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Sample records for aster microtubule dynamics

  1. Physical Basis of Large Microtubule Aster Growth

    CERN Document Server

    Ishihara, Keisuke; Mitchison, Timothy J

    2016-01-01

    Microtubule asters - radial arrays of microtubules organized by centrosomes - play a fundamental role in the spatial coordination of animal cells. The standard model of aster growth assumes a fixed number of microtubules originating from the centrosomes. However, aster morphology in this model does not scale with cell size, and we recently found evidence for non-centrosomal microtubule nucleation. Here, we combine autocatalytic nucleation and polymerization dynamics to develop a biophysical model of aster growth. Our model predicts that asters expand as traveling waves and recapitulates all major aspects of aster growth. As the nucleation rate increases, the model predicts an explosive transition from stationary to growing asters with a discontinuous jump of the growth velocity to a nonzero value. Experiments in frog egg extract confirm the main theoretical predictions. Our results suggest that asters observed in large frog and amphibian eggs are a meshwork of short, unstable microtubules maintained by autoca...

  2. Dynamics of microtubule asters in microfabricated chambers: The role of catastrophes

    Science.gov (United States)

    Faivre-Moskalenko, Cendrine; Dogterom, Marileen

    2002-01-01

    Recent in vivo as well as in vitro experiments have indicated that microtubule pushing alone is sufficient to position a microtubule-organizing center within a cell. Here, we investigate the effect of catastrophes on the dynamics of microtubule asters within microfabricated chambers that mimic the confining geometry of living cells. The use of a glass bead as the microtubule-organizing center allows us to manipulate the aster by using optical tweezers. In the case in which microtubules preexist, we show that because of microtubule buckling, repositioning almost never occurs after relocation with the optical tweezers, although initial microtubule growth always leads the aster to the geometrical center of the chamber. When a catastrophe promoter is added, we find instead that the aster is able to efficiently explore the chamber geometry even after being relocated with the optical tweezers. As predicted by theoretical calculations, the results of our in vitro experiments clearly demonstrate the need for catastrophes for proper positioning in a confining geometry. These findings correlate with recent observations of nuclear positioning in fission yeast cells. PMID:12486218

  3. The microtubule aster formation and its role in nuclear envelope assembly around the sperm chromatin in Xenopus egg extracts

    Institute of Scientific and Technical Information of China (English)

    YANG Ning; CHEN Zhongcai; LU Ping; ZHANG Chuanmao; ZHAI Zhonghe; TANG Xiaowei

    2003-01-01

    Nuclear envelope is a dynamic structure in the cell cycle. At the beginning of mitosis, nuclear envelope breaks down and its components disperse into the cytoplasm. At the end of mitosis, nuclear envelope reassembles using the dispersed components. Searching for the mechanisms of the nuclear disassembly and reassembly has for a long time been one of the key projects for cell biologists. In this report we show that microtubules take a role in the nuclear envelope assembly around the sperm chromatin in Xenopus egg extracts. Microtubule cytoskeleton has been demonstrated to take roles in the transport of intracellular membranes such as Golgi and ER vesicles. We found that the nuclear envelope assembly needs functional microtubules. At the beginning of the nuclear assembly, microtubules nucleated to form a microtubule aster around the centrosome at the base of the sperm head. Using the microtubule drug colchicine to disrupt the microtubule nucleation, nuclear envelope reassembly was seriously inhibited. If the microtubules were stabilized by taxol, another microtubule drug, the nuclear envelope reassembly was also interfered, although a significantly large aster formed around the chromatin. Based on these observations, we propose that microtubules play an important role in the nuclear envelope reassembly maybe by transporting the nuclear envelope precursors to the chromatin surfaces.

  4. Spindle pole mechanics studied in mitotic asters: dynamic distribution of spindle forces through compliant linkages.

    Science.gov (United States)

    Charlebois, Blake D; Kollu, Swapna; Schek, Henry T; Compton, Duane A; Hunt, Alan J

    2011-04-06

    During cell division, chromosomes must faithfully segregate to maintain genome integrity, and this dynamic mechanical process is driven by the macromolecular machinery of the mitotic spindle. However, little is known about spindle mechanics. For example, spindle microtubules are organized by numerous cross-linking proteins yet the mechanical properties of those cross-links remain unexplored. To examine the mechanical properties of microtubule cross-links we applied optical trapping to mitotic asters that form in mammalian mitotic extracts. These asters are foci of microtubules, motors, and microtubule-associated proteins that reflect many of the functional properties of spindle poles and represent centrosome-independent spindle-pole analogs. We observed bidirectional motor-driven microtubule movements, showing that microtubule linkages within asters are remarkably compliant (mean stiffness 0.025 pN/nm) and mediated by only a handful of cross-links. Depleting the motor Eg5 reduced this stiffness, indicating that Eg5 contributes to the mechanical properties of microtubule asters in a manner consistent with its localization to spindle poles in cells. We propose that compliant linkages among microtubules provide a mechanical architecture capable of accommodating microtubule movements and distributing force among microtubules without loss of pole integrity-a mechanical paradigm that may be important throughout the spindle.

  5. Dynamics of Microtubule Instabilities

    CERN Document Server

    Antal, T; Redner, S

    2007-01-01

    We investigate the dynamics of an idealized model of microtubule growth that evolves by: (i) attachment of guanosine triphosphate (GTP) at rate lambda, (ii) conversion of GTP to guanosine diphosphate (GDP) at rate 1, and (iii) detachment of GDP at rate mu. As a function of these rates, a microtubule can grow steadily or its length can fluctuate wildly. For mu=0, we find the exact tubule and GTP cap length distributions, and power-law length distributions of GTP and GDP islands. For mu=infinity, we argue that the time between catastrophes, where the microtubule shrinks to zero length, scales as exp(lambda). We also find the phase boundary between a growing and shrinking microtubule.

  6. Aging of dynamically stabilized microtubules

    CERN Document Server

    Ebbinghaus, M

    2009-01-01

    The microtubule network, an important part of the cytoskeleton, is constantly remodeled by alternating phases of growth and shrinkage of individual filaments. Plus-end tracking proteins (+TIPs) interact with the microtubule and in many cases alter its dynamics. While it is established that the prototypal CLIP-170 enhances microtubule stability by increasing rescues, the plus-end tracking mechanism is still under debate. We present a model for microtubule dynamics in which a rescue factor is dynamically added to the filament while growing. As a consequence, the filament shows aging behavior which should be experimentally accessible and thus allow one to exclude some hypothesized models of the inclusion of rescue factors at the microtubule plus end. Additionally, we show the strong influence of the cell geometry on the quantitative results.

  7. Visualization of microtubule growth in living platelets reveals a dynamic marginal band with multiple microtubules

    NARCIS (Netherlands)

    S. Patel-Hett (Sunita); J.L. Richardson (Jennifer); H. Schulze (Harald); K. Drabek (Ksenija); N.A. Isaac (Natasha); K. Hoffmeister (Karin); R.A. Shivdasani (Ramesh); J.C. Bulinski (J. Chloë); N.J. Galjart (Niels); J.H. Hartwig (John); J. Italiano (Joseph)

    2008-01-01

    textabstractThe marginal band of microtubules maintains the discoid shape of resting blood platelets. Although studies of platelet microtubule coil structure conclude that it is composed of a single microtubule, no investigations of its dynamics exist. In contrast to previous studies, permeabilized

  8. Dynamics and regulation of plant interphase microtubules: a comparative view.

    Science.gov (United States)

    Hashimoto, Takashi

    2003-12-01

    Microtubule and actin cytoskeletons are fundamental to a variety of cellular activities within eukaryotic organisms. Extensive information on the dynamics and functions of microtubules, as well as on their regulatory proteins, have been revealed in fungi and animals, and corresponding pictures are now slowly emerging in plants. During interphase, plant cells contain highly dynamic cortical microtubules that organize into ordered arrays, which are apparently regulated by distinct groups of microtubule regulators. Comparison with fungal and animal microtubules highlights both conserved and unique mechanisms for the regulation of the microtubule cytoskeleton in plants.

  9. Microtubules: dynamically unstable stochastic phase-switching polymers

    Science.gov (United States)

    Zakharov, P. N.; Arzhanik, V. K.; Ulyanov, E. V.; Gudimchuk, N. B.; Ataullakhanov, F. I.

    2016-08-01

    One of the simplest molecular motors, a biological microtubule, is reviewed as an example of a highly nonequilibrium molecular machine capable of stochastic transitions between slow growth and rapid disassembly phases. Basic properties of microtubules are described, and various approaches to simulating their dynamics, from statistical chemical kinetics models to molecular dynamics models using the Metropolis Monte Carlo and Brownian dynamics methods, are outlined.

  10. Deceivingly dynamic: Learning-dependent changes in stathmin and microtubules.

    Science.gov (United States)

    Uchida, Shusaku; Shumyatsky, Gleb P

    2015-10-01

    Microtubules, one of the major cytoskeletal structures, were previously considered stable and only indirectly involved in synaptic structure and function in mature neurons. However, recent evidence demonstrates that microtubules are dynamic and have an important role in synaptic structure, synaptic plasticity, and memory. In particular, learning induces changes in microtubule turnover and stability, and pharmacological manipulation of microtubule dynamics alters synaptic plasticity and long-term memory. These learning-induced changes in microtubules are controlled by the phosphoprotein stathmin, whose only known cellular activity is to negatively regulate microtubule formation. During the first eight hours following learning, changes in the phosphorylation of stathmin go through two phases causing biphasic shifts in microtubules stability/instability. These shifts, in turn, regulate memory formation by controlling in the second phase synaptic transport of the GluA2 subunit of AMPA receptors. Improper regulation of stathmin and microtubule dynamics has been observed in aged animals and in patients with Alzheimer's disease and depression. Thus, recent work on stathmin and microtubules has identified new molecular players in the early stages of memory encoding.

  11. The Role of Molecular Microtubule Motors and the Microtubule Cytoskeleton in Stress Granule Dynamics

    Directory of Open Access Journals (Sweden)

    Kristen M. Bartoli

    2011-01-01

    Full Text Available Stress granules (SGs are cytoplasmic foci that appear in cells exposed to stress-induced translational inhibition. SGs function as a triage center, where mRNAs are sorted for storage, degradation, and translation reinitiation. The underlying mechanisms of SGs dynamics are still being characterized, although many key players have been identified. The main components of SGs are stalled 48S preinitiation complexes. To date, many other proteins have also been found to localize in SGs and are hypothesized to function in SG dynamics. Most recently, the microtubule cytoskeleton and associated motor proteins have been demonstrated to function in SG dynamics. In this paper, we will discuss current literature examining the function of microtubules and the molecular microtubule motors in SG assembly, coalescence, movement, composition, organization, and disassembly.

  12. Modeling the Effects of Drug Binding on the Dynamic Instability of Microtubules

    CERN Document Server

    Hinow, Peter; Lopus, Manu; Jordan, Mary Ann; Tuszynski, Jack A

    2010-01-01

    We propose a stochastic model that accounts for the growth, catastrophe and rescue processes of steady state microtubules assembled from MAP-free tubulin. Both experimentally and theoretically we study the perturbation of microtubule dynamic instability by S-methyl-D-DM1, a synthetic derivative of the microtubule-targeted agent maytansine and a potential anticancer agent. We find that to be an effective suppressor of microtubule dynamics a drug must primarily suppress the loss of GDP tubulin from the microtubule tip.

  13. The Aster code; Code Aster

    Energy Technology Data Exchange (ETDEWEB)

    Delbecq, J.M

    1999-07-01

    The Aster code is a 2D or 3D finite-element calculation code for structures developed by the R and D direction of Electricite de France (EdF). This dossier presents a complete overview of the characteristics and uses of the Aster code: introduction of version 4; the context of Aster (organisation of the code development, versions, systems and interfaces, development tools, quality assurance, independent validation); static mechanics (linear thermo-elasticity, Euler buckling, cables, Zarka-Casier method); non-linear mechanics (materials behaviour, big deformations, specific loads, unloading and loss of load proportionality indicators, global algorithm, contact and friction); rupture mechanics (G energy restitution level, restitution level in thermo-elasto-plasticity, 3D local energy restitution level, KI and KII stress intensity factors, calculation of limit loads for structures), specific treatments (fatigue, rupture, wear, error estimation); meshes and models (mesh generation, modeling, loads and boundary conditions, links between different modeling processes, resolution of linear systems, display of results etc..); vibration mechanics (modal and harmonic analysis, dynamics with shocks, direct transient dynamics, seismic analysis and aleatory dynamics, non-linear dynamics, dynamical sub-structuring); fluid-structure interactions (internal acoustics, mass, rigidity and damping); linear and non-linear thermal analysis; steels and metal industry (structure transformations); coupled problems (internal chaining, internal thermo-hydro-mechanical coupling, chaining with other codes); products and services. (J.S.)

  14. Nonlinear Dynamics of Dipoles in Microtubules: Pseudo-Spin Model

    CERN Document Server

    Nesterov, Alexander I; Berman, Gennady P; Mavromatos, Nick E

    2016-01-01

    We perform a theoretical study of the dynamics of the electric field excitations in a microtubule by taking into consideration the realistic cylindrical geometry, dipole-dipole interactions of the tubulin-based protein heterodimers, the radial electric field produced by the solvent, and a possible degeneracy of energy states of individual heterodimers. The consideration is done in the frames of the classical pseudo-spin model. We derive the system of nonlinear dynamical ordinary differential equations of motion for interacting dipoles, and the continuum version of these equations. We obtain the solutions of these equations in the form of snoidal waves, solitons, kinks, and localized spikes. Our results will help to a better understanding of the functional properties of microtubules including the motor protein dynamics and the information transfer processes. Our considerations are based on classical dynamics. Some speculations on the role of possible quantum effects are also made.

  15. Nonlinear dynamics of dipoles in microtubules: Pseudospin model.

    Science.gov (United States)

    Nesterov, Alexander I; Ramírez, Mónica F; Berman, Gennady P; Mavromatos, Nick E

    2016-06-01

    We perform a theoretical study of the dynamics of the electric field excitations in a microtubule by taking into consideration the realistic cylindrical geometry, dipole-dipole interactions of the tubulin-based protein heterodimers, the radial electric field produced by the solvent, and a possible degeneracy of energy states of individual heterodimers. The consideration is done in the frame of the classical pseudospin model. We derive the system of nonlinear dynamical partial differential equations of motion for interacting dipoles and the continuum version of these equations. We obtain the solutions of these equations in the form of snoidal waves, solitons, kinks, and localized spikes. Our results will help to achieve a better understanding of the functional properties of microtubules including the motor protein dynamics and the information transfer processes. Our considerations are based on classical dynamics. Some speculations on the role of possible quantum effects are also made.

  16. ASTER Dunes

    Science.gov (United States)

    2000-01-01

    This image of Saudi Arabia shows a great sea of linear dunes in part of the Rub' al Khali, or the Empty Quarter. Acquired on June 25, 2000, the image covers an area 37 kilometers (23 miles) wide and 28 kilometers (17 miles) long in three bands of the reflected visible and infrared wavelength region. The dunes are yellow due to the presence of iron oxide minerals. The inter-dune area is made up of clays and silt and appears blue due to its high reflectance in band 1. The Rub' al Khali is the world's largest continuous sand desert. It covers about 650,000 square kilometers (250,966 square miles) and lies mainly in southern Saudi Arabia, though it does extend into the United Arab Emirates, Oman, and Yemen. One of the world's driest areas, it is uninhabited except for the Bedouin nomads who cross it. The first European to travel through the desert was Bertram Thomas in 1930.Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) is one of five Earth-observing instruments launched December 18, 1999, on NASA's Terra satellite. The instrument was built by Japan's Ministry of International Trade and Industry. A joint U.S./Japan science team is responsible for validation and calibration of the instrument and the data products. Dr. Anne Kahle at NASA's Jet Propulsion Laboratory, Pasadena, California, is the U.S. science team leader; Moshe Pniel of JPL is the project manager. ASTER is the only high-resolution imaging sensor on Terra. The primary goal of the ASTER mission is to obtain high-resolution image data in 14 channels over the entire land surface, as well as black and white stereo images. With revisit time of between 4 and 16 days, ASTER will provide the capability for repeat coverage of changing areas on Earth's surface.The broad spectral coverage and high spectral resolution of ASTER will provide scientists in numerous disciplines with critical information for surface mapping and monitoring dynamic conditions and temporal change. Examples of

  17. Taxol crystals can masquerade as stabilized microtubules.

    Directory of Open Access Journals (Sweden)

    Margit Foss

    Full Text Available Taxol is a potent anti-mitotic drug used in chemotherapy, angioplastic stents, and cell biology research. By binding and stabilizing microtubules, Taxol inhibits their dynamics, crucial for cell division, motility, and survival. The drug has also been reported to induce formation of asters and bundles composed of stabilized microtubules. Surprisingly, at commonly used concentrations, Taxol forms crystals that rapidly bind fluorescent tubulin subunits, generating structures with an uncanny resemblance to microtubule asters and bundles. Kinetic and topological considerations suggest that tubulin subunits, rather than microtubules, bind the crystals. This sequestration of tubulin from the subunit pool would be expected to shift the equilibrium of free to polymerized tubulin to disfavor assembly. Our results imply that some previously reported Taxol-induced asters or bundles could include or be composed of tubulin-decorated Taxol crystals. Thus, reevaluation of certain morphological, chemical, and physical properties of Taxol-treated microtubules may be necessary. Moreover, our findings suggest a novel mechanism for chemotherapy-induced cytotoxicity in non-dividing cells, with far-reaching medical implications.

  18. TOG Proteins Are Spatially Regulated by Rac-GSK3β to Control Interphase Microtubule Dynamics.

    Directory of Open Access Journals (Sweden)

    Kathryn P Trogden

    Full Text Available Microtubules are regulated by a diverse set of proteins that localize to microtubule plus ends (+TIPs where they regulate dynamic instability and mediate interactions with the cell cortex, actin filaments, and organelles. Although individual +TIPs have been studied in depth and we understand their basic contributions to microtubule dynamics, there is a growing body of evidence that these proteins exhibit cross-talk and likely function to collectively integrate microtubule behavior and upstream signaling pathways. In this study, we have identified a novel protein-protein interaction between the XMAP215 homologue in Drosophila, Mini spindles (Msps, and the CLASP homologue, Orbit. These proteins have been shown to promote and suppress microtubule dynamics, respectively. We show that microtubule dynamics are regionally controlled in cells by Rac acting to suppress GSK3β in the peripheral lamellae/lamellipodium. Phosphorylation of Orbit by GSK3β triggers a relocalization of Msps from the microtubule plus end to the lattice. Mutation of the Msps-Orbit binding site revealed that this interaction is required for regulating microtubule dynamic instability in the cell periphery. Based on our findings, we propose that Msps is a novel Rac effector that acts, in partnership with Orbit, to regionally regulate microtubule dynamics.

  19. Taxol differentially modulates the dynamics of microtubules assembled from unfractionated and purified beta-tubulin isotypes.

    Science.gov (United States)

    Derry, W B; Wilson, L; Khan, I A; Luduena, R F; Jordan, M A

    1997-03-25

    Substoichiometric binding of taxol to tubulin in microtubules potently suppresses microtubule dynamics, which appears to be the most sensitive antiproliferative mechanism of taxol. To determine whether the beta-tubulin isotype composition of a microtubule can modulate sensitivity to taxol, we measured the effects of substoichiometric ratios of taxol bound to tubulin in microtubules on the dynamics of microtubules composed of purified alphabeta(II)-, alphabeta(III)-, or alphabeta(IV)-tubulin isotypes and compared the results with the effects of taxol on microtubules assembled from unfractionated tubulin. Substoichiometric ratios of bound taxol in microtubules assembled from purified beta-tubulin isotypes or unfractionated tubulin potently suppressed the shortening rates and the lengths shortened per shortening event. Correlation of the suppression of the shortening rate with the stoichiometry of bound taxol revealed that microtubules composed of purified alphabeta(II)-, alphabeta(III)-, and alphabeta(IV)-tubulin were, respectively, 1.6-, 7.4-, and 7.2-fold less sensitive to the effects of bound taxol than microtubules assembled from unfractionated tubulin. These results indicate that taxol differentially modulates microtubule dynamics depending upon the beta-tubulin isotype composition. The results are consistent with recent studies correlating taxol resistance in tumor cells with increased levels of beta(III0- and beta(IV)-tubulin expression and suggest that altered cellular expression of beta-tubulin isotypes can be an important mechanism by which tumor cells develop resistance to taxol.

  20. Stathmin regulates microtubule dynamics and microtubule organizing center polarization in activated T cells.

    Science.gov (United States)

    Filbert, Erin L; Le Borgne, Marie; Lin, Joseph; Heuser, John E; Shaw, Andrey S

    2012-06-01

    Polarization of T cells involves reorientation of the microtubule organizing center (MTOC). Because activated ERK is localized at the immunological synapse, we investigated its role by showing that ERK activation is important for MTOC polarization. Suspecting that ERK phosphorylates a regulator of microtubules, we next focused on stathmin, a known ERK substrate. Our work indicates that during T cell activation, ERK is recruited to the synapse, allowing it to phosphorylate stathmin molecules near the immunological synapse. Supporting an important role of stathmin phosphorylation in T cell activation, we showed that T cell activation results in increased microtubule growth rate dependent on the presence of stathmin. The significance of this finding was demonstrated by results showing that CTLs from stathmin(-/-) mice displayed defective MTOC polarization and defective target cell cytolysis. These data implicate stathmin as a regulator of the microtubule network during T cell activation.

  1. Contributions of microtubule dynamic instability and rotational diffusion to kinetochore capture

    CERN Document Server

    Blackwell, Robert; Edelmaier, Christopher; Gergely, Zachary R; Flynn, Patrick J; Montes, Salvador; Crapo, Ammon; Doostan, Alireza; McIntosh, J Richard; Glaser, Matthew A; Betterton, Meredith D

    2016-01-01

    Microtubule dynamic instability allows search and capture of kinetochores during spindle formation, an important process for accurate chromosome segregation during cell division. Recent work has found that microtubule rotational diffusion about minus-end attachment points contributes to kinetochore capture in fission yeast, but the relative contributions of dynamic instability and rotational diffusion are not well understood. We have developed a biophysical model of kinetochore capture in small fission-yeast nuclei using hybrid Brownian dynamics/kinetic Monte Carlo simulation techniques. With this model, we have studied the importance of dynamic instability and microtubule rotational diffusion for kinetochore capture, both to the lateral surface of a microtubule and at or near its end. Over a range of biologically relevant parameters, microtubule rotational diffusion decreased capture time, but made a relatively small contribution compared to dynamic instability. At most, rotational diffusion reduced capture ...

  2. A plus-end raft to control microtubule dynamics and function.

    Science.gov (United States)

    Galjart, Niels; Perez, Franck

    2003-02-01

    Cells require a properly oriented and organised microtubule array to transmit positional information. Recent data have revealed a heterogeneous population of microtubule-binding proteins that accumulates mainly at distal ends of polymerising microtubules. Two mechanisms may account for this concentration: transient immobilisation, which involves association of proteins with growing ends, followed by release more proximally; and deposition at ends via a molecular motor. As with lipid rafts, protein concentration at distal ends may allow a cascade of interactions in the restricted area of a microtubule plus end. This may, in turn, control the dynamic behaviour of this cytoskeletal network and its anchoring to other structures.

  3. Paired arrangement of kinetochores together with microtubule pivoting and dynamics drive kinetochore capture in meiosis I.

    Science.gov (United States)

    Cojoc, Gheorghe; Florescu, Ana-Maria; Krull, Alexander; Klemm, Anna H; Pavin, Nenad; Jülicher, Frank; Tolić, Iva M

    2016-01-01

    Kinetochores are protein complexes on the chromosomes, whose function as linkers between spindle microtubules and chromosomes is crucial for proper cell division. The mechanisms that facilitate kinetochore capture by microtubules are still unclear. In the present study, we combine experiments and theory to explore the mechanisms of kinetochore capture at the onset of meiosis I in fission yeast. We show that kinetochores on homologous chromosomes move together, microtubules are dynamic and pivot around the spindle pole, and the average capture time is 3-4 minutes. Our theory describes paired kinetochores on homologous chromosomes as a single object, as well as angular movement of microtubules and their dynamics. For the experimentally measured parameters, the model reproduces the measured capture kinetics and shows that the paired configuration of kinetochores accelerates capture, whereas microtubule pivoting and dynamics have a smaller contribution. Kinetochore pairing may be a general feature that increases capture efficiency in meiotic cells.

  4. Microtubule dynamics control HGF-induced lung endothelial barrier enhancement.

    Directory of Open Access Journals (Sweden)

    Xinyong Tian

    Full Text Available Microtubules (MT play a vital role in many cellular functions, but their role in peripheral actin cytoskeletal dynamics which is essential for control of endothelial barrier and monolayer integrity is less understood. We have previously described the enhancement of lung endothelial cell (EC barrier by hepatocyte growth factor (HGF which was associated with Rac1-mediated remodeling of actin cytoskeleton. This study investigated involvement of MT-dependent mechanisms in the HGF-induced enhancement of EC barrier. HGF-induced Rac1 activation was accompanied by phosphorylation of stathmin, a regulator of MT dynamics. HGF also stimulated MT peripheral growth monitored by time lapse imaging and tracking analysis of EB-1-decorated MT growing tips, and increased the pool of acetylated tubulin. These effects were abolished by EC pretreatment with HGF receptor inhibitor, downregulation of Rac1 pathway, or by expression of a stathmin-S63A phosphorylation deficient mutant. Expression of stathmin-S63A abolished the HGF protective effects against thrombin-induced activation of RhoA cascade, permeability increase, and EC barrier dysfunction. These results demonstrate a novel MT-dependent mechanism of HGF-induced EC barrier regulation via Rac1/PAK1/stathmin-dependent control of MT dynamics.

  5. Microtubules as key coordinators of nuclear envelope and endoplasmic reticulum dynamics during mitosis.

    Science.gov (United States)

    Schlaitz, Anne-Lore

    2014-07-01

    During mitosis, cells comprehensively restructure their interior to promote the faithful inheritance of DNA and cytoplasmic contents. In metazoans, this restructuring entails disassembly of the nuclear envelope, redistribution of its components into the endoplasmic reticulum (ER) and eventually nuclear envelope reassembly around the segregated chromosomes. The microtubule cytoskeleton has recently emerged as a critical regulator of mitotic nuclear envelope and ER dynamics. Microtubules and associated molecular motors tear open the nuclear envelope in prophase and remove nuclear envelope remnants from chromatin. Additionally, two distinct mechanisms of microtubule-based regulation of ER dynamics operate later in mitosis. First, association of the ER with microtubules is reduced, preventing invasion of ER into the spindle area, and second, organelle membrane is actively cleared from metaphase chromosomes. However, we are only beginning to understand the role of microtubules in shaping and distributing ER and other organelles during mitosis.

  6. Dynamic microtubules regulate dendritic spine morphology and synaptic plasticity

    NARCIS (Netherlands)

    Jaworski, J.; Kapitein, L.C.; Montenegro Gouveia, S.; Dortland, B.R.; Wulf, P.S.; Grigoriev, I.; Camera, P.; Spangler, S.A.; Di Stefano, P.; Demmers, J.; Krugers, H.; Defilippi, P.; Akhmanova, A.; Hoogenraad, C.C.

    2009-01-01

    Dendritic spines are the major sites of excitatory synaptic input, and their morphological changes have been linked to learning and memory processes. Here, we report that growing microtubule plus ends decorated by the microtubule tip-tracking protein EB3 enter spines and can modulate spine morpholog

  7. Microtubule-based nanomaterials: Exploiting nature's dynamic biopolymers.

    Science.gov (United States)

    Bachand, George D; Spoerke, Erik D; Stevens, Mark J

    2015-06-01

    For more than a decade now, biomolecular systems have served as an inspiration for the development of synthetic nanomaterials and systems that are capable of reproducing many of unique and emergent behaviors of living systems. One intriguing element of such systems may be found in a specialized class of proteins known as biomolecular motors that are capable of performing useful work across multiple length scales through the efficient conversion of chemical energy. Microtubule (MT) filaments may be considered within this context as their dynamic assembly and disassembly dissipate energy, and perform work within the cell. MTs are one of three cytoskeletal filaments in eukaryotic cells, and play critical roles in a range of cellular processes including mitosis and vesicular trafficking. Based on their function, physical attributes, and unique dynamics, MTs also serve as a powerful archetype of a supramolecular filament that underlies and drives multiscale emergent behaviors. In this review, we briefly summarize recent efforts to generate hybrid and composite nanomaterials using MTs as biomolecular scaffolds, as well as computational and synthetic approaches to develop synthetic one-dimensional nanostructures that display the enviable attributes of the natural filaments.

  8. The kinesin-13 KLP10A motor regulates oocyte spindle length and affects EB1 binding without altering microtubule growth rates

    Directory of Open Access Journals (Sweden)

    Kevin K. Do

    2014-06-01

    Full Text Available Kinesin-13 motors are unusual in that they do not walk along microtubules, but instead diffuse to the ends, where they remove tubulin dimers, regulating microtubule dynamics. Here we show that Drosophila kinesin-13 klp10A regulates oocyte meiosis I spindle length and is haplo-insufficient – KLP10A, reduced by RNAi or a loss-of-function P element insertion mutant, results in elongated and mispositioned oocyte spindles, and abnormal cortical microtubule asters and aggregates. KLP10A knockdown by RNAi does not significantly affect microtubule growth rates in oocyte spindles, but, unexpectedly, EB1 binding and unbinding are slowed, suggesting a previously unobserved role for kinesin-13 in mediating EB1 binding interactions with microtubules. Kinesin-13 may regulate spindle length both by disassembling subunits from microtubule ends and facilitating EB1 binding to plus ends. We also observe an increased number of paused microtubules in klp10A RNAi knockdown spindles, consistent with a reduced frequency of microtubule catastrophes. Overall, our findings indicate that reduced kinesin-13 decreases microtubule disassembly rates and affects EB1 interactions with microtubules, rather than altering microtubule growth rates, causing spindles to elongate and abnormal cortical microtubule asters and aggregates to form.

