that the fraction of the immobile atoms is larger for S1 than for F-actin. These results suggest that F-actin actively facilitates the binding of myosin by utilizing the more frequent conformational fluctuations than those of S1. - Highlights: • We studied the internal dynamics of F-actin and myosin S1 by neutron scattering. • The correlation times of the atomic motions were smaller for F-actin than for S1. • The fraction of the immobile atoms was also smaller for F-actin than for S1. • Our results suggest that mobility of atoms in F-actin is higher than that in S1. • We propose that high flexibility of F-actin facilitates the binding of myosin

  20. Effects of morphology on the flapping dynamics of inverted flags

    Fan, Boyu; Cosse, Julia; Sader, John; Gharib, Morteza

    2014-11-01

    The behavior of inverted flags has received recent attention in the study of the interaction of flexible bodies with fluid flows. It has implications in a variety of natural phenomena, such as the fluttering of leaves in the wind. As opposed to a conventional flag, defined by a fixed leading edge and a free trailing edge, an inverted flag has a free leading edge and a fixed trailing edge. The reversed flow orientation of inverted flags has led to a surprising observation. Over a narrow range of wind speeds, they exhibit a large-amplitude flapping motion that is not present in their conventional counterparts. Our study experimentally investigates the effects of flag morphology on the flapping behavior of inverted flags. Different flags ranging from rectangles to triangles are studied in a wind tunnel to assess the underlying parameters that govern their dynamics. We observe a significant shift in the limit-cycle flapping mode that is a function of flag shape parameters. This research is supported by the Lester Lees Aeronautics Summer Undergraduate Research Fellowship and the Gordon and Betty Moore Foundation.

  1. Dynamics of Actin Cables in Polarized Growth of the Filamentous Fungus Aspergillus nidulans

    Bergs, Anna; Ishitsuka, Yuji; Evangelinos, Minoas; Nienhaus, G. U.; Takeshita, Norio

    2016-01-01

    Highly polarized growth of filamentous fungi requires a continuous supply of proteins and lipids to the hyphal tip. This transport is managed by vesicle trafficking via the actin and microtubule cytoskeletons and their associated motor proteins. Particularly, actin cables originating from the hyphal tip are essential for hyphal growth. Although, specific marker proteins have been developed to visualize actin cables in filamentous fungi, the exact organization and dynamics of actin cables has remained elusive. Here, we observed actin cables using tropomyosin (TpmA) and Lifeact fused to fluorescent proteins in living Aspergillus nidulans hyphae and studied the dynamics and regulation. GFP tagged TpmA visualized dynamic actin cables formed from the hyphal tip with cycles of elongation and shrinkage. The elongation and shrinkage rates of actin cables were similar and approximately 0.6 μm/s. Comparison of actin markers revealed that high concentrations of Lifeact reduced actin dynamics. Simultaneous visualization of actin cables and microtubules suggests temporally and spatially coordinated polymerization and depolymerization between the two cytoskeletons. Our results provide new insights into the molecular mechanism of ordered polarized growth regulated by actin cables and microtubules.

  2. Dynamics of actin cables in polarized growth of the filamentous fungus Aspergillus nidulans

    Anna eBergs

    2016-05-01

    Full Text Available Highly polarized growth of filamentous fungi requires a continuous supply of proteins and lipids to the hyphal tip. This transport is managed by vesicle trafficking via the actin and microtubule cytoskeletons and their associated motor proteins. Particularly, actin cables originating from the hyphal tip are essential for hyphal growth. Although specific marker proteins to visualize actin cables have been developed in filamentous fungi, the exact organization and dynamics of actin cables has remained elusive. Here we visualized actin cables using tropomyosin (TpmA and Lifeact fused to fluorescent proteins in Aspergillus nidulans and studied the dynamics and regulation. GFP tagged TpmA visualized dynamic actin cables formed from the hyphal tip with cycles of elongation and shrinkage. The elongation and shrinkage rates of actin cables were similar and approximately 0.6 μm/s. Comparison of actin markers revealed that high concentrations of Lifeact reduced actin dynamics. Simultaneous visualization of actin cables and microtubules suggests temporally and spatially coordinated polymerization and depolymerization between the two cytoskeletons. Our results provide new insights into the molecular mechanism of ordered polarized growth regulated by actin cables and microtubules.

  3. Investigation of flap flexibility of β-secretase using molecular dynamic simulations.

    Kumalo, Hezekiel M; Bhakat, Soumendranath; Soliman, Mahmoud E

    2016-05-01

    Flap motif and its dynamics were extensively reported in aspartate proteases, e.g. HIV proteases and plasmepsins. Herein, we report the first account of flap dynamics amongst different conformations of β-secretase using molecular dynamics simulation. Various parameters were proposed and a selected few were picked which could appropriately describe the flap motion. Three systems were studied, namely Free (BACEFree) and two ligand-bound conformations, which belonged to space groups P6122 (BACEBound1) and C2221 (BACEBound2), respectively and four parameters (distance between the flaps tip residue, Thr72 and Ser325, d1; dihedral angle, ϕ (Thr72-Asp32-Asp228-Ser325); TriCα angles, θ1 (Thr72-Asp32-Ser325), and θ2 (Thr72-Asp228-Ser325)) were proposed to understand the change in dynamics of flap domain and the extent of flap opening and closing. Analysis of, θ2, d1, θ1 and ϕ confirmed that the BACEFree adopted semi-open, open and closed conformations with slight twisting during flap opening. However, BACEBound1 (P6122) showed an adaptation to open conformation due to lack of hydrogen bond interaction between the ligand and flap tip residue. A slight flap twisting, ϕ (lateral twisting) was observed for BACEBound1 during flap opening which correlates with the opening of BACEFree. Contradictory to the BACEBound1, the BACEBound2 locked the flap in a closed conformation throughout the simulation due to formation of a stable hydrogen bond interaction between the flap tip residue and ligand. Analyses of all three systems highlight that d1, θ2 and ϕ can be precisely used to describe the extent of flap opening and closing concurrently with snapshots along the molecular dynamics trajectory across several conformations of β-secretase. PMID:26208540

  4. Coupling of the hydration water dynamics and the internal dynamics of actin detected by quasielastic neutron scattering

    Highlights: ► Quasielastic neutron scattering spectra of F-actin and G-actin were measured. ► Analysis of the samples in D2O and H2O provided the spectra of hydration water. ► The first layer hydration water around F-actin is less mobile than around G-actin. ► This difference in hydration water is in concert with the internal dynamics of actin. ► Water outside the first layer behaves bulk-like but influenced by the first layer. -- Abstract: In order to characterize dynamics of water molecules around F-actin and G-actin, quasielastic neutron scattering experiments were performed on powder samples of F-actin and G-actin, hydrated either with D2O or H2O, at hydration ratios of 0.4 and 1.0. By combined analysis of the quasielastic neutron scattering spectra, the parameter values characterizing the dynamics of the water molecules in the first hydration layer and those of the water molecules outside of the first layer were obtained. The translational diffusion coefficients (DT) of the hydration water in the first layer were found to be 1.2 × 10−5 cm2/s and 1.7 × 10−5 cm2/s for F-actin and G-actin, respectively, while that for bulk water was 2.8 × 10−5 cm2/s. The residence times were 6.6 ps and 5.0 ps for F-actin and G-actin, respectively, while that for bulk water was 0.62 ps. These differences between F-actin and G-actin, indicating that the hydration water around G-actin is more mobile than that around F-actin, are in concert with the results of the internal dynamics of F-actin and G-actin, showing that G-actin fluctuates more rapidly than F-actin. This implies that the dynamics of the hydration water is coupled to the internal dynamics of the actin molecules. The DT values of the water molecules outside of the first hydration layer were found to be similar to that of bulk water though the residence times are strongly affected by the first hydration layer. This supports the recent observation on intracellular water that shows bulk-like behavior

  5. Dynamics of Micro-Air-Vehicle with Flapping Wings

    K. Sibilski

    2004-01-01

    Full Text Available Small (approximately 6 inch long, or hand-held reconnaissance micro air vehicles (MAVs will fly inside buildings, and require hover for observation, and agility at low speeds to move in confined spaces. For this flight envelope insect-like flapping wings seem to be an optimal mode of flying. Investigation of the aerodynamics of flapping wing MAVs is very challenging. The problem involves complex unsteady, viscous flow (mainly laminar, with the moving wing generating vortices and interacting with them. At this early stage of research only a preliminary insight into the nature of the little known aerodynamics of MAVs has been obtained. This paper describes computational models for simulation of the controlled motion of a microelectromechanical flying insect – entomopter. The design of software simulation for entomopter flight (SSEF is presented. In particular, we will estimate the flight control algorithms and performance for a Micromechanical Flying Insect (MFI, a 80–100 mm (wingtip-to-wingtip device capable of sustained autonomous flight. The SSEF is an end-to-end tool composed of several modular blocks which model the wing aerodynamics and dynamics, the body dynamics, and in the future, the environment perception, control algorithms, the actuators dynamics, and the visual and inertial sensors. We present the current state of the art of its implementation, and preliminary results. 

  6. Dynamics and Regulation of Actin Cytoskeleton in Plant Cells

    Ren Haiyun

    2007-01-01

    @@ The actin cytoskeleton constituted of globular actin (G-actin) is a ubiquitous component of eukaryotic cells and plays crucial roles in diverse physiological processes in plant cells, such as cytoplasmic streaming, organelle and nucleus positioning, cell morphogenesis, cell division, tip growth, etc.

  7. When fat is not bad: the regulation of actin dynamics by phospholipid signaling molecules

    Roman ePleskot

    2014-01-01

    Full Text Available The actin cytoskeleton plays a key role in the plant morphogenesis and is involved in polar cell growth, movement of subcellular organelles, cell division, and plant defense. Organization of actin cytoskeleton undergoes dynamic remodeling in response to internal developmental cues and diverse environmental signals. This dynamic behavior is regulated by numerous actin-binding proteins that integrate various signaling pathways. Production of the signaling lipids phosphatidylinositol 4,5-bisphosphate and phosphatidic acid affects the activity and subcellular distribution of several actin-binding proteins, and typically correlates with increased actin polymerization. Here we review current knowledge of the inter-regulatory dynamics between signaling phospholipids and the actin cytoskeleton in plant cells.

  8. Actin flow and talin dynamics govern rigidity sensing in actin-integrin linkage through talin extension.

    Hirata, Hiroaki; Chiam, Keng-Hwee; Lim, Chwee Teck; Sokabe, Masahiro

    2014-10-01

    At cell-substrate adhesion sites, the linkage between actin filaments and integrin is regulated by mechanical stiffness of the substrate. Of potential molecular regulators, the linker proteins talin and vinculin are of particular interest because mechanical extension of talin induces vinculin binding with talin, which reinforces the actin-integrin linkage. For understanding the molecular and biophysical mechanism of rigidity sensing at cell-substrate adhesion sites, we constructed a simple physical model to examine a role of talin extension in the stiffness-dependent regulation of actin-integrin linkage. We show that talin molecules linking between retrograding actin filaments and substrate-bound integrin are extended in a manner dependent on substrate stiffness. The model predicts that, in adhesion complexes containing ≈30 talin links, talin is extended enough for vinculin binding when the substrate is stiffer than 1 kPa. The lifetime of talin links needs to be 2-5 s to achieve an appropriate response of talin extension against substrate stiffness. Furthermore, changes in actin velocity drastically shift the range of substrate stiffness that induces talin-vinculin binding. Our results suggest that talin extension is a key step in sensing and responding to substrate stiffness at cell adhesion sites. PMID:25142525

  9. Mammalian Fat1 cadherin regulates actin dynamics and cell–cell contact

    Tanoue, Takuji; Takeichi, Masatoshi

    2004-01-01

    Fat cadherins form a distinct subfamily of the cadherin gene superfamily, and are featured by their unusually large extracellular domain. In this work, we investigated the function of a mammalian Fat cadherin. Fat1 was localized at filopodial tips, lamellipodial edges, and cell–cell boundaries, overlapping with dynamic actin structures. RNA interference–mediated knockdown of Fat1 resulted in disorganization of cell junction–associated F-actin and other actin fibers/cables, disturbance of cell...

  10. Dynamic Chord-wise Tip Curvature on Flexible Flapping Plates

    Martin, Nathan; Gharib, Morteza

    2014-11-01

    The aerodynamic characteristics of long rectangular flapping plates are strongly influenced by the interaction between tip and edge vortices. This has led to the development of many tip actuation mechanisms to independently bend or rotate the tip towards the root of the plate in the span-wise direction. In our current work, the influence of dynamically altering the chord-wise curvature of the tip on the generation of aerodynamic forces is investigated; for this case, the two free corners of the flat plate bend towards each other. The parameters of actuation timing, maximum curvature, Reynolds number, flexibility, and tip speed are independently varied to determine their influence. These results will further the fundamental understanding of unsteady aerodynamics. This material is based upon work supported by the National Science Foundation Graduate Research Fellowship and the Gordon and Betty Moore foundation.

  11. The effect of mouse twinfilin-1 on the structure and dynamics of monomeric actin.

    Takács-Kollár, Veronika; Nyitrai, Miklós; Hild, Gábor

    2016-07-01

    The effect of twinfilin-1 on the structure and dynamics of monomeric actin was investigated with fluorescence spectroscopy and differential scanning calorimetry experiments. Fluorescence anisotropy measurements proved that G-actin and twinfilin-1 could form a complex. Due to the formation of the complexes the dissociation of the nucleotide slowed down from the nucleotide-binding pocket of actin. Fluorescence quenching experiments showed that the accessibility of the actin bound ε-ATP decreased in the presence of twinfilin-1. Temperature dependent fluorescence resonance energy transfer and differential scanning calorimetry experiments revealed that the protein matrix of actin becomes more rigid and more heat resistant in the presence of twinfilin-1. The results suggest that the nucleotide binding cleft shifted into a more closed and stable conformational state of actin in the presence of twinfilin-1. PMID:27079635

  12. TWISTED DWARF1 Mediates the Action of Auxin Transport Inhibitors on Actin Cytoskeleton Dynamics.

    Zhu, Jinsheng; Bailly, Aurelien; Zwiewka, Marta; Sovero, Valpuri; Di Donato, Martin; Ge, Pei; Oehri, Jacqueline; Aryal, Bibek; Hao, Pengchao; Linnert, Miriam; Burgardt, Noelia Inés; Lücke, Christian; Weiwad, Matthias; Michel, Max; Weiergräber, Oliver H; Pollmann, Stephan; Azzarello, Elisa; Mancuso, Stefano; Ferro, Noel; Fukao, Yoichiro; Hoffmann, Céline; Wedlich-Söldner, Roland; Friml, Jiří; Thomas, Clément; Geisler, Markus

    2016-04-01

    Plant growth and architecture is regulated by the polar distribution of the hormone auxin. Polarity and flexibility of this process is provided by constant cycling of auxin transporter vesicles along actin filaments, coordinated by a positive auxin-actin feedback loop. Both polar auxin transport and vesicle cycling are inhibited by synthetic auxin transport inhibitors, such as 1-N-naphthylphthalamic acid (NPA), counteracting the effect of auxin; however, underlying targets and mechanisms are unclear. Using NMR, we map the NPA binding surface on the Arabidopsis thaliana ABCB chaperone TWISTED DWARF1 (TWD1). We identify ACTIN7 as a relevant, although likely indirect, TWD1 interactor, and show TWD1-dependent regulation of actin filament organization and dynamics and that TWD1 is required for NPA-mediated actin cytoskeleton remodeling. The TWD1-ACTIN7 axis controls plasma membrane presence of efflux transporters, and as a consequence act7 and twd1 share developmental and physiological phenotypes indicative of defects in auxin transport. These can be phenocopied by NPA treatment or by chemical actin (de)stabilization. We provide evidence that TWD1 determines downstream locations of auxin efflux transporters by adjusting actin filament debundling and dynamizing processes and mediating NPA action on the latter. This function appears to be evolutionary conserved since TWD1 expression in budding yeast alters actin polarization and cell polarity and provides NPA sensitivity. PMID:27053424

  13. ER sheet persistence is coupled to myosin 1c–regulated dynamic actin filament arrays

    Joensuu, Merja; Belevich, Ilya; Rämö, Olli; Nevzorov, Ilya; Vihinen, Helena; Puhka, Maija; Witkos, Tomasz M.; Lowe, Martin; Vartiainen, Maria K.; Jokitalo, Eija

    2014-01-01

    The endoplasmic reticulum (ER) comprises a dynamic three-dimensional (3D) network with diverse structural and functional domains. Proper ER operation requires an intricate balance within and between dynamics, morphology, and functions, but how these processes are coupled in cells has been unclear. Using live-cell imaging and 3D electron microscopy, we identify a specific subset of actin filaments localizing to polygons defined by ER sheets and tubules and describe a role for these actin arrays in ER sheet persistence and, thereby, in maintenance of the characteristic network architecture by showing that actin depolymerization leads to increased sheet fluctuation and transformations and results in small and less abundant sheet remnants and a defective ER network distribution. Furthermore, we identify myosin 1c localizing to the ER-associated actin filament arrays and reveal a novel role for myosin 1c in regulating these actin structures, as myosin 1c manipulations lead to loss of the actin filaments and to similar ER phenotype as observed after actin depolymerization. We propose that ER-associated actin filaments have a role in ER sheet persistence regulation and thus support the maintenance of sheets as a stationary subdomain of the dynamic ER network. PMID:24523293

  14. ER sheet persistence is coupled to myosin 1c-regulated dynamic actin filament arrays.

    Joensuu, Merja; Belevich, Ilya; Rämö, Olli; Nevzorov, Ilya; Vihinen, Helena; Puhka, Maija; Witkos, Tomasz M; Lowe, Martin; Vartiainen, Maria K; Jokitalo, Eija

    2014-04-01

    The endoplasmic reticulum (ER) comprises a dynamic three-dimensional (3D) network with diverse structural and functional domains. Proper ER operation requires an intricate balance within and between dynamics, morphology, and functions, but how these processes are coupled in cells has been unclear. Using live-cell imaging and 3D electron microscopy, we identify a specific subset of actin filaments localizing to polygons defined by ER sheets and tubules and describe a role for these actin arrays in ER sheet persistence and, thereby, in maintenance of the characteristic network architecture by showing that actin depolymerization leads to increased sheet fluctuation and transformations and results in small and less abundant sheet remnants and a defective ER network distribution. Furthermore, we identify myosin 1c localizing to the ER-associated actin filament arrays and reveal a novel role for myosin 1c in regulating these actin structures, as myosin 1c manipulations lead to loss of the actin filaments and to similar ER phenotype as observed after actin depolymerization. We propose that ER-associated actin filaments have a role in ER sheet persistence regulation and thus support the maintenance of sheets as a stationary subdomain of the dynamic ER network. PMID:24523293

  15. On the wake dynamics of flapping inverted flags

    Gilmanov, Anvar; Sotiropoulos, Fotis; Cosse, Julia; Gharib, Mory

    2014-11-01

    As recently shown experimentally by Kim et al. (JFM, 2013), when a flexible flag with a fixed trailing edge (an inverted flag) is exposed to a uniform inflow it can exhibit complex structural response and rich fluid-structure interaction (FSI) dynamics. We employ a new FSI numerical method to carry out large-eddy simulation (LES) of inverted flags in the range where large-amplitude flapping instabilities have been found experimentally. The numerical method integrates the curvilinear immersed boundary (CURVIB) FSI method of Borazjani et al. (JCP, 2008) with the thin-shell, rotation-free, finite-element (FE) formulation of Stolarski et al. (Int. JNME, 2013) and is able to simulate FSI of flexible thin bodies undergoing oscillations of arbitrarily large amplitude. The dynamic Smagorinsky model is employed for subgrid scale closure and a wall model is employed for reconstructing velocity boundary conditions. Comparisons with the experimental data show that the simulations are able to capture the structural response of the flag with very good accuracy. The computed results are analyzed to elucidate the structure and dynamics of the massively separated, unsteady flow shed off the flag edges. This work is supported by the US Department of Energy and the Minnesota Supercomputing Institute.

  16. Calcium influx through CRAC channels controls actin organization and dynamics at the immune synapse

    Hartzell, Catherine A; Jankowska, Katarzyna I; Burkhardt, Janis K; Lewis, Richard S

    2016-01-01

    T cell receptor (TCR) engagement opens Ca2+ release-activated Ca2+ (CRAC) channels and triggers formation of an immune synapse between T cells and antigen-presenting cells. At the synapse, actin reorganizes into a concentric lamellipod and lamella with retrograde actin flow that helps regulate the intensity and duration of TCR signaling. We find that Ca2+ influx is required to drive actin organization and dynamics at the synapse. Calcium acts by promoting actin depolymerization and localizing actin polymerization and the actin nucleation promotion factor WAVE2 to the periphery of the lamellipod while suppressing polymerization elsewhere. Ca2+-dependent retrograde actin flow corrals ER tubule extensions and STIM1/Orai1 complexes to the synapse center, creating a self-organizing process for CRAC channel localization. Our results demonstrate a new role for Ca2+ as a critical regulator of actin organization and dynamics at the synapse, and reveal potential feedback loops through which Ca2+ influx may modulate TCR signaling. DOI: http://dx.doi.org/10.7554/eLife.14850.001 PMID:27440222

  17. Calcium influx through CRAC channels controls actin organization and dynamics at the immune synapse.

    Hartzell, Catherine A; Jankowska, Katarzyna I; Burkhardt, Janis K; Lewis, Richard S

    2016-01-01

    T cell receptor (TCR) engagement opens Ca(2+) release-activated Ca(2+) (CRAC) channels and triggers formation of an immune synapse between T cells and antigen-presenting cells. At the synapse, actin reorganizes into a concentric lamellipod and lamella with retrograde actin flow that helps regulate the intensity and duration of TCR signaling. We find that Ca(2+) influx is required to drive actin organization and dynamics at the synapse. Calcium acts by promoting actin depolymerization and localizing actin polymerization and the actin nucleation promotion factor WAVE2 to the periphery of the lamellipod while suppressing polymerization elsewhere. Ca(2+)-dependent retrograde actin flow corrals ER tubule extensions and STIM1/Orai1 complexes to the synapse center, creating a self-organizing process for CRAC channel localization. Our results demonstrate a new role for Ca(2+) as a critical regulator of actin organization and dynamics at the synapse, and reveal potential feedback loops through which Ca(2+) influx may modulate TCR signaling. PMID:27440222

  18. Flapping Dynamics of an Inverted Flexible Foil in a Uniform Axial Flow

    Gurugubelli Venkata, Pardha Saradhi; Jaiman, Rajeev K.

    2014-11-01

    This work presents a numerical study on self-induced flapping dynamics of an inverted flexible foil in uniform flow. The inverted foil considered in this study is clamped at the trailing edge and the leading edge is allowed to oscillate. A high-order coupled FSI solver based on CFEI formulation has been used to present the flapping response results for a wide range of nondimensional bending rigidity using a fixed Reynolds number of 1000 and a mass-ratio of 0.1. As a function of bending rigidity four flapping regimes have been discovered: fixed point, inverted limit-cycle oscillation, deflected flapping, and flipped flapping. The inverted foil configuration undergoes flapping motion more readily and experiences very large amplitude oscillations than the conventional foil. A wide variety of vortex wakes with a maximum of 14 vortices per oscillation cycle have been observed. The inverted limit-cycle flapping generate novel 4P +6S (14 vortices) and 2P +6S (10 vortices) wakes. On the other hand, the flipped flapping regime is characterized by a von Kármán wake. We also observe that inverted foil can extract 1000 times more energy from the surrounding fluid compared to the conventional foil.

  19. Cortactin promotes exosome secretion by controlling branched actin dynamics.

    Sinha, Seema; Hoshino, Daisuke; Hong, Nan Hyung; Kirkbride, Kellye C; Grega-Larson, Nathan E; Seiki, Motoharu; Tyska, Matthew J; Weaver, Alissa M

    2016-07-18

    Exosomes are extracellular vesicles that influence cellular behavior and enhance cancer aggressiveness by carrying bioactive molecules. The mechanisms that regulate exosome secretion are poorly understood. Here, we show that the actin cytoskeletal regulatory protein cortactin promotes exosome secretion. Knockdown or overexpression of cortactin in cancer cells leads to a respective decrease or increase in exosome secretion, without altering exosome cargo content. Live-cell imaging revealed that cortactin controls both trafficking and plasma membrane docking of multivesicular late endosomes (MVEs). Regulation of exosome secretion by cortactin requires binding to the branched actin nucleating Arp2/3 complex and to actin filaments. Furthermore, cortactin, Rab27a, and coronin 1b coordinately control stability of cortical actin MVE docking sites and exosome secretion. Functionally, the addition of purified exosomes to cortactin-knockdown cells rescued defects of those cells in serum-independent growth and invasion. These data suggest a model in which cortactin promotes exosome secretion by stabilizing cortical actin-rich MVE docking sites. PMID:27402952

  20. Computer Simulations of Mechano-Chemical Networks Choreographing Actin Dynamics in Cell Motility

    Zhuravlev, Pavel I.; Hu, Longhua; Papoian, Garegin A.

    In eukaryotic cells, cell motility is largely driven by self-assembly and growth of filamentous networks comprised of actin. Numerous proteins regulate actin network dynamics either biochemically, or through mechanical interactions. This regulation is rather complex, intricately coordinated both spatially and temporally. Although experiments in vivo and in vitro have provided a trove of structural and biochemical information about actin-based cell motility processes, experimental data is not always easy to interpret unambiguously, sometimes various interpretations being in contradiction with each other. Hence, mathematical modeling approaches are necessary for providing a physical foundation for interpreting and guiding experiments. In particular, computer simulations based on physicochemical interactions provide a systems-level description of protrusion dynamics. In this contribution, we review recent progress in modeling actin-based cell motility using detailed computer simulations. We elaborate on the way actin network dynamics is determined by the interplay between chemical reactions, mechanical feedbacks, and transport bottlenecks. We also discuss the role of inherent randomness of elementary chemical reactions in determining the dynamical behavior of the mechano-chemical network controlling actin polymerization and growth.

  1. Dendritic cell podosome dynamics does not depend on the F-actin regulator SWAP-70.

    Anne Götz

    Full Text Available In addition to classical adhesion structures like filopodia or focal adhesions, dendritic cells similar to macrophages and osteoclasts assemble highly dynamic F-actin structures called podosomes. They are involved in cellular processes such as extracellular matrix degradation, bone resorption by osteoclasts, and trans-cellular diapedesis of lymphocytes. Besides adhesion and migration, podosomes enable dendritic cells to degrade connective tissue by matrix metalloproteinases. SWAP-70 interacts with RhoGTPases and F-actin and regulates migration of dendritic cells. SWAP-70 deficient osteoclasts are impaired in F-actin-ring formation and bone resorption. In the present study, we demonstrate that SWAP-70 is not required for podosome formation and F-actin turnover in dendritic cells. Furthermore, we found that toll-like receptor 4 ligand induced podosome disassembly and podosome-mediated matrix degradation is not affected by SWAP-70 in dendritic cells. Thus, podosome formation and function in dendritic cells is independent of SWAP-70.

  2. A small molecule inhibitor of tropomyosin dissociates actin binding from tropomyosin-directed regulation of actin dynamics

    Teresa T. Bonello; Miro Janco; Jeff Hook; Alex Byun; Mark Appaduray; Irina Dedova; Sarah Hitchcock-DeGregori; Hardeman, Edna C.; Justine R. Stehn; Till Böcking; Gunning, Peter W.

    2016-01-01

    The tropomyosin family of proteins form end-to-end polymers along the actin filament. Tumour cells rely on specific tropomyosin-containing actin filament populations for growth and survival. To dissect out the role of tropomyosin in actin filament regulation we use the small molecule TR100 directed against the C terminus of the tropomyosin isoform Tpm3.1. TR100 nullifies the effect of Tpm3.1 on actin depolymerisation but surprisingly Tpm3.1 retains the capacity to bind F-actin in a cooperativ...

  3. A comparative molecular dynamics study on BACE1 and BACE2 flap flexibility.

    Kumalo, H M; Soliman, Mahmoud E

    2016-10-01

    Beta-amyloid precursor protein cleavage enzyme1 (BACE1) and beta-amyloid precursor protein cleavage enzyme2 (BACE2), members of aspartyl protease family, are close homologs and have high similarity in their protein crystal structures. However, their enzymatic properties are different, which leads to different clinical outcomes. In this study, we performed sequence analysis and all-atom molecular dynamic (MD) simulations for both enzymes in their ligand-free states in order to compare their dynamical flap behaviors. This is to enhance our understanding of the relationship between sequence, structure and the dynamics of this protein family. Sequence analysis shows that in BACE1 and BACE2, most of the ligand-binding sites are conserved, indicative of their enzymatic property as aspartyl protease members. The other conserved residues are more or less unsystematically localized throughout the structure. Herein, we proposed and applied different combined parameters to define the asymmetric flap motion; the distance, d1, between the flap tip and the flexible region; the dihedral angle, φ, to account for the twisting motion and the TriCα angle, θ2 and θ1. All four combined parameters were found to appropriately define the observed "twisting" motion during the flaps different conformational states. Additional analysis of the parameters indicated that the flaps can exist in an ensemble of conformations, i.e. closed, semi-open and open conformations for both systems. However, the behavior of the flap tips during simulations is different between BACE1 and BACE2. The BACE1 active site cavity is more spacious as compared to that of BACE2. The analysis of 10S loop and 113S loop showed a similar trend to that of flaps, with the BACE1 loops being more flexible and less stable than those of BACE2. We believe that the results, methods and perspectives highlighted in this report would assist researchers in the discovery of BACE inhibitors as potential Alzheimer's disease therapies

  4. Cdc28–Cln3 phosphorylation of Sla1 regulates actin patch dynamics in different modes of fungal growth

    Zeng, Guisheng; Wang, Yan-Ming; Wang, Yue

    2012-01-01

    A dynamic balance between targeted transport and endocytosis is critical for polarized cell growth. However, how actin-mediated endocytosis is regulated in different growth modes remains unclear. Here we report differential regulation of cortical actin patch dynamics between the yeast and hyphal growth in Candida albicans. The mechanism involves phosphoregulation of the endocytic protein Sla1 by the cyclin-dependent kinase (CDK) Cdc28–Cln3 and the actin-regulating kinase Prk1. Mutational stud...

  5. Live Cell Imaging of Actin Dynamics in the Filamentous Fungus Aspergillus nidulans.

    Schultzhaus, Zachary; Quintanilla, Laura; Hilton, Angelyn; Shaw, Brian D

    2016-04-01

    Hyphal cells of filamentous fungi grow at their tips in a method analogous to pollen tube and root hair elongation. This process, generally referred to as tip growth, requires precise regulation of the actin cytoskeleton, and characterizing the various actin structures in these cell types is currently an active area of research. Here, the actin marker Lifeact was used to document actin dynamics in the filamentous fungus Aspergillus nidulans. Contractile double rings were observed at septa, and annular clusters of puncta were seen subtending growing hyphal tips, corresponding to the well-characterized subapical endocytic collar. However, Lifeact also revealed two additional structures. One, an apical array, was dynamic on the face opposite the tip, while a subapical web was dynamic on the apical face and was located several microns behind the growth site. Each was observed turning into the other over time, implying that they could represent different localizations of the same structure, although hyphae with a subapical web grew faster than those exhibiting an apical array. The subapical web has not been documented in any filamentous fungus to date, and is separate from the networks of F-actin seen in other tip-growing organisms surrounding septa or stationary along the plasmalemma. PMID:26879694

  6. Changes in actin dynamics are involved in salicylic acid signaling pathway

    Matoušková, J.; Janda, M.; Fišer, R.; Šašek, Vladimír; Kocourková, Daniela; Burketová, Lenka; Dušková, J.; Martinec, Jan; Valentová, O.

    2014-01-01

    Roč. 223, JUN 2014 (2014), s. 36-44. ISSN 0168-9452 R&D Projects: GA ČR(CZ) GAP501/11/1654 Institutional support: RVO:61389030 Keywords : Actin dynamics * Salicylic acid * PR genes Subject RIV: CE - Biochemistry Impact factor: 3.607, year: 2014

  7. Arabidopsis Actin-Depolymerizing Factor-4 links pathogen perception, defense activation and transcription to cytoskeletal dynamics.

    Katie Porter

    Full Text Available The primary role of Actin-Depolymerizing Factors (ADFs is to sever filamentous actin, generating pointed ends, which in turn are incorporated into newly formed filaments, thus supporting stochastic actin dynamics. Arabidopsis ADF4 was recently shown to be required for the activation of resistance in Arabidopsis following infection with the phytopathogenic bacterium Pseudomonas syringae pv. tomato DC3000 (Pst expressing the effector protein AvrPphB. Herein, we demonstrate that the expression of RPS5, the cognate resistance protein of AvrPphB, was dramatically reduced in the adf4 mutant, suggesting a link between actin cytoskeletal dynamics and the transcriptional regulation of R-protein activation. By examining the PTI (PAMP Triggered Immunity response in the adf4 mutant when challenged with Pst expressing AvrPphB, we observed a significant reduction in the expression of the PTI-specific target gene FRK1 (Flg22-Induced Receptor Kinase 1. These data are in agreement with recent observations demonstrating a requirement for RPS5 in PTI-signaling in the presence of AvrPphB. Furthermore, MAPK (Mitogen-Activated Protein Kinase-signaling was significantly reduced in the adf4 mutant, while no such reduction was observed in the rps5-1 point mutation under similar conditions. Isoelectric focusing confirmed phosphorylation of ADF4 at serine-6, and additional in planta analyses of ADF4's role in immune signaling demonstrates that nuclear localization is phosphorylation independent, while localization to the actin cytoskeleton is linked to ADF4 phosphorylation. Taken together, these data suggest a novel role for ADF4 in controlling gene-for-gene resistance activation, as well as MAPK-signaling, via the coordinated regulation of actin cytoskeletal dynamics and R-gene transcription.

  8. How does the antagonism between capping and anti-capping proteins affect actin network dynamics?

    Actin-based cell motility is essential to many biological processes. We built a simplified, three-dimensional computational model and subsequently performed stochastic simulations to study the growth dynamics of lamellipodia-like branched networks. In this work, we shed light on the antagonism between capping and anti-capping proteins in regulating actin dynamics in the filamentous network. We discuss detailed mechanisms by which capping and anti-capping proteins affect the protrusion speed of the actin network and the rate of nucleation of filaments. We computed a phase diagram showing the regimes of motility enhancement and inhibition by these proteins. Our work shows that the effects of capping and anti-capping proteins are mainly transmitted by modulation of the filamentous network density and local availability of monomeric actin. We discovered that the combination of the capping/anti-capping regulatory network with nucleation-promoting proteins introduces robustness and redundancy in cell motility machinery, allowing the cell to easily achieve maximal protrusion speeds under a broader set of conditions. Finally, we discuss distributions of filament lengths under various conditions and speculate on their potential implication for the emergence of filopodia from the lamellipodial network.

  9. How does the antagonism between capping and anti-capping proteins affect actin network dynamics?

    Hu, Longhua; Papoian, Garegin A.

    2011-09-01

    Actin-based cell motility is essential to many biological processes. We built a simplified, three-dimensional computational model and subsequently performed stochastic simulations to study the growth dynamics of lamellipodia-like branched networks. In this work, we shed light on the antagonism between capping and anti-capping proteins in regulating actin dynamics in the filamentous network. We discuss detailed mechanisms by which capping and anti-capping proteins affect the protrusion speed of the actin network and the rate of nucleation of filaments. We computed a phase diagram showing the regimes of motility enhancement and inhibition by these proteins. Our work shows that the effects of capping and anti-capping proteins are mainly transmitted by modulation of the filamentous network density and local availability of monomeric actin. We discovered that the combination of the capping/anti-capping regulatory network with nucleation-promoting proteins introduces robustness and redundancy in cell motility machinery, allowing the cell to easily achieve maximal protrusion speeds under a broader set of conditions. Finally, we discuss distributions of filament lengths under various conditions and speculate on their potential implication for the emergence of filopodia from the lamellipodial network.

  10. How does the antagonism between capping and anti-capping proteins affect actin network dynamics?

    Hu Longhua; Papoian, Garegin A, E-mail: gpapoian@umd.edu [Department of Chemistry and Biochemistry, University of Maryland, College Park, MD 20742 (United States)

    2011-09-21

    Actin-based cell motility is essential to many biological processes. We built a simplified, three-dimensional computational model and subsequently performed stochastic simulations to study the growth dynamics of lamellipodia-like branched networks. In this work, we shed light on the antagonism between capping and anti-capping proteins in regulating actin dynamics in the filamentous network. We discuss detailed mechanisms by which capping and anti-capping proteins affect the protrusion speed of the actin network and the rate of nucleation of filaments. We computed a phase diagram showing the regimes of motility enhancement and inhibition by these proteins. Our work shows that the effects of capping and anti-capping proteins are mainly transmitted by modulation of the filamentous network density and local availability of monomeric actin. We discovered that the combination of the capping/anti-capping regulatory network with nucleation-promoting proteins introduces robustness and redundancy in cell motility machinery, allowing the cell to easily achieve maximal protrusion speeds under a broader set of conditions. Finally, we discuss distributions of filament lengths under various conditions and speculate on their potential implication for the emergence of filopodia from the lamellipodial network.

  11. The F-actin modifier villin regulates insulin granule dynamics and exocytosis downstream of islet cell autoantigen 512

    Hassan Mziaut

    2016-08-01

    Conclusion: Our findings show that villin controls the size of the F-actin cages restricting SGs and, thus, regulates their dynamics and availability for exocytosis. Evidence that villin acts downstream of Ica512 also indicates that SGs directly influence the remodeling properties of the cortical actin cytoskeleton for tight control of insulin secretion.

  12. Quasi-3d aerodynamic code for analyzing dynamic flap response

    Ramos García, Néstor

    is modeled using a panel method whereas the viscous part is modeled by using the integral form of the the laminar and turbulent boundary layer equations and with extensions for 3-D rotational effects. Laminar to turbulent transition can be forced with a boundary layer trip or computed with a modified...... reduced frequencies and oscillation amplitudes, and generally a good agreement is obtained. The capability of the code to simulate a trailing edge flap under steady or unsteady flow conditions has been proven. A parametric study on rotational effects induced by Coriolis and centrifugal forces in the......A computational model for predicting the aerodynamic behavior of wind turbine airfoil profiles subjected to steady and unsteady motions has been developed. The model is based on a viscous-inviscid interaction technique using strong coupling between the viscous and inviscid parts. The inviscid part...

  13. Effects of polymerization and nucleotide identity on the conformational dynamics of the bacterial actin homolog MreB

    Colavin, Alexandre; Hsin, Jen; Huang, Kerwyn Casey

    2014-01-01

    Cytoskeletal filaments drive many dynamic cellular processes, such as the regulation of shape by actin networks in eukaryotes and by the actin homolog MreB in rod-shaped bacteria. Here, we use all-atom molecular dynamics simulations to demonstrate close parallels between the conformational dynamics of actin and MreB, in which polymerization induces flattening of MreB subunits that restructures the ATP binding pocket to promote hydrolysis. We also find that ATP-bound MreB filaments are substan...

  14. The Dynamic Pollen Tube Cytoskeleton: Live Cell Studies Using Actin-Binding and Microtubule-Binding Reporter Proteins

    Alice Y. Cheung; Qiao-hong Duan; Silvia Santos Costa; Barend H.J.de Graaf; Veronica S.Di Stilio; Jose Feijo; Hen-Ming Wu

    2008-01-01

    Pollen tubes elongate within the pistil to transport sperm cells to the embryo sac for fertilization.Growth occurs exclusively at the tube apex,rendering pollen tube elongation a most dramatic polar cell growth process.A hall-mark pollen tube feature is its cytoskeleton,which comprises elaborately organized and dynamic actin microfilaments and microtubules.Pollen tube growth is dependent on the actin cytoskeleton;its organization and regulation have been exalined extensively by various approaches.including fluorescent protein labeled actin-binding proteins in live cell studies.Using the previously described GFP-NtADF1 and GFP-LIADF1, and a new actin reporter protein NtPLIM2b-GFP,we re-affirm that the predominant actin structures in elongating tobacco and lily pollen tubes are long,streaming actin cables along the pollen tube shank,and a subapical structure comprising shorter actin cables.The subapical collection of actin microfilaments undergoes dynamic changes,giving rise to the appearance of structures that range from basket-or funnel-shaped,mesh-like to a subtle ring.NtPLIM2b-GFP is used in combination with a guanine nucleotide exchange factor for the Rho GTPases,AtROP-GEF1,to illustrate the use of these actin reporter proteins to explore the linkage between the polar cell growth process and its actin cytoskeleton.Contrary to the actin cytoskeleton,microtubules appear not to play a direct role in supporting the polar cell growth process in angiosperm pollen tubes.Using a microtubule reporter protein based on the microtubule end-binding protein from Arabidopsis AtEB1,GFP-AtEB1,we show that the extensive microtubule network in elongating pollen tubes displays varying degrees of dynamics.These reporter proteins provide versatile tools to explore the functional connection between major structural and signaling components of the polar pollen tube growth process.

  15. Effect of torsional stiffness and inertia on the dynamics of low aspect ratio flapping wings

    Micro air vehicle-motivated aerodynamics in biological flight has been an important subject in the past decade. Inspired by the novel flapping wing mechanisms in insects, birds and bats, we have carried out a numerical study systematically investigating a three-dimensional flapping rigid wing with passively actuated lateral and rotational motion. Distinguishing it from the limited existing studies, this work performs a systematic examination on the effects of wing aspect ratio (AR = 1.0 to infinity), inertia (density ratio σ = 4–32), torsional stiffness (frequency ratio F = 1.5–10 and infinity) and pivot point (from chord-center to leading edge) on the dynamics response of a low AR rectangular wing under an initial zero speed flow field condition. The simulation results show that the symmetry breakdown of the flapping wing results in a forward/backward motion with a rotational pitching. When the wing reaches its stable periodic state, the induced pitching frequency is identical to its forced flapping frequency. However, depending on various kinematic and dynamic system parameters, (i.e. flapping frequency, density ratio and pitching axis), the lateral induced velocity shows a number of different oscillating frequencies. Furthermore, compared with a one degree of freedom (DoF) wing in the lateral direction only, the propulsion performance of such a two DoF wing relies very much on the magnitude of torsional stiffness adding on the pivot point, as well as its pitching axis. In all cases examined here, thrust force and moment generated by a long span wing is larger than that of a short wing, which is remarkably linked to the strong reverse von Kármán vortex street formed in the wake of a wing. (paper)

  16. Effect of torsional stiffness and inertia on the dynamics of low aspect ratio flapping wings.

    Xiao, Qing; Hu, Jianxin; Liu, Hao

    2014-03-01

    Micro air vehicle-motivated aerodynamics in biological flight has been an important subject in the past decade. Inspired by the novel flapping wing mechanisms in insects, birds and bats, we have carried out a numerical study systematically investigating a three-dimensional flapping rigid wing with passively actuated lateral and rotational motion. Distinguishing it from the limited existing studies, this work performs a systematic examination on the effects of wing aspect ratio (AR = 1.0 to infinity), inertia (density ratio σ = 4-32), torsional stiffness (frequency ratio F = 1.5-10 and infinity) and pivot point (from chord-center to leading edge) on the dynamics response of a low AR rectangular wing under an initial zero speed flow field condition. The simulation results show that the symmetry breakdown of the flapping wing results in a forward/backward motion with a rotational pitching. When the wing reaches its stable periodic state, the induced pitching frequency is identical to its forced flapping frequency. However, depending on various kinematic and dynamic system parameters, (i.e. flapping frequency, density ratio and pitching axis), the lateral induced velocity shows a number of different oscillating frequencies. Furthermore, compared with a one degree of freedom (DoF) wing in the lateral direction only, the propulsion performance of such a two DoF wing relies very much on the magnitude of torsional stiffness adding on the pivot point, as well as its pitching axis. In all cases examined here, thrust force and moment generated by a long span wing is larger than that of a short wing, which is remarkably linked to the strong reverse von Kármán vortex street formed in the wake of a wing. PMID:24434625

  17. Dynamics of actin waves on patterned substrates: a quantitative analysis of circular dorsal ruffles.

    Erik Bernitt

    Full Text Available Circular Dorsal Ruffles (CDRs have been known for decades, but the mechanism that organizes these actin waves remains unclear. In this article we systematically analyze the dynamics of CDRs on fibroblasts with respect to characteristics of current models of actin waves. We studied CDRs on heterogeneously shaped cells and on cells that we forced into disk-like morphology. We show that CDRs exhibit phenomena such as periodic cycles of formation, spiral patterns, and mutual wave annihilations that are in accord with an active medium description of CDRs. On cells of controlled morphologies, CDRs exhibit extremely regular patterns of repeated wave formation and propagation, whereas on random-shaped cells the dynamics seem to be dominated by the limited availability of a reactive species. We show that theoretical models of reaction-diffusion type incorporating conserved species capture partially the behavior we observe in our data.

  18. ATEFlap aerodynamic model, a dynamic stall model including the effects of trailing edge flap deflection

    Bergami, L.; Gaunaa, M.

    2012-02-15

    The report presents the ATEFlap aerodynamic model, which computes the unsteady lift, drag and moment on a 2D airfoil section equipped with Adaptive Trailing Edge Flap. The model captures the unsteady response related to the effects of the vorticity shed into the wake, and the dynamics of flow separation a thin-airfoil potential flow model is merged with a dynamic stall model of the Beddoes-Leishmann type. The inputs required by the model are steady data for lift, drag, and moment coefficients as function of angle of attack and flap deflection. Further steady data used by the Beddoes- Leishmann dynamic stall model are computed in an external preprocessor application, which gives the user the possibility to verify, and eventually correct, the steady data passed to the aerodynamic model. The ATEFlap aerodynamic model is integrated in the aeroelastic simulation tool HAWC2, thus al- lowing to simulate the response of a wind turbine with trailing edge flaps on the rotor. The algorithms used by the preprocessor, and by aerodynamic model are presented, and modifications to previous implementations of the aerodynamic model are briefly discussed. The performance and the validity of the model are verified by comparing the dynamic response computed by the ATEFlap with solutions from CFD simulations. (Author)

  19. Passive control of a dynamically pitching wind turbine airfoil under aeroelastic conditions using a Gurney flap

    Nikoueeyan, Pourya; Magstadt, Andrew; Strike, John; Hind, Michael; Naughton, Jonathan

    2014-11-01

    To reduce the cost of energy, wind turbine design has moved towards larger blades that are heavier and have lower relative structural stiffness compared to shorter blades. To address the lower blade stiffness, different flow control techniques have been considered. The Gurney flap, a small, low-cost and effective control method, is a promising control actuator. Wind tunnel testing has been performed on a DU97-W-300 10% flatback airfoil undergoing dynamic pitching relevant to flow conditions encountered by wind turbine blades. To mimic blade compliance, the airfoil is actively driven through a torsionally elastic element. Time-resolved surface pressure measurements have been acquired from which lift Cl and moment Cm coefficients were calculated. Changes in Cl and Cm in moderate and deep dynamic stall regimes for different Gurney flap heights were studied for different pitch drive conditions (amplitude and frequency). The results show the significant impact of compliance on the angle of attack (α) range experienced by the airfoil. Shifts in α range result in different hysteresis behavior in both Cl and Cm and demonstrate the effectiveness of the Gurney flap in modifying the aerodynamics of wind turbine blades experiencing dynamic pitching. This work supported by DOE and a gift from BP.

  20. Live imaging provides new insights on dynamic F-actin filopodia and differential endocytosis during myoblast fusion in Drosophila.

    Shruti Haralalka

    Full Text Available The process of myogenesis includes the recognition, adhesion, and fusion of committed myoblasts into multinucleate syncytia. In the larval body wall muscles of Drosophila, this elaborate process is initiated by Founder Cells and Fusion-Competent Myoblasts (FCMs, and cell adhesion molecules Kin-of-IrreC (Kirre and Sticks-and-stones (Sns on their respective surfaces. The FCMs appear to provide the driving force for fusion, via the assembly of protrusions associated with branched F-actin and the WASp, SCAR and Arp2/3 pathways. In the present study, we utilize the dorsal pharyngeal musculature that forms in the Drosophila embryo as a model to explore myoblast fusion and visualize the fusion process in live embryos. These muscles rely on the same cell types and genes as the body wall muscles, but are amenable to live imaging since they do not undergo extensive morphogenetic movement during formation. Time-lapse imaging with F-actin and membrane markers revealed dynamic FCM-associated actin-enriched protrusions that rapidly extend and retract into the myotube from different sites within the actin focus. Ultrastructural analysis of this actin-enriched area showed that they have two morphologically distinct structures: wider invasions and/or narrow filopodia that contain long linear filaments. Consistent with this, formin Diaphanous (Dia and branched actin nucleator, Arp3, are found decorating the filopodia or enriched at the actin focus, respectively, indicating that linear actin is present along with branched actin at sites of fusion in the FCM. Gain-of-function Dia and loss-of-function Arp3 both lead to fusion defects, a decrease of F-actin foci and prominent filopodia from the FCMs. We also observed differential endocytosis of cell surface components at sites of fusion, with actin reorganizing factors, WASp and SCAR, and Kirre remaining on the myotube surface and Sns preferentially taken up with other membrane proteins into early endosomes and

  1. Lamin A/C and emerin regulate MKL1-SRF activity by modulating actin dynamics.

    Ho, Chin Yee; Jaalouk, Diana E; Vartiainen, Maria K; Lammerding, Jan

    2013-05-23

    Laminopathies, caused by mutations in the LMNA gene encoding the nuclear envelope proteins lamins A and C, represent a diverse group of diseases that include Emery-Dreifuss muscular dystrophy (EDMD), dilated cardiomyopathy (DCM), limb-girdle muscular dystrophy, and Hutchison-Gilford progeria syndrome. Most LMNA mutations affect skeletal and cardiac muscle by mechanisms that remain incompletely understood. Loss of structural function and altered interaction of mutant lamins with (tissue-specific) transcription factors have been proposed to explain the tissue-specific phenotypes. Here we report in mice that lamin-A/C-deficient (Lmna(-/-)) and Lmna(N195K/N195K) mutant cells have impaired nuclear translocation and downstream signalling of the mechanosensitive transcription factor megakaryoblastic leukaemia 1 (MKL1), a myocardin family member that is pivotal in cardiac development and function. Altered nucleo-cytoplasmic shuttling of MKL1 was caused by altered actin dynamics in Lmna(-/-) and Lmna(N195K/N195K) mutant cells. Ectopic expression of the nuclear envelope protein emerin, which is mislocalized in Lmna mutant cells and also linked to EDMD and DCM, restored MKL1 nuclear translocation and rescued actin dynamics in mutant cells. These findings present a novel mechanism that could provide insight into the disease aetiology for the cardiac phenotype in many laminopathies, whereby lamin A/C and emerin regulate gene expression through modulation of nuclear and cytoskeletal actin polymerization. PMID:23644458

  2. Lamin A/C and emerin regulate MKL1/SRF activity by modulating actin dynamics

    Ho, Chin Yee; Jaalouk, Diana E.; Vartiainen, Maria K.; Lammerding, Jan

    2013-01-01

    Laminopathies, caused by mutations in the LMNA gene encoding the nuclear envelope proteins lamins A and C, represent a diverse group of diseases that include Emery-Dreifuss Muscular Dystrophy (EDMD), dilated cardiomyopathy (DCM), limb-girdle muscular dystrophy, and Hutchison-Gilford progeria syndrome (HGPS).1 The majority of LMNA mutations affect skeletal and cardiac muscle by mechanisms that remain incompletely understood. Loss of structural function and disturbed interaction of mutant lamins with (tissue-specific) transcription factors have been proposed to explain the tissue-specific phenotypes.1 We report here that lamin A/C-deficient (Lmna−/−) and Lmna N195K mutant cells have impaired nuclear translocation and downstream signaling of the mechanosensitive transcription factor megakaryoblastic leukaemia 1 (MKL1), a myocardin family member that is pivotal in cardiac development and function.2 Disturbed nucleo-cytoplasmic shuttling of MKL1 was caused by altered actin dynamics in Lmna−/− and N195K mutant cells. Ectopic expression of the nuclear envelope protein emerin, which is mislocalized in Lmna mutant cells and also linked to EDMD and DCM, restored MKL1 nuclear translocation and rescued actin dynamics in mutant cells. These findings present a novel mechanism that could provide insight into the disease etiology for the cardiac phenotype in many laminopathies, whereby lamins A/C and emerin regulate gene expression through modulation of nuclear and cytoskeletal actin polymerization. PMID:23644458

  3. Comparative dynamics of retrograde actin flow and focal adhesions: formation of nascent adhesions triggers transition from fast to slow flow.

    Antonina Y Alexandrova

    Full Text Available 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.

  4. Comparative dynamics of retrograde actin flow and focal adhesions: formation of nascent adhesions triggers transition from fast to slow flow.

    Alexandrova, Antonina Y; Arnold, Katya; Schaub, Sébastien; Vasiliev, Jury M; Meister, Jean-Jacques; Bershadsky, Alexander D; Verkhovsky, Alexander B

    2008-01-01

    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. PMID:18800171

  5. Dynamics of a pneumatic artificial muscle actuation system driving a trailing edge flap

    This study presents a time domain dynamic model of an antagonistic pneumatic artificial muscle (PAM) driven trailing edge flap (TEF) system for next generation active helicopter rotors. Active rotor concepts are currently being widely researched in the rotorcraft community as a means to provide a significant leap forward in performance through primary aircraft control, vibration mitigation and noise reduction. Recent work has shown PAMs to be a promising candidate for active rotor actuation due to their combination of high force, large stroke, light weight, and suitable bandwidth. When arranged into biologically inspired agonist/antagonist muscle pairs they can produce bidirectional torques for effectively driving a TEF. However, there are no analytical dynamic models in the literature that can accurately capture the behavior of such systems across the broad range of frequencies required for this demanding application. This work combines mechanical, pneumatic, and aerodynamic component models into a global flap system model developed for the Bell 407 rotor system. This model can accurately predict pressure, force, and flap angle response to pneumatic control valve inputs over a range of operating frequencies from 7 to 35 Hz (1/rev to 5/rev for the Bell 407) and operating pressures from 30 to 90 psi. (paper)

  6. Dynamics of a pneumatic artificial muscle actuation system driving a trailing edge flap

    Woods, Benjamin K. S.; Kothera, Curt S.; Wang, Gang; Wereley, Norman M.

    2014-09-01

    This study presents a time domain dynamic model of an antagonistic pneumatic artificial muscle (PAM) driven trailing edge flap (TEF) system for next generation active helicopter rotors. Active rotor concepts are currently being widely researched in the rotorcraft community as a means to provide a significant leap forward in performance through primary aircraft control, vibration mitigation and noise reduction. Recent work has shown PAMs to be a promising candidate for active rotor actuation due to their combination of high force, large stroke, light weight, and suitable bandwidth. When arranged into biologically inspired agonist/antagonist muscle pairs they can produce bidirectional torques for effectively driving a TEF. However, there are no analytical dynamic models in the literature that can accurately capture the behavior of such systems across the broad range of frequencies required for this demanding application. This work combines mechanical, pneumatic, and aerodynamic component models into a global flap system model developed for the Bell 407 rotor system. This model can accurately predict pressure, force, and flap angle response to pneumatic control valve inputs over a range of operating frequencies from 7 to 35 Hz (1/rev to 5/rev for the Bell 407) and operating pressures from 30 to 90 psi.

  7. Ornithine decarboxylase and extracellular polyamines regulate microvascular sprouting and actin cytoskeleton dynamics in endothelial cells

    The polyamines are essential for cancer cell proliferation during tumorigenesis. Targeted inhibition of ornithine decarboxylase (ODC), i.e. a key enzyme of polyamine biosynthesis, by α-difluoromethylornithine (DFMO) has shown anti-neoplastic activity in various experimental models. This activity has mainly been attributed to the anti-proliferative effect of DFMO in cancer cells. Here, we provide evidence that unperturbed ODC activity is a requirement for proper microvessel sprouting ex vivo as well as the migration of primary human endothelial cells. DFMO-mediated ODC inhibition was reversed by extracellular polyamine supplementation, showing that anti-angiogenic effects of DFMO were specifically related to polyamine levels. ODC inhibition was associated with an abnormal morphology of the actin cytoskeleton during cell spreading and migration. Moreover, our data suggest that de-regulated actin cytoskeleton dynamics in DFMO treated endothelial cells may be related to constitutive activation of the small GTPase CDC42, i.e. a well-known regulator of cell motility and actin cytoskeleton remodeling. These insights into the potential role of polyamines in angiogenesis should stimulate further studies testing the combined anti-tumor effect of polyamine inhibition and established anti-angiogenic therapies in vivo.

  8. Shank–cortactin interactions control actin dynamics to maintain flexibility of neuronal spines and synapses

    MacGillavry, Harold D.; Kerr, Justin M.; Kassner, Josh; Frost, Nicholas A.; Blanpied, Thomas A.

    2016-01-01

    The family of Shank scaffolding molecules (comprising Shank1, 2 and 3) are core components of the postsynaptic density (PSD) in neuronal synapses. Shanks link surface receptors to other scaffolding molecules within the PSD, as well as to the actin cytoskeleton. However, determining the function of Shank proteins in neurons has been complicated because the different Shank isoforms share a very high degree of sequence and domain homology. Therefore, to control Shank content while minimizing potential compensatory effects, a miRNA-based knockdown strategy was developed to reduce the expression of all synaptically targeted Shank isoforms simultaneously in rat hippocampal neurons. Using this approach, a strong (>75%) reduction in total Shank protein levels was achieved at individual dendritic spines, prompting an approximately 40% decrease in mushroom spine density. Furthermore, Shank knockdown reduced spine actin levels and increased sensitivity to the actin depolymerizing agent Latrunculin A. A SHANK2 mutant lacking the proline-rich cortactin-binding motif (SHANK2-ΔPRO) was unable to rescue these defects. Furthermore, Shank knockdown reduced cortactin levels in spines and increased the mobility of spine cortactin as measured by single-molecule tracking photoactivated localization microscopy, suggesting that Shank proteins recruit and stabilize cortactin at the synapse. Furthermore, it was found that Shank knockdown significantly reduced spontaneous remodelling of synapse morphology that could not be rescued by the SHANK2-ΔPRO mutant. It was concluded that Shank proteins are key intermediates between the synapse and the spine interior that, via cortactin, permit the actin cytoskeleton to dynamically regulate synapse morphology and function. PMID:26547831

  9. Enabled Negatively Regulates Diaphanous-Driven Actin Dynamics In Vitro and In Vivo

    Bilancia, C.G.; Winkelman, J.D.; Tsygankov, D.; Nowotarski, S.H.; Sees, J.A.; Comber, K.; Evans, I.; Lakhani, V.; Wood, W.; Elston, T.C.; Kovar, D.R.; Peifer, M

    2014-01-01

    Summary Actin regulators facilitate cell migration by controlling cell protrusion architecture and dynamics. As the behavior of individual actin regulators becomes clear, we must address why cells require multiple regulators with similar functions and how they cooperate to create diverse protrusions. We characterized Diaphanous (Dia) and Enabled (Ena) as a model, using complementary approaches: cell culture, biophysical analysis, and Drosophila morphogenesis. We found that Dia and Ena have di...

  10. Protocadherin FAT1 binds Ena/VASP proteins and is necessary for actin dynamics and cell polarization

    Moeller, Marcus J.; Soofi, Abdulsalam; Braun, Gerald S; Li, Xiaodong; Watzl, Carsten; Kriz, Wilhelm; Holzman, Lawrence B.

    2004-01-01

    Cell migration requires integration of cellular processes resulting in cell polarization and actin dynamics. Previous work using tools of Drosophila genetics suggested that protocadherin fat serves in a pathway necessary for determining cell polarity in the plane of a tissue. Here we identify mammalian FAT1 as a proximal element of a signaling pathway that determines both cellular polarity in the plane of the monolayer and directed actin-dependent cell motility. FAT1 is localized to the leadi...

  11. Structural dynamics and aerodynamics measurements of biologically inspired flexible flapping wings

    Flapping wing flight as seen in hummingbirds and insects poses an interesting unsteady aerodynamic problem: coupling of wing kinematics, structural dynamics and aerodynamics. There have been numerous studies on the kinematics and aerodynamics in both experimental and computational cases with both natural and artificial wings. These studies tend to ignore wing flexibility; however, observation in nature affirms that passive wing deformation is predominant and may be crucial to the aerodynamic performance. This paper presents a multidisciplinary experimental endeavor in correlating a flapping micro air vehicle wing's aeroelasticity and thrust production, by quantifying and comparing overall thrust, structural deformation and airflow of six pairs of hummingbird-shaped membrane wings of different properties. The results show that for a specific spatial distribution of flexibility, there is an effective frequency range in thrust production. The wing deformation at the thrust-productive frequencies indicates the importance of flexibility: both bending and twisting motion can interact with aerodynamic loads to enhance wing performance under certain conditions, such as the deformation phase and amplitude. By measuring structural deformations under the same aerodynamic conditions, beneficial effects of passive wing deformation can be observed from the visualized airflow and averaged thrust. The measurements and their presentation enable observation and understanding of the required structural properties for a thrust effective flapping wing. The intended passive responses of the different wings follow a particular pattern in correlation to their aerodynamic performance. Consequently, both the experimental technique and data analysis method can lead to further studies to determine the design principles for micro air vehicle flapping wings.

  12. Structural dynamics and aerodynamics measurements of biologically inspired flexible flapping wings

    Wu, P; Stanford, B K; Ifju, P G [Department of Mechanical and Aerospace Engineering, MAE-A 231, University of Florida, Gainesville, FL 32611 (United States); Saellstroem, E; Ukeiley, L, E-mail: diccidwp@ufl.edu [Department of Mechanical and Aerospace Engineering, University of Florida, Shalimar, FL 32579 (United States)

    2011-03-15

    Flapping wing flight as seen in hummingbirds and insects poses an interesting unsteady aerodynamic problem: coupling of wing kinematics, structural dynamics and aerodynamics. There have been numerous studies on the kinematics and aerodynamics in both experimental and computational cases with both natural and artificial wings. These studies tend to ignore wing flexibility; however, observation in nature affirms that passive wing deformation is predominant and may be crucial to the aerodynamic performance. This paper presents a multidisciplinary experimental endeavor in correlating a flapping micro air vehicle wing's aeroelasticity and thrust production, by quantifying and comparing overall thrust, structural deformation and airflow of six pairs of hummingbird-shaped membrane wings of different properties. The results show that for a specific spatial distribution of flexibility, there is an effective frequency range in thrust production. The wing deformation at the thrust-productive frequencies indicates the importance of flexibility: both bending and twisting motion can interact with aerodynamic loads to enhance wing performance under certain conditions, such as the deformation phase and amplitude. By measuring structural deformations under the same aerodynamic conditions, beneficial effects of passive wing deformation can be observed from the visualized airflow and averaged thrust. The measurements and their presentation enable observation and understanding of the required structural properties for a thrust effective flapping wing. The intended passive responses of the different wings follow a particular pattern in correlation to their aerodynamic performance. Consequently, both the experimental technique and data analysis method can lead to further studies to determine the design principles for micro air vehicle flapping wings.

  13. Binding of single walled carbon nanotube to WT and mutant HIV-1 proteases: analysis of flap dynamics and binding mechanism.

    Meher, Biswa Ranjan; Wang, Yixuan

    2012-09-01

    Most of the currently treated HIV-1 protease (HIV-PR) inhibitors have been prone to suffer from the mutations associated drug resistance. Therefore, it is necessary to search for potent alternatives against the drug resistance. In the current study we have tested the single-walled carbon nanotube (SWCNT) as an inhibitor in wild type (WT) as well as in three primary mutants (I50V(PR), V82A(PR) and I84V(PR)) of the HIV-1-PR through docking the SWCNT in the active site region, and then performed all-atom MD simulations for the complexes. The conformational dynamics of HIV-PR with a 20 ns trajectory reveals that the SWCNT can effectively bind to the HIV-1-PR active site and regulate the flap dynamics such as maintaining the flap-flap closed. To gain an insight into the binding affinity, we also performed the MM-PBSA based binding free energy calculations for the four HIV-PR/SWCNT complexes. It was observed that, although the binding between the SWCNT and the HIV-PR decreases due to the mutations, the SWCNTs bind to the HIV-PRs 3-5 folds stronger than the most potent HIV-1-PR inhibitor, TMC114. Remarkably, the significant interactions with binding energy higher than 1kcal/mol focus on the flap and active regions, which favors closing flap-flap and deactivating the active residues of the HIV-PR. The flap dynamics and binding strength information for HIV-PR and SWCNTs can help design SWCNT-based HIV-1-PR inhibitors. PMID:23142620

  14. Transition from a Linear to a Harmonic Potential in Collective Dynamics of a Multifilament Actin Bundle

    Schnauß, Jörg; Golde, Tom; Schuldt, Carsten; Schmidt, B. U. Sebastian; Glaser, Martin; Strehle, Dan; Händler, Tina; Heussinger, Claus; Käs, Josef A.