  9. Microtubule Dynamics and Oscillating State for Mitotic Spindle

    CERN Document Server

    Rashid-Shomali, Safura

    2010-01-01

    We present a physical mechanism that can cause the mitotic spindle to oscillate. The driving force for this mechanism emerges from the polymerization of astral microtubules interacting with the cell cortex. We show that Brownian ratchet model for growing microtubules reaching the cell cortex, mediate an effective mass to the spindle body and therefore force it to oscillate. We compare the predictions of this mechanism with the previous mechanisms which were based on the effects of motor proteins. Finally we combine the effects of microtubules polymerization and motor proteins, and present the detailed phase diagram for possible oscillating states.

  10. Dietary flavonoid fisetin binds to β-tubulin and disrupts microtubule dynamics in prostate cancer cells.

    Science.gov (United States)

    Mukhtar, Eiman; Adhami, Vaqar Mustafa; Sechi, Mario; Mukhtar, Hasan

    2015-10-28

    Microtubule targeting based therapies have revolutionized cancer treatment; however, resistance and side effects remain a major limitation. Therefore, novel strategies that can overcome these limitations are urgently needed. We made a novel discovery that fisetin, a hydroxyflavone, is a microtubule stabilizing agent. Fisetin binds to tubulin and stabilizes microtubules with binding characteristics far superior than paclitaxel. Surface plasmon resonance and computational docking studies suggested that fisetin binds to β-tubulin with superior affinity compared to paclitaxel. Fisetin treatment of human prostate cancer cells resulted in robust up-regulation of microtubule associated proteins (MAP)-2 and -4. In addition, fisetin treated cells were enriched in α-tubulin acetylation, an indication of stabilization of microtubules. Fisetin significantly inhibited PCa cell proliferation, migration, and invasion. Nudc, a protein associated with microtubule motor dynein/dynactin complex that regulates microtubule dynamics, was inhibited with fisetin treatment. Further, fisetin treatment of a P-glycoprotein overexpressing multidrug-resistant cancer cell line NCI/ADR-RES inhibited the viability and colony formation. Our results offer in vitro proof-of-concept for fisetin as a microtubule targeting agent. We suggest that fisetin could be developed as an adjuvant for treatment of prostate and other cancer types.

  11. Integrated modeling methodology for microtubule dynamics and Taxol kinetics with experimentally identifiable parameters.

    Science.gov (United States)

    Zhao, He; Sokhansanj, Bahrad A

    2007-10-01

    Microtubule dynamics play a critical role in cell function and stress response, modulating mitosis, morphology, signaling, and transport. Drugs such as paclitaxel (Taxol) can impact tubulin polymerization and affect microtubule dynamics. While theoretical methods have been previously proposed to simulate microtubule dynamics, we develop a methodology here that can be used to compare model predictions with experimental data. Our model is a hybrid of (1) a simple two-state stochastic formulation of tubulin polymerization kinetics and (2) an equilibrium approximation for the chemical kinetics of Taxol drug binding to microtubule ends. Model parameters are biologically realistic, with values taken directly from experimental measurements. Model validation is conducted against published experimental data comparing optical measurements of microtubule dynamics in cultured cells under normal and Taxol-treated conditions. To compare model predictions with experimental data requires applying a "windowing" strategy on the spatiotemporal resolution of the simulation. From a biological perspective, this is consistent with interpreting the microtubule "pause" phenomenon as at least partially an artifact of spatiotemporal resolution limits on experimental measurement.

  12. The molecular dynamics of crawling migration in microtubule-disrupted keratocytes.

    Science.gov (United States)

    Nakashima, Hitomi; Okimura, Chika; Iwadate, Yoshiaki

    2015-01-01

    Cell-crawling migration plays an essential role in complex biological phenomena. It is now generally believed that many processes essential to such migration are regulated by microtubules in many cells, including fibroblasts and neurons. However, keratocytes treated with nocodazole, which is an inhibitor of microtubule polymerization - and even keratocyte fragments that contain no microtubules - migrate at the same velocity and with the same directionality as normal keratocytes. In this study, we discovered that not only these migration properties, but also the molecular dynamics that regulate such properties, such as the retrograde flow rate of actin filaments, distributions of vinculin and myosin II, and traction forces, are also the same in nocodazole-treated keratocytes as those in untreated keratocytes. These results suggest that microtubules are not in fact required for crawling migration of keratocytes, either in terms of migrating properties or of intracellular molecular dynamics.

  13. Aurora B suppresses microtubule dynamics and limits central spindle size by locally activating KIF4A

    Science.gov (United States)

    Nunes Bastos, Ricardo; Gandhi, Sapan R.; Baron, Ryan D.; Gruneberg, Ulrike; Nigg, Erich A.

    2013-01-01

    Anaphase central spindle formation is controlled by the microtubule-stabilizing factor PRC1 and the kinesin KIF4A. We show that an MKlp2-dependent pool of Aurora B at the central spindle, rather than global Aurora B activity, regulates KIF4A accumulation at the central spindle. KIF4A phosphorylation by Aurora B stimulates the maximal microtubule-dependent ATPase activity of KIF4A and promotes its interaction with PRC1. In the presence of phosphorylated KIF4A, microtubules grew more slowly and showed long pauses in growth, resulting in the generation of shorter PRC1-stabilized microtubule overlaps in vitro. Cells expressing only mutant forms of KIF4A lacking the Aurora B phosphorylation site overextended the anaphase central spindle, demonstrating that this regulation is crucial for microtubule length control in vivo. Aurora B therefore ensures that suppression of microtubule dynamic instability by KIF4A is restricted to a specific subset of microtubules and thereby contributes to central spindle size control in anaphase. PMID:23940115

  14. Microtubule dynamics in the peripheral nervous system: A matter of balance.

    Science.gov (United States)

    Almeida-Souza, Leonardo; Timmerman, Vincent; Janssens, Sophie

    2011-11-01

    The special architecture of neurons in the peripheral nervous system, with axons extending for long distances, represents a major challenge for the intracellular transport system. Two recent studies show that mutations in the small heat shock protein HSPB1, which cause an axonal type of Charcot-Marie-Tooth (CMT) neuropathy, affect microtubule dynamics and impede axonal transport. Intriguingly, while at presymptomatic age the neurons in the mutant HSPB1 mouse show a hyperstable microtubule network, at postsymptomatic age, the microtubule network completely lost its stability as reflected by a marked decrease in tubulin acetylation levels. We here propose a model explaining the role of microtubule stabilization and tubulin acetylation in the pathogenesis of HSPB1 mutations.

  15. Push or Pull? -- Cryo-Electron Microscopy of Microtubule's Dynamic Instability and Its Roles in the Kinetochore

    Science.gov (United States)

    Wang, Hong-Wei

    2009-03-01

    Microtubule is a biopolymer made up of alpha-beta-tubulin heterodimers. The tubulin dimers assemble head-to-tail as protofilaments and about 13 protofilaments interact laterally to form a hollow cylindrical structure which is the microtubule. As the major cytoskeleton in all eukaryotic cells, microtubules have the intrinsic property to switch stochastically between growth and shrinkage phases, a phenomenon termed as their dynamic instability. Microtubule's dynamic instability is closely related to the types of nucleotide (GTP or GDP) that binds to the beta-tubulin. We have biochemically trapped two types of assembly states of tubulin with GTP or GDP bound representing the polymerizing and depolymerizing ends of microtubules respectively. Using cryo-electron microscopy, we have elucidated the structures of these intermediate assemblies, showing that tubulin protofilaments demonstrate various curvatures and form different types of lateral interactions depending on the nucleotide states of tubulin and the temperature. Our work indicates that during the microtubule's dynamic cycle, tubulin undergoes various assembly states. These states, different from the straight microtubule, lend the highly dynamic and complicated behavior of microtubules. Our study of microtubule's interaction with certain kinetochore complexes suggests that the intermediate assemblies are responsible for specific mechanical forces that are required during the mitosis or meiosis. Our discoveries strongly suggest that a microtubule is a molecular machine rather than a simple cellular scaffold.

  16. Calcium-independent disruption of microtubule dynamics by nanosecond pulsed electric fields in U87 human glioblastoma cells.

    Science.gov (United States)

    Carr, Lynn; Bardet, Sylvia M; Burke, Ryan C; Arnaud-Cormos, Delia; Leveque, Philippe; O'Connor, Rodney P

    2017-01-24

    High powered, nanosecond duration, pulsed electric fields (nsPEF) cause cell death by a mechanism that is not fully understood and have been proposed as a targeted cancer therapy. Numerous chemotherapeutics work by disrupting microtubules. As microtubules are affected by electrical fields, this study looks at the possibility of disrupting them electrically with nsPEF. Human glioblastoma cells (U87-MG) treated with 100, 10 ns, 44 kV/cm pulses at a frequency of 10 Hz showed a breakdown of their interphase microtubule network that was accompanied by a reduction in the number of growing microtubules. This effect is temporally linked to loss of mitochondrial membrane potential and independent of cellular swelling and calcium influx, two factors that disrupt microtubule growth dynamics. Super-resolution microscopy revealed microtubule buckling and breaking as a result of nsPEF application, suggesting that nsPEF may act directly on microtubules.

  17. Calcium-independent disruption of microtubule dynamics by nanosecond pulsed electric fields in U87 human glioblastoma cells

    Science.gov (United States)

    Carr, Lynn; Bardet, Sylvia M.; Burke, Ryan C.; Arnaud-Cormos, Delia; Leveque, Philippe; O’Connor, Rodney P.

    2017-01-01

    High powered, nanosecond duration, pulsed electric fields (nsPEF) cause cell death by a mechanism that is not fully understood and have been proposed as a targeted cancer therapy. Numerous chemotherapeutics work by disrupting microtubules. As microtubules are affected by electrical fields, this study looks at the possibility of disrupting them electrically with nsPEF. Human glioblastoma cells (U87-MG) treated with 100, 10 ns, 44 kV/cm pulses at a frequency of 10 Hz showed a breakdown of their interphase microtubule network that was accompanied by a reduction in the number of growing microtubules. This effect is temporally linked to loss of mitochondrial membrane potential and independent of cellular swelling and calcium influx, two factors that disrupt microtubule growth dynamics. Super-resolution microscopy revealed microtubule buckling and breaking as a result of nsPEF application, suggesting that nsPEF may act directly on microtubules. PMID:28117459

  18. Autoinhibition of TBCB regulates EB1-mediated microtubule dynamics.

    Science.gov (United States)

    Carranza, Gerardo; Castaño, Raquel; Fanarraga, Mónica L; Villegas, Juan Carlos; Gonçalves, João; Soares, Helena; Avila, Jesus; Marenchino, Marco; Campos-Olivas, Ramón; Montoya, Guillermo; Zabala, Juan Carlos

    2013-01-01

    Tubulin cofactors (TBCs) participate in the folding, dimerization, and dissociation pathways of the tubulin dimer. Among them, TBCB and TBCE are two CAP-Gly domain-containing proteins that together efficiently interact with and dissociate the tubulin dimer. In the study reported here we showed that TBCB localizes at spindle and midzone microtubules during mitosis. Furthermore, the motif DEI/M-COO(-) present in TBCB, which is similar to the EEY/F-COO(-) element characteristic of EB proteins, CLIP-170, and α-tubulin, is required for TBCE-TBCB heterodimer formation and thus for tubulin dimer dissociation. This motif is responsible for TBCB autoinhibition, and our analysis suggests that TBCB is a monomer in solution. Mutants of TBCB lacking this motif are derepressed and induce microtubule depolymerization through an interaction with EB1 associated with microtubule tips. TBCB is also able to bind to the chaperonin complex CCT containing α-tubulin, suggesting that it could escort tubulin to facilitate its folding and dimerization, recycling or degradation.

  19. A polarised population of dynamic microtubules mediates homeostatic length control in animal cells.

    Directory of Open Access Journals (Sweden)

    Remigio Picone

    Full Text Available Because physical form and function are intimately linked, mechanisms that maintain cell shape and size within strict limits are likely to be important for a wide variety of biological processes. However, while intrinsic controls have been found to contribute to the relatively well-defined shape of bacteria and yeast cells, the extent to which individual cells from a multicellular animal control their plastic form remains unclear. Here, using micropatterned lines to limit cell extension to one dimension, we show that cells spread to a characteristic steady-state length that is independent of cell size, pattern width, and cortical actin. Instead, homeostatic length control on lines depends on a population of dynamic microtubules that lead during cell extension, and that are aligned along the long cell axis as the result of interactions of microtubule plus ends with the lateral cell cortex. Similarly, during the development of the zebrafish neural tube, elongated neuroepithelial cells maintain a relatively well-defined length that is independent of cell size but dependent upon oriented microtubules. A simple, quantitative model of cellular extension driven by microtubules recapitulates cell elongation on lines, the steady-state distribution of microtubules, and cell length homeostasis, and predicts the effects of microtubule inhibitors on cell length. Together this experimental and theoretical analysis suggests that microtubule dynamics impose unexpected limits on cell geometry that enable cells to regulate their length. Since cells are the building blocks and architects of tissue morphogenesis, such intrinsically defined limits may be important for development and homeostasis in multicellular organisms.

  20. Thymoquinone disrupts the microtubule dynamics in fission yeast Schizosaccharomyces pombe

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    Nusrat Masood

    2016-11-01

    Full Text Available Mad2 deletion strain of Schizosaccharomyces pombe was found to be sensitive to thymoquinone, a signature molecule present in Nigella sativa in a dose-dependent manner. Mad2 protein is an indispensable part of mitotic spindle checkpoint complex and is required for the cell cycle arrest in response to the spindle defects. Although the expression of α tubulin was not affected in thymoquinone treated cells, but the expression of β-tubulin was reduced. Further, the absence of microtubule in thymoquinone treated cells suggests its involvement in tubulin polymerization. Molecular docking studies revealed that thymoquinone specifically binds to β-tubulin near the Taxotere binding site of Tub1 (Tubulin α-β dimer. These studies additionally showed that thymoquinone interacts with the residues present in chain B, which is an inherent part of Mad2 protein of mitotic checkpoint complex (MCC. We concluded that the thymoquinone disrupts the microtubule polymerization that leads to the requirement of spindle checkpoint protein for the cell survival.

  1. Label-Free Imaging of Single Microtubule Dynamics Using Spatial Light Interference Microscopy.

    Science.gov (United States)

    Kandel, Mikhail E; Teng, Kai Wen; Selvin, Paul R; Popescu, Gabriel

    2017-01-24

    Due to their diameter, of only 24 nm, single microtubules are extremely challenging to image without the use of extrinsic contrast agents. As a result, fluorescence tagging is the common method to visualize their motility. However, such investigation is limited by photobleaching and phototoxicity. We experimentally demonstrate the capability of combining label-free spatial light interference microscopy (SLIM) with numerical processing for imaging single microtubules in a gliding assay. SLIM combines four different intensity images to obtain the optical path length map associated with the sample. Because of the use of broadband fields, the sensitivity to path length is better than 1 nm without (temporal) averaging and better than 0.1 nm upon averaging. Our results indicate that SLIM can image the dynamics of microtubules in a full field of view, of 200 × 200 μm(2), over many hours. Modeling the microtubule transport via the diffusion-advection equation, we found that the dispersion relation yields the standard deviation of the velocity distribution, without the need for tracking individual tubes. Interestingly, during a 2 h window, the microtubules begin to decelerate, at 100 pm/s(2) over a 20 min period. Thus, SLIM is likely to serve as a useful tool for understanding molecular motor activity, especially over large time scales, where fluorescence methods are of limited utility.

  2. Alterations in ovarian cancer cell adhesion drive taxol resistance by increasing microtubule dynamics in a FAK-dependent manner.

    Science.gov (United States)

    McGrail, Daniel J; Khambhati, Niti N; Qi, Mark X; Patel, Krishan S; Ravikumar, Nithin; Brandenburg, Chandler P; Dawson, Michelle R

    2015-04-17

    Chemorefractory ovarian cancer patients show extremely poor prognosis. Microtubule-stabilizing Taxol (paclitaxel) is a first-line treatment against ovarian cancer. Despite the close interplay between microtubules and cell adhesion, it remains unknown if chemoresistance alters the way cells adhere to their extracellular environment, a process critical for cancer metastasis. To investigate this, we isolated Taxol-resistant populations of OVCAR3 and SKOV3 ovarian cancer cell lines. Though Taxol-resistant cells neither effluxed more drug nor gained resistance to other chemotherapeutics, they did display increased microtubule dynamics. These changes in microtubule dynamics coincided with faster attachment rates and decreased adhesion strength, which correlated with increased surface β1-integrin expression and decreased focal adhesion formation, respectively. Adhesion strength correlated best with Taxol-sensitivity, and was found to be independent of microtubule polymerization but dependent on focal adhesion kinase (FAK), which was up-regulated in Taxol-resistant cells. FAK inhibition also decreased microtubule dynamics to equal levels in both populations, indicating alterations in adhesive signaling are up-stream of microtubule dynamics. Taken together, this work demonstrates that Taxol-resistance dramatically alters how ovarian cancer cells adhere to their extracellular environment causing down-stream increases in microtubule dynamics, providing a therapeutic target that may improve prognosis by not only recovering drug sensitivity, but also decreasing metastasis.

  3. Tank binding kinase 1 is a centrosome-associated kinase necessary for microtubule dynamics and mitosis.

    Science.gov (United States)

    Pillai, Smitha; Nguyen, Jonathan; Johnson, Joseph; Haura, Eric; Coppola, Domenico; Chellappan, Srikumar

    2015-12-10

    TANK Binding Kinase 1 (TBK1) is a non-canonical IκB kinase that contributes to KRAS-driven lung cancer. Here we report that TBK1 plays essential roles in mammalian cell division. Specifically, levels of active phospho-TBK1 increase during mitosis and localize to centrosomes, mitotic spindles and midbody, and selective inhibition or silencing of TBK1 triggers defects in spindle assembly and prevents mitotic progression. TBK1 binds to the centrosomal protein CEP170 and to the mitotic apparatus protein NuMA, and both CEP170 and NuMA are TBK1 substrates. Further, TBK1 is necessary for CEP170 centrosomal localization and binding to the microtubule depolymerase Kif2b, and for NuMA binding to dynein. Finally, selective disruption of the TBK1-CEP170 complex augments microtubule stability and triggers defects in mitosis, suggesting that TBK1 functions as a mitotic kinase necessary for microtubule dynamics and mitosis.

  4. KATNAL1 regulation of sertoli cell microtubule dynamics is essential for spermiogenesis and male fertility.

    Directory of Open Access Journals (Sweden)

    Lee B Smith

    Full Text Available Spermatogenesis is a complex process reliant upon interactions between germ cells (GC and supporting somatic cells. Testicular Sertoli cells (SC support GCs during maturation through physical attachment, the provision of nutrients, and protection from immunological attack. This role is facilitated by an active cytoskeleton of parallel microtubule arrays that permit transport of nutrients to GCs, as well as translocation of spermatids through the seminiferous epithelium during maturation. It is well established that chemical perturbation of SC microtubule remodelling leads to premature GC exfoliation demonstrating that microtubule remodelling is an essential component of male fertility, yet the genes responsible for this process remain unknown. Using a random ENU mutagenesis approach, we have identified a novel mouse line displaying male-specific infertility, due to a point mutation in the highly conserved ATPase domain of the novel KATANIN p60-related microtubule severing protein Katanin p60 subunit A-like1 (KATNAL1. We demonstrate that Katnal1 is expressed in testicular Sertoli cells (SC from 15.5 days post-coitum (dpc and that, consistent with chemical disruption models, loss of function of KATNAL1 leads to male-specific infertility through disruption of SC microtubule dynamics and premature exfoliation of spermatids from the seminiferous epithelium. The identification of KATNAL1 as an essential regulator of male fertility provides a significant novel entry point into advancing our understanding of how SC microtubule dynamics promotes male fertility. Such information will have resonance both for future treatment of male fertility and the development of non-hormonal male contraceptives.

  5. Synergistic suppression of microtubule dynamics by discodermolide and paclitaxel in non-small cell lung carcinoma cells.

    Science.gov (United States)

    Honore, Stéphane; Kamath, Kathy; Braguer, Diane; Horwitz, Susan Band; Wilson, Leslie; Briand, Claudette; Jordan, Mary Ann

    2004-07-15

    Discodermolide is a new microtubule-targeted antimitotic drug in Phase I clinical trials that, like paclitaxel, stabilizes microtubule dynamics and enhances microtubule polymer mass in vitro and in cells. Despite their apparently similar binding sites on microtubules, discodermolide acts synergistically with paclitaxel to inhibit proliferation of A549 human lung cancer cells (L. Martello et al., Clin. Cancer Res., 6: 1978-1987, 2000). To understand their synergy, we examined the effects of the two drugs singly and in combination in A549 cells and found that, surprisingly, their antiproliferative synergy is related to their ability to synergistically inhibit microtubule dynamic instability and mitosis. The combination of discodermolide and paclitaxel at their antiproliferative IC(50)s (7 nm for discodermolide and 2 nm for paclitaxel) altered all of the parameters of dynamic instability synergistically except the time-based rescue frequency. For example, together the drugs inhibited overall microtubule dynamicity by 71%, but each drug individually inhibited dynamicity by only 24%, giving a combination index (CI) of 0.23. Discodermolide and paclitaxel also synergistically blocked cell cycle progression at G(2)-M (41, 9.6, and 16% for both drugs together, for discodermolide alone, and for paclitaxel alone, respectively; CI = 0.59), and they synergistically enhanced apoptosis (CI = 0.85). Microtubules are unique receptors for drugs. The results suggest that ligands that bind to large numbers of binding sites on an individual microtubule can interact in a poorly understood manner to synergistically suppress microtubule dynamic instability and inhibit both mitosis and cell proliferation, with important consequences for combination clinical therapy with microtubule-targeted drugs.

  6. Modelling Vegetation Cover Dynamics of the Niger Floodplain in Mali, Westafrica, Using Multitemporal MERIS Full Resolution and TERRA -ASTER Data

    Science.gov (United States)

    Seiler, Ralf

    This presentation aims at showing the potential of a combined use of multi-temporal data from two different sensors (MERIS and TERRA ASTER) for an analysis of vegetation cover changes in semi-arid environments. While MERIS data mainly provide information about the vegetation cover density, ASTER data were used to analyse soil properties -especially soil brightness and soil wetness. An algorithm is proposed that uses atmospherically corrected surface reflectance values from MERIS and ASTER measurements. These values are subsequently splitted into a signal component that is caused by the vegetation cover and the background component (triggered by soil properties) using a linear spectral unmixing approach. Vegetation cover then is described by Vegetation Indices (MGVI, NDVI / SAVI) that were calculated from the vegetation signal component. Finally, these vegetation parameter were classified for all multi-temporal MERIS data using the EM algorithm to derive the temporal behaviour of vegetation pattern at the Inland Delta. The algorithm provides, as results, a fractional vegetation cover, a vegetation density value and information on the soil type. A detailed mapping of the spatio-temporal vegetation cover patterns for the Niger Inland Delta during the period of 2002 -2005 is another outcome of this study in addition to an in-depth evaluation of the applicability of the used VIs for environments with sparse vegetation covers. Located in the western Sahel of Africa, (1330' N -17 N and 230' W -530' W), the Niger Inland Delta is one of the largest floodplains in the world. The geographic term "Niger Inland Delta" represents a vast, extremely flat area of around 40.000 km extend, which is annually inundated by water of the Niger -Bani riversystem. In contrast to its semi -arid surrounding, the delta's ecology can be described as a mosaic of permanent, periodical and episodically flooded areas. Their extend varies both in scale and time due to irregularities of amount as

  7. Microtubule Dynamicity Is More Important than Stability in Memory Formation: an In Vivo Study.

    Science.gov (United States)

    Atarod, Deyhim; Eskandari-Sedighi, Ghazaleh; Pazhoohi, Farid; Karimian, Seyed Morteza; Khajeloo, Mojtaba; Riazi, Gholam Hossein

    2015-06-01

    It has been shown that microtubule (MT) activity and dynamics can have huge impacts on synaptic plasticity and memory formation. This is mainly due to various functions of MTs in neurons; MTs are involved in dendritic spine formation, axonal transportation, neuronal polarity, and receptor trafficking. Recent studies from our group and other labs have suggested the possible role of brain MT dynamicity and activity in memory; however, there is a need for more detailed studies regarding this aspect. In this study, we have tried to evaluate the importance of microtubule dynamicity rather than stability in memory formation in vivo. In order to investigate the role of MT stability in memory formation, we treated mice with paclitaxel-a classic microtubule-stabilizing agent. We then studied the behavior of treated animals using Morris water maze (MWM) test. To measure the effect of injected paclitaxel on MT polymerization kinetics, we conducted polymerization assays on brain extracts of the same paclitaxel-treated animals. Our results show that paclitaxel treatment affects animals' memory in a negative way and treated animals behave poorly in MWM compared to control group. In addition, our kinetics studies show that MT stability is significantly increased in brain extracts from paclitaxel-treated mice, but MT dynamics is reduced. Thus, we suggest that dynamicity is a very important feature of MT protein structures, and regarding memory formation, dynamicity is more important than stability and high activity.

  8. LARG and mDia1 link Galpha12/13 to cell polarity and microtubule dynamics.

    Science.gov (United States)

    Goulimari, Polyxeni; Knieling, Helga; Engel, Ulrike; Grosse, Robert

    2008-01-01

    Regulation of cell polarity is a process observed in all cells. During directed migration, cells orientate their microtubule cytoskeleton and the microtubule-organizing-center (MTOC), which involves integrins and downstream Cdc42 and glycogen synthase kinase-3beta activity. However, the contribution of G protein-coupled receptor signal transduction for MTOC polarity is less well understood. Here, we report that the heterotrimeric Galpha(12) and Galpha(13) proteins are necessary for MTOC polarity and microtubule dynamics based on studies using Galpha(12/13)-deficient mouse embryonic fibroblasts. Cell polarization involves the Galpha(12/13)-interacting leukemia-associated RhoGEF (LARG) and the actin-nucleating diaphanous formin mDia1. Interestingly, LARG associates with pericentrin and localizes to the MTOC and along microtubule tracks. We propose that Galpha(12/13) proteins exert essential functions linking extracellular signals to microtubule dynamics and cell polarity via RhoGEF and formin activity.

  9. Erucin, the major isothiocyanate in arugula (Eruca sativa, inhibits proliferation of MCF7 tumor cells by suppressing microtubule dynamics.

    Directory of Open Access Journals (Sweden)

    Olga Azarenko

    Full Text Available Consumption of cruciferous vegetables is associated with reduced risk of various types of cancer. Isothiocyanates including sulforaphane and erucin are believed to be responsible for this activity. Erucin [1-isothiocyanato-4-(methylthiobutane], which is metabolically and structurally related to sulforaphane, is present in large quantities in arugula (Eruca sativa, Mill., kohlrabi and Chinese cabbage. However, its cancer preventive mechanisms remain poorly understood. We found that erucin inhibits proliferation of MCF7 breast cancer cells (IC50 = 28 µM in parallel with cell cycle arrest at mitosis (IC50 = 13 µM and apoptosis, by a mechanism consistent with impairment of microtubule dynamics. Concentrations of 5-15 µM erucin suppressed the dynamic instability of microtubules during interphase in the cells. Most dynamic instability parameters were inhibited, including the rates and extents of growing and shortening, the switching frequencies between growing and shortening, and the overall dynamicity. Much higher erucin concentrations were required to reduce the microtubule polymer mass. In addition, erucin suppressed dynamic instability of microtubules reassembled from purified tubulin in similar fashion. The effects of erucin on microtubule dynamics, like those of sulforaphane, are similar qualitatively to those of much more powerful clinically-used microtubule-targeting anticancer drugs, including taxanes and the vinca alkaloids. The results suggest that suppression of microtubule dynamics by erucin and the resulting impairment of critically important microtubule-dependent cell functions such as mitosis, cell migration and microtubule-based transport may be important in its cancer preventive activities.

  10. Erucin, the major isothiocyanate in arugula (Eruca sativa), inhibits proliferation of MCF7 tumor cells by suppressing microtubule dynamics.

    Science.gov (United States)

    Azarenko, Olga; Jordan, Mary Ann; Wilson, Leslie

    2014-01-01

    Consumption of cruciferous vegetables is associated with reduced risk of various types of cancer. Isothiocyanates including sulforaphane and erucin are believed to be responsible for this activity. Erucin [1-isothiocyanato-4-(methylthio)butane], which is metabolically and structurally related to sulforaphane, is present in large quantities in arugula (Eruca sativa, Mill.), kohlrabi and Chinese cabbage. However, its cancer preventive mechanisms remain poorly understood. We found that erucin inhibits proliferation of MCF7 breast cancer cells (IC50 = 28 µM) in parallel with cell cycle arrest at mitosis (IC50 = 13 µM) and apoptosis, by a mechanism consistent with impairment of microtubule dynamics. Concentrations of 5-15 µM erucin suppressed the dynamic instability of microtubules during interphase in the cells. Most dynamic instability parameters were inhibited, including the rates and extents of growing and shortening, the switching frequencies between growing and shortening, and the overall dynamicity. Much higher erucin concentrations were required to reduce the microtubule polymer mass. In addition, erucin suppressed dynamic instability of microtubules reassembled from purified tubulin in similar fashion. The effects of erucin on microtubule dynamics, like those of sulforaphane, are similar qualitatively to those of much more powerful clinically-used microtubule-targeting anticancer drugs, including taxanes and the vinca alkaloids. The results suggest that suppression of microtubule dynamics by erucin and the resulting impairment of critically important microtubule-dependent cell functions such as mitosis, cell migration and microtubule-based transport may be important in its cancer preventive activities.

  11. AKAP9, a Regulator of Microtubule Dynamics, Contributes to Blood-Testis Barrier Function.

    Science.gov (United States)

    Venkatesh, Deepak; Mruk, Dolores; Herter, Jan M; Cullere, Xavier; Chojnacka, Katarzyna; Cheng, C Yan; Mayadas, Tanya N

    2016-02-01

    The blood-testis barrier (BTB), formed between adjacent Sertoli cells, undergoes extensive remodeling to facilitate the transport of preleptotene spermatocytes across the barrier from the basal to apical compartments of the seminiferous tubules for further development and maturation into spermatozoa. The actin cytoskeleton serves unique structural and supporting roles in this process, but little is known about the role of microtubules and their regulators during BTB restructuring. The large isoform of the cAMP-responsive scaffold protein AKAP9 regulates microtubule dynamics and nucleation at the Golgi. We found that conditional deletion of Akap9 in mice after the initial formation of the BTB at puberty leads to infertility. Akap9 deletion results in marked alterations in the organization of microtubules in Sertoli cells and a loss of barrier integrity despite a relatively intact, albeit more apically localized F-actin and BTB tight junctional proteins. These changes are accompanied by a loss of haploid spermatids due to impeded meiosis. The barrier, however, progressively reseals in older Akap9 null mice, which correlates with a reduction in germ cell apoptosis and a greater incidence of meiosis. However, spermiogenesis remains defective, suggesting additional roles for AKAP9 in this process. Together, our data suggest that AKAP9 and, by inference, the regulation of the microtubule network are critical for BTB function and subsequent germ cell development during spermatogenesis.

  12. PAR-1/MARK: a kinase essential for maintaining the dynamic state of microtubules.

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    Hayashi, Kenji; Suzuki, Atsushi; Ohno, Shigeo

    2012-01-01

    The serine/threonine kinase, PAR-1, is an essential component of the evolutionary-conserved polarity-regulating system, PAR-aPKC system, which plays indispensable roles in establishing asymmetric protein distributions and cell polarity in various biological contexts (Suzuki, A. and Ohno, S. (2006). J. Cell Sci., 119: 979-987; Matenia, D. and Mandelkow, E.M. (2009). Trends Biochem. Sci., 34: 332-342). PAR-1 is also known as MARK, which phosphorylates classical microtubule-associated proteins (MAPs) and detaches MAPs from microtubules (Matenia, D. and Mandelkow, E.M. (2009). Trends Biochem. Sci., 34: 332-342). This MARK activity of PAR-1 suggests its role in microtubule (MT) dynamics, but surprisingly, only few studies have been carried out to address this issue. Here, we summarize our recent study on live imaging analysis of MT dynamics in PAR-1b-depleted cells, which clearly demonstrated the positive role of PAR-1b in maintaining MT dynamics (Hayashi, K., Suzuki, A., Hirai, S., Kurihara, Y., Hoogenraad, C.C., and Ohno, S. (2011). J. Neurosci., 31: 12094-12103). Importantly, our results further revealed the novel physiological function of PAR-1b in maintaining dendritic spine morphology in mature neurons.

  13. Enhanced dynamic instability of microtubules in a ROS free inert environment.

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    Islam, Md Sirajul; Kabir, Arif Md Rashedul; Inoue, Daisuke; Sada, Kazuki; Kakugo, Akira

    2016-04-01

    Reactive oxygen species (ROS), one of the regulators in various biological processes, have recently been suspected to modulate microtubule (MT) dynamics in cells. However due to complicated cellular environment and unavailability of any in vitro investigation, no detail is understood yet. Here, by performing simple in vitro investigations, we have unveiled the effect of ROS on MT dynamics. By studying dynamic instability of MTs in a ROS free environment and comparing with that in the presence of ROS, we disclosed that MTs showed enhanced dynamics in the ROS free environment. All the parameters that define dynamic instability of MTs e.g., growth and shrinkage rates, rescue and catastrophe frequencies were significantly affected by the presence of ROS. This work clearly reveals the role of ROS in modulating MT dynamics in vitro, and would be a great help in understanding the role of ROS in regulation of MT dynamics in cells.

  14. Regulation of Kif15 localization and motility by the C-terminus of TPX2 and microtubule dynamics

    Science.gov (United States)

    Mann, Barbara J.; Balchand, Sai K.; Wadsworth, Patricia

    2017-01-01

    Mitotic motor proteins generate force to establish and maintain spindle bipolarity, but how they are temporally and spatially regulated in vivo is unclear. Prior work demonstrated that a microtubule-associated protein, TPX2, targets kinesin-5 and kinesin-12 motors to spindle microtubules. The C-terminal domain of TPX2 contributes to the localization and motility of the kinesin-5, Eg5, but it is not known whether this domain regulates kinesin-12, Kif15. We found that the C-terminal domain of TPX2 contributes to the localization of Kif15 to spindle microtubules in cells and suppresses motor walking in vitro. Kif15 and Eg5 are partially redundant motors, and overexpressed Kif15 can drive spindle formation in the absence of Eg5 activity. Kif15-dependent bipolar spindle formation in vivo requires the C-terminal domain of TPX2. In the spindle, fluorescent puncta of GFP-Kif15 move toward the equatorial region at a rate equivalent to microtubule growth. Reduction of microtubule growth with paclitaxel suppresses GFP-Kif15 motility, demonstrating that dynamic microtubules contribute to Kif15 behavior. Our results show that the C-terminal region of TPX2 regulates Kif15 in vitro, contributes to motor localization in cells, and is required for Kif15 force generation in vivo and further reveal that dynamic microtubules contribute to Kif15 behavior in vivo. PMID:27852894

  15. Mathematical modeling of the intracellular protein dynamics: the importance of active transport along microtubules.

    Science.gov (United States)

    Szymańska, Zuzanna; Parisot, Martin; Lachowicz, Mirosław

    2014-12-21

    In this paper we propose a mathematical model of protein and mRNA transport inside a cell. The spatio-temporal model takes into account the active transport along microtubules in the cytoplasm as well as diffusion and is able to reproduce the oscillatory changes in protein concentration observed in many experimental data. In the model the protein and the mRNA interact with each other that allows us to classify the model as a simple gene regulatory network. The proposed model is generic and may be adapted to specific signaling pathways. On the basis of numerical simulations, we formulate a new hypothesis that the oscillatory dynamics is allowed by the mRNA active transport along microtubules from the nucleus to distant locations.

  16. Mechanism of dynamic reorientation of cortical microtubules due to mechanical stress

    CERN Document Server

    Muratov, Alexander

    2015-01-01

    Directional growth caused by gravitropism and corresponding bending of plant cells has been explored since 19th century, however, many aspects of mechanisms underlying the perception of gravity at the molecular level are still not well known. Perception of gravity in root and shoot gravitropisms is usually attributed to gravisensitive cells, called statocytes, which exploit sedimentation of macroscopic and heavy organelles, amyloplasts, to sense the direction of gravity. Gravity stimulus is then transduced into distal elongation zone, which is several mm far from statocytes, where it causes stretching. It is suggested that gravity stimulus is conveyed by gradients in auxin flux. We propose a theoretical model that may explain how concentration gradients and/or stretching may indirectly affect the global orientation of cortical microtubules, attached to the cell membrane and induce their dynamic reorientation perpendicular to the gradients. In turn, oriented microtubules arrays direct the growth and orientatio...

  17. The Salmonella effector SseJ disrupts microtubule dynamics when ectopically expressed in normal rat kidney cells

    Science.gov (United States)

    Raines, Sally A.; Hodgkinson, Michael R.; Dowle, Adam A.

    2017-01-01

    Salmonella effector protein SseJ is secreted by Salmonella into the host cell cytoplasm where it can then modify host cell processes. Whilst host cell small GTPase RhoA has previously been shown to activate the acyl-transferase activity of SseJ we show here an un-described effect of SseJ protein production upon microtubule dynamism. SseJ prevents microtubule collapse and this is independent of SseJ’s acyl-transferase activity. We speculate that the effects of SseJ on microtubules would be mediated via its known interactions with the small GTPases of the Rho family. PMID:28235057

  18. ATX-2, the C. elegans Ortholog of Human Ataxin-2, Regulates Centrosome Size and Microtubule Dynamics.

    Directory of Open Access Journals (Sweden)

    Michael D Stubenvoll

    2016-09-01

    Full Text Available Centrosomes are critical sites for orchestrating microtubule dynamics, and exhibit dynamic changes in size during the cell cycle. As cells progress to mitosis, centrosomes recruit more microtubules (MT to form mitotic bipolar spindles that ensure proper chromosome segregation. We report a new role for ATX-2, a C. elegans ortholog of Human Ataxin-2, in regulating centrosome size and MT dynamics. ATX-2, an RNA-binding protein, forms a complex with SZY-20 in an RNA-independent fashion. Depleting ATX-2 results in embryonic lethality and cytokinesis failure, and restores centrosome duplication to zyg-1 mutants. In this pathway, SZY-20 promotes ATX-2 abundance, which inversely correlates with centrosome size. Centrosomes depleted of ATX-2 exhibit elevated levels of centrosome factors (ZYG-1, SPD-5, γ-Tubulin, increasing MT nucleating activity but impeding MT growth. We show that ATX-2 influences MT behavior through γ-Tubulin at the centrosome. Our data suggest that RNA-binding proteins play an active role in controlling MT dynamics and provide insight into the control of proper centrosome size and MT dynamics.

  19. Visualization and analysis of microtubule dynamics using dual color-coded display of plus-end labels.

    Directory of Open Access Journals (Sweden)

    Amy K Garrison

    Full Text Available Investigating spatial and temporal control of microtubule dynamics in live cells is critical to understanding cell morphogenesis in development and disease. Tracking fluorescently labeled plus-end-tracking proteins over time has become a widely used method to study microtubule assembly. Here, we report a complementary approach that uses only two images of these labels to visualize and analyze microtubule dynamics at any given time. Using a simple color-coding scheme, labeled plus-ends from two sequential images are pseudocolored with different colors and then merged to display color-coded ends. Based on object recognition algorithms, these colored ends can be identified and segregated into dynamic groups corresponding to four events, including growth, rescue, catastrophe, and pause. Further analysis yields not only their spatial distribution throughout the cell but also provides measurements such as growth rate and direction for each labeled end. We have validated the method by comparing our results with ground-truth data derived from manual analysis as well as with data obtained using the tracking method. In addition, we have confirmed color-coded representation of different dynamic events by analyzing their history and fate. Finally, we have demonstrated the use of the method to investigate microtubule assembly in cells and provided guidance in selecting optimal image acquisition conditions. Thus, this simple computer vision method offers a unique and quantitative approach to study spatial regulation of microtubule dynamics in cells.

  20. Suppression of microtubule dynamics by discodermolide by a novel mechanism is associated with mitotic arrest and inhibition of tumor cell proliferation.

    Science.gov (United States)

    Honore, Stéphane; Kamath, Kathy; Braguer, Diane; Wilson, Leslie; Briand, Claudette; Jordan, Mary Ann

    2003-12-01

    Discodermolide is a new microtubule-targeted drug in Phase I clinical trials that inhibits tumor growth and induces G(2)-M cell cycle arrest. It is effective against paclitaxel-resistant cell lines and acts synergistically in combination with paclitaxel. Suppression of microtubule dynamics by microtubule-targeted drugs has been hypothesized to be responsible for their ability to inhibit mitotic progression and cell proliferation. To determine whether discodermolide blocks mitosis by an effect on microtubule dynamics, we analyzed the effects of discodermolide on microtubule dynamics in living A549 human lung cancer cells during interphase at concentrations that block mitosis and inhibit cell proliferation. We found that discodermolide (7-166 nM) significantly suppressed microtubule dynamic instability. At the IC(50) for proliferation (7 nM discodermolide, 72 h), overall dynamicity was reduced by 23%. The principal parameters of dynamic instability suppressed by discodermolide were the microtubule shortening rate and length shortened. In addition, discodermolide markedly increased the frequency of rescued catastrophes. At the discodermolide concentration that resulted in 50% of maximal mitotic block (83 nM, 20 h), most microtubules were completely non-dynamic, no anaphases occurred, and all spindles were abnormal. The dynamicity of the remaining dynamic microtubules was reduced by 62%. The results indicate that a principal mechanism of inhibition of cell proliferation and mitotic block by discodermolide is suppression of microtubule dynamics. Importantly, the results indicate significant additional stabilizing effects of discodermolide on microtubule dynamics as compared with those of paclitaxel that may in turn reflect differences in their binding sites and their effects on tubulin conformation.

  1. Hoxb1b controls oriented cell division, cell shape and microtubule dynamics in neural tube morphogenesis.

    Science.gov (United States)

    Zigman, Mihaela; Laumann-Lipp, Nico; Titus, Tom; Postlethwait, John; Moens, Cecilia B

    2014-02-01

    Hox genes are classically ascribed to function in patterning the anterior-posterior axis of bilaterian animals; however, their role in directing molecular mechanisms underlying morphogenesis at the cellular level remains largely unstudied. We unveil a non-classical role for the zebrafish hoxb1b gene, which shares ancestral functions with mammalian Hoxa1, in controlling progenitor cell shape and oriented cell division during zebrafish anterior hindbrain neural tube morphogenesis. This is likely distinct from its role in cell fate acquisition and segment boundary formation. We show that, without affecting major components of apico-basal or planar cell polarity, Hoxb1b regulates mitotic spindle rotation during the oriented neural keel symmetric mitoses that are required for normal neural tube lumen formation in the zebrafish. This function correlates with a non-cell-autonomous requirement for Hoxb1b in regulating microtubule plus-end dynamics in progenitor cells in interphase. We propose that Hox genes can influence global tissue morphogenesis by control of microtubule dynamics in individual cells in vivo.

  2. Opportunities within ASTERICS

    Science.gov (United States)

    van der Meer, Rob; Cimò, Giuseppe

    2016-04-01

    ASTERICS, The Astronomy ESFRI and Research Infrastructure Cluster project, brings together astronomers and astroparticle physicists of 22 institutes in Europe to help Europe's world-leading observatories work together to find common solutions to their Big Data challenges, their interoperability and scheduling, and their data access, searching for cross-cutting solutions with mutual and wide-ranging benefit to all concerned. ASTERICS is a four year project, funded through the European Union's Horizon 2020 Framework Programme. The facilities supported by ASTERICS include SKA, CTA, KM3NeT, E-ELT. ASTERICS aims to open up multi messenger astronomy to all scientists and the public through the Virtual Observatory and the citizen science work. I will draw a picture of the landscape in which ASTERICS operates and the possible interaction with the Very Large Volume Neutrino Telescope community. Attention will be given to emerging opportunities for the Neutrino community and how these can be recognised or created.

  3. Dietary flavonoid fisetin binds to β-tubulin and disrupts microtubule dynamics in prostate cancer cells

    OpenAIRE

    Mukhtar, Eiman; Adhami, Vaqar Mustafa; Sechi, Mario; Mukhtar, Hasan

    2015-01-01

    Microtubule targeting based therapies have revolutionized cancer treatment; however, resistance and side effects remain a major limitation. Therefore, novel strategies that can overcome these limitations are urgently needed. We made a novel discovery that fisetin, a hydroxyflavone, is a microtubule stabilizing agent. Fisetin binds to tubulin and stabilizes microtubules with binding characteristics far superior than paclitaxel. Surface plasmon resonance and computational docking studies sugges...

  4. Microtubule-associated protein-4 controls nanovesicle dynamics and T cell activation.

    Science.gov (United States)

    Bustos-Morán, Eugenio; Blas-Rus, Noelia; Martin-Cófreces, Noa Beatriz; Sánchez-Madrid, Francisco

    2017-04-01

    The immune synapse (IS) is a specialized structure formed at the contact area between T lymphocytes and antigen-presenting cells (APCs) that is essential for the adaptive immune response. Proper T cell activation requires its polarization towards the APC, which is highly dependent on the tubulin cytoskeleton. Microtubule-associated protein-4 (MAP4) is a microtubule (MT)-stabilizing protein that controls MTs in physiological processes, such as cell division, migration, vesicular transport or primary cilia formation. In this study, we assessed the role of MAP4 in T cell activation. MAP4 decorates the pericentrosomal area and MTs of the T cell, and it is involved in MT detyrosination and stable assembly in response to T cell activation. In addition, MAP4 prompts the timely translocation of the MT-organizing center (MTOC) towards the IS and the dynamics of signaling nanovesicles that sustains T cell activation. However, MAP4 acts as a negative regulator of other T cell activation-related signals, including diacylglycerol (DAG) production and IL2 secretion. Our data indicate that MAP4 acts as a checkpoint molecule that balances positive and negative hallmarks of T cell activation.

  5. Human TUBB3 mutations perturb microtubule dynamics, kinesin interactions, and axon guidance

    Science.gov (United States)

    Tischfield, Max A.; Baris, Hagit N.; Wu, Chen; Rudolph, Guenther; Van Maldergem, Lionel; He, Wei; Chan, Wai-Man; Andrews, Caroline; Demer, Joseph L.; Robertson, Richard L.; Mackey, David A.; Ruddle, Jonathan B.; Bird, Thomas D.; Gottlob, Irene; Pieh, Christina; Traboulsi, Elias I.; Pomeroy, Scott L.; Hunter, David G.; Soul, Janet S.; Newlin, Anna; Sabol, Louise J.; Doherty, Edward J.; de Uzcátegui, Clara E.; de Uzcátegui, Nicolas; Collins, Mary Louise Z.; Sener, Emin C.; Wabbels, Bettina; Hellebrand, Heide; Meitinger, Thomas; de Berardinis, Teresa; Magli, Adriano; Schiavi, Costantino; Pastore-Trossello, Marco; Koc, Feray; Wong, Agnes M.; Levin, Alex V.; Geraghty, Michael T.; Descartes, Maria; Flaherty, Maree; Jamieson, Robyn V.; Møller, H. U.; Meuthen, Ingo; Callen, David F.; Kerwin, Janet; Lindsay, Susan; Meindl, Alfons; Gupta, Mohan L.; Pellman, David; Engle, Elizabeth C.

    2011-01-01

    We report that eight heterozygous missense mutations in TUBB3, encoding the neuron-specific β-tubulin isotype III, result in a spectrum of human nervous system disorders we now call the TUBB3 syndromes. Each mutation causes the ocular motility disorder CFEOM3, whereas some also result in intellectual and behavioral impairments, facial paralysis, and/or later-onset axonal sensorimotor polyneuropathy. Neuroimaging reveals a spectrum of abnormalities including hypoplasia of oculomotor nerves, and dysgenesis of the corpus callosum, anterior commissure, and corticospinal tracts. A knock-in disease mouse model reveals axon guidance defects without evidence of cortical cell migration abnormalities. We show the disease-associated mutations can impair tubulin heterodimer formation in vitro, although folded mutant heterodimers can still polymerize into microtubules. Modeling each mutation in yeast tubulin demonstrates that all alter dynamic instability whereas a subset disrupts the interaction of microtubules with kinesin motors. These findings demonstrate normal TUBB3 is required for axon guidance and maintenance in mammals. PMID:20074521

  6. Analysis of dynamic instability of steady-state microtubules in vitro by video-enhanced differential interference contrast microscopy with an appendix by Emin Oroudjev.

    Science.gov (United States)

    Yenjerla, Mythili; Lopus, Manu; Wilson, Leslie

    2010-01-01

    Microtubules are major constituents of the cytoskeleton which display dynamic properties. They exhibit dynamic instability which is defined as the stochastic switching between growing and shortening at microtubule ends. Dynamic instability plays an important role in diverse cellular functions including cell migration and mitosis. Many successful antimitotic drugs and microtubule-associated proteins (MAPs) are known to modulate microtubule dynamics, and it is important to analyze the in vitro dynamic instability of microtubules to study the mechanism of action of microtubule-targeted therapeutics and MAPs. In this chapter, we describe a method to analyze the in vitro dynamic instability of microtubules at steady state using video-enhanced differential contrast (VE-DIC) microscopy in detail. In this method, microtubules are assembled to steady state at 30 degrees C with MAP-free tubulin in a slide chamber in the presence of GTP, using sea urchin axonemes as nucleating seeds. Images of microtubules are enhanced and recorded in real time by a video camera and an image processor connected to a DIC microscope which is maintained at 30 degrees C. We use two software programs to track and analyze the growing and shortening of plus or minus ends of microtubules in the real-time images recorded using VE-DIC. In this chapter, we describe the instructions to use the tracking software Real Time Measurement II (RTM II) program. The instructions to use the analysis software Microtubule Life History Analysis Procedures (MT-LHAP) in Igor Pro software have been described in detail in an appendix (Oroudjev, 2010) following this chapter.

  7. Dynamic assembly of polymer nanotube networks via kinesin powered microtubule filaments

    Science.gov (United States)

    Paxton, Walter F.; Bouxsein, Nathan F.; Henderson, Ian M.; Gomez, Andrew; Bachand, George D.

    2015-06-01

    We describe for the first time how biological nanomotors may be used to actively self-assemble mesoscale networks composed of diblock copolymer nanotubes. The collective force generated by multiple kinesin nanomotors acting on a microtubule filament is large enough to overcome the energy barrier required to extract nanotubes from polymer vesicles comprised of poly(ethylene oxide-b-butadiene) in spite of the higher force requirements relative to extracting nanotubes from lipid vesicles. Nevertheless, large-scale polymer networks were dynamically assembled by the motors. These networks displayed enhanced robustness, persisting more than 24 h post-assembly (compared to 4-5 h for corresponding lipid networks). The transport of materials in and on the polymer membranes differs substantially from the transport on analogous lipid networks. Specifically, our data suggest that polymer mobility in nanotubular structures is considerably different from planar or 3D structures, and is stunted by 1D confinement of the polymer subunits. Moreover, quantum dots adsorbed onto polymer nanotubes are completely immobile, which is related to this 1D confinement effect and is in stark contrast to the highly fluid transport observed on lipid tubules.We describe for the first time how biological nanomotors may be used to actively self-assemble mesoscale networks composed of diblock copolymer nanotubes. The collective force generated by multiple kinesin nanomotors acting on a microtubule filament is large enough to overcome the energy barrier required to extract nanotubes from polymer vesicles comprised of poly(ethylene oxide-b-butadiene) in spite of the higher force requirements relative to extracting nanotubes from lipid vesicles. Nevertheless, large-scale polymer networks were dynamically assembled by the motors. These networks displayed enhanced robustness, persisting more than 24 h post-assembly (compared to 4-5 h for corresponding lipid networks). The transport of materials in and on

  8. An antitubulin agent BCFMT inhibits proliferation of cancer cells and induces cell death by inhibiting microtubule dynamics.

    Directory of Open Access Journals (Sweden)

    Ankit Rai

    Full Text Available Using cell based screening assay, we identified a novel anti-tubulin agent (Z-5-((5-(4-bromo-3-chlorophenylfuran-2-ylmethylene-2-thioxothiazolidin-4-one (BCFMT that inhibited proliferation of human cervical carcinoma (HeLa (IC(50, 7.2 ± 1.8 µM, human breast adenocarcinoma (MCF-7 (IC(50, 10.0 ± 0.5 µM, highly metastatic breast adenocarcinoma (MDA-MB-231 (IC(50, 6.0 ± 1 µM, cisplatin-resistant human ovarian carcinoma (A2780-cis (IC(50, 5.8 ± 0.3 µM and multi-drug resistant mouse mammary tumor (EMT6/AR1 (IC(50, 6.5 ± 1 µM cells. Using several complimentary strategies, BCFMT was found to inhibit cancer cell proliferation at G2/M phase of the cell cycle apparently by targeting microtubules. In addition, BCFMT strongly suppressed the dynamics of individual microtubules in live MCF-7 cells. At its half maximal proliferation inhibitory concentration (10 µM, BCFMT reduced the rates of growing and shortening phases of microtubules in MCF-7 cells by 37 and 40%, respectively. Further, it increased the time microtubules spent in the pause (neither growing nor shortening detectably state by 135% and reduced the dynamicity (dimer exchange per unit time of microtubules by 70%. In vitro, BCFMT bound to tubulin with a dissociation constant of 8.3 ± 1.8 µM, inhibited tubulin assembly and suppressed GTPase activity of microtubules. BCFMT competitively inhibited the binding of BODIPY FL-vinblastine to tubulin with an inhibitory concentration (K(i of 5.2 ± 1.5 µM suggesting that it binds to tubulin at the vinblastine site. In cultured cells, BCFMT-treatment depolymerized interphase microtubules, perturbed the spindle organization and accumulated checkpoint proteins (BubR1 and Mad2 at the kinetochores. BCFMT-treated MCF-7 cells showed enhanced nuclear accumulation of p53 and its downstream p21, which consequently activated apoptosis in these cells. The results suggested that BCFMT inhibits proliferation of several types of cancer cells including drug

  9. Microtubules are Essential for Mitochondrial Dynamics-Fission, Fusion, and Motility- in Dictyostelium discoideum.

    Directory of Open Access Journals (Sweden)

    Laken C. Woods

    2016-03-01

    Full Text Available Mitochondrial function is dependent upon mitochondrial structure which is in turn dependent upon mitochondrial dynamics, including fission, fusion, and motility. Here we examined the relationship between mitochondrial dynamics and the cytoskeleton in Dictyostelium discoideum. Using time-lapse analysis, we quantified mitochondrial fission, fusion, and motility in the presence of cytoskeleton disrupting pharmaceuticals and the absence of the potential mitochondria-cytoskeleton linker protein, CluA. Our results indicate that microtubules are essential for mitochondrial movement, as well as fission and fusion; actin plays a less significant role, perhaps selecting the mitochondria for transport. We also suggest that CluA is not a linker protein but plays an unidentified role in mitochondrial fission and fusion. The significance of our work is to gain further insight into the role the cytoskeleton plays in mitochondrial dynamics and function. By better understanding these processes we can better appreciate the underlying mitochondrial contributions to many neurological disorders characterized by altered mitochondrial dynamics, structure, and/or function.

  10. Microtubule dynamics from mating through the first zygotic division in the budding yeast Saccharomyces cerevisiae.

    Science.gov (United States)

    Maddox, P; Chin, E; Mallavarapu, A; Yeh, E; Salmon, E D; Bloom, K

    1999-03-08

    We have used time-lapse digital imaging microscopy to examine cytoplasmic astral microtubules (Mts) and spindle dynamics during the mating pathway in budding yeast Saccharomyces cerevisiae. Mating begins when two cells of opposite mating type come into proximity. The cells arrest in the G1 phase of the cell cycle and grow a projection towards one another forming a shmoo projection. Imaging of microtubule dynamics with green fluorescent protein (GFP) fusions to dynein or tubulin revealed that the nucleus and spindle pole body (SPB) became oriented and tethered to the shmoo tip by a Mt-dependent search and capture mechanism. Dynamically unstable astral Mts were captured at the shmoo tip forming a bundle of three or four astral Mts. This bundle changed length as the tethered nucleus and SPB oscillated toward and away from the shmoo tip at growth and shortening velocities typical of free plus end astral Mts (approximately 0.5 micrometer/min). Fluorescent fiduciary marks in Mt bundles showed that Mt growth and shortening occurred primarily at the shmoo tip, not the SPB. This indicates that Mt plus end assembly/disassembly was coupled to pushing and pulling of the nucleus. Upon cell fusion, a fluorescent bar of Mts was formed between the two shmoo tip bundles, which slowly shortened (0.23 +/- 0.07 micrometer/min) as the two nuclei and their SPBs came together and fused (karyogamy). Bud emergence occurred adjacent to the fused SPB approximately 30 min after SPB fusion. During the first mitosis, the SPBs separated as the spindle elongated at a constant velocity (0.75 micrometer/min) into the zygotic bud. There was no indication of a temporal delay at the 2-micrometer stage of spindle morphogenesis or a lag in Mt nucleation by replicated SPBs as occurs in vegetative mitosis implying a lack of normal checkpoints. Thus, the shmoo tip appears to be a new model system for studying Mt plus end dynamic attachments and much like higher eukaryotes, the first mitosis after haploid

  11. FoxO regulates microtubule dynamics and polarity to promote dendrite branching in Drosophila sensory neurons.

    Science.gov (United States)

    Sears, James C; Broihier, Heather T

    2016-10-01

    The size and shape of dendrite arbors are defining features of neurons and critical determinants of neuronal function. The molecular mechanisms establishing arborization patterns during development are not well understood, though properly regulated microtubule (MT) dynamics and polarity are essential. We previously found that FoxO regulates axonal MTs, raising the question of whether it also regulates dendritic MTs and morphology. Here we demonstrate that FoxO promotes dendrite branching in all classes of Drosophila dendritic arborization (da) neurons. FoxO is required both for initiating growth of new branches and for maintaining existing branches. To elucidate FoxO function, we characterized MT organization in both foxO null and overexpressing neurons. We find that FoxO directs MT organization and dynamics in dendrites. Moreover, it is both necessary and sufficient for anterograde MT polymerization, which is known to promote dendrite branching. Lastly, FoxO promotes proper larval nociception, indicating a functional consequence of impaired da neuron morphology in foxO mutants. Together, our results indicate that FoxO regulates dendrite structure and function and suggest that FoxO-mediated pathways control MT dynamics and polarity.