    2016-03-01

    Attractive depletion forces between rodlike particles in highly crowded environments have been shown through recent modeling and experimental approaches to induce different structural and dynamic signatures depending on relative orientation between rods. For example, it has been demonstrated that the axial attraction between two parallel rods yields a linear energy potential corresponding to a constant contractile force of 0.1 pN. Here, we extend pairwise, depletion-induced interactions to a multifilament level with actin bundles, and find contractile forces up to 3 pN. Forces generated due to bundle relaxation were not constant, but displayed a harmonic potential and decayed exponentially with a mean decay time of 3.4 s. Through an analytical model, we explain these different fundamental dynamics as an emergent, collective phenomenon stemming from the additive, pairwise interactions of filaments within a bundle.

  15. Regimes of wave type patterning driven by refractory actin feedback: transition from static polarization to dynamic wave behaviour

    Patterns of waves, patches, and peaks of actin are observed experimentally in many living cells. Models of this phenomenon have been based on the interplay between filamentous actin (F-actin) and its nucleation promoting factors (NPFs) that activate the Arp2/3 complex. Here we present an alternative biologically-motivated model for F-actin-NPF interaction based on properties of GTPases acting as NPFs. GTPases (such as Cdc42, Rac) are known to promote actin nucleation, and to have active membrane-bound and inactive cytosolic forms. The model is a natural extension of a previous mathematical mini-model of small GTPases that generates static cell polarization. Like other modellers, we assume that F-actin negative feedback shapes the observed patterns by suppressing the trailing edge of NPF-generated wave-fronts, hence localizing the activity spatially. We find that our NPF-actin model generates a rich set of behaviours, spanning a transition from static polarization to single pulses, reflecting waves, wave trains, and oscillations localized at the cell edge. The model is developed with simplicity in mind to investigate the interaction between nucleation promoting factor kinetics and negative feedback. It explains distinct types of pattern initiation mechanisms, and identifies parameter regimes corresponding to distinct behaviours. We show that weak actin feedback yields static patterning, moderate feedback yields dynamical behaviour such as travelling waves, and strong feedback can lead to wave trains or total suppression of patterning. We use a recently introduced nonlinear bifurcation analysis to explore the parameter space of this model and predict its behaviour with simulations validating those results. (paper)

  16. The actin binding domain of βI-spectrin regulates the morphological and functional dynamics of dendritic spines.

    Michael W Nestor

    Full Text Available Actin microfilaments regulate the size, shape and mobility of dendritic spines and are in turn regulated by actin binding proteins and small GTPases. The βI isoform of spectrin, a protein that links the actin cytoskeleton to membrane proteins, is present in spines. To understand its function, we expressed its actin-binding domain (ABD in CA1 pyramidal neurons in hippocampal slice cultures. The ABD of βI-spectrin bundled actin in principal dendrites and was concentrated in dendritic spines, where it significantly increased the size of the spine head. These effects were not observed after expression of homologous ABDs of utrophin, dystrophin, and α-actinin. Treatment of slice cultures with latrunculin-B significantly decreased spine head size and decreased actin-GFP fluorescence in cells expressing the ABD of α-actinin, but not the ABD of βI-spectrin, suggesting that its presence inhibits actin depolymerization. We also observed an increase in the area of GFP-tagged PSD-95 in the spine head and an increase in the amplitude of mEPSCs at spines expressing the ABD of βI-spectrin. The effects of the βI-spectrin ABD on spine size and mEPSC amplitude were mimicked by expressing wild-type Rac3, a small GTPase that co-immunoprecipitates specifically with βI-spectrin in extracts of cultured cortical neurons. Spine size was normal in cells co-expressing a dominant negative Rac3 construct with the βI-spectrin ABD. We suggest that βI-spectrin is a synaptic protein that can modulate both the morphological and functional dynamics of dendritic spines, perhaps via interaction with actin and Rac3.

  17. Filopodial retraction force is generated by cortical actin dynamics and controlled by reversible tethering at the tip

    Bornschlögl, Thomas; Romero, Stéphane; Vestergaard, Christian L.;

    2013-01-01

    Filopodia are dynamic, finger-like plasma membrane protrusions that sense the mechanical and chemical surroundings of the cell. Here, we show in epithelial cells that the dynamics of filopodial extension and retraction are determined by the difference between the actin polymerization rate at the...... tip and the retrograde flow at the base of the filopodium. Adhesion of a bead to the filopodial tip locally reduces actin polymerization and leads to retraction via retrograde flow, reminiscent of a process used by pathogens to invade cells. Using optical tweezers, we show that filopodial retraction...... occurs at a constant speed against counteracting forces up to 50 pN. Our measurements point toward retrograde flow in the cortex together with frictional coupling between the filopodial and cortical actin networks as the main retraction-force generator for filopodia. The force exerted by filopodial...

  18. Exploring the Possible Role of Lysine Acetylation on Entamoeba histolytica Virulence: A Focus on the Dynamics of the Actin Cytoskeleton

    L. López-Contreras

    2013-01-01

    Full Text Available Cytoskeleton remodeling can be regulated, among other mechanisms, by lysine acetylation. The role of acetylation on cytoskeletal and other proteins of Entamoeba histolytica has been poorly studied. Dynamic rearrangements of the actin cytoskeleton are crucial for amebic motility and capping formation, processes that may be effective means of evading the host immune response. Here we report the possible effect of acetylation on the actin cytoskeleton dynamics and in vivo virulence of E. histolytica. Using western blot, immunoprecipitation, microscopy assays, and in silico analysis, we show results that strongly suggest that the increase in Aspirin-induced cytoplasm proteins acetylation reduced cell movement and capping formation, likely as a consequence of alterations in the structuration of the actin cytoskeleton. Additionally, intrahepatic inoculation of Aspirin-treated trophozoites in hamsters resulted in severe impairment of the amebic virulence. Taken together, these results suggest an important role for lysine acetylation in amebic invasiveness and virulence.

  19. Vortex dynamics and flapping patterns of the inverted flag with a bluff body

    Kim, Hyeonseong; Kim, Junyoung; Kim, Daegyoum

    2015-11-01

    Flow-induced vibration of flexible structures for energy harvesting has drawn attention recently. The inverted flag whose trailing edge is clamped and leading edge is free to move was known to self-excite in a uniform flow of both water and air. In this study, we investigated the effect of an upstream bluff body, a flat plate, on the dynamics of the downstream inverted flag. Periodic vortical structures created by an upstream bluff body make the dynamics of the inverted flag quite different from those of the inverted flag without the bluff body. We examined the motion of the inverted flag by varying the relative displacement of the inverted flag from the bluff body and their relative size. Our results show that the inverted flag can flap with higher frequency and larger amplitude with the upstream bluff body. We also compared the dynamics of the inverted flag with those of the general flag with the upstream bluff body. In order to better understand the dynamics of the flag, the analysis of the flow patterns using particle image velocimetry was also conducted.

  20. The structural dynamics of α-tropomyosin on F-actin shape the overlap complex between adjacent tropomyosin molecules.

    Lehman, William; Li, Xiaochuan Edward; Orzechowski, Marek; Fischer, Stefan

    2014-06-15

    Coiled-coil tropomyosin, localized on actin filaments in virtually all eukaryotic cells, serves as a gatekeeper regulating access of the motor protein myosin and other actin-binding proteins onto the thin filament surface. Tropomyosin's modular pseudo-repeating pattern of approximately 39 amino acid residues is designed to allow binding of the coiled-coil to successive actin subunits along thin filaments. Even though different tropomyosin isoforms contain varying numbers of repeat modules, the pseudo-repeat length, in all cases, matches that of a single actin subunit. Thus, the seven pseudo-repeats of 42nm long muscle tropomyosin bind to seven successive actin subunits along thin filaments, while simultaneously bending into a super-helical conformation that is preshaped to the actin filament helix. In order to form a continuous cable on thin filaments that is free of gaps, adjacent tropomyosin molecules polymerize head-to-tail by means of a short (∼9 residue) overlap. Several laboratories have engineered peptides to mimic the N- and C-terminal tropomyosin association and to characterize the overlap structure. All overlapping domains examined show a compact N-terminal coiled-coil inserting into a partially opened C-terminal partner, where the opposing coiled-coils at the overlap junction face each other at up to ∼90° twist angles. Here, Molecular Dynamics (MD) simulations were carried out to determine constraints on the formation of the tropomyosin overlap complex and to assess the amount of twisting exhibited by full-length tropomyosin when bound to actin. With the exception of the last 20-40 C- and N-terminal residues, we find that the average tropomyosin structure closely resembles a "canonical" model proposed in the classic work of McLachlan and Stewart, displaying perfectly symmetrical supercoil geometry matching the F-actin helix with an integral number of coiled-coil turns, a coiled-coil helical pitch of 137Å, a superhelical pitch of 770Å, and no

  1. Rac1 at the crossroad of actin dynamics and neuroinflammation in Amyotrophic Lateral Sclerosis

    Nadia D'Ambrosi

    2014-09-01

    Full Text Available Rac1 is a major player of the Rho family of small GTPases that controls multiple cell signaling pathways, such as the organization of cytoskeleton (including adhesion and motility, cell proliferation, apoptosis and activation of immune cells. In the nervous system, in particular, Rac1 GTPase plays a key regulatory function of both actin and microtubule cytoskeletal dynamics and thus it is central to axonal growth and stability, as well as dendrite and spine structural plasticity. Rac1 is also a crucial regulator of NADPH-dependent membrane oxidase (NOX, a prominent source of ROS, thus having a central role in the inflammatory response and neurotoxicity mediated by microglia cells in the nervous system. As such, alterations in Rac1 activity might well be involved in the processes that give rise to Amyotrophic Lateral Sclerosis (ALS, a complex syndrome where cytoskeletal disturbances in motor neurons and redox alterations in the inflammatory compartment play pivotal and synergic roles in the final disease outcomes. Here we will discuss the genetic and mechanistic evidence indicating the relevance of Rac1 dysregulation in the pathogenesis of ALS.

  2. Clathrin is Important for Normal Actin Dynamics and Progression of Sla2p-Containing Patches During Endocytosis in Yeast

    Newpher, Thomas M.; Lemmon, Sandra K.

    2006-01-01

    Clathrin is a major vesicle coat protein involved in receptor-mediated endocytosis. In yeast and higher eukaryotes, clathrin is recruited to the plasma membrane during the early stage of endocytosis along with clathrin-associated adaptors. As coated pits undergo maturation, a burst of actin polymerization accompanies and helps drive vesicle internalization. Here, we investigate the dynamics of clathrin relative to the early endocytic patch protein Sla2p. We find that clathrin is recruited to ...

  3. Actinic Cheilitis

    ... actinic cheilitis. Overview Actinic cheilitis, sometimes known as "farmer's lip" or "sailor's lip," is a precancerous condition ... Last Updated: 22 Dec 2008 Information for other ages: Table of Contents: Overview Who's At Risk Signs ...

  4. A flight-dynamic helicopter mathematical model with a single flap-lag-torsion main rotor

    Takahashi, Marc D.

    1990-01-01

    A mathematical model of a helicopter system with a single main rotor that includes rigid, hinge-restrained rotor blades with flap, lag, and torsion degrees of freedom is described. The model allows several hinge sequences and two offsets in the hinges. Quasi-steady Greenberg theory is used to calculate the blade-section aerodynamic forces, and inflow effects are accounted for by using three-state nonlinear dynamic inflow model. The motion of the rigid fuselage is defined by six degrees of freedom, and an optional rotor rpm degree of freedom is available. Empennage surfaces and the tail rotor are modeled, and the effect of main-rotor downwash on these elements is included. Model trim linearization, and time-integration operations are described and can be applied to a subset of the model in the rotating or nonrotating coordinate frame. A preliminary validation of the model is made by comparing its results with those of other analytical and experimental studies. This publication presents the results of research compiled in November 1989.

  5. Antiepileptic teratogen valproic acid (VPA) modulates organisation and dynamics of the actin cytoskeleton

    Walmod, P S; Skladchikova, G; Kawa, A;

    1999-01-01

    control cells and cells treated with VPA, indicating that VPA affected the cytoskeletal determinants of cell morphology. Furthermore, VPA treatment induced an increase of F-actin, and of FAK, paxillin, vinculin, and phosphotyrosine in focal adhesion complexes. These changes were accompanied by increased...... adhesion of VPA-treated cells to the extracellular matrix. Treatment with an RGD-containing peptide reducing integrin binding to components of the extracellular matrix partially reverted the motility inhibition induced by VPA, indicating that altered adhesion contributed to, but was not the sole reason for......, VPA caused a redistribution of the actin severing protein gelsolin, and left the cells unable to respond to treatment with a gelsolin-peptide known to reduce the amount of gelsolin bound to phosphatidylinositol bisphosphate (PIP2), leaving a larger amount of the protein in a potential actin binding...

  6. Actin Dynamics Regulates Voltage-Dependent Calcium-Permeable Channels of the Vicia faba Guard Cell Plasma Membrane

    Wei Zhang; Liu-Min Fan

    2009-01-01

    Free cytosolic Ca~(2+) ([Ca~(2+)]_(cyt)) is an ubiquitous second messenger in plant cell signaling, and [Ca~(2+)]_(cyt) elevation is associated with Ca~(2+)-permeable channels in the plasma membrane and endomembranes regulated by a wide range of stimuli. However, knowledge regarding Ca~(2+) channels and their regulation remains limited in planta. A type of voltage-dependent Ca~(2+)-permeable channel was identified and characterized for the Vicia faba L. guard cell plasma membrane by using patch-clamp techniques. These channels are permeable to both Ba~(2+) and Ca~(2+), and their activities can be inhibited by micromolar Gd~(3+). The unitary conductance and the reversal potential of the channels depend on the Ca~(2+) or Ba~(2+) gradients across the plasma membrane. The inward whole-cell Ca~(2+) (Ba~(2+)) current, as well as the unitary current amplitude and NP. of the single Ca~(2+) channel, increase along with the membrane hyperpolarization. Pharmacological experiments suggest that actin dynamics may serve as an upstream regulator of this type of calcium channel of the guard cell plasma membrane. Cytochalasin D, an actin polymerization blocker, activated the NP_o of these channels at the single channel level and increased the current amplitude at the whole-cell level. But these channel activations and current increments could be restrained by pretreatment with an F-actin stabilizer, phalloidin. The potential physiological significance of this regulatory mechanism is also discussed.

  7. AtFH1 formin mutation affects actin filament and microtubule dynamics in Arabidopsis thaliana

    Rosero, A.; Žárský, Viktor; Cvrčková, F.

    2013-01-01

    Roč. 64, č. 2 (2013), s. 585-597. ISSN 0022-0957 R&D Projects: GA ČR GAP305/10/0433 Institutional research plan: CEZ:AV0Z50380511 Keywords : Actin * Arabidopsis * At5g25500 Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 5.794, year: 2013

  8. Role of cyclic nucleotide-dependent actin cytoskeletal dynamics:Ca(2+](i and force suppression in forskolin-pretreated porcine coronary arteries.

    Kyle M Hocking

    Full Text Available Initiation of force generation during vascular smooth muscle contraction involves a rise in intracellular calcium ([Ca(2+]i and phosphorylation of myosin light chains (MLC. However, reversal of these two processes alone does not account for the force inhibition that occurs during relaxation or inhibition of contraction, implicating that other mechanisms, such as actin cytoskeletal rearrangement, play a role in the suppression of force. In this study, we hypothesize that forskolin-induced force suppression is dependent upon changes in actin cytoskeletal dynamics. To focus on the actin cytoskeletal changes, a physiological model was developed in which forskolin treatment of intact porcine coronary arteries (PCA prior to treatment with a contractile agonist resulted in complete suppression of force. Pretreatment of PCA with forskolin suppressed histamine-induced force generation but did not abolish [Ca(2+]i rise or MLC phosphorylation. Additionally, forskolin pretreatment reduced filamentous actin in histamine-treated tissues, and prevented histamine-induced changes in the phosphorylation of the actin-regulatory proteins HSP20, VASP, cofilin, and paxillin. Taken together, these results suggest that forskolin-induced complete force suppression is dependent upon the actin cytoskeletal regulation initiated by the phosphorylation changes of the actin regulatory proteins and not on the MLC dephosphorylation. This model of complete force suppression can be employed to further elucidate the mechanisms responsible for smooth muscle tone, and may offer cues to pathological situations, such as hypertension and vasospasm.

  9. Actinic keratosis

    Solar keratosis; Sun-induced skin changes - keratosis; Keratosis - actinic (solar) ... laser treatment called photodynamic therapy Chemical peels Skin creams such as 5-fluorouracil (5-FU) and imiquimod

  10. RNase L Interacts with Filamin A To Regulate Actin Dynamics and Barrier Function for Viral Entry

    Siddiqui, Mohammad Adnan; Dayal, Shubham; Naji, Merna; Ezelle, Heather J.; Zeng, Chun; Zhou, Aimin; Hassel, Bret A.

    2014-01-01

    ABSTRACT The actin cytoskeleton and its network of associated proteins constitute a physical barrier that viruses must circumvent to gain entry into cells for productive infection. The mechanisms by which the physical signals of infection are sensed by the host to activate an innate immune response are not well understood. The antiviral endoribonuclease RNase L is ubiquitously expressed in a latent form and activated upon binding 2-5A, a unique oligoadenylate produced during viral infections. We provide evidence that RNase L in its inactive form interacts with the actin-binding protein Filamin A to modulate the actin cytoskeleton and inhibit virus entry. Cells lacking either RNase L or Filamin A displayed increased virus entry which was exacerbated in cells lacking both proteins. RNase L deletion mutants that reduced Filamin A interaction displayed a compromised ability to restrict virus entry, supporting the idea of an important role for the RNase L-Filamin A complex in barrier function. Remarkably, both the wild type and a catalytically inactive RNase L mutant were competent to reduce virus entry when transfected into RNase L-deficient cells, indicating that this novel function of RNase L is independent of its enzymatic activity. Virus infection and RNase L activation disrupt its association with Filamin A and release RNase L to mediate its canonical nuclease-dependent antiviral activities. The dual functions of RNase L as a constitutive component of the actin cytoskeleton and as an induced mediator of antiviral signaling and effector functions provide insights into its mechanisms of antiviral activity and opportunities for the development of novel antiviral agents. PMID:25352621

  11. Toxofilin upregulates the host cortical actin cytoskeleton dynamics, facilitating Toxoplasma invasion

    Delorme-Walker, Violaine; Abrivard, Marie; Lagal, Vanessa; Anderson, Karen; Perazzi, Audrey; Gonzalez, Virginie; Page, Christopher; Chauvet, Juliette; Ochoa, Wendy; Volkmann, Niels; Hanein, Dorit; Tardieux, Isabelle

    2012-01-01

    Toxoplasma gondii, a human pathogen and a model apicomplexan parasite, actively and rapidly invades host cells. To initiate invasion, the parasite induces the formation of a parasite–cell junction, and progressively propels itself through the junction, inside a newly formed vacuole that encloses the entering parasite. Little is known about how a parasite that is a few microns in diameter overcomes the host cell cortical actin barrier to achieve the remarkably rapid process of internalization ...

  12. Dynamic actin polymerization on endosomes regulates integrin trafficking, cell adhesion and cell migration

    Duleh, Steve Niessen

    2012-01-01

    Activators of the Arp2/3 complex, termed nucleation-promoting factors (NPFs), are required for the proper spatial and temporal control of actin assembly in cells. Mammalian cells express several NPFs, each of which serve distinct functions in specific cellular processes, including N-WASP in phagocytosis and endocytosis, WAVE and JMY in cell migration, and WHAMM in ER-to-Golgi transport. Although another NPF termed WASH was recently identified, the cellular function and activity of this prot...

  13. Actin flow and talin dynamics govern rigidity sensing in actin–integrin linkage through talin extension

    Hirata, Hiroaki; Chiam, Keng-Hwee; Lim, Chwee Teck; Sokabe, Masahiro

    2014-01-01

    At cell–substrate adhesion sites, the linkage between actin filaments and integrin is regulated by mechanical stiffness of the substrate. Of potential molecular regulators, the linker proteins talin and vinculin are of particular interest because mechanical extension of talin induces vinculin binding with talin, which reinforces the actin–integrin linkage. For understanding the molecular and biophysical mechanism of rigidity sensing at cell–substrate adhesion sites, we constructed a simple ph...

  14. Antiobesity Action of ACAM by Modulating the Dynamics of Cell Adhesion and Actin Polymerization in Adipocytes.

    Murakami, Kazutoshi; Eguchi, Jun; Hida, Kazuyuki; Nakatsuka, Atsuko; Katayama, Akihiro; Sakurai, Miwa; Choshi, Haruki; Furutani, Masumi; Ogawa, Daisuke; Takei, Kohji; Otsuka, Fumio; Wada, Jun

    2016-05-01

    Coxsackie virus and adenovirus receptor-like membrane protein (CLMP) was identified as the tight junction-associated transmembrane protein of epithelial cells with homophilic binding activities. CLMP is also recognized as adipocyte adhesion molecule (ACAM), and it is upregulated in mature adipocytes in rodents and humans with obesity. Here, we present that aP2 promoter-driven ACAM transgenic mice are protected from obesity and diabetes with the prominent reduction of adipose tissue mass and smaller size of adipocytes. ACAM is abundantly expressed on plasma membrane of mature adipocytes and associated with formation of phalloidin-positive polymerized form of cortical actin (F-actin). By electron microscopy, the structure of zonula adherens with an intercellular space of ∼10-20 nm was observed with strict parallelism of the adjoining cell membranes over distances of 1-20 μm, where ACAM and γ-actin are abundantly expressed. The formation of zonula adherens may increase the mechanical strength, inhibit the adipocyte hypertrophy, and improve the insulin sensitivity. PMID:26956488

  15. Actin flow and talin dynamics govern rigidity sensing in actin–integrin linkage through talin extension

    Hirata, Hiroaki; Chiam, Keng-Hwee; Lim, Chwee Teck; Sokabe, Masahiro

    2014-01-01

    At cell–substrate adhesion sites, the linkage between actin filaments and integrin is regulated by mechanical stiffness of the substrate. Of potential molecular regulators, the linker proteins talin and vinculin are of particular interest because mechanical extension of talin induces vinculin binding with talin, which reinforces the actin–integrin linkage. For understanding the molecular and biophysical mechanism of rigidity sensing at cell–substrate adhesion sites, we constructed a simple physical model to examine a role of talin extension in the stiffness-dependent regulation of actin–integrin linkage. We show that talin molecules linking between retrograding actin filaments and substrate-bound integrin are extended in a manner dependent on substrate stiffness. The model predicts that, in adhesion complexes containing ≈30 talin links, talin is extended enough for vinculin binding when the substrate is stiffer than 1 kPa. The lifetime of talin links needs to be 2–5 s to achieve an appropriate response of talin extension against substrate stiffness. Furthermore, changes in actin velocity drastically shift the range of substrate stiffness that induces talin–vinculin binding. Our results suggest that talin extension is a key step in sensing and responding to substrate stiffness at cell adhesion sites. PMID:25142525

  16. Control of actin-based motility through localized actin binding

    A wide variety of cell biological and biomimetic systems use actin polymerization to drive motility. It has been suggested that an object such as a bacterium can propel itself by self-assembling a high concentration of actin behind it, if it is repelled by actin. However, it is also known that it is essential for the moving object to bind actin. Therefore, a key question is how the actin tail can propel an object when it both binds and repels the object. We present a physically consistent Brownian dynamics model for actin-based motility that includes the minimal components of the dendritic nucleation model and allows for both attractive and repulsive interactions between actin and a moveable disc. We find that the concentration gradient of filamentous actin generated by polymerization is sufficient to propel the object, even with moderately strong binding interactions. Additionally, actin binding can act as a biophysical cap, and may directly control motility through modulation of network growth. Overall, this mechanism is robust in that it can drive motility against a load up to a stall pressure that depends on the Young’s modulus of the actin network and can explain several aspects of actin-based motility. (paper)

  17. New compliant strain gauges for self-sensing dynamic deformation of flapping wings on miniature air vehicles

    Wissman, James; Perez-Rosado, Ariel; Edgerton, Alex; Levi, Benjamin M.; Karakas, Zeynep N.; Kujawski, Mark; Philipps, Alyssa; Papavizas, Nicholas; Fallon, Danielle; Bruck, Hugh A.; Smela, Elisabeth

    2013-08-01

    Over the past several years there has been an increasing interest in the development of miniature air vehicles (MAVs) with flapping wings. To allow these MAVs to adjust to changes in wind direction and to maximize their efficiency, it is desirable to monitor the deformation of the wing during flight. This paper presents a step in this direction, demonstrating the measurement of strain on the surface of the wing using minimally invasive compliant piezoresistive sensors. The strain gauges consisted of latex mixed with electrically conducting exfoliated graphite, and they were applied by spray coating. To calibrate the gauges, both static and dynamic testing up to 10 Hz were performed using cantilever structures. In tension the static sensitivity was a linear 0.4 Ω μɛ-1 and the gauge factor was 28; in compression, the gauge factor was -5. Although sensitivities in tension and compression differed by a factor of almost six, this was not reflected in the dynamic data, which followed the strain reversibly with little distortion. There was no attenuation with frequency, indicating a sufficiently small time constant for this application. The gauges were thin, compliant, and light enough to measure, without interference, deformations due to shape changes of the flexible wing associated with generating lift and thrust. During flapping the resistance closely tracked the generated thrust, measured on a test stand, with both signals tracing figure-8 loops as a function of wing position throughout each cycle.

  18. New compliant strain gauges for self-sensing dynamic deformation of flapping wings on miniature air vehicles

    Over the past several years there has been an increasing interest in the development of miniature air vehicles (MAVs) with flapping wings. To allow these MAVs to adjust to changes in wind direction and to maximize their efficiency, it is desirable to monitor the deformation of the wing during flight. This paper presents a step in this direction, demonstrating the measurement of strain on the surface of the wing using minimally invasive compliant piezoresistive sensors. The strain gauges consisted of latex mixed with electrically conducting exfoliated graphite, and they were applied by spray coating. To calibrate the gauges, both static and dynamic testing up to 10 Hz were performed using cantilever structures. In tension the static sensitivity was a linear 0.4 Ω με−1 and the gauge factor was 28; in compression, the gauge factor was −5. Although sensitivities in tension and compression differed by a factor of almost six, this was not reflected in the dynamic data, which followed the strain reversibly with little distortion. There was no attenuation with frequency, indicating a sufficiently small time constant for this application. The gauges were thin, compliant, and light enough to measure, without interference, deformations due to shape changes of the flexible wing associated with generating lift and thrust. During flapping the resistance closely tracked the generated thrust, measured on a test stand, with both signals tracing figure-8 loops as a function of wing position throughout each cycle. (paper)

  19. Effects of F/G-actin ratio and actin turn-over rate on NADPH oxidase activity in microglia

    Rasmussen, Izabela; Pedersen, Line Hjortshøj; Byg, Luise;

    2010-01-01

    Most in vivo studies that have addressed the role of actin dynamics in NADPH oxidase function in phagocytes have used toxins to modulate the polymerization state of actin and mostly effects on actin has been evaluated by end point measurements of filamentous actin, which says little about actin d...

  20. Effects of F/G-actin ratio and actin turn-over rate on NADPH oxidase activity in microglia

    Rasmussen Izabela; Pedersen Line H; Byg Luise; Suzuki Kazuhiro; Sumimoto Hideki; Vilhardt Frederik

    2010-01-01

    Abstract Background Most in vivo studies that have addressed the role of actin dynamics in NADPH oxidase function in phagocytes have used toxins to modulate the polymerization state of actin and mostly effects on actin has been evaluated by end point measurements of filamentous actin, which says little about actin dynamics, and without consideration for the subcellular distribution of the perturbed actin cytoskeleton. Results Here, we in addition to toxins use conditional expression of the ma...

  1. Strategies for the stabilization of longitudinal forward flapping flight revealed using a dynamically-scaled robotic fly

    The ability to regulate forward speed is an essential requirement for flying animals. Here, we use a dynamically-scaled robot to study how flapping insects adjust their wing kinematics to regulate and stabilize forward flight. The results suggest that the steady-state lift and thrust requirements at different speeds may be accomplished with quite subtle changes in hovering kinematics, and that these adjustments act primarily by altering the pitch moment. This finding is consistent with prior hypotheses regarding the relationship between body pitch and flight speed in fruit flies. Adjusting the mean stroke position of the wings is a likely mechanism for trimming the pitch moment at all speeds, whereas changes in the mean angle of attack may be required at higher speeds. To ensure stability, the flapping system requires additional pitch damping that increases in magnitude with flight speed. A compensatory reflex driven by fast feedback of pitch rate from the halteres could provide such damping, and would automatically exhibit gain scheduling with flight speed if pitch torque was regulated via changes in stroke deviation. Such a control scheme would provide an elegant solution for stabilization across a wide range of forward flight speeds. (paper)

  2. Strategies for the stabilization of longitudinal forward flapping flight revealed using a dynamically-scaled robotic fly.

    Elzinga, Michael J; van Breugel, Floris; Dickinson, Michael H

    2014-06-01

    The ability to regulate forward speed is an essential requirement for flying animals. Here, we use a dynamically-scaled robot to study how flapping insects adjust their wing kinematics to regulate and stabilize forward flight. The results suggest that the steady-state lift and thrust requirements at different speeds may be accomplished with quite subtle changes in hovering kinematics, and that these adjustments act primarily by altering the pitch moment. This finding is consistent with prior hypotheses regarding the relationship between body pitch and flight speed in fruit flies. Adjusting the mean stroke position of the wings is a likely mechanism for trimming the pitch moment at all speeds, whereas changes in the mean angle of attack may be required at higher speeds. To ensure stability, the flapping system requires additional pitch damping that increases in magnitude with flight speed. A compensatory reflex driven by fast feedback of pitch rate from the halteres could provide such damping, and would automatically exhibit gain scheduling with flight speed if pitch torque was regulated via changes in stroke deviation. Such a control scheme would provide an elegant solution for stabilization across a wide range of forward flight speeds. PMID:24855029

  3. Effects of F/G-actin ratio and actin turn-over rate on NADPH oxidase activity in microglia

    Rasmussen Izabela

    2010-09-01

    Full Text Available Abstract Background Most in vivo studies that have addressed the role of actin dynamics in NADPH oxidase function in phagocytes have used toxins to modulate the polymerization state of actin and mostly effects on actin has been evaluated by end point measurements of filamentous actin, which says little about actin dynamics, and without consideration for the subcellular distribution of the perturbed actin cytoskeleton. Results Here, we in addition to toxins use conditional expression of the major actin regulatory protein LIM kinase-1 (LIMK1, and shRNA knock-down of cofilin to modulate the cellular F/G-actin ratio in the Ra2 microglia cell line, and we use Fluorescence Recovery after Photobleaching (FRAP in β-actin-YFP-transduced cells to obtain a dynamic measure of actin recovery rates (actin turn-over rates in different F/G-actin states of the actin cytoskeleton. Our data demonstrate that stimulated NADPH oxidase function was severely impaired only at extreme actin recovery rates and F/G-actin ratios, and surprisingly, that any moderate changes of these parameters of the actin cytoskeleton invariably resulted in an increased NADPH oxidase activity. Conclusion moderate actin polymerization and depolymerization both increase the FMLP and PMA-stimulated NADPH oxidase activity of microglia, which is directly correlated with neither actin recovery rate nor F/G- actin ratio. Our results indicate that NADPH oxidase functions in an enhanced state of activity in stimulated phagocytes despite widely different states of the actin cytoskeleton.

  4. Technical advance: identification of plant actin-binding proteins by F-actin affinity chromatography

    Hu, S.; Brady, S. R.; Kovar, D. R.; Staiger, C. J.; Clark, G. B.; Roux, S. J.; Muday, G. K.

    2000-01-01

    Proteins that interact with the actin cytoskeleton often modulate the dynamics or organization of the cytoskeleton or use the cytoskeleton to control their localization. In plants, very few actin-binding proteins have been identified and most are thought to modulate cytoskeleton function. To identify actin-binding proteins that are unique to plants, the development of new biochemical procedures will be critical. Affinity columns using actin monomers (globular actin, G-actin) or actin filaments (filamentous actin, F-actin) have been used to identify actin-binding proteins from a wide variety of organisms. Monomeric actin from zucchini (Cucurbita pepo L.) hypocotyl tissue was purified to electrophoretic homogeneity and shown to be native and competent for polymerization to actin filaments. G-actin, F-actin and bovine serum albumin affinity columns were prepared and used to separate samples enriched in either soluble or membrane-associated actin-binding proteins. Extracts of soluble actin-binding proteins yield distinct patterns when eluted from the G-actin and F-actin columns, respectively, leading to the identification of a putative F-actin-binding protein of approximately 40 kDa. When plasma membrane-associated proteins were applied to these columns, two abundant polypeptides eluted selectively from the F-actin column and cross-reacted with antiserum against pea annexins. Additionally, a protein that binds auxin transport inhibitors, the naphthylphthalamic acid binding protein, which has been previously suggested to associate with the actin cytoskeleton, was eluted in a single peak from the F-actin column. These experiments provide a new approach that may help to identify novel actin-binding proteins from plants.

  5. Energetic modeling and single-molecule verification of dynamic regulation on receptor complexes by actin corrals and lipid raft domains

    Lin, Chien Y.; Huang, Jung Y.; Lo, Leu-Wei

    2014-12-01

    We developed an energetic model by integrating the generalized Langevin equation with the Cahn-Hilliard equation to simulate the diffusive behaviors of receptor proteins in the plasma membrane of a living cell. Simulation results are presented to elaborate the confinement effects from actin corrals and protein-induced lipid domains. Single-molecule tracking data of epidermal growth factor receptors (EGFR) acquired on live HeLa cells agree with the simulation results and the mechanism that controls the diffusion of single-molecule receptors is clarified. We discovered that after ligand binding, EGFR molecules move into lipid nanodomains. The transition rates between different diffusion states of liganded EGFR molecules are regulated by the lipid domains. Our method successfully captures dynamic interactions of receptors at the single-molecule level and provides insight into the functional architecture of both the diffusing EGFR molecules and their local cellular environment.

  6. Fluid dynamics of flapping aquatic flight in the bird wrasse: three-dimensional unsteady computations with fin deformation.

    Ramamurti, Ravi; Sandberg, William C; Löhner, Rainald; Walker, Jeffrey A; Westneat, Mark W

    2002-10-01

    Many fishes that swim with the paired pectoral fins use fin-stroke parameters that produce thrust force from lift in a mechanism of underwater flight. These locomotor mechanisms are of interest to behavioral biologists, biomechanics researchers and engineers. In the present study, we performed the first three-dimensional unsteady computations of fish swimming with oscillating and deforming fins. The objective of these computations was to investigate the fluid dynamics of force production associated with the flapping aquatic flight of the bird wrasse Gomphosus varius. For this computational work, we used the geometry of the wrasse and its pectoral fin, and previously measured fin kinematics, as the starting points for computational investigation of three-dimensional (3-D) unsteady fluid dynamics. We performed a 3-D steady computation and a complete set of 3-D quasisteady computations for a range of pectoral fin positions and surface velocities. An unstructured, grid-based, unsteady Navier-Stokes solver with automatic adaptive remeshing was then used to compute the unsteady flow about the wrasse through several complete cycles of pectoral fin oscillation. The shape deformation of the pectoral fin throughout the oscillation was taken from the experimental kinematics. The pressure distribution on the body of the bird wrasse and its pectoral fins was computed and integrated to give body and fin forces which were decomposed into lift and thrust. The velocity field variation on the surface of the wrasse body, on the pectoral fins and in the near-wake was computed throughout the swimming cycle. We compared our computational results for the steady, quasi-steady and unsteady cases with the experimental data on axial and vertical acceleration obtained from the pectoral fin kinematics experiments. These comparisons show that steady state computations are incapable of describing the fluid dynamics of flapping fins. Quasi-steady state computations, with correct incorporation of

  7. The structural dynamics of α-tropomyosin on F-actin shape the overlap complex between adjacent tropomyosin molecules

    Lehman, William; Li, Xiaochuan; Orzechowski, Marek; Fischer, Stefan

    2013-01-01

    Coiled-coil tropomyosin, localized on actin filaments in virtually all eukaryotic cells, serves as a gatekeeper regulating access of the motor protein myosin and other actin-binding proteins onto the thin filament surface. Tropomyosin's modular pseudo-repeating pattern of approximately 39 amino acid residues is designed to allow binding of the coiled-coil to successive actin subunits along thin filaments. Even though different tropomyosin isoforms contain varying numbers of repeat modules, th...

  8. Dynamin-2 regulates fusion pore expansion and quantal release through a mechanism that involves actin dynamics in neuroendocrine chromaffin cells.

    Arlek M González-Jamett

    Full Text Available Over the past years, dynamin has been implicated in tuning the amount and nature of transmitter released during exocytosis. However, the mechanism involved remains poorly understood. Here, using bovine adrenal chromaffin cells, we investigated whether this mechanism rely on dynamin's ability to remodel actin cytoskeleton. According to this idea, inhibition of dynamin GTPase activity suppressed the calcium-dependent de novo cortical actin and altered the cortical actin network. Similarly, expression of a small interfering RNA directed against dynamin-2, an isoform highly expressed in chromaffin cells, changed the cortical actin network pattern. Disruption of dynamin-2 function, as well as the pharmacological inhibition of actin polymerization with cytochalasine-D, slowed down fusion pore expansion and increased the quantal size of individual exocytotic events. The effects of cytochalasine-D and dynamin-2 disruption were not additive indicating that dynamin-2 and F-actin regulate the late steps of exocytosis by a common mechanism. Together our data support a model in which dynamin-2 directs actin polymerization at the exocytosis site where both, in concert, adjust the hormone quantal release to efficiently respond to physiological demands.

  9. A dynamical systems approach to actin-based motility in Listeria monocytogenes

    Hotton, S.

    2010-11-01

    A simple kinematic model for the trajectories of Listeria monocytogenes is generalized to a dynamical system rich enough to exhibit the resonant Hopf bifurcation structure of excitable media and simple enough to be studied geometrically. It is shown how L. monocytogenes trajectories and meandering spiral waves are organized by the same type of attracting set.

  10. Shallow and deep dynamic stall for flapping low Reynolds number airfoils

    Ol, Michael V. [Wright-Patterson AFB, Air Force Research Lab., Dayton, OH (United States); Bernal, Luis; Kang, Chang-Kwon; Shyy, Wei [University of Michigan, Department of Aerospace Engineering, Ann Arbor, MI (United States)

    2009-05-15

    We consider a combined experimental (based on flow visualization, direct force measurement and phase-averaged 2D particle image velocimetry in a water tunnel), computational (2D Reynolds-averaged Navier-Stokes) and theoretical (Theodorsen's formula) approach to study the fluid physics of rigid-airfoil pitch-plunge in nominally two-dimensional conditions. Shallow-stall (combined pitch-plunge) and deep-stall (pure-plunge) are compared at a reduced frequency commensurate with flapping-flight in cruise in nature. Objectives include assessment of how well attached-flow theory can predict lift coefficient even in the presence of significant separation, and how well 2D velocimetry and 2D computation can mutually validate one another. The shallow-stall case shows promising agreement between computation and experiment, while in the deep-stall case, the computation's prediction of flow separation lags that of the experiment, but eventually evinces qualitatively similar leading edge vortex size. Dye injection was found to give good qualitative match with particle image velocimetry in describing leading edge vortex formation and return to flow reattachment, and also gave evidence of strong spanwise growth of flow separation after leading-edge vortex formation. Reynolds number effects, in the range of 10,000-60,000, were found to influence the size of laminar separation in those phases of motion where instantaneous angle of attack was well below stall, but have limited effect on post-stall flowfield behavior. Discrepancy in lift coefficient time history between experiment, theory and computation was mutually comparable, with no clear failure of Theodorsen's formula. This is surprising and encouraging, especially for the deep-stall case, because the theory's assumptions are clearly violated, while its prediction of lift coefficient remains useful for capturing general trends. (orig.)

  11. Canonical and Noncanonical G-Protein Signaling Helps Coordinate Actin Dynamics To Promote Macrophage Phagocytosis of Zymosan

    Huang, Ning-Na; Becker, Steven; Boularan, Cedric; Kamenyeva, Olena; Vural, Ali; Hwang, Il-Young; Shi, Chong-Shan; Kehrl, John H

    2014-01-01

    Both chemotaxis and phagocytosis depend upon actin-driven cell protrusions and cell membrane remodeling. While chemoattractant receptors rely upon canonical G-protein signaling to activate downstream effectors, whether such signaling pathways affect phagocytosis is contentious. Here, we report that Gαi nucleotide exchange and signaling helps macrophages coordinate the recognition, capture, and engulfment of zymosan bioparticles. We show that zymosan exposure recruits F-actin, Gαi proteins, an...

  12. Actin-dependent mechanisms in AMPA receptor trafficking

    Jonathan G Hanley

    2014-11-01

    Full Text Available The precise regulation of AMPA receptor (AMPAR number and subtype at the synapse is crucial for the regulation of excitatory neurotransmission, synaptic plasticity and the consequent formation of appropriate neural circuits during learning and memory. AMPAR trafficking involves the dynamic processes of exocytosis, endocytosis and endosomal recycling, all of which involve the actin cytoskeleton. The actin cytoskeleton is highly dynamic and highly regulated by an abundance of actin-binding proteins and upstream signalling pathways that modulate actin polymerization and depolymerisation. Actin dynamics generate forces that manipulate membranes in the process of vesicle biogenesis, and also for propelling vesicles through the cytoplasm to reach their destination. In addition, trafficking mechanisms exploit more stable aspects of the actin cytoskeleton by using actin-based motor proteins to traffic vesicular cargo along actin filaments. Numerous studies have shown that actin dynamics are critical for AMPAR localization and function. The identification of actin-binding proteins that physically interact with AMPAR subunits, and research into their mode of action is starting to shed light on the mechanisms involved. Such proteins either regulate actin dynamics to modulate mechanical forces exerted on AMPAR-containing membranes, or associate with actin filaments to target or transport AMPAR-containing vesicles to specific subcellular regions. In addition, actin-regulatory proteins that do not physically interact with AMPARs may influence AMPAR trafficking by regulating the local actin environment in the dendritic spine.

  13. Actin turnover is required to prevent axon retraction driven by endogenous actomyosin contractility

    Gallo, Gianluca; Yee, Hal F.; Letourneau, Paul C.

    2002-01-01

    Growth cone motility and guidance depend on the dynamic reorganization of filamentous actin (F-actin). In the growth cone, F-actin undergoes turnover, which is the exchange of actin subunits from existing filaments. However, the function of F-actin turnover is not clear. We used jasplakinolide (jasp), a cell-permeable macrocyclic peptide that inhibits F-actin turnover, to study the role of F-actin turnover in axon extension. Treatment with jasp caused axon retraction, demonstrating that axon ...

  14. Rho, nuclear actin, and actin-binding proteins in the regulation of transcription and gene expression.

    Rajakylä, Eeva Kaisa; Vartiainen, Maria K

    2014-01-01

    Actin cytoskeleton is one of the main targets of Rho GTPases, which act as molecular switches on many signaling pathways. During the past decade, actin has emerged as an important regulator of gene expression. Nuclear actin plays a key role in transcription, chromatin remodeling, and pre-mRNA processing. In addition, the "status" of the actin cytoskeleton is used as a signaling intermediate by at least the MKL1-SRF and Hippo-pathways, which culminate in the transcriptional regulation of cytoskeletal and growth-promoting genes, respectively. Rho GTPases may therefore regulate gene expression by controlling either cytoplasmic or nuclear actin dynamics. Although the regulation of nuclear actin polymerization is still poorly understood, many actin-binding proteins, which are downstream effectors of Rho, are found in the nuclear compartment. In this review, we discuss the possible mechanisms and key proteins that may mediate the transcriptional regulation by Rho GTPases through actin. PMID:24603113

  15. Dynamic phosphoregulation of the cortical actin cytoskeleton and endocytic machinery revealed by real-time chemical genetic analysis

    Sekiya-Kawasaki, Mariko; Groen, Aaron Chris; Cope, M. Jamie T.V.; Kaksonen, Marko; Watson, Hadiya A.; Zhang, Chao; Shokat, Kevan M.; Wendland, Beverly; McDonald, Kent L.; McCaffery, J. Michael; Drubin, David G.

    2003-01-01

    We used chemical genetics to control the activity of budding yeast Prk1p, which is a protein kinase that is related to mammalian GAK and AAK1, and which targets several actin regulatory proteins implicated in endocytosis. In vivo Prk1p inhibition blocked pheromone receptor endocytosis, and caused cortical actin patches to rapidly aggregate into large clumps that contained Abp1p, Sla2p, Pan1p, Sla1p, and Ent1p. Clump formation depended on Arp2p, suggesting that this phenotype might result from...

  16. Yeast RAD2, a homolog of human XPG, plays a key role in the regulation of the cell cycle and actin dynamics

    Mi-Sun Kang

    2013-12-01

    Mutations in the human XPG gene cause Cockayne syndrome (CS and xeroderma pigmentosum (XP. Transcription defects have been suggested as the fundamental cause of CS; however, defining CS as a transcription syndrome is inconclusive. In particular, the function of XPG in transcription has not been clearly demonstrated. Here, we provide evidence for the involvement of RAD2, the Saccharomyces cerevisiae counterpart of XPG, in cell cycle regulation and efficient actin assembly following ultraviolet irradiation. RAD2 C-terminal deletion, which resembles the XPG mutation found in XPG/CS cells, caused cell growth arrest, the cell cycle stalling, a defective α-factor response, shortened lifespan, cell polarity defect, and misregulated actin-dynamics after DNA damage. Overexpression of the C-terminal 65 amino acids of Rad2p was sufficient to induce hyper-cell polarization. In addition, RAD2 genetically interacts with TPM1 during cell polarization. These results provide insights into the role of RAD2 in post-UV irradiation cell cycle regulation and actin assembly, which may be an underlying cause of XPG/CS.

  17. Dynamic boundaries in flowing fluids, from erosion sculptures to flapping wing flight

    Ristroph, Leif

    2014-01-01

    Textbook fluid mechanics addresses steady flows past fixed, rigid objects. However, Nature rarely obeys such restrictions and instead offers many fascinating situations involving the mutual influence of dynamic structures and unsteady flows. Such problems are complex because changes in shape affect flow, which in turn alters shape, and so on. Drawing inspiration from biological and geophysical flows, our Applied Math Lab attacks such fluid–structure interaction problems through tabletop exper...

  18. Dynamic Regulation of Activated Leukocyte Cell Adhesion Molecule–mediated Homotypic Cell Adhesion through the Actin CytoskeletonV⃞

    Nelissen, Judith M. D. T.; Peters, Inge M.; de Grooth, Bart G.; Van Kooyk, Yvette; Figdor, Carl G.

    2000-01-01

    Restricted expression of activated leukocyte cell adhesion molecule (ALCAM) by hematopoietic cells suggests an important role in the immune system and hematopoiesis. To get insight into the mechanisms that control ALCAM-mediated adhesion we have investigated homotypic ALCAM–ALCAM interactions. Here, we demonstrate that the cytoskeleton regulates ALCAM-mediated cell adhesion because inhibition of actin polymerization by cytochalasin D (CytD) strongly induces homotypic ALCAM–ALCAM interactions....

  19. Plekhh2, a novel podocyte protein downregulated in human focal segmental glomerulosclerosis, is involved in matrix adhesion and actin dynamics.

    Perisic, Ljubica; Lal, Mark; Hulkko, Jenny; Hultenby, Kjell; Önfelt, Björn; Sun, Ying; Dunér, Fredrik; Patrakka, Jaakko; Betsholtz, Christer; Uhlen, Mathias; Brismar, Hjalmar; Tryggvason, Karl; Wernerson, Annika; Pikkarainen, Timo

    2012-11-01

    Pleckstrin homology domain-containing, family H (with MyTH4 domain), member 2 (Plekhh2) is a 1491-residue intracellular protein highly enriched in renal glomerular podocytes for which no function has been ascribed. Analysis of renal biopsies from patients with focal segmental glomerulosclerosis revealed a significant reduction in total podocyte Plekhh2 expression compared to controls. Sequence analysis indicated a putative α-helical coiled-coil segment as the only recognizable domain within the N-terminal half of the polypeptide, while the C-terminal half contains two PH, a MyTH4, and a FERM domain. We identified a phosphatidylinositol-3-phosphate consensus-binding site in the PH1 domain required for Plekhh2 localization to peripheral regions of cell lamellipodia. The N-terminal half of Plekkh2 is not necessary for lamellipodial targeting but mediates self-association. Yeast two-hybrid screening showed that Plekhh2 directly interacts through its FERM domain with the focal adhesion protein Hic-5 and actin. Plekhh2 and Hic-5 coprecipitated and colocalized at the soles of podocyte foot processes in situ and Hic-5 partially relocated from focal adhesions to lamellipodia in Plekhh2-expressing podocytes. In addition, Plekhh2 stabilizes the cortical actin cytoskeleton by attenuating actin depolymerization. Our findings suggest a structural and functional role for Plekhh2 in the podocyte foot processes. PMID:22832517

  20. Exploring the drug resistance of V32I and M46L mutant HIV-1 protease to inhibitor TMC114: flap dynamics and binding mechanism.

    Meher, Biswa Ranjan; Wang, Yixuan

    2015-03-01

    Inhibitors of HIV-1 protease (HIV-1-pr) generally only bind to the active site of the protease. However, for some mutants such as V32I and M46L the TMC114 can bind not only to the active cavity but also to the groove of the flexible flaps. Although the second binding site suggests the higher efficiency of the drug against HIV-1-pr, the drug resistance in HIV-1-pr due to mutations cannot be ignored, which prompts us to investigate the molecular mechanisms of drug resistance and behavior of double bound TMC114 (2T) to HIV-1-pr. The conformational dynamics of HIV-1-pr and the binding of TMC114 to the WT, V32I and M46L mutants were investigated with all-atom molecular dynamic (MD) simulation. The 20 ns MD simulation shows many fascinating effects of the inhibitor binding to the WT and mutant proteases. MM-PBSA calculations explain the binding free energies unfavorable for the M46L and V32I mutants as compared to the WT. For the single binding (1T) the less binding affinity can be attributed to the entropic loss for both V32I-1T and M46L-1T. Although the second binding of TMC114 with flap does increase binding energy for the mutants (V32I-2T and M46L-2T), the considerable entropy loss results in the lower binding Gibbs free energies. Thus, binding of TMC114 in the flap region does not help much in the total gain in binding affinity of the system, which was verified from this study and thereby validating experiments. PMID:25562662

  1. Mechanics of Flapping Flight: Analytical Formulations of Unsteady Aerodynamics, Kinematic Optimization, Flight Dynamics, and Control

    Taneja, Jayant Kumar

    Electricity is an indispensable commodity to modern society, yet it is delivered via a grid architecture that remains largely unchanged over the past century. A host of factors are conspiring to topple this dated yet venerated design: developments in renewable electricity generation technology, policies to reduce greenhouse gas emissions, and advances in information technology for managing energy systems. Modern electric grids are emerging as complex distributed systems in which a portfolio of power generation resources, often incorporating fluctuating renewable resources such as wind and solar, must be managed dynamically to meet uncontrolled, time-varying demand. Uncertainty in both supply and demand makes control of modern electric grids fundamentally more challenging, and growing portfolios of renewables exacerbate the challenge. We study three electricity grids: the state of California, the province of Ontario, and the country of Germany. To understand the effects of increasing renewables, we develop a methodology to scale renewables penetration. Analyzing these grids yields key insights about rigid limits to renewables penetration and their implications in meeting long-term emissions targets. We argue that to achieve deep penetration of renewables, the operational model of the grid must be inverted, changing the paradigm from load-following supplies to supply-following loads. To alleviate the challenge of supply-demand matching on deeply renewable grids, we first examine well-known techniques, including altering management of existing supply resources, employing utility-scale energy storage, targeting energy efficiency improvements, and exercising basic demand-side management. Then, we create several instantiations of supply-following loads -- including refrigerators, heating and cooling systems, and laptop computers -- by employing a combination of sensor networks, advanced control techniques, and enhanced energy storage. We examine the capacity of each load

  2. Architecture and Connectivity Govern Actin Network Contractility.

    Ennomani, Hajer; Letort, Gaëlle; Guérin, Christophe; Martiel, Jean-Louis; Cao, Wenxiang; Nédélec, François; De La Cruz, Enrique M; Théry, Manuel; Blanchoin, Laurent

    2016-03-01

    Actomyosin contractility plays a central role in a wide range of cellular processes, including the establishment of cell polarity, cell migration, tissue integrity, and morphogenesis during development. The contractile response is variable and depends on actomyosin network architecture and biochemical composition. To determine how this coupling regulates actomyosin-driven contraction, we used a micropatterning method that enables the spatial control of actin assembly. We generated a variety of actin templates and measured how defined actin structures respond to myosin-induced forces. We found that the same actin filament crosslinkers either enhance or inhibit the contractility of a network, depending on the organization of actin within the network. Numerical simulations unified the roles of actin filament branching and crosslinking during actomyosin contraction. Specifically, we introduce the concept of "network connectivity" and show that the contractions of distinct actin architectures are described by the same master curve when considering their degree of connectivity. This makes it possible to predict the dynamic response of defined actin structures to transient changes in connectivity. We propose that, depending on the connectivity and the architecture, network contraction is dominated by either sarcomeric-like or buckling mechanisms. More generally, this study reveals how actin network contractility depends on its architecture under a defined set of biochemical conditions. PMID:26898468

  3. Disrupted dynamics of F-actin and insulin granule fusion in INS-1 832/13 beta-cells exposed to glucotoxicity: partial restoration by glucagon-like peptide 1.

    Quinault, Aurore; Gausseres, Blandine; Bailbe, Danielle; Chebbah, Nella; Portha, Bernard; Movassat, Jamileh; Tourrel-Cuzin, Cecile

    2016-08-01

    Actin dynamics in pancreatic β-cells is involved in insulin exocytosis but the molecular mechanisms of this dynamics and its role in biphasic insulin secretion in pancreatic β-cells is largely unknown. Moreover, the impact of a glucotoxic environment on the sub-cortical actin network dynamics is poorly studied. In this study, we investigate the behavior of insulin granules and the subcortical actin network dynamics in INS-1 832/13 β-cells submitted to a normal or glucotoxic environment. Our results show that glucose stimulation leads to a reorganization of the subcortical actin network with a rupture of its interactions with t-SNARE proteins (Syntaxin 1A and SNAP-25), promoting insulin secretion in INS-1 832/13 β-cells. Prolonged exposure of INS-1 832/13 β-cells to high-glucose levels (glucotoxicity) leads to the densification of the cortical actin network, which prevents its reorganization under acute glucose, and diminishes the glucose-stimulated insulin secretion, as shown by the decreased number of fusion events. The most interesting in our results is the partial restoration by GLP-1 of the insulin secretion ability from high-glucose treated INS-1 832/13 cells. This improved insulin exocytosis is associated with partial restored actin dynamics and fusion events during the two phases of the secretion, with a preferential involvement of Epac2 signaling in the first phase and a rather involvement of PKA signaling in the second phase of insulin exocytosis. All these data provide some new insights into the mechanism by which current therapeutics may be improving insulin secretion. PMID:27101990

  4. Canonical and noncanonical g-protein signaling helps coordinate actin dynamics to promote macrophage phagocytosis of zymosan.

    Huang, Ning-Na; Becker, Steven; Boularan, Cedric; Kamenyeva, Olena; Vural, Ali; Hwang, Il-Young; Shi, Chong-Shan; Kehrl, John H

    2014-11-15

    Both chemotaxis and phagocytosis depend upon actin-driven cell protrusions and cell membrane remodeling. While chemoattractant receptors rely upon canonical G-protein signaling to activate downstream effectors, whether such signaling pathways affect phagocytosis is contentious. Here, we report that Gαi nucleotide exchange and signaling helps macrophages coordinate the recognition, capture, and engulfment of zymosan bioparticles. We show that zymosan exposure recruits F-actin, Gαi proteins, and Elmo1 to phagocytic cups and early phagosomes. Zymosan triggered an increase in intracellular Ca(2+) that was partially sensitive to Gαi nucleotide exchange inhibition and expression of GTP-bound Gαi recruited Elmo1 to the plasma membrane. Reducing GDP-Gαi nucleotide exchange, decreasing Gαi expression, pharmacologically interrupting Gβγ signaling, or reducing Elmo1 expression all impaired phagocytosis, while favoring the duration that Gαi remained GTP bound promoted it. Our studies demonstrate that targeting heterotrimeric G-protein signaling offers opportunities to enhance or retard macrophage engulfment of phagocytic targets such as zymosan. PMID:25225330

  5. Pedicled perforator flaps

    Demirtas, Yener; Ozturk, Nuray; Kelahmetoglu, Osman; Demir, Ahmet

    2009-01-01

    Described in this study is a surgical concept that supports the "consider and use a pedicled perforator flap whenever possible and indicated" approach to reconstruct a particular skin defect. The operation is entirely free-style; the only principle is to obtain a pedicled perforator flap to...... reconstruct the defect. The perforators are marked with a hand-held Doppler probe and multiple flaps are designed. The appropriate flap is elevated after identifying the perforator(s). Dissection of the perforator(s) or complete incision of the flap margins are not mandatory if the flap is mobilized...... adequately to cover the defect. Defects measuring 3 x 3 cm up to 20 x 20 cm at diverse locations were successfully reconstructed in 20 of 21 patients with 26 flaps. Pedicled perforator flaps offer us reliable and satisfactory results of reconstruction at different anatomic territories of the body. It sounds...

  6. Addition of Phenylboronic Acid to Malus domestica Pollen Tubes Alters Calcium Dynamics, Disrupts Actin Filaments and Affects Cell Wall Architecture

    Fang, Kefeng; Gao, Sai; Zhang, Weiwei; Xing, Yu; Cao, Qingqin; Qin, Ling

    2016-01-01

    A key role of boron in plants is to cross-link the cell wall pectic polysaccharide rhamnogalacturonan-II (RG-II) through borate diester linkages. Phenylboronic acid (PBA) can form the same reversible ester bonds but cannot cross-link two molecules, so can be used as an antagonist to study the function of boron. This study aimed to evaluate the effect of PBA on apple (Malus domestica) pollen tube growth and the underlying regulatory mechanism. We observed that PBA caused an inhibition of pollen germination, tube growth and led to pollen tube morphological abnormalities. Fluorescent labeling, coupled with a scanning ion-selective electrode technique, revealed that PBA induced an increase in extracellular Ca2+ influx, thereby elevating the cytosolic Ca2+ concentration [Ca2+]c and disrupting the [Ca2+]c gradient, which is critical for pollen tube growth. Moreover the organization of actin filaments was severely perturbed by the PBA treatment. Immunolocalization studies and fluorescent labeling, together with Fourier-transform infrared analysis (FTIR) suggested that PBA caused an increase in the abundance of callose, de-esterified pectins and arabinogalactan proteins (AGPs) at the tip. However, it had no effect on the deposition of the wall polymers cellulose. These effects are similar to those of boron deficiency in roots and other organs, indicating that PBA can induce boron deficiency symptoms. The results provide new insights into the roles of boron in pollen tube development, which likely include regulating [Ca2+]c and the formation of the actin cytoskeleton, in addition to the synthesis and assembly of cell wall components. PMID:26886907

  7. Cellular prion protein is required for neuritogenesis: fine-tuning of multiple signaling pathways involved in focal adhesions and actin cytoskeleton dynamics

    Alleaume-Butaux A

    2013-07-01

    Full Text Available Aurélie Alleaume-Butaux,1,2 Caroline Dakowski,1,2 Mathéa Pietri,1,2 Sophie Mouillet-Richard,1,2 Jean-Marie Launay,3,4 Odile Kellermann,1,2 Benoit Schneider1,2 1INSERM, UMR-S 747, 2Paris Descartes University, Sorbonne Paris Cité, UMR-S 747, 3Public Hospital of Paris, Department of Biochemistry, INSERM UMR-S 942, Lariboisière Hospital, Paris, France; 4Pharma Research Department, Hoffmann La Roche Ltd, Basel, Switzerland Abstract: Neuritogenesis is a dynamic phenomenon associated with neuronal differentiation that allows a rather spherical neuronal stem cell to develop dendrites and axon, a prerequisite for the integration and transmission of signals. The acquisition of neuronal polarity occurs in three steps: (1 neurite sprouting, which consists of the formation of buds emerging from the postmitotic neuronal soma; (2 neurite outgrowth, which represents the conversion of buds into neurites, their elongation and evolution into axon or dendrites; and (3 the stability and plasticity of neuronal polarity. In neuronal stem cells, remodeling and activation of focal adhesions (FAs associated with deep modifications of the actin cytoskeleton is a prerequisite for neurite sprouting and subsequent neurite outgrowth. A multiple set of growth factors and interactors located in the extracellular matrix and the plasma membrane orchestrate neuritogenesis by acting on intracellular signaling effectors, notably small G proteins such as RhoA, Rac, and Cdc42, which are involved in actin turnover and the dynamics of FAs. The cellular prion protein (PrPC, a glycosylphosphatidylinositol (GPI-anchored membrane protein mainly known for its role in a group of fatal neurodegenerative diseases, has emerged as a central player in neuritogenesis. Here, we review the contribution of PrPC to neuronal polarization and detail the current knowledge on the signaling pathways fine-tuned by PrPC to promote neurite sprouting, outgrowth, and maintenance. We emphasize that Pr

  8. [Cytoskeletal actin and its associated proteins. Some examples in Protista].

    Guillén, N; Carlier, M F; Brugerolle, G; Tardieux, I; Ausseil, J

    1998-06-01

    Many processes, cell motility being an example, require cells to remodel the actin cytoskeleton in response to both intracellular and extracellular signals. Reorganization of the actin cytoskeleton involves the rapid disassembly and reassembly of actin filaments, a phenomenon regulated by the action of particular actin-binding proteins. In recent years, an interest in studying actin regulation in unicellular organisms has arisen. Parasitic protozoan are among these organisms and studies of the cytoskeleton functions of these protozoan are relevant related to either cell biology or pathogenicity. To discuss recent data in this field, a symposium concerning "Actin and actin-binding proteins in protists" was held on May 8-11 in Paris, France, during the XXXV meeting of the French Society of Protistology. As a brief summary of the symposium we report here findings concerning the in vitro actin dynamic assembly, as well as the characterization of several actin-binding proteins from the parasitic protozoan Entamoeba histolytica, Trichomonas vaginalis and Plasmodium knowlesi. In addition, localization of actin in non-pathogen protists such as Prorocentrum micans and Crypthecodinium cohnii is also presented. The data show that some actin-binding proteins facilitate organization of filaments into higher order structures as pseudopods, while others have regulatory functions, indicating very particular roles for actin-binding proteins. One of the proteins discussed during the symposium, the actin depolymerizing factor ADF, was shown to enhance the treadmilling rate of actin filaments. In vitro, ADF binds to the ADP-bound forms of G-actin and F-actin, thereby participating in and changing the rate of actin assembly. Biochemical approaches allowed the identification of a protein complex formed by HSP/C70-cap32-34 which might also be involved in depolymerization of F-actin in P. knowlesi. Molecular and cellular approaches were used to identify proteins such as ABP-120 and myosin

  9. Polarity protein Crumbs homolog-3 (CRB3) regulates ectoplasmic specialization dynamics through its action on F-actin organization in Sertoli cells

    Gao, Ying; Lui, Wing-yee; Lee, Will M.; Cheng, C. Yan

    2016-01-01

    Crumbs homolog 3 (or Crumbs3, CRB3) is a polarity protein expressed by Sertoli and germ cells at the basal compartment in the seminiferous epithelium. CRB3 also expressed at the blood-testis barrier (BTB), co-localized with F-actin, TJ proteins occludin/ZO-1 and basal ES (ectoplasmic specialization) proteins N-cadherin/β-catenin at stages IV-VII only. The binding partners of CRB3 in the testis were the branched actin polymerization protein Arp3, and the barbed end-capping and bundling protein Eps8, illustrating its possible role in actin organization. CRB3 knockdown (KD) by RNAi in Sertoli cells with an established tight junction (TJ)-permeability barrier perturbed the TJ-barrier via changes in the distribution of TJ- and basal ES-proteins at the cell-cell interface. These changes were the result of CRB3 KD-induced re-organization of actin microfilaments, in which actin microfilaments were truncated, and extensively branched, thereby destabilizing F-actin-based adhesion protein complexes at the BTB. Using Polyplus in vivo-jetPEI as a transfection medium with high efficiency for CRB3 KD in the testis, the CRB3 KD testes displayed defects in spermatid and phagosome transport, and also spermatid polarity due to a disruption of F-actin organization. In summary, CRB3 is an actin microfilament regulator, playing a pivotal role in organizing actin filament bundles at the ES. PMID:27358069

  10. Design of Insect-Scale Flapping Wing Vehicles

    Ahmed Nabawy, Mostafa Ramadan

    2015-01-01

    This thesis contributes to the state of the art in integrated design of insect-scale piezoelectric actuated flapping wing vehicles through the development of novel theoretical models for flapping wing aerodynamics and piezoelectric actuator dynamics, and integration of these models into a closed form design process.A comprehensive literature review of available engineered designs of miniature rotary and flapping wing vehicles is provided. A novel taxonomy based on wing and actuator kinematics...