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

    Institute of Scientific and Technical Information of China (English)

    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.

  13. Automated screening of microtubule growth dynamics identifies MARK2 as a regulator of leading edge microtubules downstream of Rac1 in migrating cells.

    Directory of Open Access Journals (Sweden)

    Yukako Nishimura

    Full Text Available Polarized microtubule (MT growth in the leading edge is critical to directed cell migration, and is mediated by Rac1 GTPase. To find downstream targets of Rac1 that affect MT assembly dynamics, we performed an RNAi screen of 23 MT binding and regulatory factors and identified RNAi treatments that suppressed changes in MT dynamics induced by constitutively activated Rac1. By analyzing fluorescent EB3 dynamics with automated tracking, we found that RNAi treatments targeting p150(glued, APC2, spastin, EB1, Op18, or MARK2 blocked Rac1-mediated MT growth in lamellipodia. MARK2 was the only protein whose RNAi targeting additionally suppressed Rac1 effects on MT orientation in lamellipodia, and thus became the focus of further study. We show that GFP-MARK2 rescued effects of MARK2 depletion on MT growth lifetime and orientation, and GFP-MARK2 localized in lamellipodia in a Rac1-activity-dependent manner. In a wound-edge motility assay, MARK2-depleted cells failed to polarize their centrosomes or exhibit oriented MT growth in the leading edge, and displayed defects in directional cell migration. Thus, automated image analysis of MT assembly dynamics identified MARK2 as a target regulated downstream of Rac1 that promotes oriented MT growth in the leading edge to mediate directed cell migration.

  14. plusTipTracker: Quantitative image analysis software for the measurement of microtubule dynamics.

    Science.gov (United States)

    Applegate, Kathryn T; Besson, Sebastien; Matov, Alexandre; Bagonis, Maria H; Jaqaman, Khuloud; Danuser, Gaudenz

    2011-11-01

    Here we introduce plusTipTracker, a Matlab-based open source software package that combines automated tracking, data analysis, and visualization tools for movies of fluorescently-labeled microtubule (MT) plus end binding proteins (+TIPs). Although +TIPs mark only phases of MT growth, the plusTipTracker software allows inference of additional MT dynamics, including phases of pause and shrinkage, by linking collinear, sequential growth tracks. The algorithm underlying the reconstruction of full MT trajectories relies on the spatially and temporally global tracking framework described in Jaqaman et al. (2008). Post-processing of track populations yields a wealth of quantitative phenotypic information about MT network architecture that can be explored using several visualization modalities and bioinformatics tools included in plusTipTracker. Graphical user interfaces enable novice Matlab users to track thousands of MTs in minutes. In this paper, we describe the algorithms used by plusTipTracker and show how the package can be used to study regional differences in the relative proportion of MT subpopulations within a single cell. The strategy of grouping +TIP growth tracks for the analysis of MT dynamics has been introduced before (Matov et al., 2010). The numerical methods and analytical functionality incorporated in plusTipTracker substantially advance this previous work in terms of flexibility and robustness. To illustrate the enhanced performance of the new software we thus compare computer-assembled +TIP-marked trajectories to manually-traced MT trajectories from the same movie used in Matov et al. (2010).

  15. Dynamics of centrosome translocation and microtubule organization in neocortical neurons during distinct modes of polarization.

    Science.gov (United States)

    Sakakibara, Akira; Sato, Toshiyuki; Ando, Ryota; Noguchi, Namiko; Masaoka, Makoto; Miyata, Takaki

    2014-05-01

    Neuronal migration and process formation require cytoskeletal organization and remodeling. Recent studies suggest that centrosome translocation is involved in initial axon outgrowth, while the role of centrosomal positioning is not clear. Here, we examine relations between centrosomal positioning, axonogenesis, and microtubule (MT) polarization in multipolar and bipolar neocortical neurons. We monitored dynamic movements of centrosomes and MT plus ends in migratory neurons in embryonic mouse cerebral slices. In locomoting bipolar neurons, the centrosome oriented toward the pia-directed leading process. Bipolar neurons displayed dense MT plus end dynamics in leading processes, while trailing processes showed clear bidirectional MTs. In migrating multipolar neurons, new processes emerged irrespective of centrosome localization, followed by centrosome reorientations toward the dominant process. Anterograde movements of MT plus ends occurred in growing processes and retrograde movements were observed after retraction of the distal tip. In multipolar neurons, axon formed by tangential extension of a dominant process and the centrosome oriented toward the growing axon, while in locomoting neurons, an axon formed opposite to the direction of migration and the centrosome localized to the base of the leading process. Our data suggest that MT organization may alter centrosomal localization and that centrosomal positioning does not necessarily direct process formation.

  16. Non-critical string theory formulation of microtubule dynamics and quantum aspects of brain function

    CERN Document Server

    Mavromatos, Nikolaos E

    1995-01-01

    Microtubule (MT) networks, subneural paracrystalline cytosceletal structures, seem to play a fundamental role in the neurons. We cast here the complicated MT dynamics in the form of a 1+1-dimensional non-critical string theory, thus enabling us to provide a consistent quantum treatment of MTs, including enviromental {\\em friction} effects. We suggest, thus, that the MTs are the microsites, in the brain, for the emergence of stable, macroscopic quantum coherent states, identifiable with the {\\em preconscious states}. Quantum space-time effects, as described by non-critical string theory, trigger then an {\\em organized collapse} of the coherent states down to a specific or {\\em conscious state}. The whole process we estimate to take {\\cal O}(1\\,{\\rm sec}), in excellent agreement with a plethora of experimental/observational findings. The {\\em microscopic arrow of time}, endemic in non-critical string theory, and apparent here in the self-collapse process, provides a satisfactory and simple resolution to the age...

  17. Electric fields generated by synchronized oscillations of microtubules, centrosomes and chromosomes regulate the dynamics of mitosis and meiosis

    Directory of Open Access Journals (Sweden)

    Zhao Yue

    2012-07-01

    Full Text Available Abstract Super-macromolecular complexes play many important roles in eukaryotic cells. Classical structural biological studies focus on their complicated molecular structures, physical interactions and biochemical modifications. Recent advances concerning intracellular electric fields generated by cell organelles and super-macromolecular complexes shed new light on the mechanisms that govern the dynamics of mitosis and meiosis. In this review we synthesize this knowledge to provide an integrated theoretical model of these cellular events. We suggest that the electric fields generated by synchronized oscillation of microtubules, centrosomes, and chromatin fibers facilitate several events during mitosis and meiosis, including centrosome trafficking, chromosome congression in mitosis and synapsis between homologous chromosomes in meiosis. These intracellular electric fields are generated under energy excitation through the synchronized electric oscillations of the dipolar structures of microtubules, centrosomes and chromosomes, three of the super-macromolecular complexes within an animal cell.

  18. Electric fields generated by synchronized oscillations of microtubules, centrosomes and chromosomes regulate the dynamics of mitosis and meiosis.

    Science.gov (United States)

    Zhao, Yue; Zhan, Qimin

    2012-07-02

    Super-macromolecular complexes play many important roles in eukaryotic cells. Classical structural biological studies focus on their complicated molecular structures, physical interactions and biochemical modifications. Recent advances concerning intracellular electric fields generated by cell organelles and super-macromolecular complexes shed new light on the mechanisms that govern the dynamics of mitosis and meiosis. In this review we synthesize this knowledge to provide an integrated theoretical model of these cellular events. We suggest that the electric fields generated by synchronized oscillation of microtubules, centrosomes, and chromatin fibers facilitate several events during mitosis and meiosis, including centrosome trafficking, chromosome congression in mitosis and synapsis between homologous chromosomes in meiosis. These intracellular electric fields are generated under energy excitation through the synchronized electric oscillations of the dipolar structures of microtubules, centrosomes and chromosomes, three of the super-macromolecular complexes within an animal cell.

  19. Coordination of microtubule and microfilament dynamics by Drosophila Rho1, Spire and Cappuccino.

    Science.gov (United States)

    Rosales-Nieves, Alicia E; Johndrow, James E; Keller, Lani C; Magie, Craig R; Pinto-Santini, Delia M; Parkhurst, Susan M

    2006-04-01

    The actin-nucleation factors Spire and Cappuccino (Capu) regulate the onset of ooplasmic streaming in Drosophila melanogaster. Although this streaming event is microtubule-based, actin assembly is required for its timing. It is not understood how the interaction of microtubules and microfilaments is mediated in this context. Here, we demonstrate that Capu and Spire have microtubule and microfilament crosslinking activity. The spire locus encodes several distinct protein isoforms (SpireA, SpireC and SpireD). SpireD was recently shown to nucleate actin, but the activity of the other isoforms has not been addressed. We find that SpireD does not have crosslinking activity, whereas SpireC is a potent crosslinker. We show that SpireD binds to Capu and inhibits F-actin/microtubule crosslinking, and activated Rho1 abolishes this inhibition, establishing a mechanistic basis for the regulation of Capu and Spire activity. We propose that Rho1, cappuccino and spire are elements of a conserved developmental cassette that is capable of directly mediating crosstalk between microtubules and microfilaments.

  20. Magnetic manipulation of self-assembled colloidal asters.

    Energy Technology Data Exchange (ETDEWEB)

    Snezhko, A.; Aranson, I. S. (Materials Science Division)

    2011-09-01

    Self-assembled materials must actively consume energy and remain out of equilibrium to support structural complexity and functional diversity. Here we show that a magnetic colloidal suspension confined at the interface between two immiscible liquids and energized by an alternating magnetic field dynamically self-assembles into localized asters and arrays of asters, which exhibit locomotion and shape change. By controlling a small external magnetic field applied parallel to the interface, we show that asters can capture, transport, and position target microparticles. The ability to manipulate colloidal structures is crucial for the further development of self-assembled microrobots

  1. Escape from Mitotic Arrest: An Unexpected Connection Between Microtubule Dynamics and Epigenetic Regulation of Centromeric Chromatin in Schizosaccharomyces pombe.

    Science.gov (United States)

    George, Anuja A; Walworth, Nancy C

    2015-12-01

    Accurate chromosome segregation is necessary to ensure genomic integrity. Segregation depends on the proper functioning of the centromere, kinetochore, and mitotic spindle microtubules and is monitored by the spindle assembly checkpoint (SAC). In the fission yeast Schizosaccharomyces pombe, defects in Dis1, a microtubule-associated protein that influences microtubule dynamics, lead to mitotic arrest as a result of an active SAC and consequent failure to grow at low temperature. In a mutant dis1 background (dis1-288), loss of function of Msc1, a fission yeast homolog of the KDM5 family of proteins, suppresses the growth defect and promotes normal mitosis. Genetic analysis implicates a histone deacetylase (HDAC)-linked pathway in suppression because HDAC mutants clr6-1, clr3∆, and sir2∆, though not hos2∆, also promote normal mitosis in the dis1-288 mutant. Suppression of the dis phenotype through loss of msc1 function requires the spindle checkpoint protein Mad2 and is limited by the presence of the heterochromatin-associated HP1 protein homolog Swi6. We speculate that alterations in histone acetylation promote a centromeric chromatin environment that compensates for compromised dis1 function by allowing for successful kinetochore-microtubule interactions that can satisfy the SAC. In cells arrested in mitosis by mutation of dis1, loss of function of epigenetic determinants such as Msc1 or specific HDACs can promote cell survival. Because the KDM5 family of proteins has been implicated in human cancers, an appreciation of the potential role of this family of proteins in chromosome segregation is warranted.

  2. In vitro reconstitution of dynamic microtubules interacting with actin filament networks

    NARCIS (Netherlands)

    Preciado Lopez, M.; Huber, F.; Grigoriev, Ilya; Steinmetz, M.O.; Akhmanova, Anna; Dogterom, M.; Koenderink, G.H.

    2014-01-01

    Interactions between microtubules and actin filaments (F-actin) are essential for eukaryotic cell migration, polarization, growth, and division. Although the importance of these interactions has been long recognized, the inherent complexity of the cell interior hampers a detailed mechanistic study o

  3. Dynamic behavior of GFP-CLIP-170 reveals fast protein turnover on microtubule plus ends.

    NARCIS (Netherlands)

    K.A. Drägestein (Katharina Asja); W.A. van Cappellen (Gert); J.A.J. van Haren (Jeffrey); G.D. Tsibidis (George); A.S. Akhmanova (Anna); T.A. Knoch (Tobias); F.G. Grosveld (Frank); N.J. Galjart (Niels)

    2008-01-01

    textabstractMicrotubule (MT) plus end – tracking proteins (+TIPs) specifi cally recognize the ends of growing MTs. +TIPs are involved in diverse cellular processes such as cell division, cell migration, and cell polarity. Although +TIP tracking is important for these processes, the mechanisms underl

  4. Theory of dynamic force spectroscopy for kinetochore-microtubule attachments: rupture force distribution

    CERN Document Server

    Ghanti, Dipanwita

    2016-01-01

    Application of pulling force, under force-clamp conditions, to kinetochore-microtubule attachments {\\it in-vitro} revealed a catch-bond-like behavior. In an earlier paper ({\\it Sharma et al. Phys. Biol. (2014)} the physical origin of this apparently counter-intuitive phenomenon was traced to the nature of the force-dependence of the (de-)polymerization kinetics of the microtubules. In this brief communication that work is extended to situations where the external forced is ramped up till the attachment gets ruptured. In spite of the fundamental differences in the underlying mechanisms, the trend of variation of the rupture force distribution observed in our model kinetochore-microtubule attachment with the increasing loading rate is qualitatively similar to that displayed by the catch bonds formed in some other ligand-receptor systems. Our theoretical predictions can be tested experimentally by a straightforward modification of the protocol for controlling the force in the optical trap set up that was used in...

  5. Cep70 promotes microtubule assembly in vitro by increasing microtubule elongation

    Institute of Scientific and Technical Information of China (English)

    Xingjuan Shi; Jun Wang; Yunfan Yang; Yuan Ren; Jun Zhou; Dengwen Li

    2012-01-01

    Microtubules are dynamic cytoskeletal polymers present in all eukaryotic cells,In animal cells,they are organized by the centrosome,the major microtubule-organizing center.Many centrosomal proteins act coordinately to modulate microtubule assembly and organization.Our previous work has shown that Cep70,a novel centrosomal protein regulates microtubule assembly and organization in mammalian cells.However,the molecular details remain to be investigated,in this study,we investigated the molecular mechanism of how Cep70 regulates microtubule assembly using purified proteins.Our data showed that Cep70 increased the microtubule length without affecting the microtubule number in the purified system.These results demonstrate that Cep70 could directly regulate microtubule assembly by promoting microtubule elongation instead of microtubule nucleation.

  6. ASTER DEM performance

    Science.gov (United States)

    Fujisada, H.; Bailey, G.B.; Kelly, Glen G.; Hara, S.; Abrams, M.J.

    2005-01-01

    The Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) instrument onboard the National Aeronautics and Space Administration's Terra spacecraft has an along-track stereoscopic capability using its a near-infrared spectral band to acquire the stereo data. ASTER has two telescopes, one for nadir-viewing and another for backward-viewing, with a base-to-height ratio of 0.6. The spatial resolution is 15 m in the horizontal plane. Parameters such as the line-of-sight vectors and the pointing axis were adjusted during the initial operation period to generate Level-1 data products with a high-quality stereo system performance. The evaluation of the digital elevation model (DEM) data was carried out both by Japanese and U.S. science teams separately using different DEM generation software and reference databases. The vertical accuracy of the DEM data generated from the Level-1A data is 20 m with 95% confidence without ground control point (GCP) correction for individual scenes. Geolocation accuracy that is important for the DEM datasets is better than 50 m. This appears to be limited by the spacecraft position accuracy. In addition, a slight increase in accuracy is observed by using GCPs to generate the stereo data. ?? 2005 IEEE.

  7. The acetylenic tricyclic bis(cyano enone), TBE-31, targets microtubule dynamics and cell polarity in migrating cells.

    Science.gov (United States)

    Chan, Eddie; Saito, Akira; Honda, Tadashi; Di Guglielmo, Gianni M

    2016-04-01

    Cell migration is dependent on the microtubule network for structural support as well as for the proper delivery and positioning of polarity proteins at the leading edge of migrating cells. Identification of drugs that target cytoskeletal-dependent cell migration and protein transport in polarized migrating cells is important in understanding the cell biology of normal and tumor cells and can lead to new therapeutic targets in disease processes. Here, we show that the tricyclic compound TBE-31 directly binds to tubulin and interferes with microtubule dynamics, as assessed by end binding 1 (EB1) live cell imaging. Interestingly, this interference is independent of in vitro tubulin polymerization. Using immunofluorescence microscopy, we also observed that TBE-31 interferes with the polarity of migratory cells. The polarity proteins Rac1, IQGAP and Tiam1 were localized at the leading edge of DMSO-treated migrating cell, but were observed to be in multiple protrusions around the cell periphery of TBE-31-treated cells. Finally, we observed that TBE-31 inhibits the migration of Rat2 fibroblasts with an IC50 of 0.75 μM. Taken together, our results suggest that the inhibition of cell migration by TBE-31 may result from the improper maintenance of cell polarity of migrating cells.

  8. NASA ASTER Level 1T

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — The Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) is an advanced multispectral imager that was launched on board NASA's Terra spacecraft in...

  9. Cell cycle-dependent microtubule-based dynamic transport of cytoplasmic dynein in mammalian cells.

    Directory of Open Access Journals (Sweden)

    Takuya Kobayashi

    Full Text Available BACKGROUND: Cytoplasmic dynein complex is a large multi-subunit microtubule (MT-associated molecular motor involved in various cellular functions including organelle positioning, vesicle transport and cell division. However, regulatory mechanism of the cell-cycle dependent distribution of dynein has not fully been understood. METHODOLOGY/PRINCIPAL FINDINGS: Here we report live-cell imaging of cytoplasmic dynein in HeLa cells, by expressing multifunctional green fluorescent protein (mfGFP-tagged 74-kDa intermediate chain (IC74. IC74-mfGFP was successfully incorporated into functional dynein complex. In interphase, dynein moved bi-directionally along with MTs, which might carry cargos such as transport vesicles. A substantial fraction of dynein moved toward cell periphery together with EB1, a member of MT plus end-tracking proteins (+TIPs, suggesting +TIPs-mediated transport of dynein. In late-interphase and prophase, dynein was localized at the centrosomes and the radial MT array. In prometaphase and metaphase, dynein was localized at spindle MTs where it frequently moved from spindle poles toward chromosomes or cell cortex. +TIPs may be involved in the transport of spindle dyneins. Possible kinetochore and cortical dyneins were also observed. CONCLUSIONS AND SIGNIFICANCE: These findings suggest that cytoplasmic dynein is transported to the site of action in preparation for the following cellular events, primarily by the MT-based transport. The MT-based transport may have greater advantage than simple diffusion of soluble dynein in rapid and efficient transport of the limited concentration of the protein.

  10. The influence of Aster x salignus Willd. Invasion on the diversity of soil yeast communities

    Science.gov (United States)

    Glushakova, A. M.; Kachalkin, A. V.; Chernov, I. Yu.

    2016-07-01

    The annual dynamics of yeast communities were studied in the soddy-podzolic soil under the thickets of Aster x salignus Willd., one of the widespread invasive plant species in central Russia. Yeast groups in the soils under continuous aster thickets were found to differ greatly from the yeast communities in the soils under the adjacent indigenous meadow vegetation. In both biotopes the same species ( Candida vartiovaarae, Candida sake, and Cryptococcus terreus) are dominants. However, in the soils under indigenous grasses, eurybiontic yeasts Rhodotorula mucilaginosa, which almost never occur in the soil under aster, are widespread. In the soil under aster, the shares of other typical epiphytic and pedobiontic yeast fungi (ascomycetic species Wickerhamomyces aniomalus, Barnettozyma californica and basidiomycetic species Cystofilobasidium macerans, Guehomyces pullulans) significantly increase. Thus, the invasion of Aster x salignus has a clear effect on soil yeast complexes reducing their taxonomic and ecological diversity.

  11. Models for microtubule cargo transport coupling the Langevin equation to stochastic stepping motor dynamics: Caring about fluctuations.

    Science.gov (United States)

    Bouzat, Sebastián

    2016-01-01

    One-dimensional models coupling a Langevin equation for the cargo position to stochastic stepping dynamics for the motors constitute a relevant framework for analyzing multiple-motor microtubule transport. In this work we explore the consistence of these models focusing on the effects of the thermal noise. We study how to define consistent stepping and detachment rates for the motors as functions of the local forces acting on them in such a way that the cargo velocity and run-time match previously specified functions of the external load, which are set on the base of experimental results. We show that due to the influence of the thermal fluctuations this is not a trivial problem, even for the single-motor case. As a solution, we propose a motor stepping dynamics which considers memory on the motor force. This model leads to better results for single-motor transport than the approaches previously considered in the literature. Moreover, it gives a much better prediction for the stall force of the two-motor case, highly compatible with the experimental findings. We also analyze the fast fluctuations of the cargo position and the influence of the viscosity, comparing the proposed model to the standard one, and we show how the differences on the single-motor dynamics propagate to the multiple motor situations. Finally, we find that the one-dimensional character of the models impede an appropriate description of the fast fluctuations of the cargo position at small loads. We show how this problem can be solved by considering two-dimensional models.

  12. MVL-PLA2, a snake venom phospholipase A2, inhibits angiogenesis through an increase in microtubule dynamics and disorganization of focal adhesions.

    Directory of Open Access Journals (Sweden)

    Amine Bazaa

    Full Text Available Integrins are essential protagonists of the complex multi-step process of angiogenesis that has now become a major target for the development of anticancer therapies. We recently reported and characterized that MVL-PLA2, a novel phospholipase A2 from Macrovipera lebetina venom, exhibited anti-integrin activity. In this study, we show that MVL-PLA2 also displays potent anti-angiogenic properties. This phospholipase A2 inhibited adhesion and migration of human microvascular-endothelial cells (HMEC-1 in a dose-dependent manner without being cytotoxic. Using Matrigel and chick chorioallantoic membrane assays, we demonstrated that MVL-PLA2, as well as its catalytically inactivated form, significantly inhibited angiogenesis both in vitro and in vivo. We have also found that the actin cytoskeleton and the distribution of alphav beta3 integrin, a critical regulator of angiogenesis and a major component of focal adhesions, were disturbed after MVL-PLA2 treatment. In order to further investigate the mechanism of action of this protein on endothelial cells, we analyzed the dynamic instability behavior of microtubules in living endothelial cells. Interestingly, we showed that MVL-PLA2 significantly increased microtubule dynamicity in HMEC-1 cells by 40%. We propose that the enhancement of microtubule dynamics may explain the alterations in the formation of focal adhesions, leading to inhibition of cell adhesion and migration.

  13. Mammalian diaphanous-related formin 1 regulates GSK3β-dependent microtubule dynamics required for T cell migratory polarization.

    Directory of Open Access Journals (Sweden)

    Baoxia Dong

    Full Text Available The mammalian diaphanous-related formin (mDia1, a Rho-regulated cytoskeletal modulator, has been shown to promote T lymphocyte chemotaxis and interaction with antigen presenting cells, but the mechanisms underpinning mDia1 roles in these processes have not been defined. Here we show that mDia1(-/- T cells exhibit impaired lymphocyte function-associated antigen 1 (LFA-1-mediated T cell adhesion, migration and in vivo trafficking. These defects are associated with impaired microtubule (MT polarization and stabilization, altered MT dynamics and reduced peripheral clustering of the MT plus-end-protein, adenomatous polyposis coli (APC in migrating T cells following LFA-1-engagement. Loss of mDia1 also leads to impaired inducible inactivation of the glycogen synthase kinase (GSK 3β as well as hyperphosphorylation and reduced levels of APC in migrating T cells. These findings identify essential roles for the mDia1 formin in modulating GSK3β-dependent MT contributions to induction of T-cell polarity, adhesion and motility.

  14. Mammalian diaphanous-related formin 1 regulates GSK3β-dependent microtubule dynamics required for T cell migratory polarization.

    Science.gov (United States)

    Dong, Baoxia; Zhang, Steven S; Gao, Wen; Su, Haichun; Chen, Jun; Jin, Fuzi; Bhargava, Ajay; Chen, Xiequn; Jorgensen, Lars; Alberts, Arthur S; Zhang, Jinyi; Siminovitch, Katherine A

    2013-01-01

    The mammalian diaphanous-related formin (mDia1), a Rho-regulated cytoskeletal modulator, has been shown to promote T lymphocyte chemotaxis and interaction with antigen presenting cells, but the mechanisms underpinning mDia1 roles in these processes have not been defined. Here we show that mDia1(-/-) T cells exhibit impaired lymphocyte function-associated antigen 1 (LFA-1)-mediated T cell adhesion, migration and in vivo trafficking. These defects are associated with impaired microtubule (MT) polarization and stabilization, altered MT dynamics and reduced peripheral clustering of the MT plus-end-protein, adenomatous polyposis coli (APC) in migrating T cells following LFA-1-engagement. Loss of mDia1 also leads to impaired inducible inactivation of the glycogen synthase kinase (GSK) 3β as well as hyperphosphorylation and reduced levels of APC in migrating T cells. These findings identify essential roles for the mDia1 formin in modulating GSK3β-dependent MT contributions to induction of T-cell polarity, adhesion and motility.

  15. Recruitment of EB1, a Master Regulator of Microtubule Dynamics, to the Surface of the Theileria annulata Schizont

    KAUST Repository

    Woods, Kerry L.

    2013-05-09

    The apicomplexan parasite Theileria annulata transforms infected host cells, inducing uncontrolled proliferation and clonal expansion of the parasitized cell population. Shortly after sporozoite entry into the target cell, the surrounding host cell membrane is dissolved and an array of host cell microtubules (MTs) surrounds the parasite, which develops into the transforming schizont. The latter does not egress to invade and transform other cells. Instead, it remains tethered to host cell MTs and, during mitosis and cytokinesis, engages the cell\\'s astral and central spindle MTs to secure its distribution between the two daughter cells. The molecular mechanism by which the schizont recruits and stabilizes host cell MTs is not known. MT minus ends are mostly anchored in the MT organizing center, while the plus ends explore the cellular space, switching constantly between phases of growth and shrinkage (called dynamic instability). Assuming the plus ends of growing MTs provide the first point of contact with the parasite, we focused on the complex protein machinery associated with these structures. We now report how the schizont recruits end-binding protein 1 (EB1), a central component of the MT plus end protein interaction network and key regulator of host cell MT dynamics. Using a range of in vitro experiments, we demonstrate that T. annulata p104, a polymorphic antigen expressed on the schizont surface, functions as a genuine EB1-binding protein and can recruit EB1 in the absence of any other parasite proteins. Binding strictly depends on a consensus SxIP motif located in a highly disordered C-terminal region of p104. We further show that parasite interaction with host cell EB1 is cell cycle regulated. This is the first description of a pathogen-encoded protein to interact with EB1 via a bona-fide SxIP motif. Our findings provide important new insight into the mode of interaction between Theileria and the host cell cytoskeleton. 2013 Woods et al.

  16. Recruitment of EB1, a master regulator of microtubule dynamics, to the surface of the Theileria annulata schizont.

    Directory of Open Access Journals (Sweden)

    Kerry L Woods

    2013-05-01

    Full Text Available The apicomplexan parasite Theileria annulata transforms infected host cells, inducing uncontrolled proliferation and clonal expansion of the parasitized cell population. Shortly after sporozoite entry into the target cell, the surrounding host cell membrane is dissolved and an array of host cell microtubules (MTs surrounds the parasite, which develops into the transforming schizont. The latter does not egress to invade and transform other cells. Instead, it remains tethered to host cell MTs and, during mitosis and cytokinesis, engages the cell's astral and central spindle MTs to secure its distribution between the two daughter cells. The molecular mechanism by which the schizont recruits and stabilizes host cell MTs is not known. MT minus ends are mostly anchored in the MT organizing center, while the plus ends explore the cellular space, switching constantly between phases of growth and shrinkage (called dynamic instability. Assuming the plus ends of growing MTs provide the first point of contact with the parasite, we focused on the complex protein machinery associated with these structures. We now report how the schizont recruits end-binding protein 1 (EB1, a central component of the MT plus end protein interaction network and key regulator of host cell MT dynamics. Using a range of in vitro experiments, we demonstrate that T. annulata p104, a polymorphic antigen expressed on the schizont surface, functions as a genuine EB1-binding protein and can recruit EB1 in the absence of any other parasite proteins. Binding strictly depends on a consensus SxIP motif located in a highly disordered C-terminal region of p104. We further show that parasite interaction with host cell EB1 is cell cycle regulated. This is the first description of a pathogen-encoded protein to interact with EB1 via a bona-fide SxIP motif. Our findings provide important new insight into the mode of interaction between Theileria and the host cell cytoskeleton.

  17. External electric field effects on the mechanical properties of the αβ-tubulin dimer of microtubules: a molecular dynamics study.

    Science.gov (United States)

    Saeidi, H R; Lohrasebi, A; Mahnam, K

    2014-08-01

    The mechanical properties of the αβ-tubulin dimer of microtubules was modeled by using the molecular dynamics (MD) simulation method. The effect on the mechanical properties of the dimer of the existence and nonexistence of an applied electric field, either constant or periodic, was studied. Since there are charged or polar groups in the dimer structure, the electric field can interact with the dimer. The elastic constant and Young's modulus of the dimer were decreased when the dimer was exposed to a constant electric field of 0.03 V/nm. Furthermore, applying an oscillating electric field in the 1 GHz range to the dimer increased the elastic constant and Young's modulus of the dimer. These parameters were related to dimer rigidity and, consequently, in this frequency range, the application of electric fields may affect the function of microtubules.

  18. SAS-4 is recruited to a dynamic structure in newly forming centrioles that is stabilized by the gamma-tubulin-mediated addition of centriolar microtubules.

    Science.gov (United States)

    Dammermann, Alexander; Maddox, Paul S; Desai, Arshad; Oegema, Karen

    2008-02-25

    Centrioles are surrounded by pericentriolar material (PCM), which is proposed to promote new centriole assembly by concentrating gamma-tubulin. Here, we quantitatively monitor new centriole assembly in living Caenorhabditis elegans embryos, focusing on the conserved components SAS-4 and SAS-6. We show that SAS-4 and SAS-6 are coordinately recruited to the site of new centriole assembly and reach their maximum levels during S phase. Centriolar SAS-6 is subsequently reduced by a mechanism intrinsic to the early assembly pathway that does not require progression into mitosis. Centriolar SAS-4 remains in dynamic equilibrium with the cytoplasmic pool until late prophase, when it is stably incorporated in a step that requires gamma-tubulin and microtubule assembly. These results indicate that gamma-tubulin in the PCM stabilizes the nascent daughter centriole by promoting microtubule addition to its outer wall. Such a mechanism may help restrict new centriole assembly to the vicinity of preexisting parent centrioles that recruit PCM.

  19. Structural investigations into the binding mode of novel neolignans Cmp10 and Cmp19 microtubule stabilizers by in silico molecular docking, molecular dynamics, and binding free energy calculations.

    Science.gov (United States)

    Tripathi, Shubhandra; Kumar, Akhil; Kumar, B Sathish; Negi, Arvind S; Sharma, Ashok

    2016-06-01

    Microtubule stabilizers provide an important mode of treatment via mitotic cell arrest of cancer cells. Recently, we reported two novel neolignans derivatives Cmp10 and Cmp19 showing anticancer activity and working as microtubule stabilizers at micromolar concentrations. In this study, we have explored the binding site, mode of binding, and stabilization by two novel microtubule stabilizers Cmp10 and Cmp19 using in silico molecular docking, molecular dynamics (MD) simulation, and binding free energy calculations. Molecular docking studies were performed to explore the β-tubulin binding site of Cmp10 and Cmp19. Further, MD simulations were used to probe the β-tubulin stabilization mechanism by Cmp10 and Cmp19. Binding affinity was also compared for Cmp10 and Cmp19 using binding free energy calculations. Our docking results revealed that both the compounds bind at Ptxl binding site in β-tubulin. MD simulation studies showed that Cmp10 and Cmp19 binding stabilizes M-loop (Phe272-Val288) residues of β-tubulin and prevent its dynamics, leading to a better packing between α and β subunits from adjacent tubulin dimers. In addition, His229, Ser280 and Gln281, and Arg278, Thr276, and Ser232 were found to be the key amino acid residues forming H-bonds with Cmp10 and Cmp19, respectively. Consequently, binding free energy calculations indicated that Cmp10 (-113.655 kJ/mol) had better binding compared to Cmp19 (-95.216 kJ/mol). This study provides useful insight for better understanding of the binding mechanism of Cmp10 and Cmp19 and will be helpful in designing novel microtubule stabilizers.

  20. Persistence Length of Stable Microtubules

    Science.gov (United States)

    Hawkins, Taviare; Mirigian, Matthew; Yasar, M. Selcuk; Ross, Jennifer

    2011-03-01

    Microtubules are a vital component of the cytoskeleton. As the most rigid of the cytoskeleton filaments, they give shape and support to the cell. They are also essential for intracellular traffic by providing the roadways onto which organelles are transported, and they are required to reorganize during cellular division. To perform its function in the cell, the microtubule must be rigid yet dynamic. We are interested in how the mechanical properties of stable microtubules change over time. Some ``stable'' microtubules of the cell are recycled after days, such as in the axons of neurons or the cilia and flagella. We measured the persistence length of freely fluctuating taxol-stabilized microtubules over the span of a week and analyzed them via Fourier decomposition. As measured on a daily basis, the persistence length is independent of the contour length. Although measured over the span of the week, the accuracy of the measurement and the persistence length varies. We also studied how fluorescently-labeling the microtubule affects the persistence length and observed that a higher labeling ratio corresponded to greater flexibility. National Science Foundation Grant No: 0928540 to JLR.

  1. Recent developments in seismic analysis in the code Aster; Les developpements recents en analyse sismique dans le code aster

    Energy Technology Data Exchange (ETDEWEB)

    Guihot, P.; Devesa, G.; Dumond, A.; Panet, M.; Waeckel, F.

    1996-12-31

    Progress in the field of seismic qualification and design methods made these last few years allows physical phenomena actually in play to be better considered, while cutting down the conservatism associated with some simplified design methods. So following the change in methods and developing the most advantageous ones among them contributes to the process of the seismic margins assessment and the preparation of new design tools for future series. In this paper, the main developments and improvements in methods which have been made these last two years in the Code Aster, in order to improve seismic calculation methods and seismic margin assessment are presented. The first development relates to making the MISS3D soil structure interaction code available, thanks to an interface made with the Code Aster. The second relates to the possibility of making modal basis time calculations on multi-supported structures by considering local non linearities like impact, friction or squeeze fluid forces. Recent developments in random dynamics and postprocessing devoted to earthquake designs are then mentioned. Three applications of these developments are then ut forward. The first application relates to a test case for soil structure interaction design using MISS3D-Aster coupling. The second is a test case for a multi-supported structure. The last application, more for manufacturing, refers to seismic qualification of Main Live Steam stop valves. First results of the independent validation of the Code Aster seismic design functionalities, which provide and improve the quality of software, are also recalled. (authors). 11 refs.

  2. Modeling microtubule oscillations

    DEFF Research Database (Denmark)

    Jobs, E.; Wolf, D.E.; Flyvbjerg, H.

    1997-01-01

    Synchronization of molecular reactions in a macroscopic volume may cause the volume's physical properties to change dynamically and thus reveal much about the reactions. As an example, experimental time series for so-called microtubule oscillations are analyzed in terms of a minimal model...... for this complex polymerization-depolymerization cycle. The model reproduces well the qualitatively different time series that result from different experimental conditions, and illuminates the role and importance of individual processes in the cycle. Simple experiments are suggested that can further test...... and define the model and the polymer's reaction cycle....

  3. Kinesin-12 motors cooperate to suppress microtubule catastrophes and drive the formation of parallel microtubule bundles.

    Science.gov (United States)

    Drechsler, Hauke; McAinsh, Andrew D

    2016-03-22

    Human Kinesin-12 (hKif15) plays a crucial role in assembly and maintenance of the mitotic spindle. These functions of hKif15 are partially redundant with Kinesin-5 (Eg5), which can cross-link and drive the extensile sliding of antiparallel microtubules. Although both motors are known to be tetramers, the functional properties of hKif15 are less well understood. Here we reveal how single or multiple Kif15 motors can cross-link, transport, and focus the plus-ends of intersecting microtubules. During transport, Kif15 motors step simultaneously along both microtubules with relative microtubule transport driven by a velocity differential between motor domain pairs. Remarkably, this differential is affected by the underlying intersection geometry: the differential is low on parallel and extreme on antiparallel microtubules where one motor domain pair becomes immobile. As a result, when intersecting microtubules are antiparallel, canonical transport of one microtubule along the other is allowed because one motor is firmly attached to one microtubule while it is stepping on the other. When intersecting microtubules are parallel, however, Kif15 motors can drive (biased) parallel sliding because the motor simultaneously steps on both microtubules that it cross-links. These microtubule rearrangements will focus microtubule plus-ends and finally lead to the formation of parallel bundles. At the same time, Kif15 motors cooperate to suppress catastrophe events at polymerizing microtubule plus-ends, raising the possibility that Kif15 motors may synchronize the dynamics of bundles that they have assembled. Thus, Kif15 is adapted to operate on parallel microtubule substrates, a property that clearly distinguishes it from the other tetrameric spindle motor, Eg5.

  4. The use of compressive sensing and peak detection in the reconstruction of microtubules length time series in the process of dynamic instability.

    Science.gov (United States)

    Mahrooghy, Majid; Yarahmadian, Shantia; Menon, Vineetha; Rezania, Vahid; Tuszynski, Jack A

    2015-10-01

    Microtubules (MTs) are intra-cellular cylindrical protein filaments. They exhibit a unique phenomenon of stochastic growth and shrinkage, called dynamic instability. In this paper, we introduce a theoretical framework for applying Compressive Sensing (CS) to the sampled data of the microtubule length in the process of dynamic instability. To reduce data density and reconstruct the original signal with relatively low sampling rates, we have applied CS to experimental MT lament length time series modeled as a Dichotomous Markov Noise (DMN). The results show that using CS along with the wavelet transform significantly reduces the recovery errors comparing in the absence of wavelet transform, especially in the low and the medium sampling rates. In a sampling rate ranging from 0.2 to 0.5, the Root-Mean-Squared Error (RMSE) decreases by approximately 3 times and between 0.5 and 1, RMSE is small. We also apply a peak detection technique to the wavelet coefficients to detect and closely approximate the growth and shrinkage of MTs for computing the essential dynamic instability parameters, i.e., transition frequencies and specially growth and shrinkage rates. The results show that using compressed sensing along with the peak detection technique and wavelet transform in sampling rates reduces the recovery errors for the parameters.

  5. Catechol-o-methyltransferase expression and 2-methoxyestradiol affect microtubule dynamics and modify steroid receptor signaling in leiomyoma cells.

    Directory of Open Access Journals (Sweden)

    Salama A Salama

    Full Text Available CONTEXT: Development of optimal medicinal treatments of uterine leiomyomas represents a significant challenge. 2-Methoxyestradiol (2ME is an endogenous estrogen metabolite formed by sequential action of CYP450s and catechol-O-methyltransferase (COMT. Our previous study demonstrated that 2ME is a potent antiproliferative, proapoptotic, antiangiogenic, and collagen synthesis inhibitor in human leiomyomas cells (huLM. OBJECTIVES: Our objectives were to investigate whether COMT expression, by the virtue of 2ME formation, affects the growth of huLM, and to explore the cellular and molecular mechanisms whereby COMT expression or treatment with 2ME affect these cells. RESULTS: Our data demonstrated that E(2-induced proliferation was less pronounced in cells over-expressing COMT or treated with 2ME (500 nM. This effect on cell proliferation was associated with microtubules stabilization and diminution of estrogen receptor alpha (ERalpha and progesterone receptor (PR transcriptional activities, due to shifts in their subcellular localization and sequestration in the cytoplasm. In addition, COMT over expression or treatment with 2ME reduced the expression of hypoxia-inducible factor -1alpha (HIF-1 alpha and the basal level as well as TNF-alpha-induced aromatase (CYP19 expression. CONCLUSIONS: COMT over expression or treatment with 2ME stabilize microtubules, ameliorates E(2-induced proliferation, inhibits ERalpha and PR signaling, and reduces HIF-1 alpha and CYP19 expression in human uterine leiomyoma cells. Thus, microtubules are a candidate target for treatment of uterine leiomyomas. In addition, the naturally occurring microtubule-targeting agent 2ME represents a potential new therapeutic for uterine leiomyomas.

  6. Microtubule heterogeneity of Ornithogalum umbellatum ovary epidermal cells: non-stable cortical microtubules and stable lipotubuloid microtubules

    Directory of Open Access Journals (Sweden)

    Maria Kwiatkowska

    2011-07-01

    Full Text Available Lipotubuloids, structures containing lipid bodies and microtubules, are described in ovary epidermalcells of Ornithogalum umbellatum. Microtubules of lipotubuloids can be fixed in electron microscope fixativecontaining only buffered OsO4 or in glutaraldehyde with OsO4 post-fixation, or in a mixture of OsO4 and glutaraldehyde[1]. None of these substances fixes cortical microtubules of ovary epidermis of this plant which ischaracterized by dynamic longitudinal growth. However, cortical microtubules can be fixed with cold methanolaccording immunocytological methods with the use of b-tubulin antibodies and fluorescein. The existence ofcortical microtubules has also been evidenced by EM observations solely after the use of taxol, microtubulestabilizer, and fixation in a glutaraldehyde/OsO4 mixture. These microtubules mostly lie transversely, sometimesobliquely, and rarely parallel to the cell axis. Staining, using Ruthenium Red and silver hexamine, has revealedthat lipotubuloid microtubules surface is covered with polysaccharides. The presumption has been made thatthe presence of a polysaccharide layer enhances the stability of lipotubuloid microtubules.

  7. Potent antiproliferative cembrenoids accumulate in tobacco upon infection with Rhodococcus fascians and trigger unusual microtubule dynamics in human glioblastoma cells.

    Directory of Open Access Journals (Sweden)

    Aminata P Nacoulma

    Full Text Available AIMS: Though plant metabolic changes are known to occur during interactions with bacteria, these were rarely challenged for pharmacologically active compounds suitable for further drug development. Here, the occurrence of specific chemicals with antiproliferative activity against human cancer cell lines was evidenced in hyperplasia (leafy galls induced when plants interact with particular phytopathogens, such as the Actinomycete Rhodococcus fascians. METHODS: We examined leafy galls fraction F3.1.1 on cell proliferation, cell division and cytoskeletal disorganization of human cancer cell lines using time-lapse videomicroscopy imaging, combined with flow cytometry and immunofluorescence analysis. We determined the F3.1.1-fraction composition by gas chromatography coupled to mass spectrometry. RESULTS: The leafy galls induced on tobacco by R. fascians yielded fraction F3.1.1 which inhibited proliferation of glioblastoma U373 cells with an IC50 of 4.5 µg/mL, F.3.1.1 was shown to increase cell division duration, cause nuclear morphological deformations and cell enlargement, and, at higher concentrations, karyokinesis defects leading to polyploidization and apoptosis. F3.1.1 consisted of a mixture of isomers belonging to the cembrenoids. The cellular defects induced by F3.1.1 were caused by a peculiar cytoskeletal disorganization, with the occurrence of fragmented tubulin and strongly organized microtubule aggregates within the same cell. Colchicine, paclitaxel, and cembrene also affected U373 cell proliferation and karyokinesis, but the induced microtubule rearrangement was very different from that provoked by F3.1.1. Altogether our data indicate that the cembrenoid isomers in F3.1.1 have a unique mode of action and are able to simultaneously modulate microtubule polymerization and stability.

  8. Effect of microtubule-associated protein tau in dynamics of single-headed motor proteins KIF1A

    CERN Document Server

    Sparacino, J; Lamberti, P W

    2013-01-01

    Intracellular transport based on molecular motors and its regulation are crucial to the functioning of cells. Filamentary tracks of the cells are abundantly decorated with non-motile microtubule-associated proteins, such as tau. Motivated by experiments on kinesin-tau interactions [Dixit et al. Science 319, 1086 (2008)] we developed a stochastic model of interacting single-headed motor proteins KIF1A that also takes into account the interactions between motor proteins and tau molecules. Our model reproduce experimental observations and predicts significant effects of tau on bound time and run length which suggest an important role of tau in regulation of kinesin-based transport.

  9. Chronological Reorganization of Microtubules, Actin Microfilaments, and Chromatin during the First Cell Cycle in Swamp Buffalo (Bubalus bubalis Embryos

    Directory of Open Access Journals (Sweden)

    Vibuntita Chankitisakul

    2010-01-01

    Full Text Available This paper aimed to study the dynamics of early embryonic development, in terms of redistribution of cytoskeleton (microtubules, actin microfilaments and chromatin configurations during the first cell cycle in swamp buffalo embryos. Oocytes were matured and fertilized in vitro, and they were fixed at various time points after IVF. At 6 h after IVF, 44.4% matured oocytes were penetrated by spermatozoa. Partial ZP digestion, however, did not improve fertilization rate compared to control (P>.05. At 12 h after IVF, the fertilized oocytes progressed to the second meiotic division and formed the female pronucleus simultaneously with the paternal chromatin continued to decondense. A sperm aster was observed radiating from the base of the decondensing sperm head. At 18 h after IVF, most presumptive zygotes had reached the pronuclear stage. The sperm aster was concurrently enlarged to assist the migration and apposition of pronuclei. Cell cleavage was facilitated by microfilaments and firstly observed by 30 h after IVF. In conclusion, the cytoskeleton actively involves with the process of fertilization and cleavage in swamp buffalo oocytes. The centrosomal material is paternally inherited. Fertilization failure is predominantly caused by poor sperm penetration. However, partial digestion of ZP did not improve fertilization rate.

  10. Terpenoid Glucosides from Aster Smithianus

    Institute of Scientific and Technical Information of China (English)

    GUO Shou-jun; ZHAO Xin-hua; YANG Yong-li; CHENG Dong-liang

    2005-01-01

    A diterpenoid trisaccharide, smithoside A, and a saponin, smithoside B, as well as six known compounds, apigenin, apigenin-7-O-glucoside, daucosterol, shanzhiside methyl ester, 8-acetylshanzhiside methyl ester and acteoside, were isolated from Aster smithianus. On the basis of spectral evidence and chemical analytical results, smithosides A and B were identified as pimar-15(16)-β-ene-8β,11α-diol-3β-O-β-D-glucopyranosyl(1→3)-[β-glucopyranosyl(1→2)]-β-D-glucopyranoside and 2β,3β,16β,21β,23-pentahydroxy-12-ene-28 oleanoic acid lactone-3-O-β-D-glucopyranosyl(1→3)-β-D-glucopyranoside, respectively.

  11. Pharmacologic reductions of total tau levels; implications for the role of microtubule dynamics in regulating tau expression

    Directory of Open Access Journals (Sweden)

    Dickey Chad A

    2006-07-01

    Full Text Available Abstract The microtubule-associated protein tau (MAPT is a pathological component of several neurodegenerative diseases and clinical dementias. Here, we have investigated the effects of a series of commercially available FDA-approved compounds and natural products on total tau protein levels using a cell-based approach that allows for the rapid and efficient measurement of changes in protein expression. Results The compounds that reduced tau largely fell within 3 functional categories with the largest percentage being microtubule regulators. Several of these candidates were validated in both a human neuroglioma and a human neuroblastoma cell line. While these drugs lead to a rapid reduction in tau protein levels, a selective decrease in MAPT mRNA expression was also observed. Conclusion These findings suggest that the identified compounds that reduce tau levels may act either through direct effects on the MAPT promoter itself or by altering a feedback transcriptional mechanism regulating MAPT transcription. This is particularly interesting in light of recent evidence suggesting that MAPT 5' UTR mutations in late-onset PD and PSP cases alter the expression of tau mRNA. In fact, one of the compounds we identified, rotenone, has been used extensively to model PD in rodents. These observations may provide key insights into the mechanism of tau turnover within the neuron while also providing the first evidence that selectively reducing tau protein levels may be possible using compounds that are FDA-approved for other uses.

  12. Wave->Diffusion Transition in Microtubules

    OpenAIRE

    2005-01-01

    In this paper the heat transport in microtubules (MT) is investigated. When the dimension of the structure is of the order of the de Broglie wave length the transport phenomena must be analyzed within quantum mechanics. In this paper we developed the Dirac type thermal equation for MT .The solution of the equation-the temperature fields for electrons can be wave type or diffusion type depending on the dynamics of the scattering. Key words: Microtubules ultrashort laser pulses, Dirac thermal e...

  13. Microtubules, Tubulins and Associated Proteins.

    Science.gov (United States)

    Raxworthy, Michael J.

    1988-01-01

    Reviews much of what is known about microtubules, which are biopolymers consisting predominantly of subunits of the globular protein, tubulin. Describes the functions of microtubules, their structure and assembly, microtube associated proteins, and microtubule-disrupting agents. (TW)

  14. A mutation of the fission yeast EB1 overcomes negative regulation by phosphorylation and stabilizes microtubules

    Energy Technology Data Exchange (ETDEWEB)

    Iimori, Makoto; Ozaki, Kanako [Graduate School of Biostudies, Kyoto University, Kitashirakawa-Oiwake cho, Sakyo ku, Kyoto, 606-8502 (Japan); Chikashige, Yuji [Kobe Advanced ICT Research Center, National Institute of Information and Communications Technology, Kobe, 651-2492 (Japan); Habu, Toshiyuki [Graduate School of Biostudies, Kyoto University, Kitashirakawa-Oiwake cho, Sakyo ku, Kyoto, 606-8502 (Japan); Radiation Biology Center, Kyoto University, Yoshida-Konoe cho, Sakyo ku, Kyoto, 606-8501 (Japan); Hiraoka, Yasushi [Kobe Advanced ICT Research Center, National Institute of Information and Communications Technology, Kobe, 651-2492 (Japan); Graduate School of Frontier Biosciences, Osaka University, 1-3 Yamadaoka, Suita, 565-0871 (Japan); Maki, Takahisa; Hayashi, Ikuko [Graduate School of Nanobioscience, Yokohama City University, Tsurumi, Yokohama, 230-0045 (Japan); Obuse, Chikashi [Graduate School of Life Science, Hokkaido University, Sapporo 001-0021 (Japan); Matsumoto, Tomohiro, E-mail: tmatsumo@house.rbc.kyoto-u.ac.jp [Graduate School of Biostudies, Kyoto University, Kitashirakawa-Oiwake cho, Sakyo ku, Kyoto, 606-8502 (Japan); Radiation Biology Center, Kyoto University, Yoshida-Konoe cho, Sakyo ku, Kyoto, 606-8501 (Japan)

    2012-02-01

    Mal3 is a fission yeast homolog of EB1, a plus-end tracking protein (+ TIP). We have generated a mutation (89R) replacing glutamine with arginine in the calponin homology (CH) domain of Mal3. Analysis of the 89R mutant in vitro has revealed that the mutation confers a higher affinity to microtubules and enhances the intrinsic activity to promote the microtubule-assembly. The mutant Mal3 is no longer a + TIP, but binds strongly the microtubule lattice. Live cell imaging has revealed that while the wild type Mal3 proteins dissociate from the tip of the growing microtubules before the onset of shrinkage, the mutant Mal3 proteins persist on microtubules and reduces a rate of shrinkage after a longer pausing period. Consequently, the mutant Mal3 proteins cause abnormal elongation of microtubules composing the spindle and aster. Mal3 is phosphorylated at a cluster of serine/threonine residues in the linker connecting the CH and EB1-like C-terminal motif domains. The phosphorylation occurs in a microtubule-dependent manner and reduces the affinity of Mal3 to microtubules. We propose that because the 89R mutation is resistant to the effect of phosphorylation, it can associate persistently with microtubules and confers a stronger stability of microtubules likely by reinforcing the cylindrical structure. -- Highlights: Black-Right-Pointing-Pointer We characterize a mutation (mal3-89R) in fission yeast homolog of EB1. Black-Right-Pointing-Pointer The mutation enhances the activity to assemble microtubules. Black-Right-Pointing-Pointer Mal3 is phosphorylated in a microtubule-dependent manner. Black-Right-Pointing-Pointer The phosphorylation negatively regulates the Mal3 activity.

  15. Microtubule stabilization reduces scarring and causes axon regeneration after spinal cord injury

    NARCIS (Netherlands)

    F. Hellal (Farida); A. Hurtado (Andres); J. Ruschel (Jörg); K.C. Flynn (Kevin); C.J. Laskowski (Claudia); M. Umlauf (Martina); L.C. Kapitein (Lukas); D. Strikis (Dinara); V. Lemmon (Vance); J. Bixby (John); C.C. Hoogenraad (Casper); F. Bradke (Frank)

    2011-01-01

    textabstractHypertrophic scarring and poor intrinsic axon growth capacity constitute major obstacles for spinal cord repair. These processes are tightly regulated by microtubule dynamics. Here, moderate microtubule stabilization decreased scar formation after spinal cord injury in rodents through va

  16. Digital elevation modelling using ASTER stereo imagery.

    Science.gov (United States)

    Forkuo, Eric Kwabena

    2010-04-01

    Digital elevation model (DEM) in recent times has become an integral part of national spatial data infrastructure of many countries world-wide due to its invaluable importance. Although DEMs are mostly generated from contours maps, stereo aerial photographs and air-borne and terrestrial laser scanning, the stereo interpretation and auto-correlation from satellite image stereo-pairs such as with SPOT, IRS, and relatively new ASTER imagery is also an effective means of producing DEM data. In this study, terrain elevation data were derived by applying photogrammetric process to ASTER stereo imagery. Also, the quality ofDEMs produced from ASTER stereo imagery was analysed by comparing it with DEM produced from topographic map at a scale of 1:50,000. While analyzing the vertical accuracy of the generated ASTER DEM, fifty ground control points were extracted from the map and overlaid on the DEM. Results indicate that a root-mean-square error in elevation of +/- 14 m was achieved with ASTER stereo image data of good quality. The horizontal accuracy obtained from the ground control points was 14.77, which is within the acceptable range of +/- 7m to +/- 25 m. The generated (15 m) DEM was compared with a 20m, 25m, and a 30 m pixel DEM to the original map. In all, the results proved that, the 15 m DEM conform to the original map DEM than the others. Overall, this analysis proves that, the generated digital terrain model, DEM is acceptable.

  17. Effects of silver ions (Ag+) on contractile ring function and microtubule dynamics during first cleavage in Ilyanassa obsoleta

    Science.gov (United States)

    Conrad, A. H.; Stephens, A. P.; Paulsen, A. Q.; Schwarting, S. S.; Conrad, G. W.; Spooner, B. S. (Principal Investigator)

    1994-01-01

    The terminal phase of cell division involves tight constriction of the cleavage furrow contractile ring, stabilization/elongation of the intercellular bridge, and final separation of the daughter cells. At first cleavage, the fertilized eggs of the mollusk, Ilyanassa obsoleta, form two contractile rings at right angles to each other in the same cytoplasm that constrict to tight necks and partition the egg into a trefoil shape. The cleavage furrow contractile ring (CF) normally constricts around many midbody microtubules (MTs) and results in cleavage; the polar lobe constriction contractile ring (PLC) normally constricts around very few MTs and subsequently relaxes without cleavage. In the presence of Ag+ ions, the PLC 1) begins MT-dependent rapid constriction sooner than controls, 2) encircles more MTs than control egg PLCs, 3) elongates much more than control PLCs, and 4) remains tightly constricted and effectively cleaves the polar lobe from the egg. If Ag(+)-incubated eggs are returned to normal seawater at trefoil, tubulin fluorescence disappears from the PLC neck and the neck relaxes. If nocodazole, a drug that depolymerizes MTs, is added to Ag(+)-incubated eggs during early PLC constriction, the PLC is not stabilized and eventually relaxes. However, if nocodazole is added to Ag(+)-incubated eggs at trefoil, tubulin fluorescence disappears from the PLC neck but the neck remains constricted. These results suggest that Ag+ accelerates and gradually stabilizes the PLC constriction by a mechanism that is initially MT-dependent, but that progressively becomes MT-independent.

  18. Measurement of Binding Force between Microtubule-Associated Protein and Microtubule

    Institute of Scientific and Technical Information of China (English)

    XU Chun-Hua; GUO Hong-Lian; QU E; LI Zhao-Lin; YUAN Ming; CHENG Bing-Ying; ZHANG Dao-Zhong

    2007-01-01

    Microtubule-associated proteins (MAPs) are important proteins in cells. They can regulate the organization,dynamics and function of microtubules. We measure the binding force between microtubule and a new plant MAP, i.e. AtMAP65-1, by dual-optical tweezers. The force is obtained to be 14.6±3.5 pN from the data statistics and analysis. This force measurement is helpful to understand the function and mechanism of MAPs from the mechanical point of view and lays the groundwork for future measurements of the mechanical properties of other biological macro-molecules.

  19. Fission yeast mtr1p regulates interphase microtubule cortical dwell-time

    Directory of Open Access Journals (Sweden)

    Frédérique Carlier-Grynkorn

    2014-06-01

    Full Text Available The microtubule cytoskeleton plays important roles in cell polarity, motility and division. Microtubules inherently undergo dynamic instability, stochastically switching between phases of growth and shrinkage. In cells, some microtubule-associated proteins (MAPs and molecular motors can further modulate microtubule dynamics. We present here the fission yeast mtr1+, a new regulator of microtubule dynamics that appears to be not a MAP or a motor. mtr1-deletion (mtr1Δ primarily results in longer microtubule dwell-time at the cell tip cortex, suggesting that mtr1p acts directly or indirectly as a destabilizer of microtubules. mtr1p is antagonistic to mal3p, the ortholog of mammalian EB1, which stabilizes microtubules. mal3Δ results in short microtubules, but can be partially rescued by mtr1Δ, as the double mutant mal3Δ mtr1Δ exhibits longer microtubules than mal3Δ single mutant. By sequence homology, mtr1p is predicted to be a component of the ribosomal quality control complex. Intriguingly, deletion of a predicted ribosomal gene, rps1801, also resulted in longer microtubule dwell-time similar to mtr1Δ. The double-mutant mal3Δ rps1801Δ also exhibits longer microtubules than mal3Δ single mutant alone. Our study suggests a possible involvement of mtr1p and the ribosome complex in modulating microtubule dynamics.