  11. Stimulation of Actin Polymerization by Filament Severing

    Carlsson, A E

    2005-01-01

    The extent and dynamics of actin polymerization in solution are calculated as functions of the filament severing rate, using a simple model of in vitro polymerization. The model is solved by both analytic theory and stochastic-growth simulation. The results show that severing essentially always enhances actin polymerization by freeing up barbed ends, if barbed-end cappers are present. Severing has much weaker effects if only pointed-end cappers are present. In the early stages of polymerizati...

  12. Computational Study of the Binding Mechanism of Actin-Depolymerizing Factor 1 with Actin in Arabidopsis thaliana.

    Du, Juan; Wang, Xue; Dong, Chun-Hai; Yang, Jian Ming; Yao, Xiao Jun

    2016-01-01

    Actin is a highly conserved protein. It plays important roles in cellular function and exists either in the monomeric (G-actin) or polymeric form (F-actin). Members of the actin-depolymerizing factor (ADF)/cofilin protein family bind to both G-actin and F-actin and play vital roles in actin dynamics by manipulating the rates of filament polymerization and depolymerization. It has been reported that the S6D and R98A/K100A mutants of actin-depolymerizing factor 1 (ADF1) in Arabidopsis thaliana decreased the binding affinity of ADF for the actin monomer. To investigate the binding mechanism and dynamic behavior of the ADF1-actin complex, we constructed a homology model of the AtADF1-actin complex based on the crystal structure of AtADF1 and the twinfilin C-terminal ADF-H domain in a complex with a mouse actin monomer. The model was then refined for subsequent molecular dynamics simulations. Increased binding energy of the mutated system was observed using the Molecular Mechanics Generalized Born Surface Area and Poisson-Boltzmann Surface Area (MM-GB/PBSA) methods. To determine the residues that make decisive contributions to the ADF1 actin-binding affinity, per-residue decomposition and computational alanine scanning analyses were performed, which provided more detailed information on the binding mechanism. Root-mean-square fluctuation and principal component analyses confirmed that the S6D and R98A/K100A mutants induced an increased conformational flexibility. The comprehensive molecular insight gained from this study is of great importance for understanding the binding mechanism of ADF1 and G-actin. PMID:27414648

  13. The platysma myocutaneous flap.

    Baur, Dale A; Williams, Jonathan; Alakaily, Xena

    2014-08-01

    Reconstructing defects of the oral mucosa or skin of the lower one-third of the face can be accomplished by a variety of techniques. This article presents two versions of the platysma myocutaneous flap, which is a reliable, axial pattern, pedicled flap capable of providing excellent one-stage reconstruction of such defects. As discussed herein, the superiorly based and posteriorly based versions of the flap have wide application in the oral and facial region. Also provided is a review of other uses of this flap in head and neck surgery. PMID:24958382

  14. Propeller TAP flap

    Thomsen, Jørn Bo; Bille, Camilla; Wamberg, Peter;

    2013-01-01

    The aim of this study was to examine if a propeller thoracodorsal artery perforator (TAP) flap can be used for breast reconstruction. Fifteen women were reconstructed using a propeller TAP flap, an implant, and an ADM. Preoperative colour Doppler ultrasonography was used for patient selection to...... major complications needing additional surgery. One flap was lost due to a vascular problem. Breast reconstruction can be performed by a propeller TAP flap without cutting the descending branch of the thoracodorsal vessels. However, the authors would recommend that a small cuff of muscle is left around...

  15. Energetic modeling and single-molecule verification of dynamic regulation on receptor protein diffusion by actin corrals and lipid raft domains receptor

    Lin, Chien Yu; Huang, Jung Y.; Lo, Leu-Wei

    2015-03-01

    To faithfully estimate a signal that varies in both space and time, the optimization strategy used by a live cell is to organize a collection of distributed and mobile receptors into a mobile active clustering. However, living eukaryotic cells are highly heterogeneous and stochastically dynamic. It is therefore important to develop an energetic model based on fundamental laws to verify that the underlying processes are energetically favorable. We developed an energetic model based on the generalized Langevin equation and the Cahn-Hilliard equation to simulate the diffusive behaviors of receptor proteins in the plasma membrane with a hierarchical structure of actin corrals, lipid domains, and receptor proteins. Single-molecule tracking data of EGFR acquired on live HeLa cells agrees with the simulation results. We discovered that after ligand binding, EGFR molecules move into lipid nanodomains. The transition rates between different diffusion states of liganded EGFR molecules are regulated by the lipid domains. Our method captures both the sensitivity of single-molecule processes, statistic accuracy of data analysis, and the hierarchical structure of plasma membranes.

  16. TccP2-mediated subversion of actin dynamics by EPEC 2 – a distinct evolutionary lineage of enteropathogenic Escherichia coli

    Whale, Andrew D.; Hernandes, Rodrigo T.; Ooka, Tadasuke; Beutin, Lothar; Schüller, Stephanie; Garmendia, Junkal; Crowther, Lynette; Vieira, Mônica A. M.; Ogura, Yoshitoshi; Krause, Gladys; Phillips, Alan D.; Tania A. T. Gomes; Hayashi, Tetsuya; Frankel, Gad

    2007-01-01

    Enteropathogenic Escherichia coli (EPEC) is a major cause of infantile diarrhoea in developing countries. While colonizing the gut mucosa, EPEC triggers extensive actin-polymerization activity at the site of intimate bacterial attachment, which is mediated by avid interaction between the outer-membrane adhesin intimin and the type III secretion system (T3SS) effector Tir. The prevailing dogma is that actin polymerization by EPEC is achieved following tyrosine phosphorylation of Tir, recruitme...

  17. Probing GFP-actin diffusion in living cells using fluorescence correlation spectroscopy

    The cytoskeleton of eukaryotic cells is continuously remodeled by polymerization and depolymerization of actin. Consequently, the relative content of polymerized filamentous actin (F-actin) and monomeric globular actin (G-actin) is subject to temporal and spatial fluctuations. Since fluorescence correlation spectroscopy (FCS) can measure the diffusion of fluorescently labeled actin it seems likely that FCS allows us to determine the dynamics and hence indirectly the structural properties of the cytoskeleton components with high spatial resolution. To this end we investigate the FCS signal of GFP-actin in living Dictyostelium discoideum cells and explore the inherent spatial and temporal signatures of the actin cytoskeleton. Using the free green fluorescent protein (GFP) as a reference, we find that actin diffusion inside cells is dominated by G-actin and slower than diffusion in diluted cell extract. The FCS signal in the dense cortical F-actin network near the cell membrane is probed using the cytoskeleton protein LIM and is found to be slower than cytosolic G-actin diffusion. Furthermore, we show that polymerization of the cytoskeleton induced by Jasplakinolide leads to a substantial decrease of G-actin diffusion. Pronounced fluctuations in the distribution of the FCS correlation curves can be induced by latrunculin, which is known to induce actin waves. Our work suggests that the FCS signal of GFP-actin in combination with scanning or spatial correlation techniques yield valuable information about the local dynamics and concomitant cytoskeletal properties

  18. The rhombic bilobed flap, a simple, geometrically designed flap

    Yoshiaki Sakamoto

    2014-01-01

    Full Text Available We describe a combination of the common rhomboid flap and bilobed flap and provide an example of its use. The rhombic bilobed flap is simple to use and is associated with fewer complications, such as pin-cushioning and standing cone deformities, while minimizing the risk of skin necrosis and tension on the flap.

  19. A novel method to study the electrodynamic behavior of actin filaments. Evidence for cable-like properties of actin.

    Lin, E C; Cantiello, H. F.

    1993-01-01

    Actin, one of the most abundant intracellular proteins, forms long linear polyelectrolytic polymers in solution. A novel technique to handle single actin filaments in solution was developed that allows the study of ionic currents elicited along the surface of electrically stimulated actin filaments. Electrical currents were observed about the polymer's surface under both high (100 mM KCl) and low (1 mM KCl) ionic strength conditions. The data are consistent with a dynamic behavior of the coun...

  20. Aerodynamic flight performance in flap-gliding birds and bats.

    Muijres, Florian T; Henningsson, Per; Stuiver, Melanie; Hedenström, Anders

    2012-08-01

    Many birds use a flight mode called undulating or flap-gliding flight, where they alternate between flapping and gliding phases, while only a few bats make use of such a flight mode. Among birds, flap-gliding is commonly used by medium to large species, where it is regarded to have a lower energetic cost than continuously flapping flight. Here, we introduce a novel model for estimating the energetic flight economy of flap-gliding animals, by determining the lift-to-drag ratio for flap-gliding based on empirical lift-to-drag ratio estimates for continuous flapping flight and for continuous gliding flight, respectively. We apply the model to flight performance data of the common swift (Apus apus) and of the lesser long-nosed bat (Leptonycteris yerbabuenae). The common swift is a typical flap-glider while-to the best of our knowledge-the lesser long-nosed bat does not use flap-gliding. The results show that, according to the model, the flap-gliding common swift saves up to 15% energy compared to a continuous flapping swift, and that this is primarily due to the exceptionally high lift-to-drag ratio in gliding flight relative to that in flapping flight for common swifts. The lesser long-nosed bat, on the other hand, seems not to be able to reduce energetic costs by flap-gliding. The difference in relative costs of flap-gliding flight between the common swift and the lesser long-nosed bat can be explained by differences in morphology, flight style and wake dynamics. The model presented here proves to be a valuable tool for estimating energetic flight economy in flap-gliding animals. The results show that flap-gliding flight that is naturally used by common swifts is indeed the most economic one of the two flight modes, while this is not the case for the non-flap-gliding lesser long-nosed bat. PMID:22726811

  1. Sequence and comparative genomic analysis of actin-related proteins.

    Muller, Jean; Oma, Yukako; Vallar, Laurent; Friederich, Evelyne; Poch, Olivier; Winsor, Barbara

    2005-12-01

    Actin-related proteins (ARPs) are key players in cytoskeleton activities and nuclear functions. Two complexes, ARP2/3 and ARP1/11, also known as dynactin, are implicated in actin dynamics and in microtubule-based trafficking, respectively. ARP4 to ARP9 are components of many chromatin-modulating complexes. Conventional actins and ARPs codefine a large family of homologous proteins, the actin superfamily, with a tertiary structure known as the actin fold. Because ARPs and actin share high sequence conservation, clear family definition requires distinct features to easily and systematically identify each subfamily. In this study we performed an in depth sequence and comparative genomic analysis of ARP subfamilies. A high-quality multiple alignment of approximately 700 complete protein sequences homologous to actin, including 148 ARP sequences, allowed us to extend the ARP classification to new organisms. Sequence alignments revealed conserved residues, motifs, and inserted sequence signatures to define each ARP subfamily. These discriminative characteristics allowed us to develop ARPAnno (http://bips.u-strasbg.fr/ARPAnno), a new web server dedicated to the annotation of ARP sequences. Analyses of sequence conservation among actins and ARPs highlight part of the actin fold and suggest interactions between ARPs and actin-binding proteins. Finally, analysis of ARP distribution across eukaryotic phyla emphasizes the central importance of nuclear ARPs, particularly the multifunctional ARP4. PMID:16195354

  2. Actin is required for IFT regulation in Chlamydomonas reinhardtii.

    Avasthi, Prachee; Onishi, Masayuki; Karpiak, Joel; Yamamoto, Ryosuke; Mackinder, Luke; Jonikas, Martin C; Sale, Winfield S; Shoichet, Brian; Pringle, John R; Marshall, Wallace F

    2014-09-01

    Assembly of cilia and flagella requires intraflagellar transport (IFT), a highly regulated kinesin-based transport system that moves cargo from the basal body to the tip of flagella [1]. The recruitment of IFT components to basal bodies is a function of flagellar length, with increased recruitment in rapidly growing short flagella [2]. The molecular pathways regulating IFT are largely a mystery. Because actin network disruption leads to changes in ciliary length and number, actin has been proposed to have a role in ciliary assembly. However, the mechanisms involved are unknown. In Chlamydomonas reinhardtii, conventional actin is found in both the cell body and the inner dynein arm complexes within flagella [3, 4]. Previous work showed that treating Chlamydomonas cells with the actin-depolymerizing compound cytochalasin D resulted in reversible flagellar shortening [5], but how actin is related to flagellar length or assembly remains unknown. Here we utilize small-molecule inhibitors and genetic mutants to analyze the role of actin dynamics in flagellar assembly in Chlamydomonas reinhardtii. We demonstrate that actin plays a role in IFT recruitment to basal bodies during flagellar elongation and that when actin is perturbed, the normal dependence of IFT recruitment on flagellar length is lost. We also find that actin is required for sufficient entry of IFT material into flagella during assembly. These same effects are recapitulated with a myosin inhibitor, suggesting that actin may act via myosin in a pathway by which flagellar assembly is regulated by flagellar length. PMID:25155506

  3. Folding in and out: passive morphing in flapping wings.

    Stowers, Amanda K; Lentink, David

    2015-04-01

    We present a new mechanism for passive wing morphing of flapping wings inspired by bat and bird wing morphology. The mechanism consists of an unactuated hand wing connected to the arm wing with a wrist joint. Flapping motion generates centrifugal accelerations in the hand wing, forcing it to unfold passively. Using a robotic model in hover, we made kinematic measurements of unfolding kinematics as functions of the non-dimensional wingspan fold ratio (2-2.5) and flapping frequency (5-17 Hz) using stereo high-speed cameras. We find that the wings unfold passively within one to two flaps and remain unfolded with only small amplitude oscillations. To better understand the passive dynamics, we constructed a computer model of the unfolding process based on rigid body dynamics, contact models, and aerodynamic correlations. This model predicts the measured passive unfolding within about one flap and shows that unfolding is driven by centrifugal acceleration induced by flapping. The simulations also predict that relative unfolding time only weakly depends on flapping frequency and can be reduced to less than half a wingbeat by increasing flapping amplitude. Subsequent dimensional analysis shows that the time required to unfold passively is of the same order of magnitude as the flapping period. This suggests that centrifugal acceleration can drive passive unfolding within approximately one wingbeat in small and large wings. Finally, we show experimentally that passive unfolding wings can withstand impact with a branch, by first folding and then unfolding passively. This mechanism enables flapping robots to squeeze through clutter without sophisticated control. Passive unfolding also provides a new avenue in morphing wing design that makes future flapping morphing wings possibly more energy efficient and light-weight. Simultaneously these results point to possible inertia driven, and therefore metabolically efficient, control strategies in bats and birds to morph or recover

  4. Steric effects induce geometric remodeling of actin bundles in filopodia

    Dobramysl, Ulrich; Erban, Radek

    2016-01-01

    Filopodia are ubiquitous fingerlike protrusions, spawned by many eukaryotic cells, to probe and interact with their environments. Polymerization dynamics of actin filaments, comprising the structural core of filopodia, largely determine their instantaneous lengths and overall lifetimes. The polymerization reactions at the filopodial tip require transport of G-actin, which enter the filopodial tube from the filopodial base and diffuse toward the filament barbed ends near the tip. Actin filaments are mechanically coupled into a tight bundle by cross-linker proteins. Interestingly, many of these proteins are relatively short, restricting the free diffusion of cytosolic G-actin throughout the bundle and, in particular, its penetration into the bundle core. To investigate the effect of steric restrictions on G-actin diffusion by the porous structure of filopodial actin filament bundle, we used a particle-based stochastic simulation approach. We discovered that excluded volume interactions result in partial and the...

  5. Histamine activates p38 MAP kinase and alters local lamellipodia dynamics, reducing endothelial barrier integrity and eliciting central movement of actin fibers.

    Adderley, Shaquria P; Lawrence, Curtis; Madonia, Eyong; Olubadewo, Joseph O; Breslin, Jerome W

    2015-07-01

    The role of the actin cytoskeleton in endothelial barrier function has been debated for nearly four decades. Our previous investigation revealed spontaneous local lamellipodia in confluent endothelial monolayers that appear to increase overlap at intercellular junctions. We tested the hypothesis that the barrier-disrupting agent histamine would reduce local lamellipodia protrusions and investigated the potential involvement of p38 mitogen-activated protein (MAP) kinase activation and actin stress fiber formation. Confluent monolayers of human umbilical vein endothelial cells (HUVEC) expressing green fluorescent protein-actin were studied using time-lapse fluorescence microscopy. The protrusion and withdrawal characteristics of local lamellipodia were assessed before and after addition of histamine. Changes in barrier function were determined using electrical cell-substrate impedance sensing. Histamine initially decreased barrier function, lamellipodia protrusion frequency, and lamellipodia protrusion distance. A longer time for lamellipodia withdrawal and reduced withdrawal distance and velocity accompanied barrier recovery. After barrier recovery, a significant number of cortical fibers migrated centrally, eventually resembling actin stress fibers. The p38 MAP kinase inhibitor SB203580 attenuated the histamine-induced decreases in barrier function and lamellipodia protrusion frequency. SB203580 also inhibited the histamine-induced decreases in withdrawal distance and velocity, and the subsequent actin fiber migration. These data suggest that histamine can reduce local lamellipodia protrusion activity through activation of p38 MAP kinase. The findings also suggest that local lamellipodia have a role in maintaining endothelial barrier integrity. Furthermore, we provide evidence that actin stress fiber formation may be a reaction to, rather than a cause of, reduced endothelial barrier integrity. PMID:25948734

  6. Flapping of Insectile Wings

    Huang, Yangyang; Kanso, Eva

    2015-11-01

    Insects use flight muscles attached at the base of the wings to produce impressive wing flapping frequencies. Yet the effects of muscle stiffness on the performance of insect wings remain unclear. Here, we construct an insectile wing model, consisting of two rigid wings connected at their base by an elastic torsional spring and submerged in an oscillatory flow. The wing system is free to rotate and flap. We first explore the extent to which the flyer can withstand roll perturbations, then study its flapping behavior and performance as a function of spring stiffness. We find an optimal range of spring stiffness that results in large flapping amplitudes, high force generation and good storage of elastic energy. We conclude by conjecturing that insects may select and adjust the muscle spring stiffness to achieve desired movement. These findings may have significant implications on the design principles of wings in micro air-vehicles.

  7. Control of Flap Vortices

    Greenblatt, David

    2005-01-01

    A wind tunnel investigation was carried out on a semi-span wing model to assess the feasibility of controlling vortices emanating from outboard flaps and tip-flaps by actively varying the degree of boundary layer separation. Separation was varied by means of perturbations produced from segmented zero-efflux oscillatory blowing slots, while estimates of span loadings and vortex sheet strengths were obtained by integrating wing surface pressures. These estimates were used as input to inviscid rollup relations as a means of predicting changes to the vortex characteristics resulting from the perturbations. Surveys of flow in the wake of the outboard and tip-flaps were made using a seven-hole probe, from which the vortex characteristics were directly deduced. Varying the degree of separation had a marked effect on vortex location, strength, tangential velocity, axial velocity and size for both outboard and tip-flaps. Qualitative changes in vortex characteristics were well predicted by the inviscid rollup relations, while the failure to account for viscosity was presumed to be the main reason for observed discrepancies. Introducing perturbations near the outboard flap-edges or on the tip-flap exerted significant control over vortices while producing negligible lift excursions.

  8. Quantifying Blood Flow in the DIEP Flap: An Ultrasonographic Study

    Pennington, David G.

    2014-01-01

    Background: The maximum weight of tissue that a single perforator can perfuse remains an important question in reconstructive microsurgery. An empirically based equation, known as the flap viability index (FVI), has been established to determine what weight of tissue will survive on one or more perforators. The equation is FVI = Sum d(n)^4/W, where d is the internal diameter of each perforator and W is the final weight of the flap. It has been shown that if FVI exceeds 10, total flap survival is likely, but if under 10, partial flap necrosis is probable. The aim of this study was to measure absolute flow rates in deep inferior epigastric perforator (DIEP) flap pedicles and assess correlation with the determinants of the FVI, perforator diameter and flap weight. Methods: Color Doppler ultrasound was used to quantify arterial flow in 10 consecutive DIEP flap pedicles 24 hours after anastomosis. Results: In single-perforator DIEP flaps, flow rate was highly correlated with perforator diameter (r = 0.82, P = 0.01). Mean arterial flow rate was significantly reduced in DIEP flaps with 2 or more perforators (6 vs 38 cm3/min; P perforator size is a critical factor in optimizing blood flow in perforator-based free tissue transfer. Further research is required to understand the flow dynamics of perforator flaps based on multiple perforators. However, surgeons should be cognizant that a single large perforator may have substantially higher flow rates than multiple small perforators. Routine FVI calculation is recommended to ensure complete flap survival. PMID:25426345

  9. Perforator Flaps for Perineal Reconstructions

    Niranjan, Niri S.

    2006-01-01

    Whenever there is soft tissue loss from the perineum there are many options for reconstruction. These include allowing the wound to heal by secondary intention and the use of local random or axial pattern flaps, regional flaps, or free flaps. The axial skin flap can be defined as a flap based on known constant vessels of the subcutaneous tissue and its vena comitantes. The perforator flap on the other hand is a randomly selected perforator consisting of an artery with vena comitantes, which p...

  10. Orientational Order of the Lamellipodial Actin Network as Demonstrated in Living Motile CellsV⃞

    Alexander B. Verkhovsky; Chaga, Oleg Y.; Schaub, Sébastien; Svitkina, Tatyana M.; Meister, Jean-Jacques; Borisy, Gary G.

    2003-01-01

    Lamellipodia of crawling cells represent both the motor for cell advance and the primary building site for the actin cytoskeleton. The organization of actin in the lamellipodium reflects actin dynamics and is of critical importance for the mechanism of cell motility. In previous structural studies, the lamellipodial actin network was analyzed primarily by electron microscopy (EM). An understanding of lamellipodial organization would benefit significantly if the EM data were complemented and p...

  11. Cell Elasticity Is Regulated by the Tropomyosin Isoform Composition of the Actin Cytoskeleton

    Jalilian, Iman; Heu, Celine; Cheng, Hong; Freittag, Hannah; Desouza, Melissa; Justine R. Stehn; Bryce, Nicole S.; Whan, Renee M.; Hardeman, Edna C.; Fath, Thomas; Schevzov, Galina; Gunning, Peter W.

    2015-01-01

    The actin cytoskeleton is the primary polymer system within cells responsible for regulating cellular stiffness. While various actin binding proteins regulate the organization and dynamics of the actin cytoskeleton, the proteins responsible for regulating the mechanical properties of cells are still not fully understood. In the present study, we have addressed the significance of the actin associated protein, tropomyosin (Tpm), in influencing the mechanical properties of cells. Tpms belong to...

  12. Cooperation between actin-binding proteins of invasive Salmonella: SipA potentiates SipC nucleation and bundling of actin

    Emma J McGhie; Hayward, Richard D.; Koronakis, Vassilis

    2001-01-01

    Pathogen-induced remodelling of the host cell actin cytoskeleton drives internalization of invasive Salmon ella by non-phagocytic intestinal epithelial cells. Two Salmonella actin-binding proteins are involved in internalization: SipC is essential for the process, while SipA enhances its efficiency. Using purified SipC and SipA proteins in in vitro assays of actin dynamics and F-actin bundling, we demonstrate that SipA stimulates substantially SipC-mediated nucleation of actin polymerization....

  13. The Evolution of Perforator Flaps

    Khan, Farah N.; Spiegel, Aldona J.

    2006-01-01

    Perforator flaps have recently become ubiquitous in the field of plastic surgery. To understand and appreciate their unique nature, it is necessary to compare and contrast them with the development of other types of flaps. A complete yet abridged version of the history of flap surgery is presented in this article. Beginning with Sushruta's Indian cheek flap method for nasal reconstruction, a trip through time and space is taken to highlight the milestones leading to the evolution of the perfo...

  14. The ubiquitin C-terminal hydrolase UCH-L1 promotes bacterial invasion by altering the dynamics of the actin cytoskeleton

    Basseres, Eugene; Coppotelli, Giuseppe; Pfirrmann, Thorsten;

    2010-01-01

    Invasion of eukaryotic target cells by pathogenic bacteria requires extensive remodelling of the membrane and actin cytoskeleton. Here we show that the remodelling process is regulated by the ubiquitin C-terminal hydrolase UCH-L1 that promotes the invasion of epithelial cells by Listeria monocyto...... findings highlight a previously unrecognized involvement of the ubiquitin cycle in bacterial entry. UCH-L1 is highly expressed in malignant cells that may therefore be particularly susceptible to invasion by bacteria-based drug delivery systems.......Invasion of eukaryotic target cells by pathogenic bacteria requires extensive remodelling of the membrane and actin cytoskeleton. Here we show that the remodelling process is regulated by the ubiquitin C-terminal hydrolase UCH-L1 that promotes the invasion of epithelial cells by Listeria...

  15. Membrane waves driven by forces from actin filaments

    Membrane waves propagating along the cell circumference in a top down view have been observed with several eukaryotic cells (Döbereiner et al 2006 Phys. Rev. Lett. 97 10; Machacek and Danuser 2006 Biophys. J. 90 1439–52). We present a mathematical model reproducing these traveling membrane undulations during lamellipodial motility of cells on flat substrates. The model describes the interplay of pushing forces exerted by actin polymerization on the membrane, pulling forces of attached actin filaments on the cell edge, contractile forces powered by molecular motors across the actin gel and resisting membrane tension. The actin filament network in the bulk of lamellipodia obeys gel flow equations. We investigated in particular the dependence of wave properties on gel parameters and found that inhibition of myosin motors abolishes waves in some cells but not in others in agreement with experimental observations. The model provides a unifying mechanism explaining the dynamics of actin-based motility in a variety of systems. (paper)

  16. The Actin-Binding Protein α-Adducin Is Required for Maintaining Axon Diameter.

    Leite, Sérgio Carvalho; Sampaio, Paula; Sousa, Vera Filipe; Nogueira-Rodrigues, Joana; Pinto-Costa, Rita; Peters, Luanne Laurel; Brites, Pedro; Sousa, Mónica Mendes

    2016-04-19

    The actin-binding protein adducin was recently identified as a component of the neuronal subcortical cytoskeleton. Here, we analyzed mice lacking adducin to uncover the function of this protein in actin rings. α-adducin knockout mice presented progressive axon enlargement in the spinal cord and optic and sciatic nerves, followed by axon degeneration and loss. Using stimulated emission depletion super-resolution microscopy, we show that a periodic subcortical actin cytoskeleton is assembled in every neuron type inspected including retinal ganglion cells and dorsal root ganglia neurons. In neurons devoid of adducin, the actin ring diameter increased, although the inter-ring periodicity was maintained. In vitro, the actin ring diameter adjusted as axons grew, suggesting the lattice is dynamic. Our data support a model in which adducin activity is not essential for actin ring assembly and periodicity but is necessary to control the diameter of both actin rings and axons and actin filament growth within rings. PMID:27068466

  17. Pattern formation in polymerising actin flocks: spirals, spots and waves without nonlinear chemistry

    Goff, Thomas Le; Marenduzzo, Davide

    2016-01-01

    We propose a model solely based on actin treadmilling and polymerisation which describes many characteristic states of actin wave formation: spots, spirals and travelling waves. In our model, as in experiments on cell recovering motility following actin depolymerisation, we choose an isotropic low density initial condition; polymerisation of actin filaments then raises the density towards the Onsager threshold where they align. We show that this alignment, in turn, destabilizes the isotropic phase and generically induces transient actin spots or spirals as part of the dynamical pathway towards a polarized phase which can either be uniform or consist of a series of actin-wave trains (flocks). Our results uncover a universal route to actin wave formation in the absence of any system specific nonlinear biochemistry, and it may help understand the mechanism underlying the observation of actin spots and waves in vivo. They also suggest a minimal setup to design similar patterns in vitro.

  18. Computational analysis of viscoelastic properties of crosslinked actin networks.

    Taeyoon Kim

    2009-07-01

    Full Text Available Mechanical force plays an important role in the physiology of eukaryotic cells whose dominant structural constituent is the actin cytoskeleton composed mainly of actin and actin crosslinking proteins (ACPs. Thus, knowledge of rheological properties of actin networks is crucial for understanding the mechanics and processes of cells. We used Brownian dynamics simulations to study the viscoelasticity of crosslinked actin networks. Two methods were employed, bulk rheology and segment-tracking rheology, where the former measures the stress in response to an applied shear strain, and the latter analyzes thermal fluctuations of individual actin segments of the network. It was demonstrated that the storage shear modulus (G' increases more by the addition of ACPs that form orthogonal crosslinks than by those that form parallel bundles. In networks with orthogonal crosslinks, as crosslink density increases, the power law exponent of G' as a function of the oscillation frequency decreases from 0.75, which reflects the transverse thermal motion of actin filaments, to near zero at low frequency. Under increasing prestrain, the network becomes more elastic, and three regimes of behavior are observed, each dominated by different mechanisms: bending of actin filaments, bending of ACPs, and at the highest prestrain tested (55%, stretching of actin filaments and ACPs. In the last case, only a small portion of actin filaments connected via highly stressed ACPs support the strain. We thus introduce the concept of a 'supportive framework,' as a subset of the full network, which is responsible for high elasticity. Notably, entropic effects due to thermal fluctuations appear to be important only at relatively low prestrains and when the average crosslinking distance is comparable to or greater than the persistence length of the filament. Taken together, our results suggest that viscoelasticity of the actin network is attributable to different mechanisms depending on

  19. Flap-edge aeroacoustic measurements and predictions

    Brooks, Thomas F.; Humphreys, William M.

    2003-03-01

    An aeroacoustic model test has been conducted to investigate the mechanisms of sound generation on high-lift wing configurations. This paper presents an analysis of flap side-edge noise, which is often the most dominant source. A model of a main element wing section with a half-span flap was tested at low speeds of up to a Mach number of 0.17, corresponding to a wing chord Reynolds number of approximately 1.7 million. Results are presented for flat (or blunt), flanged, and round flap-edge geometries, with and without boundary-layer tripping, deployed at both moderate and high flap angles. The acoustic database is obtained from a small aperture directional array (SADA) of microphones, which was constructed to electronically steer to different regions of the model and to obtain farfield noise spectra and directivity from these regions. The basic flap-edge aerodynamics is established by static surface pressure data, as well as by computational fluid dynamics (CFD) calculations and simplified edge flow analyses. Distributions of unsteady pressure sensors over the flap allow the noise source regions to be defined and quantified via cross-spectral diagnostics using the SADA output. It is found that shear layer instability and related pressure scatter is the primary noise mechanism. For the flat edge flap, two noise prediction methods based on unsteady-surface-pressure measurements are evaluated and compared to measured noise. One is a new causality spectral approach developed here. The other is a new application of an edge-noise scatter prediction method. The good comparisons for both approaches suggest that the prediction models capture much of the physics. Areas of disagreement appear to reveal when the assumed edge noise mechanism does not fully define the noise production. For the different edge conditions, extensive spectra and directivity are presented. The complexity of the directivity results demonstrate the strong role of edge source geometry and frequency in

  20. Flap Edge Aeroacoustic Measurements and Predictions

    Brooks, Thomas F.; Humphreys, William M., Jr.