  20. Estimation of broadband emissivity (8-12 um) from ASTER data by using RM-NN.

    Science.gov (United States)

    Mao, K B; Ma, Y; Shen, X Y; Li, B P; Li, C Y; Li, Z L

    2012-08-27

    Land surface window emissivity is a key parameter for estimating the longwave radiative budget. The combined radiative transfer model (RM) with neural network (NN) algorithm is utilized to directly estimate the window (8-12 um) emissivity from the brightness temperature of the Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) with 90 m spatial resolution. Although the estimation accuracy is very high when the broadband emissivity is estimated from AST05 (ASTER Standard Data Product) by using regression method, the accuracy of AST05 is about ± 0.015 for 86 spectra which is determined by the atmosphere correction for ASTER 1B data. The MODTRAN 4 is used to simulate the process of radiance transfer, and the broadband emissivity is directly estimated from the brightness temperature of ASTER 1B data at satellite. The comparison analysis indicates that the RM-NN is more competent to estimate broadband emissivity than other method when the brightness temperatures of band 11, 12, 13, 14 are made as input nodes of dynamic neural network. The estimation average accuracy is about 0.009, and the estimation results are not sensitive to instrument noise. The RM-NN is applied to extract broadband emissivity from an image of ASTER 1B data in China, and the comparison against a classification based multiple bands with 15 m spatial resolution shows that the estimation results from RM-NN are very good.

  1. Using a deformable mirror to generate custom laser guidestar asterisms: simulation and laboratory results

    Science.gov (United States)

    Norton, Andrew P.; Srinath, Srikar; Gavel, Donald; Kupke, Renate; Dillon, Daren

    2014-08-01

    It is possible to create custom laser guidestar (LGS) asterisms from a single beam by using a deformable mirror to pattern the phase of the outgoing laser guidestar beam. This avoids the need for multiple laser launch assemblies, and in principle would allow one to position the multiple LGS spots in any desired arrangement around the science target, as well as dynamically rotate the LGS pattern on-sky and control the distribution of intensity in each spot. Simulations and laboratory experiments indicate that a PTT111 and PTT489 IrisAO MEMS deformable mirror and a Hamamatsu X8267 spatial light modulator may have applications for creating small LGS asterisms for biological imaging with adaptive optics. For astronomy applications, the phase values required to produce the "3+1" laser guidestar asterism of Keck's Next Generation AO system is also investigated.

  2. The dual specificity phosphatase Cdc14B bundles and stabilizes microtubules

    Energy Technology Data Exchange (ETDEWEB)

    Plumley, Hyekyung [ORNL; Liu, Yie [ORNL; Gomez, Marla V [ORNL; Wang, Yisong [ORNL

    2005-01-01

    The Cdc14 dual-specificity phosphatases regulate key events in the eukaryotic cell cycle. However, little is known about the function of mammalian CDC14B family members. Here, we demonstrate that subcellular localization of CDC14B protein is cell cycle regulated. CDC14B can bind, bundle, and stabilize microtubules in vitro independently of its catalytic activity. Basic amino acid residues within the nucleolar targeting domain are important for both retaining CDC14B in the nucleolus and preventing microtubule bundling. Overexpression of CDC14B resulted in the formation of cytoplasmic CDC14B and microtubule bundles in interphase cells. These microtubule bundles were resistant to microtubule depolymerization reagents and enriched in acetylated -tubulin. Expression of cytoplasmic forms of CDC14B impaired microtubule nucleation from the microtubule organization center. CDC14B is thus a novel microtubule-bundling and -stabilizing protein, whose regulated subcellular localization may help modulate spindle and microtubule dynamics in mitosis.

  3. In vitro assembly of plant tubulin in the absence of microtubule-stabilizing reagents

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    The assembly of microtubules is essential for physiological functions of microtubules. Addition of microtubule-stabilizing reagents or microtubule "seeds" is usually necessary for plant tubulin assembly in vitro, which hinders the investigation of plant microtubule dynamics. In the present note, highly purified plant tubulins have been obtained from lily pollen, a non-microtubule-stabilizing reagent or microtubule "seed" system for plant tubulin assembly has been established and the analysis of plant tubulin assembly performed. Experiment results showed that purified tubulin polymerized in vitro, and a typical microtubule structure was observed with electron microscopy. The kinetics curve of tubulin assembly exhibited typical "parabola". The presence of taxol significantly altered the character of plant tubulin assembly, including that abnormal microtubules were assembled and the critical concentration for plant tubulin assembly was decreased exceedingly from 3 mg/mL in the absence of taxol to 0.043 mg/mL in the presence of taxol.

  4. CLIP-170 facilitates the formation of kinetochore-microtubule attachments.

    Science.gov (United States)

    Tanenbaum, Marvin E; Galjart, Niels; van Vugt, Marcel A T M; Medema, René H

    2006-01-11

    CLIP-170 is a microtubule 'plus end tracking' protein involved in several microtubule-dependent processes in interphase. At the onset of mitosis, CLIP-170 localizes to kinetochores, but at metaphase, it is no longer detectable at kinetochores. Although RNA interference (RNAi) experiments have suggested an essential role for CLIP-170 during mitosis, the molecular function of CLIP-170 in mitosis has not yet been revealed. Here, we used a combination of high-resolution microscopy and RNAi-mediated depletion to study the function of CLIP-170 in mitosis. We found that CLIP-170 dynamically localizes to the outer most part of unattached kinetochores and to the ends of growing microtubules. In addition, we provide evidence that a pool of CLIP-170 is transported along kinetochore-microtubules by the dynein/dynactin complex. Interference with CLIP-170 expression results in defective chromosome congression and diminished kinetochore-microtubule attachments, but does not detectibly affect microtubule dynamics or kinetochore-microtubule stability. Taken together, our results indicate that CLIP-170 facilitates the formation of kinetochore-microtubule attachments, possibly through direct capture of microtubules at the kinetochore.

  5. Microtubules Are Essential for Guard-Cell Function in Vicia and Arabidopsis

    Institute of Scientific and Technical Information of China (English)

    William Eisinger; David Ehrhardt; Winslow Briggs

    2012-01-01

    Radially arranged cortical microtubules are a prominent feature of guard cells.Guard cells expressing GFPtubulin showed consistent changes in the appearance of microtubules when stomata opened or closed.Guard cells showed fewer microtubule structures as stomata closed,whether induced by transfer to darkness,ABA,hydrogen peroxide,or sodium hydrogen carbonate.Guard cells kept in the dark (closed stomata) showed increases in microtubule structures and stomatal aperture on light treatment.GFP-EB1,marking microtubule growing plus ends,showed no change in number of plus ends or velocity of assembly on stomatal closure.Since the number of growing plus ends and the rate of plus-end growth did not change when microtubule structure numbers declined,microtubule instability and/or rearrangement must be responsible for the apparent loss of microtubules.Guard cells with closed stomata showed more cytosolic GFP-fluorescence than those with open stomata as cortical microtubules became disassembled,although with a large net loss in total fluorescence.Microtubule-targeted drugs blocked guard-cell function in Vicia and Arabidopsis.Oryzalin disrupted guard-cell microtubules and prevented stomatal opening and taxol stabilized guard-cell microtubules and delayed stomatal closure.Gas exchange measurements indicated that the transgenes for fluorescent-labeled proteins did not disrupt normal stomatal function.These dynamic changes in guard-cell microtubules combined with our inhibitor studies provide evidence for an active role of microtubules in guard-cell function.

  6. Microtubule stability and MAPI B upregulation control neuritogenesis in CAD cells

    Institute of Scientific and Technical Information of China (English)

    Wen LI; Jin-tang XIA; Yue FENG

    2006-01-01

    Aim: To study the role of microtubule dynamics and microtubule associated protein 1B (MAP1B) in regulation of the neurite extension in CAD catecholaminergic neuronal cell line. Methods: The neuritogenesis of the CAD cells was abolished by inhibiting microtubule polymerization with nocodazole and by blocking microtubule depolymerization with taxol. MAP1B and tubulin protein expression was detected by Western blot. Immunofluorescent staining of tubulins was observed by fluorescent and confocal microscopy. Results: Microtubule dynamics was essential for CAD neurite extension. Dosage analysis revealed that neurite extension was much more sensitive to nocodazole than to taxol, suggesting a functional requirement for highly active microtubule assembly. A remarkable upregulation of MAP1B protein was detected during neurite extension accompanied with increased microtubule stability. Conclusion: Upregulation of MAP1B leads to the stabilization of newly formed microtubules in the developing neurites, which in turn promotes neurite extension.

  7. Alignment of Microtubule Imagery

    CERN Document Server

    Yu, Feiyang; Bakker, Erwin M

    2011-01-01

    This work discusses preliminary work aimed at simulating and visualizing the growth process of a tiny structure inside the cell---the microtubule. Difficulty of recording the process lies in the fact that the tissue preparation method for electronic microscopes is highly destructive to live cells. Here in this paper, our approach is to take pictures of microtubules at different time slots and then appropriately combine these images into a coherent video. Experimental results are given on real data.

  8. Two New Diterpene Acetylxylosides From Aster Veitchianus

    Institute of Scientific and Technical Information of China (English)

    Er Wei LI; Kun GAO; Zhong Jian JIA

    2004-01-01

    Two new acetylxylosides, ent-manool-13-O-β-D-2'-acetylxylopyranoside(1)and ent-manool-13-O-β-D-2',4'-diacetylxylopyranoside(2)were isolated from Aster veitchianus.Their structures were elucidated by spectroscopic methods.

  9. Thermal monitoring using an ASTER image

    Science.gov (United States)

    Costantino, Domenica; Angelini, Maria Giuseppa

    2016-10-01

    The research is focused on the study of the applicability of remote sensing techniques (specifically using ASTER data) for marine environmental analysis, relative to the determination of the surface temperatures. Using a multitemporal approach, two images (classified as level-1B), acquired in August 2000 and in 2005, were considered. The thermal maps were realized by means of the emissivity spectral normalization method, defining the thermal gradients in the area under investigation. The spatial and temporal anomalies related to the temperature distribution were highlighted; these anomalies represent an important parameter for the identification of probable groundwater pollution and soil contamination. To define the map of the surface temperature using thermal infrared ASTER channels, a numerical model dedicated to ASTER data was implemented during the experimentation. This model is based on the principle of interpolation of the type "least square interpolation (linear)," and it implies a reduction in the number of unknowns to obtain an acceptable solution to the problem. This experimental model has provided good results in the phase of implementation and in the tests on a synthetic image that were simulated in the laboratory. However, further verifications and modifications are necessary for the processing of real ASTER images.

  10. Biological Information Processing in Single Microtubules

    Science.gov (United States)

    2014-03-05

    the time for reviewing instructions, searching existing data sources, gathering and maintaining the data needed, and completing and reviewing the...generated by synchronized oscillations of microtubules, centrosomes and chromosomes regulate the dynamics of mitosis and meiosis , Yue Zhao and Qimin

  11. S. pombe kinesins-8 promote both nucleation and catastrophe of microtubules.

    Directory of Open Access Journals (Sweden)

    Muriel Erent

    Full Text Available The kinesins-8 were originally thought to be microtubule depolymerases, but are now emerging as more versatile catalysts of microtubule dynamics. We show here that S. pombe Klp5-436 and Klp6-440 are non-processive plus-end-directed motors whose in vitro velocities on S. pombe microtubules at 7 and 23 nm s(-1 are too slow to keep pace with the growing tips of dynamic interphase microtubules in living S. pombe. In vitro, Klp5 and 6 dimers exhibit a hitherto-undescribed combination of strong enhancement of microtubule nucleation with no effect on growth rate or catastrophe frequency. By contrast in vivo, both Klp5 and Klp6 promote microtubule catastrophe at cell ends whilst Klp6 also increases the number of interphase microtubule arrays (IMAs. Our data support a model in which Klp5/6 bind tightly to free tubulin heterodimers, strongly promoting the nucleation of new microtubules, and then continue to land as a tubulin-motor complex on the tips of growing microtubules, with the motors then dissociating after a few seconds residence on the lattice. In vivo, we predict that only at cell ends, when growing microtubule tips become lodged and their growth slows down, will Klp5/6 motor activity succeed in tracking growing microtubule tips. This mechanism would allow Klp5/6 to detect the arrival of microtubule tips at cells ends and to amplify the intrinsic tendency for microtubules to catastrophise in compression at cell ends. Our evidence identifies Klp5 and 6 as spatial regulators of microtubule dynamics that enhance both microtubule nucleation at the cell centre and microtubule catastrophe at the cell ends.

  12. Do prokaryotes contain microtubules?

    Science.gov (United States)

    Bermudes, D.; Hinkle, G.; Margulis, L.

    1994-01-01

    In eukaryotic cells, microtubules are 24-nm-diameter tubular structures composed of a class of conserved proteins called tubulin. They are involved in numerous cell functions including ciliary motility, nerve cell elongation, pigment migration, centrosome formation, and chromosome movement. Although cytoplasmic tubules and fibers have been observed in bacteria, some with diameters similar to those of eukaryotes, no homologies to eukaryotic microtubules have been established. Certain groups of bacteria including azotobacters, cyanobacteria, enteric bacteria, and spirochetes have been frequently observed to possess microtubule-like structures, and others, including archaebacteria, have been shown to be sensitive to drugs that inhibit the polymerization of microtubules. Although little biochemical or molecular biological information is available, the differences observed among these prokaryotic structures suggest that their composition generally differs among themselves as well as from that of eukaryotes. We review the distribution of cytoplasmic tubules in prokaryotes, even though, in all cases, their functions remain unknown. At least some tend to occur in cells that are large, elongate, and motile, suggesting that they may be involved in cytoskeletal functions, intracellular motility, or transport activities comparable to those performed by eukaryotic microtubules. In Escherichia coli, the FtsZ protein is associated with the formation of a ring in the division zone between the newly forming offspring cells. Like tubulin, FtsZ is a GTPase and shares with tubulin a 7-amino-acid motif, making it a promising candidate in which to seek the origin of tubulins.

  13. Pseudomonas aeruginosa exotoxin Y-mediated tau hyperphosphorylation impairs microtubule assembly in pulmonary microvascular endothelial cells.

    Directory of Open Access Journals (Sweden)

    Ron Balczon

    Full Text Available Pseudomonas aeruginosa uses a type III secretion system to introduce the adenylyl and guanylyl cyclase exotoxin Y (ExoY into the cytoplasm of endothelial cells. ExoY induces Tau hyperphosphorylation and insolubility, microtubule breakdown, barrier disruption and edema, although the mechanism(s responsible for microtubule breakdown remain poorly understood. Here we investigated both microtubule behavior and centrosome activity to test the hypothesis that ExoY disrupts microtubule dynamics. Fluorescence microscopy determined that infected pulmonary microvascular endothelial cells contained fewer microtubules than control cells, and further studies demonstrated that the microtubule-associated protein Tau was hyperphosphorylated following infection and dissociated from microtubules. Disassembly/reassembly studies determined that microtubule assembly was disrupted in infected cells, with no detectable effects on either microtubule disassembly or microtubule nucleation by centrosomes. This effect of ExoY on microtubules was abolished when the cAMP-dependent kinase phosphorylation site (Ser-214 on Tau was mutated to a non-phosphorylatable form. These studies identify Tau in microvascular endothelial cells as the target of ExoY in control of microtubule architecture following pulmonary infection by Pseudomonas aeruginosa and demonstrate that phosphorylation of tau following infection decreases microtubule assembly.

  14. Kinetochore-Dependent Microtubule Rescue Ensures Their Efficient and Sustained Interactions in Early Mitosis

    Science.gov (United States)

    Gandhi, Sapan R.; Gierliński, Marek; Mino, Akihisa; Tanaka, Kozo; Kitamura, Etsushi; Clayton, Lesley; Tanaka, Tomoyuki U.

    2011-01-01

    Summary How kinetochores regulate microtubule dynamics to ensure proper kinetochore-microtubule interactions is unknown. Here, we studied this during early mitosis in Saccharomyces cerevisiae. When a microtubule shrinks and its plus end reaches a kinetochore bound to its lateral surface, the microtubule end attempts to tether the kinetochore. This process often fails and, responding to this failure, microtubule rescue (conversion from shrinkage to growth) occurs, preventing kinetochore detachment from the microtubule end. This rescue is promoted by Stu2 transfer (ortholog of vertebrate XMAP215/ch-TOG) from the kinetochore to the microtubule end. Meanwhile, microtubule rescue distal to the kinetochore is also promoted by Stu2, which is transported by a kinesin-8 motor Kip3 along the microtubule from the kinetochore. Microtubule extension following rescue facilitates interaction with other widely scattered kinetochores, diminishing long delays in collecting the complete set of kinetochores by microtubules. Thus, kinetochore-dependent microtubule rescue ensures efficient and sustained kinetochore-microtubule interactions in early mitosis. PMID:22075150

  15. CYLD Regulates Noscapine Activity in Acute Lymphoblastic Leukemia via a Microtubule-Dependent Mechanism.

    Science.gov (United States)

    Yang, Yunfan; Ran, Jie; Sun, Lei; Sun, Xiaodong; Luo, Youguang; Yan, Bing; Tala; Liu, Min; Li, Dengwen; Zhang, Lei; Bao, Gang; Zhou, Jun

    2015-01-01

    Noscapine is an orally administrable drug used worldwide for cough suppression and has recently been demonstrated to disrupt microtubule dynamics and possess anticancer activity. However, the molecular mechanisms regulating noscapine activity remain poorly defined. Here we demonstrate that cylindromatosis (CYLD), a microtubule-associated tumor suppressor protein, modulates the activity of noscapine both in cell lines and in primary cells of acute lymphoblastic leukemia (ALL). Flow cytometry and immunofluorescence microscopy reveal that CYLD increases the ability of noscapine to induce mitotic arrest and apoptosis. Examination of cellular microtubules as well as in vitro assembled microtubules shows that CYLD enhances the effect of noscapine on microtubule polymerization. Microtubule cosedimentation and fluorescence titration assays further reveal that CYLD interacts with microtubule outer surface and promotes noscapine binding to microtubules. These findings thus demonstrate CYLD as a critical regulator of noscapine activity and have important implications for ALL treatment.

  16. Microtubule doublets are double-track railways for intraflagellar transport trains.

    Science.gov (United States)

    Stepanek, Ludek; Pigino, Gaia

    2016-05-06

    The cilium is a large macromolecular machine that is vital for motility, signaling, and sensing in most eukaryotic cells. Its conserved core structure, the axoneme, contains nine microtubule doublets, each comprising a full A-microtubule and an incomplete B-microtubule. However, thus far, the function of this doublet geometry has not been understood. We developed a time-resolved correlative fluorescence and three-dimensional electron microscopy approach to investigate the dynamics of intraflagellar transport (IFT) trains, which carry ciliary building blocks along microtubules during the assembly and disassembly of the cilium. Using this method, we showed that each microtubule doublet is used as a bidirectional double-track railway: Anterograde IFT trains move along B-microtubules, and retrograde trains move along A-microtubules. Thus, the microtubule doublet geometry provides direction-specific rails to coordinate bidirectional transport of ciliary components.

  17. Multiscale Polar Theory of Microtubule and Motor-Protein Assemblies

    Science.gov (United States)

    Gao, Tong; Blackwell, Robert; Glaser, Matthew A.; Betterton, M. D.; Shelley, Michael J.

    2015-01-01

    Microtubules and motor proteins are building blocks of self-organized subcellular biological structures such as the mitotic spindle and the centrosomal microtubule array. These same ingredients can form new “bioactive” liquid-crystalline fluids that are intrinsically out of equilibrium and which display complex flows and defect dynamics. It is not yet well understood how microscopic activity, which involves polarity-dependent interactions between motor proteins and microtubules, yields such larger-scale dynamical structures. In our multiscale theory, Brownian dynamics simulations of polar microtubule ensembles driven by cross-linking motors allow us to study microscopic organization and stresses. Polarity sorting and cross-link relaxation emerge as two polar-specific sources of active destabilizing stress. On larger length scales, our continuum Doi-Onsager theory captures the hydrodynamic flows generated by polarity-dependent active stresses. The results connect local polar structure to flow structures and defect dynamics. PMID:25679909

  18. The ability to induce microtubule acetylation is a general feature of formin proteins.

    Directory of Open Access Journals (Sweden)

    Susan F Thurston

    Full Text Available Cytoplasmic microtubules exist as distinct dynamic and stable populations within the cell. Stable microtubules direct and maintain cell polarity and it is thought that their stabilization is dependent on coordinative organization between the microtubule network and the actin cytoskeleton. A growing body of work suggests that some members of the formin family of actin remodeling proteins also regulate microtubule organization and stability. For example, we showed previously that expression of the novel formin INF1 is sufficient to induce microtubule stabilization and tubulin acetylation, but not tubulin detyrosination. An important issue with respect to the relationship between formins and microtubules is the determination of which formin domains mediate microtubule stabilization. INF1 has a distinct microtubule-binding domain at its C-terminus and the endogenous INF1 protein is associated with the microtubule network. Surprisingly, the INF1 microtubule-binding domain is not essential for INF1-induced microtubule acetylation. We show here that expression of the isolated FH1 + FH2 functional unit of INF1 is sufficient to induce microtubule acetylation independent of the INF1 microtubule-binding domain. It is not yet clear whether or not microtubule stabilization is a general property of all mammalian formins; therefore we expressed constitutively active derivatives of thirteen of the fifteen mammalian formin proteins in HeLa and NIH3T3 cells and measured their effects on stress fiber formation, MT organization and MT acetylation. We found that expression of the FH1 + FH2 unit of the majority of mammalian formins is sufficient to induce microtubule acetylation. Our results suggest that the regulation of microtubule acetylation is likely a general formin activity and that the FH2 should be thought of as a dual-function domain capable of regulating both actin and microtubule networks.

  19. Effect of hexylene glycol-altered microtubule distributions on cytokinesis and polar lobe formation in fertilized eggs of Ilyanassa obsoleta

    Science.gov (United States)

    Conrad, A. H.; Stephens, A. P.; Conrad, G. W.; Spooner, B. S. (Principal Investigator)

    1994-01-01

    Some effects of gravity on early morphogenesis are correlated with microtubule locations within cells. During first cleavage in Ilyanassa obsoleta embryos, a transitory polar lobe constriction forms and then relaxes, allowing the polar lobe to merge with one daughter cell. If the polar lobe is equally divided or removed, morphogenesis is severely disrupted. To examine microtuble locations during early Ilyanassa development, eggs were fixed and stained for polymerized alpha-tubulin during first cleavage. The mitotic apparatus assembles at the animal pole. The cleavage furrow forms between the asters, constricting to a stabilized intercellular bridge encircling midbody-bound microtubules, whereas the polar lobe constriction forms below and parallel to the spindle, constricting to a transitory intercellular bridge encircling no detectable microtubules. At metaphase an alpha-tubulin epitope is distributed throughout the spindle, whereas a beta-tubulin epitope is present predominantly in the asters. Incubation in hexylene glycol, a drug that increases microtubule polymerization, during mitosis causes the polar lobe constriction to tighten around polymerized alpha-tubulin and remain stably constricted. If hexylene glycol is removed, alpha-tubulin staining disappears from the polar lobe constriction, which relaxes, whereas microtubules remain in the cleavage furrow, which remains constricted. These observations suggest that asymmetric distribution of microtubules affects early Ilyanassa cleavage patterns, and that continued presence of microtubules extending through an intercellular bridge is important for stabilization of the bridge constriction prior to completion of cytokinesis. These data provide the basis for further analysis of the role of microtubules in possible microgravity disruptions of Ilyanassa development.

  20. Depolymerizing kinesins Kip3 and MCAK shape cellular microtubule architecture by differential control of catastrophe.

    Science.gov (United States)

    Gardner, Melissa K; Zanic, Marija; Gell, Christopher; Bormuth, Volker; Howard, Jonathon

    2011-11-23

    Microtubules are dynamic filaments whose ends alternate between periods of slow growth and rapid shortening as they explore intracellular space and move organelles. A key question is how regulatory proteins modulate catastrophe, the conversion from growth to shortening. To study this process, we reconstituted microtubule dynamics in the absence and presence of the kinesin-8 Kip3 and the kinesin-13 MCAK. Surprisingly, we found that, even in the absence of the kinesins, the microtubule catastrophe frequency depends on the age of the microtubule, indicating that catastrophe is a multistep process. Kip3 slowed microtubule growth in a length-dependent manner and increased the rate of aging. In contrast, MCAK eliminated the aging process. Thus, both kinesins are catastrophe factors; Kip3 mediates fine control of microtubule length by narrowing the distribution of maximum lengths prior to catastrophe, whereas MCAK promotes rapid restructuring of the microtubule cytoskeleton by making catastrophe a first-order random process.

  1. A novel role for aquaporin-5 in enhancing microtubule organization and stability.

    Directory of Open Access Journals (Sweden)

    Venkataramana K Sidhaye

    Full Text Available Aquaporin-5 (AQP5 is a water-specific channel located on the apical surface of airway epithelial cells. In addition to regulating transcellular water permeability, AQP5 can regulate paracellular permeability, though the mechanisms by which this occurs have not been determined. Microtubules also regulate paracellular permeability. Here, we report that AQP5 promotes microtubule assembly and helps maintain the assembled microtubule steady state levels with slower turnover dynamics in cells. Specifically, reduced levels of AQP5 correlated with lower levels of assembled microtubules and decreased paracellular permeability. In contrast, overexpression of AQP5 increased assembly of microtubules, with evidence of increased MT stability, and promoted the formation of long straight microtubules in the apical domain of the epithelial cells. These findings indicate that AQP5-mediated regulation of microtubule dynamics modulates airway epithelial barrier properties and epithelial function.

  2. PACSIN1, a Tau-interacting protein, regulates axonal elongation and branching by facilitating microtubule instability.

    Science.gov (United States)

    Liu, Yingying; Lv, Kaosheng; Li, Zenglong; Yu, Albert C H; Chen, Jianguo; Teng, Junlin

    2012-11-16

    Tau is a major member of the neuronal microtubule-associated proteins. It promotes tubulin assembly and stabilizes axonal microtubules. Previous studies have demonstrated that Tau forms cross-bridges between microtubules, with some particles located on cross-bridges, suggesting that some proteins interact with Tau and might be involved in regulating Tau-related microtubule dynamics. This study reports that PACSIN1 interacts with Tau in axon. PACSIN1 blockade results in impaired axonal elongation and a higher number of primary axonal branches in mouse dorsal root ganglia neurons, which is induced by increasing the binding ability of Tau to microtubules. In PACSIN1-blocked dorsal root ganglia neurons, a greater amount of Tau is inclined to accumulate in the central domain of growth cones, and it promotes the stability of the microtubule network. Taken together, these results suggest that PACSIN1 is an important Tau binding partner in regulating microtubule dynamics and forming axonal plasticity.

  3. Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER)

    Data.gov (United States)

    National Aeronautics and Space Administration — This dataset represents multiple products archived at the Land Processes DAAC for ASTER (Advanced Spaceborne Thermal Emission and Reflection Radiometer) aboard the...

  4. Microtubule's conformational cap

    DEFF Research Database (Denmark)

    Chretien, D.; Janosi, I.; Taveau, J.C.

    1999-01-01

    The molecular mechanisms that allow elongation of the unstable microtubule lattice remain unclear. It is usually thought that the GDP-liganded tubulin lattice is capped by a small layer of GTP- or GDP-P(i)-liganded molecules, the so called "GTP-cap". Here, we point-out that the elastic properties...

  5. The Microtubule Regulatory Protein Stathmin Is Required to Maintain the Integrity of Axonal Microtubules in Drosophila.

    Directory of Open Access Journals (Sweden)

    Jason E Duncan

    Full Text Available Axonal transport, a form of long-distance, bi-directional intracellular transport that occurs between the cell body and synaptic terminal, is critical in maintaining the function and viability of neurons. We have identified a requirement for the stathmin (stai gene in the maintenance of axonal microtubules and regulation of axonal transport in Drosophila. The stai gene encodes a cytosolic phosphoprotein that regulates microtubule dynamics by partitioning tubulin dimers between pools of soluble tubulin and polymerized microtubules, and by directly binding to microtubules and promoting depolymerization. Analysis of stai function in Drosophila, which has a single stai gene, circumvents potential complications with studies performed in vertebrate systems in which mutant phenotypes may be compensated by genetic redundancy of other members of the stai gene family. This has allowed us to identify an essential function for stai in the maintenance of the integrity of axonal microtubules. In addition to the severe disruption in the abundance and architecture of microtubules in the axons of stai mutant Drosophila, we also observe additional neurological phenotypes associated with loss of stai function including a posterior paralysis and tail-flip phenotype in third instar larvae, aberrant accumulation of transported membranous organelles in stai deficient axons, a progressive bang-sensitive response to mechanical stimulation reminiscent of the class of Drosophila mutants used to model human epileptic seizures, and a reduced adult lifespan. Reductions in the levels of Kinesin-1, the primary anterograde motor in axonal transport, enhance these phenotypes. Collectively, our results indicate that stai has an important role in neuronal function, likely through the maintenance of microtubule integrity in the axons of nerves of the peripheral nervous system necessary to support and sustain long-distance axonal transport.