    2000-01-01

    An aeroacoustic model test has been conducted to investigate the mechanisms of sound generation on high-lift wing configurations. This paper presents an analysis of flap side-edge noise, which is often the most dominant source. A model of a main element wing section with a half-span flap was tested at low speeds of up to a Mach number of 0.17, corresponding to a wing chord Reynolds number of approximately 1.7 million. Results are presented for flat (or blunt), flanged, and round flap-edge geometries, with and without boundary-layer tripping, deployed at both moderate and high flap angles. The acoustic database is obtained from a Small Aperture Directional Array (SADA) of microphones, which was constructed to electronically steer to different regions of the model and to obtain farfield noise spectra and directivity from these regions. The basic flap-edge aerodynamics is established by static surface pressure data, as well as by Computational Fluid Dynamics (CFD) calculations and simplified edge flow analyses. Distributions of unsteady pressure sensors over the flap allow the noise source regions to be defined and quantified via cross-spectral diagnostics using the SADA output. It is found that shear layer instability and related pressure scatter is the primary noise mechanism. For the flat edge flap, two noise prediction methods based on unsteady surface pressure measurements are evaluated and compared to measured noise. One is a new causality spectral approach developed here. The other is a new application of an edge-noise scatter prediction method. The good comparisons for both approaches suggest that much of the physics is captured by the prediction models. Areas of disagreement appear to reveal when the assumed edge noise mechanism does not fully define the noise production. For the different edge conditions, extensive spectra and directivity are presented. Significantly, for each edge configuration, the spectra for different flow speeds, flap angles, and

  1. Hydrodynamic schooling of flapping swimmers

    Becker, Alexander D.; Masoud, Hassan; Newbolt, Joel W.; Shelley, Michael; Ristroph, Leif

    2015-10-01

    Fish schools and bird flocks are fascinating examples of collective behaviours in which many individuals generate and interact with complex flows. Motivated by animal groups on the move, here we explore how the locomotion of many bodies emerges from their flow-mediated interactions. Through experiments and simulations of arrays of flapping wings that propel within a collective wake, we discover distinct modes characterized by the group swimming speed and the spatial phase shift between trajectories of neighbouring wings. For identical flapping motions, slow and fast modes coexist and correspond to constructive and destructive wing-wake interactions. Simulations show that swimming in a group can enhance speed and save power, and we capture the key phenomena in a mathematical model based on memory or the storage and recollection of information in the flow field. These results also show that fluid dynamic interactions alone are sufficient to generate coherent collective locomotion, and thus might suggest new ways to characterize the role of flows in animal groups.

  2. Flapping wing aerodynamics: from insects to vertebrates.

    Chin, Diana D; Lentink, David

    2016-04-01

    More than a million insects and approximately 11,000 vertebrates utilize flapping wings to fly. However, flapping flight has only been studied in a few of these species, so many challenges remain in understanding this form of locomotion. Five key aerodynamic mechanisms have been identified for insect flight. Among these is the leading edge vortex, which is a convergent solution to avoid stall for insects, bats and birds. The roles of the other mechanisms - added mass, clap and fling, rotational circulation and wing-wake interactions - have not yet been thoroughly studied in the context of vertebrate flight. Further challenges to understanding bat and bird flight are posed by the complex, dynamic wing morphologies of these species and the more turbulent airflow generated by their wings compared with that observed during insect flight. Nevertheless, three dimensionless numbers that combine key flow, morphological and kinematic parameters - the Reynolds number, Rossby number and advance ratio - govern flapping wing aerodynamics for both insects and vertebrates. These numbers can thus be used to organize an integrative framework for studying and comparing animal flapping flight. Here, we provide a roadmap for developing such a framework, highlighting the aerodynamic mechanisms that remain to be quantified and compared across species. Ultimately, incorporating complex flight maneuvers, environmental effects and developmental stages into this framework will also be essential to advancing our understanding of the biomechanics, movement ecology and evolution of animal flight. PMID:27030773

  3. Pathogenic microbes manipulate cofilin activity to subvert actin cytoskeleton.

    Zheng, Kai; Kitazato, Kaio; Wang, Yifei; He, Zhendan

    2016-09-01

    Actin-depolymerizing factor (ADF)/cofilin proteins are key players in controlling the temporal and spatial extent of actin dynamics, which is crucial for mediating host-pathogen interactions. Pathogenic microbes have evolved molecular mechanisms to manipulate cofilin activity to subvert the actin cytoskeletal system in host cells, promoting their internalization into the target cells, modifying the replication niche and facilitating their intracellular and intercellular dissemination. The study of how these pathogens exploit cofilin pathways is crucial for understanding infectious disease and providing potential targets for drug therapies. PMID:25853495

  4. Papilla Preservation Flap as Aesthetic Consideration in Periodontal Flap Surgery

    Sandra Olivia; Natalina Natalina; Felix Hartono

    2013-01-01

    Flap surgery is treatment for periodontal disease with alveolar bone destruction. Surgical periodontal flap with conventional incision will result in gingival recession and loss of interdental papillae after treatment. Dilemma arises in areas required high aesthetic value or regions with a fixed denture. It is challenging to perform periodontal flap with good aesthetic results and minimal gingival recession. This case report aimed to inform and to explain the work procedures, clinical and rad...

  5. Mechanical force-induced polymerization and depolymerization of F-actin at water/solid interfaces

    Zhang, Xueqiang; Hu, Xiuyuan; Lei, Haozhi; Hu, Jun; Zhang, Yi

    2016-03-01

    Actin molecules are among the three main cytoskeleton proteins of cells and undergo rapid cycling to regulate critical processes such as endocytosis, cytokinesis, cell polarity, and cell morphogenesis. Although extensive studies have been carried out on the dynamics as well as biological functions of actin polymerization and depolymerization both in vivo and in vitro, the molecular mechanisms by which cells sense and respond to mechanical signals are not fully understood. In particular, little attention has been paid to the effect of a physical force that is exerted directly on the actin cytoskeleton. In this paper, we have explored how the mechanical force affects the actin polymerization and depolymerization behaviors at water/solid interfaces using an atomic force microscope (AFM) operated in liquid. By raster scanning an AFM probe on a substrate surface with a certain load, it was found that actin monomers could polymerize into filaments without the help of actin related proteins (ARPs). Further study indicated that actin monomers were inclined to form filaments only under a small scanning load. The polymerized actin filaments would be depolymerized when the mechanical force was stronger. A possible mechanism has been suggested to explain the mechanical force induced actin polymerization.Actin molecules are among the three main cytoskeleton proteins of cells and undergo rapid cycling to regulate critical processes such as endocytosis, cytokinesis, cell polarity, and cell morphogenesis. Although extensive studies have been carried out on the dynamics as well as biological functions of actin polymerization and depolymerization both in vivo and in vitro, the molecular mechanisms by which cells sense and respond to mechanical signals are not fully understood. In particular, little attention has been paid to the effect of a physical force that is exerted directly on the actin cytoskeleton. In this paper, we have explored how the mechanical force affects the actin

  6. Numerical simulation of self-propulsion of flapping flexible plates

    Lu, Xi-Yun; Hua, Ru-Nan; Fluid Mechanics Team

    2012-11-01

    Self-propulsion of flapping flexible plates has been studied numerically by means of an immersed boundary-lattice Boltzmann method for the fluid dynamics around the plate and a finite element method for the deformable flapping motion of the plate. Both the two- and three-dimensional flexible plates are investigated to reveal the propulsion properties and their differences. As a result of the fluid-plate interaction, three typical movement regimes have been identified and can be briefly described as forward, backward, and irregular movements which mainly depend on the flapping amplitude and bending rigidity. It is found that there exists an optimal range of the bending rigidity for large propulsive speed or high efficiency in the forward movement regime, consistent with the observation and measurement of swimming and flying animals. The results obtained in this study provide physical insights into understanding of the propulsive mechanisms of the flapping fin or wing of animals.

  7. Flap Conformations in HIV-1 Protease are Altered by Mutations

    Fanucci, Gail; Blackburn, Mandy; Veloro, Angelo; Galiano, Luis; Fangu, Ding; Simmerling, Carlos

    2009-03-01

    HIV-1 protease (PR) is an enzyme that is a major drug target in the treatment of AIDS. Although the structure and function of HIV-1 PR have been studied for over 20 years, questions remain regarding the conformations and dynamics of the β-hairpin turns (flaps) that cover the active site cavity. Distance measurements with pulsed EPR spectroscopy of spin labeled constructs of HIV-1 PR have been used to characterize the flap conformations in the apo and inhibitor bound states. From the most probably distances and the breadth of the distance distribution profiles from analysis of the EPR data, insights regarding the flap conformations and flexibility are gained. The EPR results clearly show how drug pressure selected mutations alter the average conformation of the flaps and the degree of opening of the flaps. Molecular dynamics simulations successfully regenerate the experimentally determined distance distribution profiles, and more importantly, provide structural models for full interpretation of the EPR results. By combining experiment and theory to understand the role that altered flap flexibility/conformations play in the mechanism of drug resistance, key insights are gained toward the rational development of new inhibitors of this important enzyme.

  8. A Miniature Controllable Flapping Wing Robot

    Arabagi, Veaceslav Gheorghe

    The agility and miniature size of nature's flapping wing fliers has long baffled researchers, inspiring biological studies, aerodynamic simulations, and attempts to engineer their robotic replicas. Flapping wing flight is characterized by complex reciprocating wing kinematics, transient aerodynamic effects, and very small body lengths. These characteristics render robotic flapping wing aerial vehicles ideal for surveillance and defense applications, search and rescue missions, and environment monitoring, where their ability to hover and high maneuverability is immensely beneficial. One of the many difficulties in creating flapping wing based miniature robotic aerial vehicles lies in generating a proper wing trajectory that would result in sufficient lift forces for hovering and maneuvering. Since design of a flapping wing system is a balance between overall weight and the number of actuated inputs, we take the approach of having minimal controlled inputs, allowing passive behavior wherever possible. Hence, we propose a completely passive wing pitch reversal design that relies on wing inertial dynamics, an elastic energy storage mechanism, and low Reynolds number aerodynamic effects. Theoretical models, compiling previous research on piezoelectric actuators, four-bar transmissions, and aerodynamics effects, are developed and used as basis for a complete numerical simulation. Limitations of the model are discussed in comparison to experimental results obtained from a working prototype of the proposed passive pitch reversal flapping wing mechanism. Given that the mechanism is under-actuated, methods to control lift force generation by actively varying system parameters are proposed, discussed, and tested experimentally. A dual wing aerial platform is developed based on the passive pitch reversal wing concept. Design considerations are presented, favoring controllability and structural rigidity of the final platform. Finite element analysis and experimental

  9. Exploring the drug resistance of V32I and M46L mutant HIV-1 protease to inhibitor TMC114: flap dynamics and binding mechanism

    Meher, Biswa Ranjan; Wang, Yixuan

    2014-01-01

    Inhibitors of HIV-1 protease (HIV-1-pr) generally only bind to the active site of the protease. However, for some mutants such as V32I and M46L the TMC114 can bind not only to the active cavity also to the groove of the flexible flaps. Although the second binding site suggests the higher efficiency of the drug against HIV-1-pr, the drug resistance in HIV-1-pr due to mutations cannot be ignored, which prompts us to investigate the molecular mechanisms of drug resistance and behavior of double ...

  10. Smart dynamic rotor control using active flaps on a small-scale wind turbine: aeroelastic modeling and comparison with wind tunnel measurements

    Barlas, Thanasis K.; van Wingerden, W.; Hulskamp, A.W.;

    2013-01-01

    using the aeroelastic tool, load predictions are compared with the wind tunnel measurements, and similar control concepts are compared and evaluated in the numerical environment. Conclusions regarding evaluation of the performance of smart rotor concepts for wind turbines are drawn from this threefold......In this paper, the proof of concept of a smart rotor is illustrated by aeroelastic simulations on a small-scale rotor and comparison with wind tunnel experiments. The application of advanced feedback controllers using actively deformed flaps in the wind tunnel measurements is shown to alleviate...

  11. Flag flapping in a channel

    Alben, Silas; Shoele, Kourosh; Mittal, Rajat; Jha, Sourabh; Glezer, Ari

    2015-11-01

    We study the flapping of a flag in an inviscid channel flow. We focus especially on how quantities vary with channel spacing. As the channel walls move inwards towards the flag, heavier flags become more unstable, while light flags' stability is less affected. We use a vortex sheet model to compute large-amplitude flapping, and find that the flag undergoes a series of jumps to higher flapping modes as the channel walls are moved towards the flag. Meanwhile, the drag on the flag and the energy lost to the wake first rise as the walls become closer, then drop sharply as the flag moves to a higher flapping mode.

  12. Keystone flaps in coloured skin: Flap technology for the masses?

    Satish P Bhat

    2013-01-01

    Full Text Available Introduction: Viscoelastic properties of skin in coloured ethnic groups are less favourable compared to Caucasians for executing Keystone flaps. Keystone flaps have so far been evaluated and reported only in Caucasians. The potential of Keystone flaps in a coloured ethnic group is yet unknown. Aim: This article reviews the experience to reconstruct skin defects presenting in a coloured ethnic group, by using Keystone flaps, with a review of existing literature. Design: Uncontrolled case series. Materials and Methods: This retrospective review involves 55 consecutive Keystone flaps used from 2009 to 2012, for skin defects in various locations. Patient demographic data, medical history, co-morbidity, surgical indication, defect features, complications, and clinical outcomes are evaluated and presented. Results: In this population group with Fitzpatrick type 4 and 5 skin, the average patient age was 35.73. Though 60% of flaps (33/55 in the series involved specific risk factors, only two flaps failed. Though seven flaps had complications, sound healing was achieved by suitable intervention giving a success rate of 96.36%. Skin grafts were needed in only four cases. Conclusions: Keystone flaps achieve primary wound healing for a wide spectrum of defects with an acceptable success rate in a coloured skin population with unfavorable biophysical properties. By avoiding conventional local flaps and at times even microsurgical flaps, good aesthetic outcome is achieved without additional skin grafts or extensive operative time. All advantages seen in previous studies were verified. These benefits can be most appreciated in coloured populations, with limited resources and higher proportion of younger patients and unfavorable defects.

  13. Locomotion of a flapping flexible plate

    Hua, Ru-Nan; Zhu, Luoding; Lu, Xi-Yun

    2013-12-01

    The locomotion of a flapping flexible plate in a viscous incompressible stationary fluid is numerically studied by an immersed boundary-lattice Boltzmann method for the fluid and a finite element method for the plate. When the leading-edge of the flexible plate is forced to heave sinusoidally, the entire plate starts to move freely as a result of the fluid-structure interaction. Mechanisms underlying the dynamics of the plate are elucidated. Three distinct states of the plate motion are identified and can be described as forward, backward, and irregular. Which state to occur depends mainly on the heaving amplitude and the bending rigidity of the plate. In the forward motion regime, analysis of the dynamic behaviors of the flapping flexible plate indicates that a suitable degree of flexibility can improve the propulsive performance. Moreover, there exist two kinds of vortex streets in the downstream of the plate which are normal and deflected wake. Further the forward motion is compared with the flapping-based locomotion of swimming and flying animals. The results obtained in the present study are found to be consistent with the relevant observations and measurements and can provide some physical insights into the understanding of the propulsive mechanisms of swimming and flying animals.

  14. Managing Flap Vortices via Separation Control

    Greenblatt, David

    2006-01-01

    A pilot study was conducted on a flapped semi-span model to investigate the concept and viability of near-wake vortex management by means of boundary layer separation control. Passive control was achieved using a simple fairing and active control was achieved via zero mass-flux blowing slots. Vortex sheet strength, estimated by integrating surface pressures, was used to predict vortex characteristics based on inviscid rollup relations and vortices trailing the flaps were mapped using a seven-hole probe. Separation control was found to have a marked effect on vortex location, strength, tangential velocity, axial velocity and size over a wide range of angles of attack and control conditions. In general, the vortex trends were well predicted by the inviscid rollup relations. Manipulation of the separated flow near the flap edges exerted significant control over either outboard or inboard edge vortices while producing small lift and moment excursions. Unsteady surface pressures indicated that dynamic separation and attachment control can be exploited to perturb vortices at wavelengths shorter than a typical wingspan. In summary, separation control has the potential for application to time-independent or time-dependent wake alleviation schemes, where the latter can be deployed to minimize adverse effects on ride-quality and dynamic structural loading.

  15. The role of the cofilin-actin rod stress response in neurodegenerative diseases uncovers potential new drug targets

    Munsie, Lise N.; Truant, Ray

    2012-01-01

    The cofilin-actin rod stress response is an actin cytoskeletal dynamic arrest that occurs in cells under a variety of stress conditions. Upon stress, the rapidly activated cofilin saturates actin filaments causing them to bundle into rod structures in either the nucleus or cytoplasm, halting actin polymerization and thus freeing ATP. Importantly, these rods dissociate quickly following relief of the transient stress. The rods form inappropriately in neurons involved in the progression of Alzh...

  16. Antenna mechanism of length control of actin cables

    Mohapatra, Lishibanya; Kondev, Jane

    2014-01-01

    Actin cables are linear cytoskeletal structures that serve as tracks for myosin-based intracellular transport of vesicles and organelles in both yeast and mammalian cells. In a yeast cell undergoing budding, cables are in constant dynamic turnover yet some cables grow from the bud neck toward the back of the mother cell until their length roughly equals the diameter of the mother cell. This raises the question: how is the length of these cables controlled? Here we describe a novel molecular mechanism for cable length control inspired by recent experimental observations in cells. This antenna mechanism involves three key proteins: formins, which polymerize actin, Smy1 proteins, which bind formins and inhibit actin polymerization, and myosin motors, which deliver Smy1 to formins, leading to a length-dependent actin polymerization rate. We compute the probability distribution of cable lengths as a function of several experimentally tuneable parameters such as the formin-binding affinity of Smy1 and the concentra...

  17. The Actin Cytoskeleton as a Therapeutic Target for the Prevention of Relapse to Methamphetamine Use.

    Young, Erica J; Briggs, Sherri B; Miller, Courtney A

    2015-01-01

    A high rate of relapse is a defining characteristic of substance use disorder for which few treatments are available. Exposure to environmental cues associated with previous drug use can elicit relapse by causing the involuntary retrieval of deeply engrained associative memories that trigger a strong motivation to seek out drugs. Our lab is focused on identifying and disrupting mechanisms that support these powerful consolidated memories, with the goal of developing therapeutics. A particularly promising mechanism is regulation of synaptic dynamics by actin polymerization within dendritic spines. Emerging evidence indicates that memory is supported by structural and functional plasticity dendritic spines, for which actin polymerization is critical, and that prior drug use increases both spine and actin dynamics. Indeed we have found that inhibiting amygdala (AMY) actin polymerization immediately or twenty-four hours prior to testing disrupted methamphetamine (METH)-associated memories, but not food reward or fear memories. Furthermore, METH training increased AMY spine density which was reversed by actin depolymerization treatment. Actin dynamics were also shifted to a more dynamic state by METH training. While promising, actin polymerization inhibitors are not a viable therapeutic, as a multitude of peripheral process (e.g. cardiac function) rely on dynamic actin. For this reason, we have shifted our focus upstream of actin polymerization to nonmuscle myosin II. We and others have demonstrated that myosin IIb imparts a mechanical force that triggers spine actin polymerization in response to synaptic stimulation. Similar to an actin depolymerizing compound, pre-test inhibition of myosin II ATPase activity in the AMY produced a rapid and lasting disruption of drug-seeking behavior. While many questions remain, these findings indicate that myosin II represents a potential therapeutic avenue to target the actin cytoskeleton and disrupt the powerful, extinction

  18. Regulation of T cell receptor signaling by the actin cytoskeleton and poroelastic cytoplasm

    Beemiller, Peter; Krummel, Matthew F.

    2013-01-01

    Summary The actin cytoskeleton plays essential roles in modulating T-cell activation. Most models of T-cell receptor (TCR) triggering, signalosome assembl, y and immune synapse formation invoke actin-dependent mechanisms. As T cells are constitutively motile cells, TCR triggering and signaling occur against a cytoskeletal backdrop that is constantly remodeling. While the interplay between actin dynamics and TCR signaling have been the focus of research for many years, much of the work in T cells has considered actin largely for its ‘scaffolding’ function. We examine the roles of the actin cytoskeleton in TCR signaling and immune synapse formation with an emphasis on how poroelasticity, an ensemble feature of actin dynamics with the cytosol, relates to how T cells respond to stimulation. PMID:24117819

  19. Regulation of T-cell receptor signaling by the actin cytoskeleton and poroelastic cytoplasm.

    Beemiller, Peter; Krummel, Matthew F

    2013-11-01

    The actin cytoskeleton plays essential roles in modulating T-cell activation. Most models of T-cell receptor (TCR) triggering signalosome assembly and immune synapse formation invoke actin-dependent mechanisms. As T cells are constitutively motile cells, TCR triggering and signaling occur against a cytoskeletal backdrop that is constantly remodeling. While the interplay between actin dynamics and TCR signaling have been the focus of research for many years, much of the work in T cells has considered actin largely for its 'scaffolding' function. We examine the roles of the actin cytoskeleton in TCR signaling and immune synapse formation with an emphasis on how poroelasticity, an ensemble feature of actin dynamics with the cytosol, relates to how T cells respond to stimulation. PMID:24117819

  20. Direct Microtubule-Binding by Myosin-10 Orients Centrosomes toward Retraction Fibers and Subcortical Actin Clouds.

    Kwon, Mijung; Bagonis, Maria; Danuser, Gaudenz; Pellman, David

    2015-08-10

    Positioning of centrosomes is vital for cell division and development. In metazoan cells, spindle positioning is controlled by a dynamic pool of subcortical actin that organizes in response to the position of retraction fibers. These actin "clouds" are proposed to generate pulling forces on centrosomes and mediate spindle orientation. However, the motors that pull astral microtubules toward these actin structures are not known. Here, we report that the unconventional myosin, Myo10, couples actin-dependent forces from retraction fibers and subcortical actin clouds to centrosomes. Myo10-mediated centrosome positioning requires its direct microtubule binding. Computational image analysis of large microtubule populations reveals a direct effect of Myo10 on microtubule dynamics and microtubule-cortex interactions. Myo10's role in centrosome positioning is distinct from, but overlaps with, that of dynein. Thus, Myo10 plays a key role in integrating the actin and microtubule cytoskeletons to position centrosomes and mitotic spindles. PMID:26235048

  1. Arp2/3-mediated F-actin formation controls regulated exocytosis in vivo

    Tran, Duy T.; Masedunskas, Andrius; Weigert, Roberto; Ten Hagen, Kelly G.

    2015-01-01

    The actin cytoskeleton plays crucial roles in many cellular processes, including regulated secretion. However, the mechanisms controlling F-actin dynamics in this process are largely unknown. Through 3D time-lapse imaging in a secreting organ, we show that F-actin is actively disassembled along the apical plasma membrane at the site of secretory vesicle fusion and re-assembled directionally on vesicle membranes. Moreover, we show that fusion pore formation and PIP2 redistribution precedes act...

  2. Forces generated during actin-based propulsion: A direct measurement by micromanipulation

    Marcy, Yann; Prost, Jacques; Carlier, Marie-France; Sykes, Cécile

    2004-01-01

    Dynamic actin networks generate forces for numerous types of movements such as lamellipodia protrusion or the motion of endocytic vesicles. The actin-based propulsive movement of Listeria monocytogenes or of functionalized microspheres have been extensively used as model systems to identify the biochemical components that are necessary for actin-based motility. However, quantitative force measurements are required to elucidate the mechanism of force generation, which is still under debate. To...

  3. Regulation of T cell receptor signaling by the actin cytoskeleton and poroelastic cytoplasm

    Beemiller, Peter; Krummel, Matthew F.

    2013-01-01

    The actin cytoskeleton plays essential roles in modulating T-cell activation. Most models of T-cell receptor (TCR) triggering, signalosome assembl, y and immune synapse formation invoke actin-dependent mechanisms. As T cells are constitutively motile cells, TCR triggering and signaling occur against a cytoskeletal backdrop that is constantly remodeling. While the interplay between actin dynamics and TCR signaling have been the focus of research for many years, much of the work in T cells has ...

  4. The role of actin networks in cellular mechanosensing

    Azatov, Mikheil

    Physical processes play an important role in many biological phenomena, such as wound healing, organ development, and tumor metastasis. During these processes, cells constantly interact with and adapt to their environment by exerting forces to mechanically probe the features of their surroundings and generating appropriate biochemical responses. The mechanisms underlying how cells sense the physical properties of their environment are not well understood. In this thesis, I present my studies to investigate cellular responses to the stiffness and topography of the environment. In order to sense the physical properties of their environment, cells dynamically reorganize the structure of their actin cytoskeleton, a dynamic network of biopolymers, altering the shape and spatial distribution of protein assemblies. Several observations suggest that proteins that crosslink actin filaments may play an important role in cellular mechanosensitivity. Palladin is an actin-crosslinking protein that is found in the lamellar actin network, stress fibers and focal adhesions, cellular structures that are critical for mechanosensing of the physical environment. By virtue of its close interactions with these structures in the cell, palladin may play an important role in cell mechanics. However, the role of actin crosslinkers in general, and palladin in particular, in cellular force generation and mechanosensing is not well known. I have investigated the role of palladin in regulating the plasticity of the actin cytoskeleton and cellular force generation in response to alterations in substrate stiffness. I have shown that the expression levels of palladin modulate the forces exerted by cells and their ability to sense substrate stiffness. Perturbation experiments also suggest that palladin levels in cells altered myosin motor activity. These results suggest that the actin crosslinkers, such as palladin, and myosin motors coordinate for optimal cell function and to prevent aberrant

  5. Characterization of ring-like F-actin structure as a mechanical partner for spindle positioning in mitosis.

    Huan Lu

    Full Text Available Proper spindle positioning and orientation are essential for accurate mitosis which requires dynamic interactions between microtubule and actin filament (F-actin. Although mounting evidence demonstrates the role of F-actin in cortical cytoskeleton dynamics, it remains elusive as to the structure and function of F-actin-based networks in spindle geometry. Here we showed a ring-like F-actin structure surrounding the mitotic spindle which forms since metaphase and maintains in MG132-arrested metaphase HeLa cells. This cytoplasmic F-actin structure is relatively isotropic and less dynamic. Our computational modeling of spindle position process suggests a possible mechanism by which the ring-like F-actin structure can regulate astral microtubule dynamics and thus mitotic spindle orientation. We further demonstrated that inhibiting Plk1, Mps1 or Myosin, and disruption of microtubules or F-actin polymerization perturbs the formation of the ring-like F-actin structure and alters spindle position and symmetric division. These findings reveal a previously unrecognized but important link between mitotic spindle and ring-like F-actin network in accurate mitosis and enables the development of a method to theoretically illustrate the relationship between mitotic spindle and cytoplasmic F-actin.

  6. Feedback Interactions of Polymerized Actin with the Cell Membrane: Waves, Pulses, and Oscillations

    Carlsson, Anders

    Polymerized filaments of the protein actin have crucial functions in cell migration, and in bending the cell membrane to drive endocytosis or the formation of protrusions. The nucleation and polymerization of actin filaments are controlled by upstream agents in the cell membrane, including nucleation-promoting factors (NPFs) that activate the Arp2/3 complex to form new branches on pre-existing filaments. But polymerized actin (F-actin) also feeds back on the assembly of NPFs. We explore the effects of the resulting feedback loop of F-actin and NPFs on two phenomena: actin pulses that drive endocytosis in yeast, and actin waves traveling along the membrane of several cell types. In our model of endocytosis in yeast, the actin network is grown explicitly in three dimensions, exerts a negative feedback interaction on localized patch of NPFs in the membrane, and bends the membrane by exerting a distribution of forces. This model explains observed actin and NPF pulse dynamics, and the effects of several interventions including i) NPF mutations, ii) inhibition of actin polymerization, and iii) deletion of a protein that allows F-actin to bend the cell membrane. The model predicts that mutation of the active region of an NPF will enhance the accumulation of that NPF, and we confirm this prediction by quantitative fluorescence microscopy. For actin waves, we treat a similar model, with NPFs distributed over a larger region of the cell membrane. This model naturally generates actin waves, and predicts a transition from wave behavior to spatially localized oscillations when NPFs are confined to a small region. We also predict a transition from waves to static polarization as the negative-feedback coupling between F-actin and the NPFs is reduced. Supported by NIGMS Grant R01 GM107667.

  7. The Formin Diaphanous Regulates Myoblast Fusion through Actin Polymerization and Arp2/3 Regulation.

    Su Deng

    2015-08-01

    Full Text Available The formation of multinucleated muscle cells through cell-cell fusion is a conserved process from fruit flies to humans. Numerous studies have shown the importance of Arp2/3, its regulators, and branched actin for the formation of an actin structure, the F-actin focus, at the fusion site. This F-actin focus forms the core of an invasive podosome-like structure that is required for myoblast fusion. In this study, we find that the formin Diaphanous (Dia, which nucleates and facilitates the elongation of actin filaments, is essential for Drosophila myoblast fusion. Following cell recognition and adhesion, Dia is enriched at the myoblast fusion site, concomitant with, and having the same dynamics as, the F-actin focus. Through analysis of Dia loss-of-function conditions using mutant alleles but particularly a dominant negative Dia transgene, we demonstrate that reduction in Dia activity in myoblasts leads to a fusion block. Significantly, no actin focus is detected, and neither branched actin regulators, SCAR or WASp, accumulate at the fusion site when Dia levels are reduced. Expression of constitutively active Dia also causes a fusion block that is associated with an increase in highly dynamic filopodia, altered actin turnover rates and F-actin distribution, and mislocalization of SCAR and WASp at the fusion site. Together our data indicate that Dia plays two roles during invasive podosome formation at the fusion site: it dictates the level of linear F-actin polymerization, and it is required for appropriate branched actin polymerization via localization of SCAR and WASp. These studies provide new insight to the mechanisms of cell-cell fusion, the relationship between different regulators of actin polymerization, and invasive podosome formation that occurs in normal development and in disease.

  8. Active Control of Long Bridges Using Flaps

    Hansen, H. I.; Thoft-Christensen, Palle

    The main problem in designing ultra-long span suspension bridges is flutter. A solution to this problem might be to introduce an active flap control system to increase the flutter wind velocity. The investigated flap control system consists of flaps integrated in the bridge girder so each flap is...... different flap configurations for a bridge section model by using aerodynamic derivatives for a flat plate. The example shows that different flap configurations can either increase or decrease the flutter wind velocity. for optimal flap configurations flutter will not occur....

  9. State transitions of actin cortices in vitro and in vivo

    Tan, Tzer Han; Keren, Kinneret; Mackintosh, Fred; Schmidt, Christoph; Fakhri, Nikta

    Most animal cells are enveloped by a thin layer of actin cortex which governs the cell mechanics. A functional cortex must be rigid to provide mechanical support while being flexible to allow for rapid restructuring events such as cell division. To satisfy these requirements, the actin cortex is highly dynamic with fast actin turnover and myosin-driven contractility. The regulatory mechanism responsible for the transition between a mechanically stable state and a restructuring state is not well understood. Here, we develop a technique to map the dynamics of reconstituted actin cortices in emulsion droplets using IR fluorescent single-walled carbon nanotubes (SWNTs). By increasing crosslinker concentration, we find that a homogeneous cortex transitions to an intermediate state with broken rotational symmetry and a globally contractile state which further breaks translational symmetry. We apply this new dynamic mapping technique to cortices of live starfish oocytes in various developmental stages. To identify the regulatory mechanism for steady state transitions, we subject the oocytes to actin and myosin disrupting drugs.