  6. Acentrosomal Microtubule Assembly in Mitosis: The Where, When, and How.

    Science.gov (United States)

    Meunier, Sylvain; Vernos, Isabelle

    2016-02-01

    In mitosis the cell assembles the bipolar spindle, a microtubule (MT)-based apparatus that segregates the duplicated chromosomes into two daughter cells. Most animal cells enter mitosis with duplicated centrosomes that provide an active source of dynamic MTs. However, it is now established that spindle assembly relies on the nucleation of acentrosomal MTs occurring around the chromosomes after nuclear envelope breakdown, and on pre-existing microtubules. Where chromosome-dependent MT nucleation occurs, when MT amplification takes place and how the two pathways function are still key questions that generate some controversies. We reconcile the data and present an integrated model accounting for acentrosomal microtubule assembly in the dividing cell.

  7. Studying neuronal microtubule organization and microtubule-associated proteins using single molecule localization microscopy

    NARCIS (Netherlands)

    Chazeau, Anaël; Katrukha, Eugene A; Hoogenraad, Casper C; Kapitein, Lukas C

    2016-01-01

    The formation and maintenance of highly polarized neurons critically depends on the proper organization of the microtubule (MT) cytoskeleton. In axons, MTs are uniformly oriented with their plus-end pointing outward whereas in mature dendrites MTs have mixed orientations. MT organization and dynamic

  8. [Disruption of organization of mitotic microtubules in root meristem cells of Allium cepa induced by chloral hydrate].

    Science.gov (United States)

    Smirnova, E A; Svetlitskaia, O M; Chentsov, Iu S

    2002-01-01

    Data are presented on the effect of chlorahydrate on microtubule organization in the root meristem of Allium cepa. Our studies show that an incomplete preprophase band commonly appears during G2-prophase transition, yet the major effect is the lack of perinuclear microtubules, leading to inhibition of the prophase spindle formation and transition to C-mitosis. Upon chloralhydrate treatment of metaphase cells, we found cells with chromosomes regularly aligned within the metaphase plate and differently disorganized mitotic spindles. Concurrently, C-metaphase cells with remnants of kinetochore fibers were present. In addition, normal bipolar and abnormal irregular types of chromosome segregation were detected, this representing multipolar and diffuse anaphases. The major difference between them is the presence of polar microtubules during multipolar anaphase, and their lacking during diffuse anaphase. Alternatively, microtubule clusters between segregated groups of chromosomes are typical for cells with diffuse anaphase. During bipolar anaphase, excessive aster-like microtubules emanate from the spindle poles, and in telophase accessory phragmoplasts are observed at the cell periphery. The formation of incomplete phragmoplasts was observed after normal bipolar and abnormal chromosome segregation. We conclude that chloralhydrate may affect the nuclear surface capability to initiate the growth of perinuclear microtubules, thus blocking the prophase spindle formation. It also disturbs the spatial interaction between microtubules, which is crucial for the formation and functioning of various microtubular systems (preprophase band, spindle and phragmoplast).

  9. The microtubule plus-end-tracking protein CLIP-170 associates with the spermatid manchette and is essential for spermatogenesis.

    NARCIS (Netherlands)

    A.S. Akhmanova (Anna); A.L. Mausset-Bonnefont (Anne-Laure); W.A. van Cappellen (Gert); N. Keijzer (Nanda); C.C. Hoogenraad (Casper); T. Stepanova (Tatiana); K. Drabek (Ksenija); J. van der Wees (Jacqueline); M. Mommaas (Mieke); J. Onderwater (Jos); H. van der Meulen (Hans); M.E. Tanenbaum (Marvin); R.H. Medema (Rene); J.W. Hoogerbrugge (Jos); J.T.M. Vreeburg (Jan); E.J. Uringa; J.A. Grootegoed (Anton); F.G. Grosveld (Frank); N.J. Galjart (Niels)

    2005-01-01

    textabstractCLIP-170 is a microtubule "plus-end-tracking protein" implicated in the control of microtubule dynamics, dynactin localization, and the linking of endosomes to microtubules. To investigate the function of mouse CLIP-170, we generated CLIP-170 knockout and GFP-CLIP-170 knock-in alleles. R

  10. The microtubule plus-end-tracking protein CLIP-170 associates with the spermatid manchette and is essential for spermatogenesis

    NARCIS (Netherlands)

    Akhmanova, A.S.; Mausset-Bonnefont, A.-L.; Cappellen, W. van; Keijzer, N.; Hoogenraad, C.C.; Stepanova, T.; Drabek, K.; Wees, J. van der; Mommaas, M.; Onderwater, J.; Meulen, H. van der; Tanenbaum, M.E.; Medema, R.H.; Hoogerbrugge, J.; Vreeburg, J.; Uringa, E.-J.; Grootegoed, J.A.; Grosveld, F.; Galjart, N.

    2005-01-01

    CLIP-170 is a microtubule "plus-end-tracking protein" implicated in the control of microtubule dynamics, dynactin localization, and the linking of endosomes to microtubules. To investigate the function of mouse CLIP-170, we generated CLIP-170 knockout and GFP-CLIP-170 knock-in alleles. Residual CLIP

  11. Electrostatically biased binding of kinesin to microtubules.

    Directory of Open Access Journals (Sweden)

    Barry J Grant

    2011-11-01

    Full Text Available The minimum motor domain of kinesin-1 is a single head. Recent evidence suggests that such minimal motor domains generate force by a biased binding mechanism, in which they preferentially select binding sites on the microtubule that lie ahead in the progress direction of the motor. A specific molecular mechanism for biased binding has, however, so far been lacking. Here we use atomistic Brownian dynamics simulations combined with experimental mutagenesis to show that incoming kinesin heads undergo electrostatically guided diffusion-to-capture by microtubules, and that this produces directionally biased binding. Kinesin-1 heads are initially rotated by the electrostatic field so that their tubulin-binding sites face inwards, and then steered towards a plus-endwards binding site. In tethered kinesin dimers, this bias is amplified. A 3-residue sequence (RAK in kinesin helix alpha-6 is predicted to be important for electrostatic guidance. Real-world mutagenesis of this sequence powerfully influences kinesin-driven microtubule sliding, with one mutant producing a 5-fold acceleration over wild type. We conclude that electrostatic interactions play an important role in the kinesin stepping mechanism, by biasing the diffusional association of kinesin with microtubules.

  12. Globally visualizing the microtubule-dependent transport behaviors of influenza virus in live cells.

    Science.gov (United States)

    Liu, Shu-Lin; Zhang, Li-Juan; Wang, Zhi-Gang; Zhang, Zhi-Ling; Wu, Qiu-Mei; Sun, En-Ze; Shi, Yun-Bo; Pang, Dai-Wen

    2014-04-15

    Understanding the microtubule-dependent behaviors of viruses in live cells is very meaningful for revealing the mechanisms of virus infection and endocytosis. Herein, we used a quantum dots-based single-particle tracking technique to dynamically and globally visualize the microtubule-dependent transport behaviors of influenza virus in live cells. We found that the intersection configuration of microtubules can interfere with the transport behaviors of the virus in live cells, which lead to the changing and long-time pausing of the transport behavior of viruses. Our results revealed that most of the viruses moved along straight microtubules rapidly and unidirectionally from the cell periphery to the microtubule organizing center (MTOC) near the bottom of the cell, and the viruses were confined in the grid of microtubules near the top of the cell and at the MTOC near the bottom of the cell. These results provided deep insights into the influence of entire microtubule geometry on the virus infection.

  13. Tubulin bond energies and microtubule biomechanics determined from nanoindentation in silico

    CERN Document Server

    Kononova, Olga; Theisen, Kelly E; Marx, Kenneth A; Dima, Ruxandra I; Ataullakhanov, Fazly I; Grishchuk, Ekaterina L; Barsegov, Valeri

    2015-01-01

    Microtubules, the primary components of the chromosome segregation machinery, are stabilized by longitudinal and lateral non-covalent bonds between the tubulin subunits. However, the thermodynamics of these bonds and the microtubule physico-chemical properties are poorly understood. Here, we explore the biomechanics of microtubule polymers using multiscale computational modeling and nanoindentations in silico of a contiguous microtubule fragment. A close match between the simulated and experimental force-deformation spectra enabled us to correlate the microtubule biomechanics with dynamic structural transitions at the nanoscale. Our mechanical testing revealed that the compressed MT behaves as a system of rigid elements interconnected through a network of lateral and longitudinal elastic bonds. The initial regime of continuous elastic deformation of the microtubule is followed by the transition regime, during which the microtubule lattice undergoes discrete structural changes, which include first the reversib...

  14. A Chimeric Cetuximab-Functionalized Corona as a Potent Delivery System for Microtubule-Destabilizing Nanocomplexes to Hepatocellular Carcinoma Cells: A Focus on EGFR and Tubulin Intracellular Dynamics.

    Science.gov (United States)

    Poojari, Radhika; Kini, Sudarshan; Srivastava, Rohit; Panda, Dulal

    2015-11-01

    In this study, we have developed microtubule destabilizing agents combretastatin A4 (CA4) or 2-methoxyestradiol (2ME) encapsulated poly(d,l-lactide-co-glycolide)-b-poly(ethylene glycol) (PLGA-b-PEG) nanocomplexes for targeted delivery to human hepatocellular carcinoma (HCC) cells. An epidermal growth factor receptor (EGFR) is known to be overexpressed in HCC cells. Therefore, the targeting moiety cetuximab (Cet), an anti-EGFR chimeric monoclonal antibody, is functionalized on the surface of these diblock copolymeric coronas. Cetuximab is associated with the extracellular domain of the EGFR; therefore, the uptake of the cetuximab conjugated nanocomplexes occurred efficiently in EGFR overexpressing HCC cells indicating potent internalization of the complex. The cetuximab targeted-PLGA-b-PEG nanocomplexes encapsulating CA4 or 2ME strongly inhibited phospho-EGFR expression, depolymerized microtubules, produced spindle abnormalities, stalled mitosis, and induced apoptosis in Huh7 cells compared to the free drugs, CA4 or 2ME. Further, the combinatorial strategy of targeted nanocomplexes, Cet-PLGA-b-PEG-CA4 NP and Cet-PLGA-b-PEG-2ME NP, significantly reduced the migration of Huh7 cells, and markedly enhanced the anticancer effects of the microtubule-targeted drugs in Huh7 cells compared to the free drugs, CA4 or 2ME. The results indicated that EGFR receptor-mediated internalization via cetuximab facilitated enhanced uptake of the nanocomplexes leading to potent anticancer efficacy in Huh7 cells. Cetuximab-functionalized PLGA-b-PEG nanocomplexes possess a strong potential for the targeted delivery of CA4 or 2ME in EGFR overexpressed HCC cells, and the strategy may be useful for selectively targeting microtubules in these cells.

  15. A thermodynamic model of microtubule assembly and disassembly.

    Directory of Open Access Journals (Sweden)

    Bernard M A G Piette

    Full Text Available Microtubules are self-assembling polymers whose dynamics are essential for the normal function of cellular processes including chromosome separation and cytokinesis. Therefore understanding what factors effect microtubule growth is fundamental to our understanding of the control of microtubule based processes. An important factor that determines the status of a microtubule, whether it is growing or shrinking, is the length of the GTP tubulin microtubule cap. Here, we derive a Monte Carlo model of the assembly and disassembly of microtubules. We use thermodynamic laws to reduce the number of parameters of our model and, in particular, we take into account the contribution of water to the entropy of the system. We fit all parameters of the model from published experimental data using the GTP tubulin dimer attachment rate and the lateral and longitudinal binding energies of GTP and GDP tubulin dimers at both ends. Also we calculate and incorporate the GTP hydrolysis rate. We have applied our model and can mimic published experimental data, which formerly suggested a single layer GTP tubulin dimer microtubule cap, to show that these data demonstrate that the GTP cap can fluctuate and can be several microns long.

  16. A thermodynamic model of microtubule assembly and disassembly.

    Science.gov (United States)

    Piette, Bernard M A G; Liu, Junli; Peeters, Kasper; Smertenko, Andrei; Hawkins, Timothy; Deeks, Michael; Quinlan, Roy; Zakrzewski, Wojciech J; Hussey, Patrick J

    2009-08-11

    Microtubules are self-assembling polymers whose dynamics are essential for the normal function of cellular processes including chromosome separation and cytokinesis. Therefore understanding what factors effect microtubule growth is fundamental to our understanding of the control of microtubule based processes. An important factor that determines the status of a microtubule, whether it is growing or shrinking, is the length of the GTP tubulin microtubule cap. Here, we derive a Monte Carlo model of the assembly and disassembly of microtubules. We use thermodynamic laws to reduce the number of parameters of our model and, in particular, we take into account the contribution of water to the entropy of the system. We fit all parameters of the model from published experimental data using the GTP tubulin dimer attachment rate and the lateral and longitudinal binding energies of GTP and GDP tubulin dimers at both ends. Also we calculate and incorporate the GTP hydrolysis rate. We have applied our model and can mimic published experimental data, which formerly suggested a single layer GTP tubulin dimer microtubule cap, to show that these data demonstrate that the GTP cap can fluctuate and can be several microns long.

  17. Analysis of Soybean Microtubule Persistence Length; New Evidence on the Correlation between Structural Composition and Mechanical Properties

    Science.gov (United States)

    Shojania Feizabadi, Mitra; Winton, Carly; Barrientos, Jimmy

    2012-02-01

    Recent studies on microtubules composed of different β tubulin isotypes indicate their different functionality in terms of their dynamical behavior or the mechanism of their interaction with chemotherapeutic drugs. Along these lines, the result of our recent study measuring the rigidity of neural and non-neural samples of microtubules with different β tubulin isotype compositions suggests that the distinguished mechanical properties of microtubules, such as rigidity, may also be associated with the different distribution of their β tubulin isotypes. In our current study, we have reported the persistence length of a single soybean microtubule. This plant microtubule has a structural composition different from that of mammalian microtubules. Under the same experimental methods of measurement, the soybean microtubules showed a different persistence length as compared to the value of the persistence length that we estimated in the study of both single Bovine Brain and MCF7 microtubules.

  18. Insights from an erroneous kinetochore-microtubule attachment state.

    Science.gov (United States)

    Cane, Stuart; McGilvray, Philip T; Maresca, Thomas J

    2013-01-01

    Faithful distribution of the genome requires that sister kinetochores, which assemble on each chromatid during cell division, interact with dynamic microtubules from opposite spindle poles in a configuration called chromosome biorientation. Biorientation produces tension that increases the affinity of kinetochores for microtubules via ill-defined mechanisms. Non-bioriented kinetochore-microtubule (kt-MT) interactions are prevalent but short-lived due to an error correction pathway that reduces the affinity of kinetochores for microtubules. Interestingly, incorrect kt-MT interactions can be stabilized by experimentally applying force to misoriented chromosomes. Here, a live-cell force assay is utilized to characterize the molecular composition of a common type of improper kt-MT attachment. Our force-related studies are also discussed in the context of current models for tension-dependent stabilization of kt-MT interactions.

  19. Mechanical Models of Microtubule Bundle Collapse in Alzheimer's Disease

    Science.gov (United States)

    Sendek, Austin; Singh, Rajiv; Cox, Daniel

    2013-03-01

    Amyloid-beta aggregates initiate Alzheimer's disease, and downstream trigger degradation of tau proteins that act as microtubule bundle stabilizers and mechanical spacers. Currently it is unclear which of tau cutting by proteases, tau phosphorylation, or tau aggregation are responsible for cytoskeleton degradation., We construct a percolation simulation of the microtubule bundle using a molecular spring model for the taus and including depletion force attraction between microtubules and membrane/actin cytoskeletal surface tension. The simulation uses a fictive molecular dynamics to model the motion of the individual microtubules within the bundle as a result of random tau removal, and calculates the elastic modulus of the bundle as the tau concentration falls. We link the tau removal steps to kinetic tau steps in various models of tau degradation. Supported by US NSF Grant DMR 1207624

  20. Monitoring volcanic threats using ASTER satellite data

    Science.gov (United States)

    Duda, K.A.; Wessels, R.; Ramsey, M.; Dehn, J.

    2008-01-01

    This document summarizes ongoing activities associated with a research project funded by the National Aeronautics and Space Administration (NASA) focusing on volcanic change detection through the use of satellite imagery. This work includes systems development as well as improvements in data analysis methods. Participating organizations include the NASA Land Processes Distributed Active Archive Center (LP DAAC) at the U.S. Geological Survey (USGS) Center for Earth Resources Observation and Science (EROS), the Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) Science Team, the Alaska Volcano Observatory (AVO) at the USGS Alaska Science Center, the Jet Propulsion Laboratory/California Institute of Technology (JPL/CalTech), the University of Pittsburgh, and the University of Alaska Fairbanks. ?? 2007 IEEE.

  1. Mechanical modulation of cardiac microtubules.

    Science.gov (United States)

    White, Ed

    2011-07-01

    Microtubules are a major component of the cardiac myocyte cytoskeleton. Interventions that alter it may influence cardiac mechanical and electrical activity by disrupting the trafficking of proteins to and from the surface membrane by molecular motors such as dynein, which use microtubules as tracks to step along. Free tubulin dimers may transfer GTP to the α-subunits of G-proteins, thus an increase in free tubulin could increase the activity of G-proteins; evidence for and against such a role exists. There is more general agreement that microtubules act as compression-resisting structures within myocytes, influencing visco-elasticity of myocytes and increasing resistance to shortening when proliferated and resisting deformation from longitudinal shear stress. In response to pressure overload, there can be post-translational modifications resulting in more stable microtubules and an increase in microtubule density. This is accompanied by contractile dysfunction of myocytes which can be reversed by microtubule disruption. There are reports of mechanically induced changes in electrical activity that are dependent upon microtubules, but at present, a consensus is lacking on whether disruption or proliferation would be beneficial in the prevention of arrhythmias. Microtubules certainly play a role in the response of cardiac myocytes to mechanical stimulation, the exact nature and significance of this role is still to be fully determined.

  2. Efficiency of organelle capture by microtubules as a function of centrosome nucleation capacity: general theory and the special case of polyspermia.

    Directory of Open Access Journals (Sweden)

    Ivan V Maly

    Full Text Available Transport of organelles along microtubules is essential for the cell metabolism and morphogenesis. The presented analysis derives the probability that an organelle of a given size comes in contact with the microtubule aster. The question is asked how this measure of functionality of the microtubule aster is controlled by the centrosome. A quantitative model is developed to address this question. It is shown that for the given set of cellular parameters, such as size and total tubulin content, a centrosome nucleation capacity exists that maximizes the probability of the organelle capture. The developed general model is then applied to the capture of the female pronucleus by microtubules assembled on the sperm centrosome, following physiologically polyspermic fertilization. This application highlights an unintuitive reflection of nonlinearity of the nucleated polymerization of the cellular pool of tubulin. The prediction that the sperm centrosome should lower its nucleation capacity in the face of the competition from the other sperm is a stark illustration of the new optimality principle. Overall, the model calls attention to the capabilities of the centrosomal pathway of regulation of the transport-related functionality of the microtubule cytoskeleton. It establishes a quantitative and conceptual framework that can guide experiment design and interpretation.

  3. Estimating Coastal Lagoon Tidal Flooding and Repletion with Multidate ASTER Thermal Imagery

    Directory of Open Access Journals (Sweden)

    Thomas R. Allen

    2012-10-01

    Full Text Available Coastal lagoons mix inflowing freshwater and tidal marine waters in complex spatial patterns. This project sought to detect and measure temperature and spatial variability of flood tides for a constricted coastal lagoon using multitemporal remote sensing. Advanced Spaceborne Thermal Emission Radiometer (ASTER thermal infrared data provided estimates of surface temperature for delineation of repletion zones in portions of Chincoteague Bay, Virginia. ASTER high spatial resolution sea-surface temperature imagery in conjunction with in situ observations and tidal predictions helped determine the optimal seasonal data for analyses. The selected time series ASTER satellite data sets were analyzed at different tidal phases and seasons in 2004–2006. Skin surface temperatures of ocean and estuarine waters were differentiated by flood tidal penetration and ebb flows. Spatially variable tidal flood penetration was evaluated using discrete seed-pixel area analysis and time series Principal Components Analysis. Results from these techniques provide spatial extent and variability dynamics of tidal repletion, flushing, and mixing, important factors in eutrophication assessment, water quality and resource monitoring, and application of hydrodynamic modeling for coastal estuary science and management.

  4. ASTER Level 1B Registered Radiance at the Sensor

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — The Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) is an advanced multispectral imager that was launched on board NASA's Terra spacecraft in...

  5. ASTER Global Emissivity Dataset 100-meter V003 - AG100

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — The Advanced Spaceborne Thermal Emission and Reflection radiometer Global Emissivity Database (ASTER GED) was developed by the National Aeronautics and Space...

  6. Stabilizing versus destabilizing the microtubules: a double-edge sword for an effective cancer treatment option?

    Science.gov (United States)

    Fanale, Daniele; Bronte, Giuseppe; Passiglia, Francesco; Calò, Valentina; Castiglia, Marta; Di Piazza, Florinda; Barraco, Nadia; Cangemi, Antonina; Catarella, Maria Teresa; Insalaco, Lavinia; Listì, Angela; Maragliano, Rossella; Massihnia, Daniela; Perez, Alessandro; Toia, Francesca; Cicero, Giuseppe; Bazan, Viviana

    2015-01-01

    Microtubules are dynamic and structural cellular components involved in several cell functions, including cell shape, motility, and intracellular trafficking. In proliferating cells, they are essential components in the division process through the formation of the mitotic spindle. As a result of these functions, tubulin and microtubules are targets for anticancer agents. Microtubule-targeting agents can be divided into two groups: microtubule-stabilizing, and microtubule-destabilizing agents. The former bind to the tubulin polymer and stabilize microtubules, while the latter bind to the tubulin dimers and destabilize microtubules. Alteration of tubulin-microtubule equilibrium determines the disruption of the mitotic spindle, halting the cell cycle at the metaphase-anaphase transition and, eventually, resulting in cell death. Clinical application of earlier microtubule inhibitors, however, unfortunately showed several limits, such as neurological and bone marrow toxicity and the emergence of drug-resistant tumor cells. Here we review several natural and synthetic microtubule-targeting agents, which showed antitumor activity and increased efficacy in comparison to traditional drugs in various preclinical and clinical studies. Cryptophycins, combretastatins, ombrabulin, soblidotin, D-24851, epothilones and discodermolide were used in clinical trials. Some of them showed antiangiogenic and antivascular activity and others showed the ability to overcome multidrug resistance, supporting their possible use in chemotherapy.

  7. Stabilizing versus Destabilizing the Microtubules: A Double-Edge Sword for an Effective Cancer Treatment Option?

    Science.gov (United States)

    Fanale, Daniele; Bronte, Giuseppe; Passiglia, Francesco; Calò, Valentina; Castiglia, Marta; Di Piazza, Florinda; Barraco, Nadia; Cangemi, Antonina; Catarella, Maria Teresa; Insalaco, Lavinia; Listì, Angela; Maragliano, Rossella; Massihnia, Daniela; Perez, Alessandro; Toia, Francesca; Cicero, Giuseppe; Bazan, Viviana

    2015-01-01

    Microtubules are dynamic and structural cellular components involved in several cell functions, including cell shape, motility, and intracellular trafficking. In proliferating cells, they are essential components in the division process through the formation of the mitotic spindle. As a result of these functions, tubulin and microtubules are targets for anticancer agents. Microtubule-targeting agents can be divided into two groups: microtubule-stabilizing, and microtubule-destabilizing agents. The former bind to the tubulin polymer and stabilize microtubules, while the latter bind to the tubulin dimers and destabilize microtubules. Alteration of tubulin-microtubule equilibrium determines the disruption of the mitotic spindle, halting the cell cycle at the metaphase-anaphase transition and, eventually, resulting in cell death. Clinical application of earlier microtubule inhibitors, however, unfortunately showed several limits, such as neurological and bone marrow toxicity and the emergence of drug-resistant tumor cells. Here we review several natural and synthetic microtubule-targeting agents, which showed antitumor activity and increased efficacy in comparison to traditional drugs in various preclinical and clinical studies. Cryptophycins, combretastatins, ombrabulin, soblidotin, D-24851, epothilones and discodermolide were used in clinical trials. Some of them showed antiangiogenic and antivascular activity and others showed the ability to overcome multidrug resistance, supporting their possible use in chemotherapy. PMID:26484003

  8. Stabilizing versus Destabilizing the Microtubules: A Double-Edge Sword for an Effective Cancer Treatment Option?

    Directory of Open Access Journals (Sweden)

    Daniele Fanale

    2015-01-01

    Full Text Available Microtubules are dynamic and structural cellular components involved in several cell functions, including cell shape, motility, and intracellular trafficking. In proliferating cells, they are essential components in the division process through the formation of the mitotic spindle. As a result of these functions, tubulin and microtubules are targets for anticancer agents. Microtubule-targeting agents can be divided into two groups: microtubule-stabilizing, and microtubule-destabilizing agents. The former bind to the tubulin polymer and stabilize microtubules, while the latter bind to the tubulin dimers and destabilize microtubules. Alteration of tubulin-microtubule equilibrium determines the disruption of the mitotic spindle, halting the cell cycle at the metaphase-anaphase transition and, eventually, resulting in cell death. Clinical application of earlier microtubule inhibitors, however, unfortunately showed several limits, such as neurological and bone marrow toxicity and the emergence of drug-resistant tumor cells. Here we review several natural and synthetic microtubule-targeting agents, which showed antitumor activity and increased efficacy in comparison to traditional drugs in various preclinical and clinical studies. Cryptophycins, combretastatins, ombrabulin, soblidotin, D-24851, epothilones and discodermolide were used in clinical trials. Some of them showed antiangiogenic and antivascular activity and others showed the ability to overcome multidrug resistance, supporting their possible use in chemotherapy.

  9. Fifteen Years of ASTER Data on NASA's Terra Platform

    Science.gov (United States)

    Abrams, M.; Tsu, H.

    2014-12-01

    The Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) is one of five instruments operating on NASA's Terra platform. Launched in 1999, ASTER has been acquiring data for 15 years. ASTER is a joint project between Japan's Ministry of Economy, Trade and Industry; and US NASA. Data processing and distribution are done by both organizations; a joint science team helps to define mission priorities. ASTER acquires ~550 images per day, with a 60 km swath width. A daytime acquisition is three visible bands and a backward-looking stereo band with 15 m resolution, six SWIR bands with 30 m resolution, and 5 TIR bands with 90 m resolution. Nighttime TIR-only data are routinely collected. The stereo capability has allowed the ASTER project to produce a global Digital Elevation Model (GDEM) data set, covering the earth's land surfaces from 83 degrees north to 83 degrees south, with 30 m data postings. This is the only (near-) global DEM available to all users at no charge; to date, over 28 million 1-by-1 degree DEM tiles have been distributed. As a general-purpose imaging instrument, ASTER-acquired data are used in numerous scientific disciplines, including: land use/land cover, urban monitoring, urban heat island studies, wetlands studies, agriculture monitoring, forestry, etc. Of particular emphasis has been the acquisition and analysis of data for natural hazard and disaster applications. We have been systematically acquiring images for 15,000 valley glaciers through the USGS Global Land Ice Monitoring from Space Project. The recently published Randolph Glacier Inventory, and the GLIMS book, both relied heavily on ASTER data as the basis for glaciological and climatological studies. The ASTER Volcano Archive is a unique on-line archive of thousands of daytime and nighttime ASTER images of ~1500 active glaciers, along with a growing archive of Landsat images. ASTER was scheduled to target active volcanoes at least 4 times per year, and more frequently for

  10. Xenopus oocyte wound healing as a model system for analysis of microtubule-actin interactions.

    Science.gov (United States)

    Zhang, Tong; Mandato, Craig A

    2007-01-01

    Microtubule-actin interactions are fundamental to many cellular processes such as cytokinesis and cellular locomotion. Investigating the mechanism of microtubule-actin interactions is the key to understand the cellular morphogenesis and related pathological processes. The abundance and highly dynamic nature of microtubules and F-actin raise a serious challenge when trying to distinguish between the real and fortuitous interactions within a cell. Xenopus oocyte wound model represents an ideal system to study microtubule-actin interactions as well as microtubule-dependent control of the actin polymerization. Here, we describe a series of cytoskeleton specific treatments in Xenopus oocyte wound healing experiments and use confocal fluorescence microscopy to analyze fixed oocytes to examine microtubule-actin interactions.

  11. Polyamine sharing between tubulin dimers favours microtubule nucleation and elongation via facilitated diffusion.

    Directory of Open Access Journals (Sweden)

    Alain Mechulam

    2009-01-01

    Full Text Available We suggest for the first time that the action of multivalent cations on microtubule dynamics can result from facilitated diffusion of GTP-tubulin to the microtubule ends. Facilitated diffusion can promote microtubule assembly, because, upon encountering a growing nucleus or the microtubule wall, random GTP-tubulin sliding on their surfaces will increase the probability of association to the target sites (nucleation sites or MT ends. This is an original explanation for understanding the apparent discrepancy between the high rate of microtubule elongation and the low rate of tubulin association at the microtubule ends in the viscous cytoplasm. The mechanism of facilitated diffusion requires an attraction force between two tubulins, which can result from the sharing of multivalent counterions. Natural polyamines (putrescine, spermidine, and spermine are present in all living cells and are potent agents to trigger tubulin self-attraction. By using an analytical model, we analyze the implication of facilitated diffusion mediated by polyamines on nucleation and elongation of microtubules. In vitro experiments using pure tubulin indicate that the promotion of microtubule assembly by polyamines is typical of facilitated diffusion. The results presented here show that polyamines can be of particular importance for the regulation of the microtubule network in vivo and provide the basis for further investigations into the effects of facilitated diffusion on cytoskeleton dynamics.