  10. Hippocampal Dendritic Spines Are Segregated Depending on Their Actin Polymerization.

    Domínguez-Iturza, Nuria; Calvo, María; Benoist, Marion; Esteban, José Antonio; Morales, Miguel

    2016-01-01

    Dendritic spines are mushroom-shaped protrusions of the postsynaptic membrane. Spines receive the majority of glutamatergic synaptic inputs. Their morphology, dynamics, and density have been related to synaptic plasticity and learning. The main determinant of spine shape is filamentous actin. Using FRAP, we have reexamined the actin dynamics of individual spines from pyramidal hippocampal neurons, both in cultures and in hippocampal organotypic slices. Our results indicate that, in cultures, the actin mobile fraction is independently regulated at the individual spine level, and mobile fraction values do not correlate with either age or distance from the soma. The most significant factor regulating actin mobile fraction was the presence of astrocytes in the culture substrate. Spines from neurons growing in the virtual absence of astrocytes have a more stable actin cytoskeleton, while spines from neurons growing in close contact with astrocytes show a more dynamic cytoskeleton. According to their recovery time, spines were distributed into two populations with slower and faster recovery times, while spines from slice cultures were grouped into one population. Finally, employing fast lineal acquisition protocols, we confirmed the existence of loci with high polymerization rates within the spine. PMID:26881098

  11. The Role of Actin Cytoskeleton in Memory Formation in Amygdala.

    Lamprecht, Raphael

    2016-01-01

    The central, lateral and basolateral amygdala (BLA) nuclei are essential for the formation of long-term memories including emotional and drug-related memories. Studying cellular and molecular mechanisms of memory in amygdala may lead to better understanding of how memory is formed and of fear and addiction-related disorders. A challenge is to identify molecules activated by learning that subserve cellular changes needed for memory formation and maintenance in amygdala. Recent studies show that activation of synaptic receptors during fear and drug-related learning leads to alteration in actin cytoskeleton dynamics and structure in amygdala. Such changes in actin cytoskeleton in amygdala are essential for fear and drug-related memories formation. Moreover, the actin cytoskeleton subserves, after learning, changes in neuronal morphogenesis and glutamate receptors trafficking in amygdala. These cellular events are involved in fear and drug-related memories formation. Actin polymerization is also needed for the maintenance of drug-associated memories in amygdala. Thus, the actin cytoskeleton is a key mediator between receptor activation during learning and cellular changes subserving long-term memory (LTM) in amygdala. The actin cytoskeleton may serve as a target for pharmacological treatment of fear memory associated with fear and anxiety disorders and drug addiction to prevent the debilitating consequences of these diseases. PMID:27065800

  12. Progresses in studies of nuclear actin

    ZHU Xiaojuan; ZENG Xianlu; SONG Zhaoxia; HAO Shui

    2004-01-01

    Actin is a protein abundant in cells. Recently, it has been proved to be universally existent in the nuclei of many cell types. Actin and actin-binding proteins, as well as actin-related proteins, are necessary for the mediation of the conformation and function of nuclear actin, including the transformation of actin between unpolymerized and polymerized, chroinatin remodeling, regulation of gene expression and RNA processing as well as RNA transportation. In this paper, we summarized the progresses in the research of nu clear actin.

  13. Flap Edge Noise Reduction Fins

    Khorrami, Mehdi R. (Inventor); Choudhan, Meelan M. (Inventor)

    2015-01-01

    A flap of the type that is movably connected to an aircraft wing to provide control of an aircraft in flight includes opposite ends, wherein at least a first opposite end includes a plurality of substantially rigid, laterally extending protrusions that are spaced apart to form a plurality of fluidly interconnected passageways. The passageways have openings adjacent to upper and lower sides of the flap, and the passageways include a plurality of bends such that high pressure fluid flows from a high pressure region to a low pressure region to provide a boundary condition that inhibits noise resulting from airflow around the end of the flap.

  14. Characterization of engineered actin binding proteins that control filament assembly and structure.

    Crista M Brawley

    Full Text Available BACKGROUND: Eukaryotic cells strictly regulate the structure and assembly of their actin filament networks in response to various stimuli. The actin binding proteins that control filament assembly are therefore attractive targets for those who wish to reorganize actin filaments and reengineer the cytoskeleton. Unfortunately, the naturally occurring actin binding proteins include only a limited set of pointed-end cappers, or proteins that will block polymerization from the slow-growing end of actin filaments. Of the few that are known, most are part of large multimeric complexes that are challenging to manipulate. METHODOLOGY/PRINCIPAL FINDINGS: We describe here the use of phage display mutagenesis to generate of a new class of binding protein that can be targeted to the pointed-end of actin. These proteins, called synthetic antigen binders (sABs, are based on an antibody-like scaffold where sequence diversity is introduced into the binding loops using a novel "reduced genetic code" phage display library. We describe effective strategies to select and screen for sABs that ensure the generated sABs bind to the pointed-end surface of actin exclusively. CONCLUSIONS/SIGNIFICANCE: From our set of pointed-end binders, we identify three sABs with particularly useful properties to systematically probe actin dynamics: one protein that caps the pointed end, a second that crosslinks actin filaments, and a third that severs actin filaments and promotes disassembly.

  15. Genome-wide RNAi screen for nuclear actin reveals a network of cofilin regulators.

    Dopie, Joseph; Rajakylä, Eeva K; Joensuu, Merja S; Huet, Guillaume; Ferrantelli, Evelina; Xie, Tiao; Jäälinoja, Harri; Jokitalo, Eija; Vartiainen, Maria K

    2015-07-01

    Nuclear actin plays an important role in many processes that regulate gene expression. Cytoplasmic actin dynamics are tightly controlled by numerous actin-binding proteins, but regulation of nuclear actin has remained unclear. Here, we performed a genome-wide RNA interference (RNAi) screen in Drosophila cells to identify proteins that influence either nuclear polymerization or import of actin. We validate 19 factors as specific hits, and show that Chinmo (known as Bach2 in mammals), SNF4Aγ (Prkag1 in mammals) and Rab18 play a role in nuclear localization of actin in both fly and mammalian cells. We identify several new regulators of cofilin activity, and characterize modulators of both cofilin kinases and phosphatase. For example, Chinmo/Bach2, which regulates nuclear actin levels also in vivo, maintains active cofilin by repressing the expression of the kinase Cdi (Tesk in mammals). Finally, we show that Nup98 and lamin are candidates for regulating nuclear actin polymerization. Our screen therefore reveals new aspects of actin regulation and links nuclear actin to many cellular processes. PMID:26021350

  16. Altered Cell Mechanics from the Inside: Dispersed Single Wall Carbon Nanotubes Integrate with and Restructure Actin

    Mohammad F. Islam

    2012-05-01

    Full Text Available With a range of desirable mechanical and optical properties, single wall carbon nanotubes (SWCNTs are a promising material for nanobiotechnologies. SWCNTs also have potential as biomaterials for modulation of cellular structures. Previously, we showed that highly purified, dispersed SWCNTs grossly alter F-actin inside cells. F-actin plays critical roles in the maintenance of cell structure, force transduction, transport and cytokinesis. Thus, quantification of SWCNT-actin interactions ranging from molecular, sub-cellular and cellular levels with both structure and function is critical for developing SWCNT-based biotechnologies. Further, this interaction can be exploited, using SWCNTs as a unique actin-altering material. Here, we utilized molecular dynamics simulations to explore the interactions of SWCNTs with actin filaments. Fluorescence lifetime imaging microscopy confirmed that SWCNTs were located within ~5 nm of F-actin in cells but did not interact with G-actin. SWCNTs did not alter myosin II sub-cellular localization, and SWCNT treatment in cells led to significantly shorter actin filaments. Functionally, cells with internalized SWCNTs had greatly reduced cell traction force. Combined, these results demonstrate direct, specific SWCNT alteration of F-actin structures which can be exploited for SWCNT-based biotechnologies and utilized as a new method to probe fundamental actin-related cellular processes and biophysics.

  17. A Potential Yeast Actin Allosteric Conduit Dependent on Hydrophobic Core Residues Val-76 and Trp-79*

    Wen, Kuo-Kuang; McKane, Melissa; Stokasimov, Ema; Fields, Jonathon; Rubenstein, Peter A.

    2010-01-01

    Intramolecular allosteric interactions responsible for actin conformational regulation are largely unknown. Previous work demonstrated that replacing yeast actin Val-76 with muscle actin Ile caused decreased nucleotide exchange. Residue 76 abuts Trp-79 in a six-residue linear array beginning with Lys-118 on the surface and ending with His-73 in the nucleotide cleft. To test if altering the degree of packing of these two residues would affect actin dynamics, we constructed V76I, W79F, and W79Y...

  18. The Actin Binding Protein Adseverin Regulates Osteoclastogenesis

    Hassanpour, Siavash; Jiang, Hongwei; Wang, Yongqiang; Kuiper, Johannes W. P.; Glogauer, Michael

    2014-01-01

    Adseverin (Ads), a member of the Gelsolin superfamily of actin binding proteins, regulates the actin cytoskeleton architecture by severing and capping existing filamentous actin (F-actin) strands and nucleating the assembly of new F-actin filaments. Ads has been implicated in cellular secretion, exocytosis and has also been shown to regulate chondrogenesis and megakaryoblastic leukemia cell differentiation. Here we report for the first time that Ads is involved in regulating osteoclastogenesi...

  19. Inhibiting actin depolymerization enhances osteoblast differentiation and bone formation in human stromal stem cells

    Chen, Li; Shi, Kaikai; Frary, Charles Edward;

    2015-01-01

    Remodeling of the actin cytoskeleton through actin dynamics is involved in a number of biological processes, but its role in human stromal (skeletal) stem cells (hMSCs) differentiation is poorly understood. In the present study, we demonstrated that stabilizing actin filaments by inhibiting gene...... expression of the two main actin depolymerizing factors (ADFs): Cofilin 1 (CFL1) and Destrin (DSTN) in hMSCs, enhanced cell viability and differentiation into osteoblastic cells (OB) in vitro, as well as heterotopic bone formation in vivo. Similarly, treating hMSC with Phalloidin, which is known to stabilize...... polymerized actin filaments, increased hMSCs viability and OB differentiation. Conversely, Cytocholasin D, an inhibitor of actin polymerization, reduced cell viability and inhibited OB differentiation of hMSC. At a molecular level, preventing Cofilin phosphorylation through inhibition of LIM domain kinase 1...

  20. Distribution of G-actin is Related to Root Hair Growth of Wheat

    He, Xue; Liu, Yi-Min; Wang, Wei; LI Yan

    2006-01-01

    • Background and Aims Actin distribution in root hair tips is a controversial topic. Although the relationship between Ca2+ gradient and actin dynamics in plant tip-growth has been a focus of study, there is still little direct evidence on the exact relationship in root hair tip-growth.

  1. The freestyle pedicle perforator flap

    Gunnarsson, Gudjon Leifur; Jackson, Ian T; Westvik, Tormod S; Thomsen, Jorn Bo

    2015-01-01

    not widely performed by the general plastic surgeons. The aim of this paper is to present the simplicity of pedicled perforator flap reconstruction of moderate-sized defects of the extremities and torso. METHODS: We retrospectively reviewed the charts of 34 patients reconstructed using 34 freestyle...... aspects in the context of current literature. RESULTS: The reconstructive goals were achieved in all cases without any total flap loss or major complications. Minor complications occurred in 7/34 (21 %) cases consisting of venous congestion leading to distal tip necrosis or epidermolysis; partial flap...... loss was significant in 4 cases, however never more than 10 % of the total flap size. Reconstruction was performed on the lower limb in 13 cases, upper limb in 12, and 9 cases were on the truncus. The angle of rotation was 90° in 21 cases and 180° in 13 cases. The most common indication was...

  2. Modeling of Airfoil Trailing Edge Flap with Immersed Boundary Method

    Zhu, Wei Jun; Shen, Wen Zhong; Sørensen, Jens Nørkær

    2011-01-01

    The present work considers incompressible flow over a 2D airfoil with a deformable trailing edge. The aerodynamic characteristics of an airfoil with a trailing edge flap is numerically investigated using computational fluid dynamics. A novel hybrid immersed boundary (IB) technique is applied to...... simulate the moving part of the trailing edge. Over the main fixed part of the airfoil the Navier-Stokes (NS) equations are solved using a standard body-fitted finite volume technique whereas the moving trailing edge flap is simulated with the immersed boundary method on a curvilinear mesh. The obtained...

  3. Differential effects of LifeAct-GFP and actin-GFP on cell mechanics assessed using micropipette aspiration

    Sliogeryte, Kristina; Stephen D Thorpe; Wang, Zhao; Thompson, Clare L.; Gavara, Nuria; Knight, Martin M.

    2016-01-01

    The actin cytoskeleton forms a dynamic structure involved in many fundamental cellular processes including the control of cell morphology, migration and biomechanics. Recently LifeAct-GFP (green fluorescent protein) has been proposed for visualising actin structure and dynamics in live cells as an alternative to actin-GFP which has been shown to affect cell mechanics. Here we compare the two approaches in terms of their effect on cellular mechanical behaviour. Human mesenchymal stem cells (hM...

  4. Modified osteomyocutaneous iliac crest flaps transplantation.

    Liu, Jun; Song, Dajiang; Li, Jinsong; Xu, Jian; Lv, Hongbin

    2015-04-01

    The paper aims to improve the operative technique of osteomyocutaneous iliac crest flap harvesting, further minimise morbidity of donor site, and improve the effect of recipient site reconstruction. From March 2005 to March 2011, 55 cases of osteomyocutaneous iliac crest flap harvested by different methods were performed to reconstruct the defects of the extremities. Twenty-nine cases were reconstructed with a traditional deep circumflex iliac artery osteomusculocutaneous flap. Twenty-six cases were repaired with modified osteomyocutaneous iliac crest flaps. In 29 cases with a traditional DCIA osteomusculocutaneous flap, two cases showed the injured lateral femoral cutaneous nerve. Flapnecrosis was significant in two cases. Arterial compromise occurred in one case 5 days after operation completion and led to flap failure. Three flaps developed postoperative venous congestion, but only one flap received re-exploration. In the other two cases, some stitches were removed for decompression. All three flaps survived. In two cases, marginal flap necrosis occurred, but no secondary skin grafting was required. In 26 cases with modified flap transplantation, one case showed the injured lateral femoral cutaneous nerve. All flaps survived totally. Osseous integration was achieved in all 55 cases in 3 ∼ 9 months after operation. The modified osteomyocutaneous iliac crest flap technique enhances flap safety, provides the additional advantages of reducing donor-site morbidity, and improves the recipient-site contour. PMID:25001367

  5. Regulation of water flow by actin-binding protein-induced actin gelatin.

    Ito, T.; Suzuki, A.; Stossel, T. P.

    1992-01-01

    Actin filaments inhibit osmotically driven water flow (Ito, T., K.S. Zaner, and T.P. Stossel. 1987. Biophys. J. 51: 745-753). Here we show that the actin gelation protein, actin-binding protein (ABP), impedes both osmotic shrinkage and swelling of an actin filament solution and reduces markedly the concentration of actin filaments required for this inhibition. These effects depend on actin filament immobilization, because the ABP concentration that causes initial impairment of water flow by a...

  6. Four-winged flapping flyer in forward flight

    Godoy-Diana, Ramiro; Centeno, Mariana; Weinreb, Alexis; Thiria, Benjamin

    2015-01-01

    We study experimentally a four-winged flapping flyer with chord-wise flexible wings in a self-propelled setup. For a given physical configuration of the flyer (i.e. fixed distance between the forewing and hindwing pairs and fixed wing flexibility), we explore the kinematic parameter space constituted by the flapping frequency and the forewing-hindwing phase lag. Cruising speed and consumed electric power measurements are performed for each point in the $(f,\\varphi)$ parameter space and allow us to discuss the problem of performance and efficiency in four-winged flapping flight. We show that different phase-lags are needed for the system to be optimised for fastest flight or lowest energy consumption. A conjecture of the underlying mechanism is proposed in terms of the coupled dynamics of the forewing-hindwing phase lag and the deformation kinematics of the flexible wings.

  7. Interaction of Two Flapping Flags in Axial Flow

    Gunter, Amy-Lee; Fayed, Mohamed; Abderrahmane, Hamid Ait; Paidoussis, Michael P.; Ng, Hoi Dick

    2010-11-01

    The flapping of two parallel flags in axial low turbulence flow is investigated experimentally inside a small scale wind tunnel test section. The variables of the problem are the size and flexural rigidity of the flags, and the distance that separates the two flags. The flow velocity represents the control parameter that governs the coupling and flapping mode of the flags. Two flapping modes, in-phase and out-of-phase modes, were observed in the experiment. Image processing technique was used and the time series of a given point on the flag edge was analyzed. The stability condition of the flags was obtained and compared to the recent theoretical models. The dynamics of the coupling between the two flags is also studied.

  8. Chord-wise Tip Actuation on Flexible Flapping Plates

    Martin, Nathan; Gharib, Morteza

    2015-11-01

    The aerodynamic characteristics of low aspect ratio flapping plates are strongly influenced by the interaction between tip and edge vortices. This has led to the development of tip actuation mechanisms which bend the tip towards the root of the plate in the span-wise direction during oscillation to investigate its impact. In our current work, a tip actuation mechanism to bend a flat plate's two free corners towards one another in the chord-wise direction is developed using a shape memory alloy. The aerodynamic forces and resulting flow field are investigated from dynamically altering the tip chord-wise curvature while flapping. The frequency of oscillation, stroke angle, flexibility, and tip actuation timing are independently varied to determine their individual effects. These results will further the fundamental understanding of flapping wing aerodynamics. This material is based upon work supported by the National Science Foundation Graduate Research Fellowship under Grant No. DGE 1144469.

  9. A Robust Actin Filaments Image Analysis Framework.

    Alioscha-Perez, Mitchel; Benadiba, Carine; Goossens, Katty; Kasas, Sandor; Dietler, Giovanni; Willaert, Ronnie; Sahli, Hichem

    2016-08-01

    The cytoskeleton is a highly dynamical protein network that plays a central role in numerous cellular physiological processes, and is traditionally divided into three components according to its chemical composition, i.e. actin, tubulin and intermediate filament cytoskeletons. Understanding the cytoskeleton dynamics is of prime importance to unveil mechanisms involved in cell adaptation to any stress type. Fluorescence imaging of cytoskeleton structures allows analyzing the impact of mechanical stimulation in the cytoskeleton, but it also imposes additional challenges in the image processing stage, such as the presence of imaging-related artifacts and heavy blurring introduced by (high-throughput) automated scans. However, although there exists a considerable number of image-based analytical tools to address the image processing and analysis, most of them are unfit to cope with the aforementioned challenges. Filamentous structures in images can be considered as a piecewise composition of quasi-straight segments (at least in some finer or coarser scale). Based on this observation, we propose a three-steps actin filaments extraction methodology: (i) first the input image is decomposed into a 'cartoon' part corresponding to the filament structures in the image, and a noise/texture part, (ii) on the 'cartoon' image, we apply a multi-scale line detector coupled with a (iii) quasi-straight filaments merging algorithm for fiber extraction. The proposed robust actin filaments image analysis framework allows extracting individual filaments in the presence of noise, artifacts and heavy blurring. Moreover, it provides numerous parameters such as filaments orientation, position and length, useful for further analysis. Cell image decomposition is relatively under-exploited in biological images processing, and our study shows the benefits it provides when addressing such tasks. Experimental validation was conducted using publicly available datasets, and in osteoblasts grown in

  10. A Robust Actin Filaments Image Analysis Framework

    Alioscha-Perez, Mitchel; Benadiba, Carine; Goossens, Katty; Kasas, Sandor; Dietler, Giovanni; Willaert, Ronnie; Sahli, Hichem

    2016-01-01

    The cytoskeleton is a highly dynamical protein network that plays a central role in numerous cellular physiological processes, and is traditionally divided into three components according to its chemical composition, i.e. actin, tubulin and intermediate filament cytoskeletons. Understanding the cytoskeleton dynamics is of prime importance to unveil mechanisms involved in cell adaptation to any stress type. Fluorescence imaging of cytoskeleton structures allows analyzing the impact of mechanical stimulation in the cytoskeleton, but it also imposes additional challenges in the image processing stage, such as the presence of imaging-related artifacts and heavy blurring introduced by (high-throughput) automated scans. However, although there exists a considerable number of image-based analytical tools to address the image processing and analysis, most of them are unfit to cope with the aforementioned challenges. Filamentous structures in images can be considered as a piecewise composition of quasi-straight segments (at least in some finer or coarser scale). Based on this observation, we propose a three-steps actin filaments extraction methodology: (i) first the input image is decomposed into a ‘cartoon’ part corresponding to the filament structures in the image, and a noise/texture part, (ii) on the ‘cartoon’ image, we apply a multi-scale line detector coupled with a (iii) quasi-straight filaments merging algorithm for fiber extraction. The proposed robust actin filaments image analysis framework allows extracting individual filaments in the presence of noise, artifacts and heavy blurring. Moreover, it provides numerous parameters such as filaments orientation, position and length, useful for further analysis. Cell image decomposition is relatively under-exploited in biological images processing, and our study shows the benefits it provides when addressing such tasks. Experimental validation was conducted using publicly available datasets, and in osteoblasts

  11. Robotics in microsurgery: use of a surgical robot to perform a free flap in a pig.

    Katz, Ryan D; Rosson, Gedge D; Taylor, Jesse A; Singh, Navin K

    2005-01-01

    We present the concept that a surgical robot may be used to successfully perform a free flap. To study different microsurgical techniques, a porcine free flap model was developed in our laboratory. Dissection of the free flap model and isolation of the vessels were completed under traditional loupe magnification. The da Vinci robot was then used to perform vessel adventitiectomy and microanastomoses. The model was observed for 4 h postoperatively, noting flap color, temperature, capillary refill, and Doppler signal. At the end of this period, the flap was noted to be viable; anastomoses were evaluated and found to be grossly and microscopically patent. Advantages conferred by the da Vinci robot include elimination of tremor, scalable movements, fully articulating instruments with six degrees of spatial freedom, and a dynamic three-dimensional visualization system. Drawbacks include the cost and the absence of true microsurgical instruments. PMID:16178007

  12. Boolean gates on actin filaments

    Siccardi, Stefano; Tuszynski, Jack A.; Adamatzky, Andrew

    2016-01-01

    Actin is a globular protein which forms long polar filaments in the eukaryotic cytoskeleton. Actin networks play a key role in cell mechanics and cell motility. They have also been implicated in information transmission and processing, memory and learning in neuronal cells. The actin filaments have been shown to support propagation of voltage pulses. Here we apply a coupled nonlinear transmission line model of actin filaments to study interactions between voltage pulses. To represent digital information we assign a logical TRUTH value to the presence of a voltage pulse in a given location of the actin filament, and FALSE to the pulse's absence, so that information flows along the filament with pulse transmission. When two pulses, representing Boolean values of input variables, interact, then they can facilitate or inhibit further propagation of each other. We explore this phenomenon to construct Boolean logical gates and a one-bit half-adder with interacting voltage pulses. We discuss implications of these findings on cellular process and technological applications.

  13. EhCoactosin stabilizes actin filaments in the protist parasite Entamoeba histolytica.

    Nitesh Kumar

    2014-09-01

    Full Text Available Entamoeba histolytica is a protist parasite that is the causative agent of amoebiasis, and is a highly motile organism. The motility is essential for its survival and pathogenesis, and a dynamic actin cytoskeleton is required for this process. EhCoactosin, an actin-binding protein of the ADF/cofilin family, participates in actin dynamics, and here we report our studies of this protein using both structural and functional approaches. The X-ray crystal structure of EhCoactosin resembles that of human coactosin-like protein, with major differences in the distribution of surface charges and the orientation of terminal regions. According to in vitro binding assays, full-length EhCoactosin binds both F- and G-actin. Instead of acting to depolymerize or severe F-actin, EhCoactosin directly stabilizes the polymer. When EhCoactosin was visualized in E. histolytica cells using either confocal imaging or total internal reflectance microscopy, it was found to colocalize with F-actin at phagocytic cups. Over-expression of this protein stabilized F-actin and inhibited the phagocytic process. EhCoactosin appears to be an unusual type of coactosin involved in E. histolytica actin dynamics.

  14. Epidermolysis Bullosa Simplex-Type Mutations Alter the Dynamics of the Keratin Cytoskeleton and Reveal a Contribution of Actin to the Transport of Keratin SubunitsD⃞V⃞

    Werner, Nicola Susann; Windoffer, Reinhard; Strnad, Pavel; Grund, Christine; Leube, Rudolf Eberhard; Magin, Thomas Michael

    2004-01-01

    Dominant keratin mutations cause epidermolysis bullosa simplex by transforming keratin (K) filaments into aggregates. As a first step toward understanding the properties of mutant keratins in vivo, we stably transfected epithelial cells with an enhanced yellow fluorescent protein-tagged K14R125C mutant. K14R125C became localized as aggregates in the cell periphery and incorporated into perinuclear keratin filaments. Unexpectedly, keratin aggregates were in dynamic equilibrium with soluble sub...

  15. Experience in Reconstruction for Small Digital Defects With Free Flaps.

    Hung, Min-Hsiang; Huang, Kuo-Feng; Chiu, Haw-Yen; Chao, Wai-Nang

    2016-03-01

    Traumatic injuries to the digits resulting in soft tissue or bone loss require reconstruction. Traditionally, local flaps, such as homodigital flaps, heterodigital flaps, pedicled flaps, or distant flaps, are used for digital resurfacing. However, free tissue transfers can be used in selected patients. In this study, we present the use of different free flaps including groin skin flaps, groin osteocutaneous flaps, groin chimeric flaps, second dorsal metacarpal artery flaps, and partial toe flaps for digital reconstruction. A total of 19 digits were treated with 16 free flaps in our hospital. Of the flaps used, 5 were free groin skin flaps, 4 were free partial toe flaps, 3 were free groin chimeric flaps, 2 were free groin osteocutaneous flaps, and 2 were free second dorsal metacarpal artery flaps. The average flap size was 4.7 × 2.0 cm (range, 1.5 × 1 to 5 × 4 cm), and the average operative time was 6.0 hours (range, 4-9 hours). All flaps survived without partial or total necrosis. In conclusion, the free flap is a reliable and safe alternative for digital reconstruction. Moreover, the free groin flap provides not only a chimeric pattern for multiple fingers coverage but also an osteocutaneous pattern for thumb lengthening. The free second dorsal metacarpal artery flap provides a tenocutaneous pattern for tendon reconstruction and soft tissue coverage simultaneously, and the free partial toe flap is an excellent alternative for pulp reconstruction in terms of aesthetic appearance and functional outcome. PMID:26808771

  16. Flapping propulsion with tip pitch control

    Huera-Huarte, Francisco; Gharib, Morteza

    2014-11-01

    The effect of flexibility in the propulsion performance and efficiency of oscillating pitching foils has received a large amount of attention in the past years. Scientists have used simplified robotic models that mimic the kinematics of flying and swimming animals, in order to get inspiration to build more efficient engineering systems. Compliance is one of the aspects that has received more attention, as it seems to be a common feature in nature's flyers and swimmers. Active or passive control elements are also common in nature. We will show how thrust generation in a pitching fin, can be greatly affected by controlling the tip pitch motion dynamically and independently of the fin itself. This is in fact a controlled local change of curvature of the end of the fin. A robotic system has been designed in a way that not only flapping amplitudes and frequencies can be controlled, but also the amplitudes and frequencies of the tip and the phase difference between the tip and the fin. We measured thrust forces and the vortex dynamics in the near wake of the system, by using planar DPIV (Digital Particle Image Velocimetry) in a wide variety of flapping situations with tip control. Funding from Spanish Ministry of Science through Grant DPI2012-37904 is gratefully acknowledged.

  17. Actin-myosin network is required for proper assembly of influenza virus particles

    Actin filaments are known to play a central role in cellular dynamics. After polymerization of actin, various actin-crosslinking proteins including non-muscle myosin II facilitate the formation of spatially organized actin filament networks. The actin-myosin network is highly expanded beneath plasma membrane. The genome of influenza virus (vRNA) replicates in the cell nucleus. Then, newly synthesized vRNAs are nuclear-exported to the cytoplasm as ribonucleoprotein complexes (vRNPs), followed by transport to the beneath plasma membrane where virus particles assemble. Here, we found that, by inhibiting actin-myosin network formation, the virus titer tends to be reduced and HA viral spike protein is aggregated on the plasma membrane. These results indicate that the actin-myosin network plays an important role in the virus formation. - Highlights: • Actin-myosin network is important for the influenza virus production. • HA forms aggregations at the plasma membrane in the presence of blebbistatin. • M1 is recruited to the budding site through the actin-myosin network

  18. Actin-myosin network is required for proper assembly of influenza virus particles

    Kumakura, Michiko; Kawaguchi, Atsushi, E-mail: ats-kawaguchi@md.tsukuba.ac.jp; Nagata, Kyosuke, E-mail: knagata@md.tsukuba.ac.jp

    2015-02-15

    Actin filaments are known to play a central role in cellular dynamics. After polymerization of actin, various actin-crosslinking proteins including non-muscle myosin II facilitate the formation of spatially organized actin filament networks. The actin-myosin network is highly expanded beneath plasma membrane. The genome of influenza virus (vRNA) replicates in the cell nucleus. Then, newly synthesized vRNAs are nuclear-exported to the cytoplasm as ribonucleoprotein complexes (vRNPs), followed by transport to the beneath plasma membrane where virus particles assemble. Here, we found that, by inhibiting actin-myosin network formation, the virus titer tends to be reduced and HA viral spike protein is aggregated on the plasma membrane. These results indicate that the actin-myosin network plays an important role in the virus formation. - Highlights: • Actin-myosin network is important for the influenza virus production. • HA forms aggregations at the plasma membrane in the presence of blebbistatin. • M1 is recruited to the budding site through the actin-myosin network.

  19. Cell stress promotes the association of phosphorylated HspB1 with F-actin.

    Joseph P Clarke

    Full Text Available Previous studies have suggested that the small heat shock protein, HspB1, has a direct influence on the dynamics of cytoskeletal elements, in particular, filamentous actin (F-actin polymerization. In this study we have assessed the influence of HspB1 phosphorylation on its interaction(s with F-actin. We first determined the distribution of endogenous non-phosphorylated HspB1, phosphorylated HspB1 and F-actin in neuroendocrine PC12 cells by immunocytochemistry and confocal microscopy. We then investigated a potential direct interaction between HspB1 with F-actin by precipitating F-actin directly with biotinylated phalloidin followed by Western analyses; the reverse immunoprecipitation of HspB1 was also carried out. The phosphorylation influence of HspB1 in this interaction was investigated by using pharmacologic inhibition of p38 MAPK. In control cells, HspB1 interacts with F-actin as a predominantly non-phosphorylated protein, but subsequent to stress there is a redistribution of HspB1 to the cytoskeletal fraction and a significantly increased association of pHspB1 with F-actin. Our data demonstrate HspB1 is found in a complex with F-actin both in phosphorylated and non-phosphorylated forms, with an increased association of pHspB1 with F-actin after heat stress. Overall, our study combines both cellular and biochemical approaches to show cellular localization and direct demonstration of an interaction between endogenous HspB1 and F-actin using methodolgy that specifically isolates F-actin.

  20. Experimental Study of Wake / Flap Interaction Noise and the Reduction of Flap Side Edge Noise

    Hutcheson, Florence V.; Stead, Daniel J.; Plassman, Gerald E.

    2016-01-01

    The effects of the interaction of a wake with a half-span flap on radiated noise are examined. The incident wake is generated by bars of various widths and lengths or by a simplified landing gear model. Single microphone and phased array measurements are used to isolate the effects of the wake interaction on the noise radiating from the flap side edge and flap cove regions. The effects on noise of the wake generator's geometry and relative placement with respect to the flap are assessed. Placement of the wake generators upstream of the flap side edge is shown to lead to the reduction of flap side edge noise by introducing a velocity deficit and likely altering the instabilities in the flap side edge vortex system. Significant reduction in flap side edge noise is achieved with a bar positioned directly upstream of the flap side edge. The noise reduction benefit is seen to improve with increased bar width, length and proximity to the flap edge. Positioning of the landing gear model upstream of the flap side edge also leads to decreased flap side edge noise. In addition, flap cove noise levels are significantly lower than when the landing gear is positioned upstream of the flap mid-span. The impact of the local flow velocity on the noise radiating directly from the landing gear is discussed. The effects of the landing gear side-braces on flap side edge, flap cove and landing gear noise are shown.

  1. Spiral actin-polymerization waves can generate amoeboidal cell crawling

    Amoeboidal cell crawling on solid substrates is characterized by protrusions that seemingly appear randomly along the cell periphery and drive the cell forward. For many cell types, it is known that the protrusions result from polymerization of the actin cytoskeleton. However, little is known about how the formation of protrusions is triggered and whether the appearance of subsequent protrusions is coordinated. Recently, the spontaneous formation of actin-polymerization waves was observed. These waves have been proposed to orchestrate the cytoskeletal dynamics during cell crawling. Here, we study the impact of cytoskeletal polymerization waves on cell migration using a phase-field approach. In addition to directionally moving cells, we find states reminiscent of amoeboidal cell crawling. In this framework, new protrusions are seen to emerge from a nucleation process, generating spiral actin waves in the cell interior. Nucleation of new spirals does not require noise, but occurs in a state that is apparently displaying spatio-temporal chaos. (paper)

  2. Calcium-actin waves and oscillations of cellular membranes.

    Veksler, Alex; Gov, Nir S

    2009-09-16

    We propose a mechanism for the formation of membrane oscillations and traveling waves, which arise due to the coupling between the actin cytoskeleton and the calcium flux through the membrane. In our model, the fluid cell membrane has a mobile but constant population of proteins with a convex spontaneous curvature, which act as nucleators of actin polymerization and adhesion. Such a continuum model couples the forces of cell-substrate adhesion, actin polymerization, membrane curvature, and the flux of calcium through the membrane. Linear stability analysis shows that sufficiently strong coupling among the calcium, membrane, and protein dynamics may induce robust traveling waves on the membrane. This result was checked for a reduced feedback scheme and is compared to the results without the effects of calcium, where permanent phase separation without waves or oscillations is obtained. The model results are compared to the published observations of calcium waves in cell membranes, and a number of testable predictions are proposed. PMID:19751660

  3. Cortactin Adopts a Globular Conformation and Bundles Actin into Sheets

    Cowieson, Nathan P.; King, Gordon; Cookson, David; Ross, Ian; Huber, Thomas; Hume, David A.; Kobe, Bostjan; Martin, Jennifer L. (Queensland); (Aust. Synch.)

    2008-08-21

    Cortactin is a filamentous actin-binding protein that plays a pivotal role in translating environmental signals into coordinated rearrangement of the cytoskeleton. The dynamic reorganization of actin in the cytoskeleton drives processes including changes in cell morphology, cell migration, and phagocytosis. In general, structural proteins of the cytoskeleton bind in the N-terminal region of cortactin and regulatory proteins in the C-terminal region. Previous structural studies have reported an extended conformation for cortactin. It is therefore unclear how cortactin facilitates cross-talk between structural proteins and their regulators. In the study presented here, circular dichroism, chemical cross-linking, and small angle x-ray scattering are used to demonstrate that cortactin adopts a globular conformation, thereby bringing distant parts of the molecule into close proximity. In addition, the actin bundling activity of cortactin is characterized, showing that fully polymerized actin filaments are bundled into sheet-like structures. We present a low resolution structure that suggests how the various domains of cortactin interact to coordinate its array of binding partners at sites of actin branching.

  4. Antenna Mechanism of Length Control of Actin Cables.

    Lishibanya Mohapatra

    2015-06-01

    Full Text Available Actin cables are linear cytoskeletal structures that serve as tracks for myosin-based intracellular transport of vesicles and organelles in both yeast and mammalian cells. In a yeast cell undergoing budding, cables are in constant dynamic turnover yet some cables grow from the bud neck toward the back of the mother cell until their length roughly equals the diameter of the mother cell. This raises the question: how is the length of these cables controlled? Here we describe a novel molecular mechanism for cable length control inspired by recent experimental observations in cells. This "antenna mechanism" involves three key proteins: formins, which polymerize actin, Smy1 proteins, which bind formins and inhibit actin polymerization, and myosin motors, which deliver Smy1 to formins, leading to a length-dependent actin polymerization rate. We compute the probability distribution of cable lengths as a function of several experimentally tuneable parameters such as the formin-binding affinity of Smy1 and the concentration of myosin motors delivering Smy1. These results provide testable predictions of the antenna mechanism of actin-cable length control.

  5. Novel actin-like filament structure from Clostridium tetani.

    Popp, David; Narita, Akihiro; Lee, Lin Jie; Ghoshdastider, Umesh; Xue, Bo; Srinivasan, Ramanujam; Balasubramanian, Mohan K; Tanaka, Toshitsugu; Robinson, Robert C

    2012-06-15

    Eukaryotic F-actin is constructed from two protofilaments that gently wind around each other to form a helical polymer. Several bacterial actin-like proteins (Alps) are also known to form F-actin-like helical arrangements from two protofilaments, yet with varied helical geometries. Here, we report a unique filament architecture of Alp12 from Clostridium tetani that is constructed from four protofilaments. Through fitting of an Alp12 monomer homology model into the electron microscopy data, the filament was determined to be constructed from two antiparallel strands, each composed of two parallel protofilaments. These four protofilaments form an open helical cylinder separated by a wide cleft. The molecular interactions within single protofilaments are similar to F-actin, yet interactions between protofilaments differ from those in F-actin. The filament structure and assembly and disassembly kinetics suggest Alp12 to be a dynamically unstable force-generating motor involved in segregating the pE88 plasmid, which encodes the lethal tetanus toxin, and thus a potential target for drug design. Alp12 can be repeatedly cycled between states of polymerization and dissociation, making it a novel candidate for incorporation into fuel-propelled nanobiopolymer machines. PMID:22514279

  6. Free thin paraumbilical perforator-based flaps.

    Koshima, I; Moriguchi, T; Soeda, S; Tanaka, H; Umeda, N

    1992-07-01

    A free paraumbilical perforator-based flap fed by a muscle perforator from the inferior deep epigastric artery and with no muscle was used in 13 patients. Among them, a free thin paraumbilical perforator-based flap with a thin layer of fat, to protect the subdermal plexus of the vessels, was used in seven patients. The dominant pedicle perforator of this thin flap is usually located around the umbilicus and a large flap can be obtained. Its critical length-to-breath ratio is considered to be 4:3. The advantages of this flap are a long and large vascular pedicle, rare postoperative abdominal herniation, little bulkiness of the flap, and a relatively large skin territory. The disadvantages are technical difficulties in dissection of the perforator and anatomical variation in the location of the perforator. We believe this flap largely overcomes the problems of the conventional rectus abdominis musculocutaneous flap. PMID:1386718

  7. Identification of Actin-Binding Proteins from Maize Pollen

    Staiger, C.J.

    2004-01-13

    Specific Aims--The goal of this project was to gain an understanding of how actin filament organization and dynamics are controlled in flowering plants. Specifically, we proposed to identify unique proteins with novel functions by investigating biochemical strategies for the isolation and characterization of actin-binding proteins (ABPs). In particular, our hunt was designed to identify capping proteins and nucleation factors. The specific aims included: (1) to use F-actin affinity chromatography (FAAC) as a general strategy to isolate pollen ABPs (2) to produce polyclonal antisera and perform subcellular localization in pollen tubes (3) to isolate cDNA clones for the most promising ABPs (4) to further purify and characterize ABP interactions with actin in vitro. Summary of Progress By employing affinity chromatography on F-actin or DNase I columns, we have identified at least two novel ABPs from pollen, PrABP80 (gelsolin-like) and ZmABP30, We have also cloned and expressed recombinant protein, as well as generated polyclonal antisera, for 6 interesting ABPs from Arabidopsis (fimbrin AtFIM1, capping protein a/b (AtCP), adenylyl cyclase-associated protein (AtCAP), AtCapG & AtVLN1). We performed quantitative analyses of the biochemical properties for two of these previously uncharacterized ABPs (fimbrin and capping protein). Our studies provide the first evidence for fimbrin activity in plants, demonstrate the existence of barbed-end capping factors and a gelsolin-like severing activity, and provide the quantitative data necessary to establish and test models of F-actin organization and dynamics in plant cells.

  8. Steady-state nuclear actin levels are determined by export competent actin pool.

    Skarp, Kari-Pekka; Huet, Guillaume; Vartiainen, Maria K

    2013-10-01

    A number of studies in the last decade have irrevocably promoted actin into a fully fledged member of the nuclear compartment, where it, among other crucial tasks, facilitates transcription and chromatin remodeling. Changes in nuclear actin levels have been linked to different cellular processes: decreased nuclear actin to quiescence and increased nuclear actin to differentiation. Importin 9 and exportin 6 transport factors are responsible for the continuous nucleocytoplasmic shuttling of actin, but the mechanisms, which result in modulated actin levels, have not been characterized. We find that in cells growing under normal growth conditions, the levels of nuclear actin vary considerably from cell to cell. To understand the basis for this, we have extensively quantified several cellular parameters while at the same time recording the import and export rates of green fluorescent protein (GFP)-tagged actin. Surprisingly, our dataset shows that the ratio of nuclear to cytoplasmic fluorescence intensity, but not nuclear shape, size, cytoplasm size, or their ratio, correlates negatively with both import and export rate of actin. This suggests that high-nuclear actin content is maintained by both diminished import and export. The high nuclear actin containing cells still show high mobility of actin, but it is not export competent, suggesting increased binding of actin to nuclear complexes. Creation of such export incompetent actin pool would ensure enough actin is retained in the nucleus and make it available for the various nuclear functions described for actin. PMID:23749625

  9. Efficient flapping flight of pterosaurs

    Strang, Karl Axel

    In the late eighteenth century, humans discovered the first pterosaur fossil remains and have been fascinated by their existence ever since. Pterosaurs exploited their membrane wings in a sophisticated manner for flight control and propulsion, and were likely the most efficient and effective flyers ever to inhabit our planet. The flapping gait is a complex combination of motions that sustains and propels an animal in the air. Because pterosaurs were so large with wingspans up to eleven meters, if they could have sustained flapping flight, they would have had to achieve high propulsive efficiencies. Identifying the wing motions that contribute the most to propulsive efficiency is key to understanding pterosaur flight, and therefore to shedding light on flapping flight in general and the design of efficient ornithopters. This study is based on published results for a very well-preserved specimen of Coloborhynchus robustus, for which the joints are well-known and thoroughly described in the literature. Simplifying assumptions are made to estimate the characteristics that can not be inferred directly from the fossil remains. For a given animal, maximizing efficiency is equivalent to minimizing power at a given thrust and speed. We therefore aim at finding the flapping gait, that is the joint motions, that minimize the required flapping power. The power is computed from the aerodynamic forces created during a given wing motion. We develop an unsteady three-dimensional code based on the vortex-lattice method, which correlates well with published results for unsteady motions of rectangular wings. In the aerodynamic model, the rigid pterosaur wing is defined by the position of the bones. In the aeroelastic model, we add the flexibility of the bones and of the wing membrane. The nonlinear structural behavior of the membrane is reduced to a linear modal decomposition, assuming small deflections about the reference wing geometry. The reference wing geometry is computed for

  10. Effects of leading-edge flap oscillation on unsteady delta wing flow and rock control

    Kandil, Osama A.; Salman, Ahmed A.

    1991-01-01

    The isolated and interdisciplinary problems of unsteady fluid dynamics and rigid-body dynamics and control of delta wings with and without leading-edge flap oscillation are considered. For the fluid dynamics problem, the unsteady, compressible, thin-layer Navier-Stokes (NS) equations, which are written relative to a moving frame of reference, are solved along with the unsteady, linearized, Navier-displacement (ND) equations. The NS equations are solved for the flowfield using an implicit finite-volume scheme. The ND equations are solved for the grid deformation, if the leading-edge flaps oscillate, using an ADI scheme. For the dynamics and control problem, the Euler equation of rigid-body rolling motion for a wing and its flaps are solved interactively with the fluid dynamics equations for the wing-rock motion and subsequently for its control. A four-stage Runge-Kutta scheme is used to explicitly integrate the dynamics equation.

  11. A new twist on gyroscopic sensing: body rotations lead to torsion in flapping, flexing insect wings.

    Eberle, A L; Dickerson, B H; Reinhall, P G; Daniel, T L

    2015-03-01

    Insects perform fast rotational manoeuvres during flight. While two insect orders use flapping halteres (specialized organs evolved from wings) to detect body dynamics, it is unknown how other insects detect rotational motions. Like halteres, insect wings experience gyroscopic forces when they are flapped and rotated and recent evidence suggests that wings might indeed mediate reflexes to body rotations. But, can gyroscopic forces be detected using only changes in the structural dynamics of a flapping, flexing insect wing? We built computational and robotic models to rotate a flapping wing about an axis orthogonal to flapping. We recorded high-speed video of the model wing, which had a flexural stiffness similar to the wing of the Manduca sexta hawkmoth, while flapping it at the wingbeat frequency of Manduca (25 Hz). We compared the three-dimensional structural dynamics of the wing with and without a 3 Hz, 10° rotation about the yaw axis. Our computational model revealed that body rotation induces a new dynamic mode: torsion. We verified our result by measuring wing tip displacement, shear strain and normal strain of the robotic wing. The strains we observed could stimulate an insect's mechanoreceptors and trigger reflexive responses to body rotations. PMID:25631565

  12. The IpaC carboxyterminal effector domain mediates Src-dependent actin polymerization during Shigella invasion of epithelial cells

    Mounier, Joëlle; Popoff, Michel R.; Enninga, Jost; Frame, Margaret C; Sansonetti, Philippe J.; Van Nhieu, Guy Tran

    2009-01-01

    Shigella, the causative agent of bacillary dysentery, invades epithelial cells by locally reorganizing the actin cytoskeleton. Shigella invasion requires actin polymerization dependent on the Src tyrosine kinase and a functional bacterial type III secretion (T3S) apparatus. Using dynamic as well as immunofluorescence microscopy, we show that the T3S translocon component IpaC allows the recruitment of the Src kinase required for actin polymerization at bacterial entry sites during the initial ...

  13. The IpaC Carboxyterminal Effector Domain Mediates Src-Dependent Actin Polymerization during Shigella Invasion of Epithelial Cells

    Mounier, Joëlle; Popoff, Michel R.; Enninga, Jost; Frame, Margaret C; Sansonetti, Philippe J.; Van Nhieu, Guy Tran

    2009-01-01

    Shigella, the causative agent of bacillary dysentery, invades epithelial cells by locally reorganizing the actin cytoskeleton. Shigella invasion requires actin polymerization dependent on the Src tyrosine kinase and a functional bacterial type III secretion (T3S) apparatus. Using dynamic as well as immunofluorescence microscopy, we show that the T3S translocon component IpaC allows the recruitment of the Src kinase required for actin polymerization at bacterial entry sites during the initial ...

  14. Uncertainty Analysis for a Jet Flap Airfoil

    Green, Lawrence L.; Cruz, Josue

    2006-01-01

    An analysis of variance (ANOVA) study was performed to quantify the potential uncertainties of lift and pitching moment coefficient calculations from a computational fluid dynamics code, relative to an experiment, for a jet flap airfoil configuration. Uncertainties due to a number of factors including grid density, angle of attack and jet flap blowing coefficient were examined. The ANOVA software produced a numerical model of the input coefficient data, as functions of the selected factors, to a user-specified order (linear, 2-factor interference, quadratic, or cubic). Residuals between the model and actual data were also produced at each of the input conditions, and uncertainty confidence intervals (in the form of Least Significant Differences or LSD) for experimental, computational, and combined experimental / computational data sets were computed. The LSD bars indicate the smallest resolvable differences in the functional values (lift or pitching moment coefficient) attributable solely to changes in independent variable, given just the input data points from selected data sets. The software also provided a collection of diagnostics which evaluate the suitability of the input data set for use within the ANOVA process, and which examine the behavior of the resultant data, possibly suggesting transformations which should be applied to the data to reduce the LSD. The results illustrate some of the key features of, and results from, the uncertainty analysis studies, including the use of both numerical (continuous) and categorical (discrete) factors, the effects of the number and range of the input data points, and the effects of the number of factors considered simultaneously.

  15. Actomyosin contraction, aggregation and traveling waves in a treadmilling actin array

    Oelz, Dietmar; Mogilner, Alex

    2016-04-01

    We use perturbation theory to derive a continuum model for the dynamic actomyosin bundle/ring in the regime of very strong crosslinking. Actin treadmilling is essential for contraction. Linear stability analysis and numerical solutions of the model equations reveal that when the actin treadmilling is very slow, actin and myosin aggregate into equidistantly spaced peaks. When treadmilling is significant, actin filament of one polarity are distributed evenly, while filaments of the opposite polarity develop a shock wave moving with the treadmilling velocity. Myosin aggregates into a sharp peak surfing the crest of the actin wave. Any actomyosin aggregation diminishes contractile stress. The easiest way to maintain higher contraction is to upregulate the actomyosin turnover which destabilizes nontrivial patterns and stabilizes the homogeneous actomyosin distributions. We discuss the model's implications for the experiment.

  16. Stability of actin-lysozyme complexes formed in cystic fibrosis disease.

    Mohammadinejad, Sarah; Ghamkhari, Behnoush; Abdolmaleki, Sarah

    2016-08-21

    Finding the conditions for destabilizing actin-lysozyme complexes is of biomedical importance in preventing infections in cystic fibrosis. In this manuscript, the effects of different charge-mutants of lysozyme and salt concentration on the stability of actin-lysozyme complexes are studied using Langevin dynamics simulation. A coarse-grained model of F-actin is used in which both its twist and bending rigidities are considered. We observe that the attraction between F-actins is stronger in the presence of wild-type lysozymes relative to the mutated lysozymes of lower charges. By calculating the potential of mean force between F-actins, we conclude that the stability of actin-lysozyme complexes is decreased by reducing the charge of lysozyme mutants. The distributions of different lysozyme charge-mutants show that wild-type (+9e) lysozymes are mostly accumulated in the center of triangles formed by three adjacent F-actins, while lysozyme mutants of charges +7e and +5e occupy the bridging regions between F-actins. Low-charge mutants of lysozyme (+3e) distribute uniformly around F-actins. A rough estimate of the electrostatic energy for these different distributions proves that the distribution in which lysozymes reside in the center of triangles leads to more stable complexes. Also our results in the presence of a salt suggest that at physiological salt concentration of airway, F-actin complexes are not formed by charge-reduced mutants of lysozyme. The findings are interesting because if we can design charge-reduced lysozyme mutants with considerable antibacterial activity, they are not sequestered inside F-actin aggregates and can play their role as antibacterial agents against airway infection. PMID:27436705

  17. Alpha-herpesvirus infection induces the formation of nuclear actin filaments.

    Feierbach, Becket; Piccinotti, Silvia; Bisher, Margaret; Denk, Winfried; Enquist, Lynn W

    2006-08-01

    Herpesviruses are large double-stranded DNA viruses that replicate in the nuclei of infected cells. Spatial control of viral replication and assembly in the host nucleus is achieved by the establishment of nuclear compartments that serve to concentrate viral and host factors. How these compartments are established and maintained remains poorly understood. Pseudorabies virus (PRV) is an alpha-herpesvirus often used to study herpesvirus invasion and spread in the nervous system. Here, we report that PRV and herpes simplex virus type 1 infection of neurons results in formation of actin filaments in the nucleus. Filamentous actin is not found in the nucleus of uninfected cells. Nuclear actin filaments appear physically associated with the viral capsids, as shown by serial block-face scanning electron micropscopy and confocal microscopy. Using a green fluorescent protein-tagged viral capsid protein (VP26), we show that nuclear actin filaments form prior to capsid assembly and are required for the efficient formation of viral capsid assembly sites. We find that actin polymerization dynamics (e.g., treadmilling) are not necessary for the formation of these sites. Green fluorescent protein-VP26 foci co-localize with the actin motor myosin V, suggesting that viral capsids travel along nuclear actin filaments using myosin-based directed transport. Viral transcription, but not viral DNA replication, is required for actin filament formation. The finding that infection, by either PRV or herpes simplex virus type 1, results in formation of nuclear actin filaments in neurons, and that PRV infection of an epithelial cell line results in a similar phenotype is evidence that F-actin plays a conserved role in herpesvirus assembly. Our results suggest a mechanism by which assembly domains are organized within infected cells and provide insight into how the viral infectious cycle and host actin cytoskeleton are integrated to promote the infection process. PMID:16933992

  18. Membrane Supply and Demand Regulates F-Actin in a Cell Surface Reservoir.

    Figard, Lauren; Wang, Mengyu; Zheng, Liuliu; Golding, Ido; Sokac, Anna Marie

    2016-05-01

    Cells store membrane in surface reservoirs of pits and protrusions. These membrane reservoirs facilitate cell shape change and buffer mechanical stress, but we do not know how reservoir dynamics are regulated. During cellularization, the first cytokinesis in Drosophila embryos, a reservoir of microvilli unfolds to fuel cleavage furrow ingression. We find that regulated exocytosis adds membrane to the reservoir before and during unfolding. Dynamic F-actin deforms exocytosed membrane into microvilli. Single microvilli extend and retract in ∼20 s, while the overall reservoir is depleted in sync with furrow ingression over 60-70 min. Using pharmacological and genetic perturbations, we show that exocytosis promotes microvillar F-actin assembly, while furrow ingression controls microvillar F-actin disassembly. Thus, reservoir F-actin and, consequently, reservoir dynamics are regulated by membrane supply from exocytosis and membrane demand from furrow ingression. PMID:27165556

  19. The need for higher-order averaging in the stability analysis of hovering, flapping-wing flight

    Because of the relatively high flapping frequency associated with hovering insects and flapping wing micro-air vehicles (FWMAVs), dynamic stability analysis typically involves direct averaging of the time-periodic dynamics over a flapping cycle. However, direct application of the averaging theorem may lead to false conclusions about the dynamics and stability of hovering insects and FWMAVs. Higher-order averaging techniques may be needed to understand the dynamics of flapping wing flight and to analyze its stability. We use second-order averaging to analyze the hovering dynamics of five insects in response to high-amplitude, high-frequency, periodic wing motion. We discuss the applicability of direct averaging versus second-order averaging for these insects. (paper)

  20. The need for higher-order averaging in the stability analysis of hovering, flapping-wing flight.

    Taha, Haithem E; Tahmasian, Sevak; Woolsey, Craig A; Nayfeh, Ali H; Hajj, Muhammad R

    2015-01-01

    Because of the relatively high flapping frequency associated with hovering insects and flapping wing micro-air vehicles (FWMAVs), dynamic stability analysis typically involves direct averaging of the time-periodic dynamics over a flapping cycle. However, direct application of the averaging theorem may lead to false conclusions about the dynamics and stability of hovering insects and FWMAVs. Higher-order averaging techniques may be needed to understand the dynamics of flapping wing flight and to analyze its stability. We use second-order averaging to analyze the hovering dynamics of five insects in response to high-amplitude, high-frequency, periodic wing motion. We discuss the applicability of direct averaging versus second-order averaging for these insects. PMID:25561166

  1. The Actin-Binding Protein α-Adducin Is Required for Maintaining Axon Diameter

    Sérgio Carvalho Leite

    2016-04-01

    Full Text Available The actin-binding protein adducin was recently identified as a component of the neuronal subcortical cytoskeleton. Here, we analyzed mice lacking adducin to uncover the function of this protein in actin rings. α-adducin knockout mice presented progressive axon enlargement in the spinal cord and optic and sciatic nerves, followed by axon degeneration and loss. Using stimulated emission depletion super-resolution microscopy, we show that a periodic subcortical actin cytoskeleton is assembled in every neuron type inspected including retinal ganglion cells and dorsal root ganglia neurons. In neurons devoid of adducin, the actin ring diameter increased, although the inter-ring periodicity was maintained. In vitro, the actin ring diameter adjusted as axons grew, suggesting the lattice is dynamic. Our data support a model in which adducin activity is not essential for actin ring assembly and periodicity but is necessary to control the diameter of both actin rings and axons and actin filament growth within rings.

  2. Actinic Keratoses: A Comprehensive Update

    Ibrahim, Sherrif F.; Brown, Marc D.

    2009-01-01

    Actinic keratoses are common intra-epidermal neoplasms that lie on a continuum with squamous cell carcinoma. Tightly linked to ultraviolet irradiation, they occur in areas of chronic sun exposure, and early treatment of these lesions may prevent their progression to invasive disease. A large variety of effective treatment modalities exist, and the optimal therapeutic choice is dependent on a variety of patient- and physician-associated variables. Many established and more recent approaches ar...

  3. The possibility for use of venous flaps in plastic surgery

    The use of venous flaps is controversial. The mechanism of perfusion of venous flaps is still not fully understood. The research was conducted on 56 white rats. In our experimental work we studied two different models of venous flaps: pedicled venous flap (PVF) and pedicled arterialized venous flap (PAVF). Our results showed that postoperative congestion was present in all flaps. However 66.7% of all pedicled venous flaps and 100% of all pedicled arterialized venous flaps eventually survived. Histological examination revealed that postoperatively the blood flow in the skin of the pedicled arterialized venous flap became «re-reversed» again; there were no differences between mechanism of survival of venous flaps and other flaps. On the 7-14th day in the skin of all flaps were processes of neoangiogenesis and proliferation. Hence the best scenario for the clinical use of venous flaps unfolds when both revascularization and skin coverage are required

  4. The possibility for use of venous flaps in plastic surgery

    Baytinger, V. F.; Kurochkina, O. S.; Selianinov, K. V.; Baytinger, A. V.; Dzyuman, A. N.

    2015-11-01

    The use of venous flaps is controversial. The mechanism of perfusion of venous flaps is still not fully understood. The research was conducted on 56 white rats. In our experimental work we studied two different models of venous flaps: pedicled venous flap (PVF) and pedicled arterialized venous flap (PAVF). Our results showed that postoperative congestion was present in all flaps. However 66.7% of all pedicled venous flaps and 100% of all pedicled arterialized venous flaps eventually survived. Histological examination revealed that postoperatively the blood flow in the skin of the pedicled arterialized venous flap became «re-reversed» again; there were no differences between mechanism of survival of venous flaps and other flaps. On the 7-14th day in the skin of all flaps were processes of neoangiogenesis and proliferation. Hence the best scenario for the clinical use of venous flaps unfolds when both revascularization and skin coverage are required.

  5. The possibility for use of venous flaps in plastic surgery

    Baytinger, V. F., E-mail: baitinger@mail.tomsknet.ru; Kurochkina, O. S., E-mail: kurochkinaos@yandex.ru; Selianinov, K. V.; Baytinger, A. V. [Research Institute of Microsurgery, Tomsk (Russian Federation); Dzyuman, A. N. [Siberian State Medical University, Tomsk (Russian Federation)

    2015-11-17

    The use of venous flaps is controversial. The mechanism of perfusion of venous flaps is still not fully understood. The research was conducted on 56 white rats. In our experimental work we studied two different models of venous flaps: pedicled venous flap (PVF) and pedicled arterialized venous flap (PAVF). Our results showed that postoperative congestion was present in all flaps. However 66.7% of all pedicled venous flaps and 100% of all pedicled arterialized venous flaps eventually survived. Histological examination revealed that postoperatively the blood flow in the skin of the pedicled arterialized venous flap became «re-reversed» again; there were no differences between mechanism of survival of venous flaps and other flaps. On the 7-14th day in the skin of all flaps were processes of neoangiogenesis and proliferation. Hence the best scenario for the clinical use of venous flaps unfolds when both revascularization and skin coverage are required.

  6. Modelling phagosomal lipid networks that regulate actin assembly

    Schwarz Roland

    2008-12-01

    Full Text Available Abstract Background When purified phagosomes are incubated in the presence of actin under appropriate conditions, microfilaments start growing from the membrane in a process that is affected by ATP and the lipid composition of the membrane. Isolated phagosomes are metabolically active organelles that contain enzymes and metabolites necessary for lipid interconversion. Hence, addition of ATP, lipids, and actin to the system alter the steady-state composition of the phagosomal membrane at the same time that the actin nucleation is initiated. Our aim was to model all these processes in parallel. Results We compiled detailed experimental data on the effects of different lipids and ATP on actin nucleation and we investigated experimentally lipid interconversion and ATP metabolism in phagosomes by using suitable radioactive compounds. In a first step, a complex lipid network interconnected by chemical reactions catalyzed by known enzymes was modelled in COPASI (Complex Pathway Simulator. However, several lines of experimental evidence indicated that only the phosphatidylinositol branch of the network was active, an observation that dramatically reduced the number of parameters in the model. The results also indicated that a lipid network-independent ATP-consuming activity should be included in the model. When this activity was introduced, the set of differential equations satisfactorily reproduced the experimental data. On the other hand, a molecular mechanism connecting membrane lipids, ATP, and the actin nucleation process is still missing. We therefore adopted a phenomenological (black-box approach to represent the empirical observations. We proposed that lipids and ATP influence the dynamic interconversion between active and inactive actin nucleation sites. With this simple model, all the experimental data were satisfactorily fitted with a single positive parameter per lipid and ATP. Conclusion By establishing an active 'dialogue' between an

  7. Mesoscopic model of actin-based propulsion.

    Jie Zhu

    Full Text Available Two theoretical models dominate current understanding of actin-based propulsion: microscopic polymerization ratchet model predicts that growing and writhing actin filaments generate forces and movements, while macroscopic elastic propulsion model suggests that deformation and stress of growing actin gel are responsible for the propulsion. We examine both experimentally and computationally the 2D movement of ellipsoidal beads propelled by actin tails and show that neither of the two models can explain the observed bistability of the orientation of the beads. To explain the data, we develop a 2D hybrid mesoscopic model by reconciling these two models such that individual actin filaments undergoing nucleation, elongation, attachment, detachment and capping are embedded into the boundary of a node-spring viscoelastic network representing the macroscopic actin gel. Stochastic simulations of this 'in silico' actin network show that the combined effects of the macroscopic elastic deformation and microscopic ratchets can explain the observed bistable orientation of the actin-propelled ellipsoidal beads. To test the theory further, we analyze observed distribution of the curvatures of the trajectories and show that the hybrid model's predictions fit the data. Finally, we demonstrate that the model can explain both concave-up and concave-down force-velocity relations for growing actin networks depending on the characteristic time scale and network recoil. To summarize, we propose that both microscopic polymerization ratchets and macroscopic stresses of the deformable actin network are responsible for the force and movement generation.

  8. From pollen actin to crop male sterility

    2000-01-01

    Actin plays an important role in the life activity of animal and plant cells. Pollen cells have plenty of actin whose structure and characteristics are very similar to the animal actin. The nucleotide sequence and amino acid sequence of plant actin gene are very similar to those of the animal gene. The content of pollen actin from male sterile plants is much more lower than that from its maintainer plants. The expression of actin gene is organ-specific during the plant development. The expression quantity of actin gene in pollen is much more higher than those from root, stem and leaf. The expression plasmid of the anti-sense actin gene was constructed, transferred to the protoplasts of wheat and tomato to inhibit the expression of actin gene in pollen and thus the male sterile plants of wheat and tomato were obtained. The actin in pollens from the transgenic plants was reduced significantly, whereas the pistil was not affected. This study might pave a new way to breeding male sterile lines for the application of hybrid vigor of wheat and tomato.

  9. Effects of torsion frequencies on rotor performance and structural loads with trailing edge flap

    The effects of variation of blade torsion frequency on rotor performance and structural loads are investigated for a 1/rev active flap rotor and baseline rotor (no active control). The UH-60A four-bladed articulated main rotor is studied at a high-speed forward flight condition. The torsion frequencies are varied by modifying the spanwise torsional stiffness of the blade and/or the pitch link stiffness. First, a parametric/optimization study on the flap deployment schedule is carried out using lifting-line comprehensive analysis for the soft, baseline, and stiff rotor configurations, and then a higher fidelity coupled computational fluid dynamics–computational structural dynamics analysis is carried out for the optimal flap deployment. It is shown that with the soft rotor there is degradation in performance—of about 6% with respect to the baseline rotor in the case where the flaps are not activated, and of about 1% if flap deflections are applied. On the other hand, for the stiff rotor there is a slight improvement in performance of about 2.3% when the flaps are not activated, and no appreciable change in the case where active flap deflections are applied. It appears that the peak performance achievable with using active flaps on a baseline stiffness rotor cannot be further improved significantly by varying the torsional frequencies. Variation of torsion frequency does not appear to have a significant influence on blade torsion moments and pitch link loads, although the 1/rev flap activation examined has an important role. (paper)

  10. Aerodynamic response of an airfoil section undergoing pitch motion and trailing edge flap deflection: a comparison of simulation methods

    Bergami, Leonardo; Riziotis, Vasilis A.; Gaunaa, Mac

    2015-01-01

    generated by the airfoil undergoing harmonic pitching motions and harmonic flap deflections. The unsteady aerodynamic coefficients exhibit significant variations over the corresponding steady-state values. The dynamic characteristics of the unsteady response are predicted with an excellent agreement among...