  12. Aster Global dem Version 3, and New Aster Water Body Dataset

    Science.gov (United States)

    Abrams, M.

    2016-06-01

    In 2016, the US/Japan ASTER (Advanced Spaceborne Thermal Emission and Reflection Radiometer) project released Version 3 of the Global DEM (GDEM). This 30 m DEM covers the earth's surface from 82N to 82S, and improves on two earlier versions by correcting some artefacts and filling in areas of missing DEMs by the acquisition of additional data. The GDEM was produced by stereocorrelation of 2 million ASTER scenes and operation on a pixel-by-pixel basis: cloud screening; stacking data from overlapping scenes; removing outlier values, and averaging elevation values. As previously, the GDEM is packaged in ~ 23,000 1 x 1 degree tiles. Each tile has a DEM file, and a NUM file reporting the number of scenes used for each pixel, and identifying the source for fill-in data (where persistent clouds prevented computation of an elevation value). An additional data set was concurrently produced and released: the ASTER Water Body Dataset (AWBD). This is a 30 m raster product, which encodes every pixel as either lake, river, or ocean; thus providing a global inland and shore-line water body mask. Water was identified through spectral analysis algorithms and manual editing. This product was evaluated against the Shuttle Water Body Dataset (SWBD), and the Landsat-based Global Inland Water (GIW) product. The SWBD only covers the earth between about 60 degrees north and south, so it is not a global product. The GIW only delineates inland water bodies, and does not deal with ocean coastlines. All products are at 30 m postings.

  13. Termination of Protofilament Elongation by Eribulin Induces Lattice Defects that Promote Microtubule Catastrophes

    NARCIS (Netherlands)

    Doodhi, Harinath; Prota, Andrea E; Rodríguez-García, Ruddi; Xiao, Hui; Custar, Daniel W; Bargsten, Katja; Katrukha, Eugene A; Hilbert, Manuel; Hua, Shasha; Jiang, Kai; Grigoriev, Ilya; Yang, Chia-Ping H; Cox, David; Horwitz, Susan Band; Kapitein, Lukas C; Akhmanova, Anna; Steinmetz, Michel O

    2016-01-01

    Microtubules are dynamic polymers built of tubulin dimers that attach in a head-to-tail fashion to form protofilaments, which further associate laterally to form a tube. Asynchronous elongation of individual protofilaments can potentially lead to an altered microtubule-end structure that promotes su

  14. ASTER satellite observations for international disaster management

    Science.gov (United States)

    Duda, K.A.; Abrams, M.

    2012-01-01

    When lives are threatened or lost due to catastrophic disasters, and when massive financial impacts are experienced, international emergency response teams rapidly mobilize to provide urgently required support. Satellite observations of affected areas often provide essential insight into the magnitude and details of the impacts. The large cost and high complexity of developing and operating satellite flight and ground systems encourages international collaboration in acquiring imagery for such significant global events in order to speed delivery of critical information to help those affected, and optimize spectral, spatial, and temporal coverage of the areas of interest. The International Charter-Space and Major Disasters was established to enable such collaboration in sensor tasking during times of crisis and is often activated in response to calls for assistance from authorized users. Insight is provided from a U.S. perspective into sensor support for Charter activations and other disaster events through a description of the Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER), which has been used to support emergency situations for over a decade through its expedited tasking and near real-time data delivery capabilities. Examples of successes achieved and challenges encountered in international collaboration to develop related systems and fulfill tasking requests suggest operational considerations for new missions as well as areas for future enhancements.

  15. The octarepeat region of hamster PrP (PrP51-91) enhances the formation of microtubule and antagonize Cu~(2+)-induced microtubule-disrupting activity

    Institute of Scientific and Technical Information of China (English)

    Xiaoli Li; Chenfang Dong; Song Shi; Guirong Wang; Yuan Li; Xin Wang; Qi Shi; Chan Tian; Ruimin Zhou; Chen Gao; Xiaoping Dong

    2009-01-01

    Prion protein (PrP) is considered to associate with microtubule and its major component, tubulin. In the present study, octarepeat region of PrP (PrP51-91) was expressed in prokaryotic-expressing system. Using GST pull-down assay and co-immunoprecipitation, the mol-ecular interaction between PrP51-91 and tubulin was observed. Our data also demonstrated that PrP51-91 could efficiently stimulate microtubule assembly in vitro, indicating a potential effect of PrP on microtu-bule dynamics. Moreover, PrP51-91 was confirmed to be able to antagonize Cu~(2+)-induced microtubule-disrupt-ing activity in vivo, partially protecting against Cu~(2+) intoxication to culture cells and stabilize cellular micro-tubule structure. The association of the octarepeat region of PrP with tubulin may further provide insight into the biological function of PrP in the neurons.

  16. Crowding of molecular motors determines microtubule depolymerization

    CERN Document Server

    Reese, Louis; Frey, Erwin

    2011-01-01

    Assembly and disassembly dynamics of microtubules (MTs) is tightly controlled by MT associated proteins. Here, we investigate how plus-end-directed depolymerases of the kinesin-8 family regulate MT depolymerization dynamics. Employing an individual-based model, we reproduce experimental findings. Moreover, crowding is identified as the key regulatory mechanism of depolymerization dynamics. Our analysis gives two qualitatively distinct regimes. For motor densities above a particular threshold, a macroscopic traffic jam emerges at the plus-end and the MT dynamics become independent of the motor concentration. Below this threshold, microscopic traffic jams at the tip arise which cancel out the effect of the depolymerization kinetics such that the depolymerization speed is solely determined by the motor density. Because this density changes over the MT length, length-dependent regulation is possible. Remarkably, motor cooperativity does not affect the depolymerization speed but only the end-residence time of depo...

  17. ASTER Imaging and Analysis of Glacier Hazards

    Science.gov (United States)

    Kargel, Jeffrey; Furfaro, Roberto; Kaser, Georg; Leonard, Gregory; Fink, Wolfgang; Huggel, Christian; Kääb, Andreas; Raup, Bruce; Reynolds, John; Wolfe, David; Zapata, Marco

    Most scientific attention to glaciers, including ASTER and other satellite-derived applications in glacier science, pertains to their roles in the following seven functions: (1) as signposts of climate change (Kaser et al. 1990; Williams and Ferrigno 1999, 2002; Williams et al. 2008; Kargel et al. 2005; Oerlemans 2005), (2) as natural reservoirs of fresh water (Yamada and Motoyama 1988; Yang and Hu 1992; Shiyin et al. 2003; Juen et al. 2007), (3) as contributors to sea-level change (Arendt et al. 2002), (4) as sources of hydropower (Reynolds 1993); much work also relates to the basic science of glaciology, especially (5) the physical phenomeno­logy of glacier flow processes and glacier change (DeAngelis and Skvarca 2003; Berthier et al. 2007; Rivera et al. 2007), (6) glacial geomorphology (Bishop et al. 1999, 2003), and (7) the technology required to acquire and analyze satellite images of glaciers (Bishop et al. 1999, 2000, 2003, 2004; Quincey et al. 2005, 2007; Raup et al. 2000, 2006ab; Khalsa et al. 2004; Paul et al. 2004a, b). These seven functions define the important areas of glaciological science and technology, yet a more pressing issue in parts of the world is the direct danger to people and infrastructure posed by some glaciers (Trask 2005; Morales 1969; Lliboutry et al. 1977; Evans and Clague 1988; Xu and Feng 1989; Reynolds 1993, 1998, 1999; Yamada and Sharma 1993; Hastenrath and Ames 1995; Mool 1995; Ames 1998; Chikita et al. 1999; Williams and Ferrigno 1999; Richardson and Reynolds 2000a, b; Zapata 2002; Huggel et al. 2002, 2004; Xiangsong 1992; Kääb et al. 2003, 2005, 2005c; Salzmann et al. 2004; Noetzli et al. 2006).

  18. CSE1L/CAS, a microtubule-associated protein, inhibits taxol (paclitaxel)-induced apoptosis but enhances cancer cell apoptosis induced by various chemotherapeutic drugs.

    Science.gov (United States)

    Liao, Ching-Fong; Luo, Shue-Fen; Shen, Tzu-Yun; Lin, Chin-Huang; Chien, Jung-Tsun; Du, Shin-Yi; Jiang, Ming-Chung

    2008-03-31

    CSE1L/CAS, a microtubule-associated, cellular apoptosis susceptibility protein, is highly expressed in various cancers. Microtubules are the target of paclitaxel-induced apoptosis. We studied the effects of increased or reduced CAS expression on cancer cell apoptosis induced by chemotherapeutic drugs including paclitaxel. Our results showed that CAS overexpression enhanced apoptosis induced by doxorubicin, 5-fluorouracil, cisplatin, and tamoxifen, but inhibited paclitaxel-induced apoptosis of cancer cells. Reductions in CAS produced opposite results. CAS overexpression enhanced p53 accumulation induced by doxorubicin, 5-fluorouracil, cisplatin, tamoxifen, and etoposide. CAS was associated with alpha-tubulin and beta-tubulin and enhanced the association between alpha-tubulin and beta-tubulin. Paclitaxel can induce G2/M phase cell cycle arrest and microtubule aster formation during apoptosis induction, but CAS overexpression reduced paclitaxel-induced G2/M phase cell cycle arrest and microtubule aster formation. Our results indicate that CAS may play an important role in regulating the cytotoxicities of chemotherapeutic drugs used in cancer chemotherapy against cancer cells.

  19. Kinetochore-microtubule attachment is sufficient to satisfy the human spindle assembly checkpoint.

    Science.gov (United States)

    Etemad, Banafsheh; Kuijt, Timo E F; Kops, Geert J P L

    2015-12-01

    The spindle assembly checkpoint (SAC) is a genome surveillance mechanism that protects against aneuploidization. Despite profound progress on understanding mechanisms of its activation, it remains unknown what aspect of chromosome-spindle interactions is monitored by the SAC: kinetochore-microtubule attachment or the force generated by dynamic microtubules that signals stable biorientation of chromosomes? To answer this, we uncoupled these two processes by expressing a non-phosphorylatable version of the main microtubule-binding protein at kinetochores (HEC1-9A), causing stabilization of incorrect kinetochore-microtubule attachments despite persistent activity of the error-correction machinery. The SAC is fully functional in HEC1-9A-expressing cells, yet cells in which chromosomes cannot biorient but are stably attached to microtubules satisfy the SAC and exit mitosis. SAC satisfaction requires neither intra-kinetochore stretching nor dynamic microtubules. Our findings support the hypothesis that in human cells the end-on interactions of microtubules with kinetochores are sufficient to satisfy the SAC without the need for microtubule-based pulling forces.

  20. Termination of Protofilament Elongation by Eribulin Induces Lattice Defects that Promote Microtubule Catastrophes.

    Science.gov (United States)

    Doodhi, Harinath; Prota, Andrea E; Rodríguez-García, Ruddi; Xiao, Hui; Custar, Daniel W; Bargsten, Katja; Katrukha, Eugene A; Hilbert, Manuel; Hua, Shasha; Jiang, Kai; Grigoriev, Ilya; Yang, Chia-Ping H; Cox, David; Horwitz, Susan Band; Kapitein, Lukas C; Akhmanova, Anna; Steinmetz, Michel O

    2016-07-11

    Microtubules are dynamic polymers built of tubulin dimers that attach in a head-to-tail fashion to form protofilaments, which further associate laterally to form a tube. Asynchronous elongation of individual protofilaments can potentially lead to an altered microtubule-end structure that promotes sudden depolymerization, termed catastrophe [1-4]. However, how the dynamics of individual protofilaments relates to overall growth persistence has remained unclear. Here, we used the microtubule targeting anti-cancer drug Eribulin [5-7] to explore the consequences of stalled protofilament elongation on microtubule growth. Using X-ray crystallography, we first revealed that Eribulin binds to a site on β-tubulin that is required for protofilament plus-end elongation. Based on the structural information, we engineered a fluorescent Eribulin molecule. We demonstrate that single Eribulin molecules specifically interact with microtubule plus ends and are sufficient to either trigger a catastrophe or induce slow and erratic microtubule growth in the presence of EB3. Interestingly, we found that Eribulin increases the frequency of EB3 comet "splitting," transient events where a slow and erratically progressing comet is followed by a faster comet. This observation possibly reflects the "healing" of a microtubule lattice. Because EB3 comet splitting was also observed in control microtubules in the absence of any drugs, we propose that Eribulin amplifies a natural pathway toward catastrophe by promoting the arrest of protofilament elongation.

  1. Microtubule affinity-regulating kinase 4: structure, function, and regulation.

    Science.gov (United States)

    Naz, Farha; Anjum, Farah; Islam, Asimul; Ahmad, Faizan; Hassan, Md Imtaiyaz

    2013-11-01

    MAP/Microtubule affinity-regulating kinase 4 (MARK4) belongs to the family of serine/threonine kinases that phosphorylate the microtubule-associated proteins (MAP) causing their detachment from the microtubules thereby increasing microtubule dynamics and facilitating cell division, cell cycle control, cell polarity determination, cell shape alterations, etc. The MARK4 gene encodes two alternatively spliced isoforms, L and S that differ in their C-terminal region. These isoforms are differentially regulated in human tissues including central nervous system. MARK4L is a 752-residue-long polypeptide that is divided into three distinct domains: (1) protein kinase domain (59-314), (2) ubiquitin-associated domain (322-369), and (3) kinase-associated domain (703-752) plus 54 residues (649-703) involved in the proper folding and function of the enzyme. In addition, residues 65-73 are considered to be the ATP-binding domain and Lys88 is considered as ATP-binding site. Asp181 has been proposed to be the active site of MARK4 that is activated by phosphorylation of Thr214 side chain. The isoform MARK4S is highly expressed in the normal brain and is presumably involved in neuronal differentiation. On the other hand, the isoform MARK4L is upregulated in hepatocarcinoma cells and gliomas suggesting its involvement in cell cycle. Several biological functions are also associated with MARK4 including microtubule bundle formation, nervous system development, and positive regulation of programmed cell death. Therefore, MARK4 is considered as the most suitable target for structure-based rational drug design. Our sequence, structure- and function-based analysis should be helpful for better understanding of mechanisms of regulation of microtubule dynamics and MARK4 associated diseases.

  2. On the Nature and Shape of Tubulin Trails: Implications on Microtubule Self-Organization

    CERN Document Server

    Glade, Nicolas

    2012-01-01

    Microtubules, major elements of the cell skeleton are, most of the time, well organized in vivo, but they can also show self-organizing behaviors in time and/or space in purified solutions in vitro. Theoretical studies and models based on the concepts of collective dynamics in complex systems, reaction-diffusion processes and emergent phenomena were proposed to explain some of these behaviors. In the particular case of microtubule spatial self-organization, it has been advanced that microtubules could behave like ants, self-organizing by 'talking to each other' by way of hypothetic (because never observed) concentrated chemical trails of tubulin that are expected to be released by their disassembling ends. Deterministic models based on this idea yielded indeed like-looking spatio-temporal self-organizing behaviors. Nevertheless the question remains of whether microscopic tubulin trails produced by individual or bundles of several microtubules are intense enough to allow microtubule self-organization at a macr...

  3. Oxidative stress decreases microtubule growth and stability in ventricular myocytes

    OpenAIRE

    Drum, BML; Yuan, C.; Li, L; Liu, Q.; Wordeman, L; Santana, LF

    2016-01-01

    © 2016 Elsevier Ltd.Microtubules (MTs) have many roles in ventricular myocytes, including structural stability, morphological integrity, and protein trafficking. However, despite their functional importance, dynamic MTs had never been visualized in living adult myocytes. Using adeno-associated viral vectors expressing the MT-associated protein plus end binding protein 3 (EB3) tagged with EGFP, we were able to perform live imaging and thus capture and quantify MT dynamics in ventricular myocyt...

  4. Microtubule assembly affects bone mass by regulating both osteoblast and osteoclast functions: stathmin deficiency produces an osteopenic phenotype in mice.

    Science.gov (United States)

    Liu, Hongbin; Zhang, Rongrong; Ko, Seon-Yle; Oyajobi, Babatunde O; Papasian, Christopher J; Deng, Hong-Wen; Zhang, Shujun; Zhao, Ming

    2011-09-01

    Cytoskeleton microtubules regulate various cell signaling pathways that are involved in bone cell function. We recently reported that inhibition of microtubule assembly by microtubule-targeting drugs stimulates osteoblast differentiation and bone formation. To further elucidate the role of microtubules in bone homeostasis, we characterized the skeletal phenotype of mice null for stathmin, an endogenous protein that inhibits microtubule assembly. In vivo micro-computed tomography (µCT) and histology revealed that stathmin deficiency results in a significant reduction of bone mass in adult mice concurrent with decreased osteoblast and increased osteoclast numbers in bone tissues. Phenotypic analyses of primary calvarial cells and bone marrow cells showed that stathmin deficiency inhibited osteoblast differentiation and induced osteoclast formation. In vitro overexpression studies showed that increased stathmin levels enhanced osteogenic differentiation of preosteoblast MC3T3-E1 cells and mouse bone marrow-derived cells and attenuated osteoclast formation from osteoclast precursor Raw264.7 cells and bone marrow cells. Results of immunofluorescent studies indicated that overexpression of stathmin disrupted radial microtubule filaments, whereas deficiency of stathmin stabilized the microtubule network structure in these bone cells. In addition, microtubule-targeting drugs that inhibit microtubule assembly and induce osteoblast differentiation lost these effects in the absence of stathmin. Collectively, these results suggest that stathmin, which alters microtubule dynamics, plays an essential role in maintenance of postnatal bone mass by regulating both osteoblast and osteoclast functions in bone. \\

  5. ASTER Global DEM contribution to GEOSS demonstrates open data sharing

    Science.gov (United States)

    Sohre, T.; Duda, K. A.; Meyer, D. J.; Behnke, J.; Nasa Esdis Lp Daac

    2010-12-01

    The Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) remote sensing instrument on the Terra spacecraft has been acquiring images of Earth since launch in 1999. Throughout this time data products have been openly available to the general public through sites in the U.S. and Japan. As the ASTER mission matured, a spatially broad and temporally deep data archive was gradually established. With this extensive accumulation of Earth observations, it became possible to create a new global digital elevation product, the ASTER Global Digital Elevation Model (GDEM), using multi-temporal data, resulting in over 22,000 static 10 X 10 tiles. The ASTER GDEM was contributed by Japan’s Ministry of Economy Trade and Industry (METI) and the U.S. National Aeronautics and Space Administration (NASA) to the Global Earth Observation System of Systems (GEOSS) for distribution at no cost to users. As such, both METI and NASA desired to understand the uses of the ASTER GDEM, expressed as one of the GEOSS applications themes: disasters, health, energy, climate, water, weather, ecosystems, agriculture or biodiversity. This required both the registration of users, and restrictions on redistribution, to capture the intended use in terms of the GEOSS themes. The ASTER GDEM was made available to users worldwide via electronic download from the Earth Remote Sensing Data Analysis Center (ERSDAC) of Japan and from NASA’s Land Processes Distributed Active Archive Center (LP DAAC). During the first three months after product release, over 4 million GDEM tiles were distributed from the LP DAAC and ERSDAC. The ASTER GDEM release generated nearly 20,000 new user registrations in the NASA EOS ClearingHOuse (ECHO)/WIST and the ERSDAC systems. By the end of 2009, over 6.5 Million GDEM tiles were distributed by the LP DAAC and ERSDAC. Users have requested tiles over specific areas of interest as well as the entire dataset for global research. Intense global interest in the GDEM

  6. CHROMITITE PROSPECTING USING LANDSAT TM AND ASTER REMOTE SENSING DATA

    Directory of Open Access Journals (Sweden)

    A. Beiranvand Pour

    2015-10-01

    Full Text Available Studying the ophiolite complexes using multispectral remote sensing satellite data are interesting because of high diversity of minerals and the source of podiform chromitites. This research developed an approach to discriminate lithological units and detecting host rock of chromitite bodies within ophiolitic complexes using the Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER and Landsat Thematic Mapper (TM satellite data. Three main ophiolite complexes located in south of Iran have been selected for the study. Spectral transform techniques, including minimum noise fraction (MNF and specialized band ratio were employed to detect different rock units and the identification of high-potential areas of chromite ore deposits within ophiolitic complexes. A specialized band ratio (4/1, 4/5, 4/7 of ASTER, MNF components and Spectral Angle Mapper (SAM on ASTER and Landsat TM data were used to distinguish ophiolitic rock units. Results show that the specialized band ratio was able to identify different rock units and serpentinized dunite as host rock of chromitites within ophiolitic complexes, appropriately. MNF components of ASTER and Landsat TM data were suitable to distinguish ophiolitic rock complexes at a regional scale. The integration of SAM and Feature Level Fusion (FLF used in this investigation discriminated the ophiolitic rock units and prepared detailed geological map for the study area. Accordingly, high potential areas (serpentinite dunite were identified in the study area for chromite exploration targets.The approach used in this research offers the image processing techniques as a robust, reliable, fast and cost-effective method for detecting serpentinized dunite as host rock of chromitite bodies within vast ophiolite complexes using ASTER and Landsat TM satellite data.

  7. Molecular mechanisms of antitumor activity of taxanes. I. Interaction of docetaxel with microtubules

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    Sabina Tabaczar

    2010-11-01

    Full Text Available Docetaxel (Taxotere, a new semisynthetic taxoid, is a mitotic inhibitor, widely used in monotherapy or in combination with other anticancer drugs against many types of cancer. The structure and dynamics of microtubules as the main target for docetaxel activity inside the cell and the taxane-binding site on β-tubulin are discussed. Microtubules are highly dynamic assemblies of α- and β-tubulin. They readily polymerize and depolymerize in cells and these dynamic behaviours are crucial to cell mitosis. Microtubule instability is attributed to their capability to hydrolyze GTP to GDP, which causes their depolymerization. Addition of new α-, β-tubulin heterodimer bound to GTP leads to tubulin polymerization, which increases the length of the microtubule. Docetaxel alters the polymerization dynamics of microtubules, which causes blockage of cell mitosis, and consequently induces apoptotic and non-apoptotic cell death. Docetaxel specifically acts on the S, M and G2 phases of the cell cycle. This paper reviews the current state of knowledge related to the molecular mechanisms of docetaxel action on the cell cycle and microtubule dynamics. In addition, a brief survey of the present state of research on the new generation (2nd and 3rd of taxanes is presented.

  8. Mechanism for the catastrophe-promoting activity of the microtubule destabilizer Op18/stathmin.

    Science.gov (United States)

    Gupta, Kamlesh K; Li, Chunlei; Duan, Aranda; Alberico, Emily O; Kim, Oleg V; Alber, Mark S; Goodson, Holly V

    2013-12-17

    Regulation of microtubule dynamic instability is crucial for cellular processes, ranging from mitosis to membrane transport. Stathmin (also known as oncoprotein 18/Op18) is a prominent microtubule destabilizer that acts preferentially on microtubule minus ends. Stathmin has been studied intensively because of its association with multiple types of cancer, but its mechanism of action remains controversial. Two models have been proposed. One model is that stathmin promotes microtubule catastrophe indirectly, and does so by sequestering tubulin; the other holds that stathmin alters microtubule dynamics by directly destabilizing growing microtubules. Stathmin's sequestration activity is well established, but the mechanism of any direct action is mysterious because stathmin binds to microtubules very weakly. To address these issues, we have studied interactions between stathmin and varied tubulin polymers. We show that stathmin binds tightly to Dolastatin-10 tubulin rings, which mimic curved tubulin protofilaments, and that stathmin depolymerizes stabilized protofilament-rich polymers. These observations lead us to propose that stathmin promotes catastrophe by binding to and acting upon protofilaments exposed at the tips of growing microtubules. Moreover, we suggest that stathmin's minus-end preference results from interactions between stathmin's N terminus and the surface of α-tubulin that is exposed only at the minus end. Using computational modeling of microtubule dynamics, we show that these mechanisms could account for stathmin's observed activities in vitro, but that both the direct and sequestering activities are likely to be relevant in a cellular context. Taken together, our results suggest that stathmin can promote catastrophe by direct action on protofilament structure and interactions.

  9. Microtubule-microtubule sliding by kinesin-1 is essential for normal cytoplasmic streaming in Drosophila oocytes.

    Science.gov (United States)

    Lu, Wen; Winding, Michael; Lakonishok, Margot; Wildonger, Jill; Gelfand, Vladimir I

    2016-08-23

    Cytoplasmic streaming in Drosophila oocytes is a microtubule-based bulk cytoplasmic movement. Streaming efficiently circulates and localizes mRNAs and proteins deposited by the nurse cells across the oocyte. This movement is driven by kinesin-1, a major microtubule motor. Recently, we have shown that kinesin-1 heavy chain (KHC) can transport one microtubule on another microtubule, thus driving microtubule-microtubule sliding in multiple cell types. To study the role of microtubule sliding in oocyte cytoplasmic streaming, we used a Khc mutant that is deficient in microtubule sliding but able to transport a majority of cargoes. We demonstrated that streaming is reduced by genomic replacement of wild-type Khc with this sliding-deficient mutant. Streaming can be fully rescued by wild-type KHC and partially rescued by a chimeric motor that cannot move organelles but is active in microtubule sliding. Consistent with these data, we identified two populations of microtubules in fast-streaming oocytes: a network of stable microtubules anchored to the actin cortex and free cytoplasmic microtubules that moved in the ooplasm. We further demonstrated that the reduced streaming in sliding-deficient oocytes resulted in posterior determination defects. Together, we propose that kinesin-1 slides free cytoplasmic microtubules against cortically immobilized microtubules, generating forces that contribute to cytoplasmic streaming and are essential for the refinement of posterior determinants.

  10. Atomic-resolution structure of the CAP-Gly domain of dynactin on polymeric microtubules determined by magic angle spinning NMR spectroscopy.

    Science.gov (United States)

    Yan, Si; Guo, Changmiao; Hou, Guangjin; Zhang, Huilan; Lu, Xingyu; Williams, John Charles; Polenova, Tatyana

    2015-11-24

    Microtubules and their associated proteins perform a broad array of essential physiological functions, including mitosis, polarization and differentiation, cell migration, and vesicle and organelle transport. As such, they have been extensively studied at multiple levels of resolution (e.g., from structural biology to cell biology). Despite these efforts, there remain significant gaps in our knowledge concerning how microtubule-binding proteins bind to microtubules, how dynamics connect different conformational states, and how these interactions and dynamics affect cellular processes. Structures of microtubule-associated proteins assembled on polymeric microtubules are not known at atomic resolution. Here, we report a structure of the cytoskeleton-associated protein glycine-rich (CAP-Gly) domain of dynactin motor on polymeric microtubules, solved by magic angle spinning NMR spectroscopy. We present the intermolecular interface of CAP-Gly with microtubules, derived by recording direct dipolar contacts between CAP-Gly and tubulin using double rotational echo double resonance (dREDOR)-filtered experiments. Our results indicate that the structure adopted by CAP-Gly varies, particularly around its loop regions, permitting its interaction with multiple binding partners and with the microtubules. To our knowledge, this study reports the first atomic-resolution structure of a microtubule-associated protein on polymeric microtubules. Our approach lays the foundation for atomic-resolution structural analysis of other microtubule-associated motors.

  11. A Stochastic Multiscale Model That Explains the Segregation of Axonal Microtubules and Neurofilaments in Neurological Diseases.

    Directory of Open Access Journals (Sweden)

    Chuan Xue

    2015-08-01

    Full Text Available The organization of the axonal cytoskeleton is a key determinant of the normal function of an axon, which is a long thin projection of a neuron. Under normal conditions two axonal cytoskeletal polymers, microtubules and neurofilaments, align longitudinally in axons and are interspersed in axonal cross-sections. However, in many neurotoxic and neurodegenerative disorders, microtubules and neurofilaments segregate apart from each other, with microtubules and membranous organelles clustered centrally and neurofilaments displaced to the periphery. This striking segregation precedes the abnormal and excessive neurofilament accumulation in these diseases, which in turn leads to focal axonal swellings. While neurofilament accumulation suggests an impairment of neurofilament transport along axons, the underlying mechanism of their segregation from microtubules remains poorly understood for over 30 years. To address this question, we developed a stochastic multiscale model for the cross-sectional distribution of microtubules and neurofilaments in axons. The model describes microtubules, neurofilaments and organelles as interacting particles in a 2D cross-section, and is built upon molecular processes that occur on a time scale of seconds or shorter. It incorporates the longitudinal transport of neurofilaments and organelles through this domain by allowing stochastic arrival and departure of these cargoes, and integrates the dynamic interactions of these cargoes with microtubules mediated by molecular motors. Simulations of the model demonstrate that organelles can pull nearby microtubules together, and in the absence of neurofilament transport, this mechanism gradually segregates microtubules from neurofilaments on a time scale of hours, similar to that observed in toxic neuropathies. This suggests that the microtubule-neurofilament segregation can be a consequence of the selective impairment of neurofilament transport. The model generates the

  12. The Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) after fifteen years: Review of global products

    Science.gov (United States)

    Abrams, Michael; Tsu, Hiroji; Hulley, Glynn; Iwao, Koki; Pieri, David; Cudahy, Tom; Kargel, Jeffrey

    2015-06-01

    The Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) is a 15-channel imaging instrument operating on NASA's Terra satellite. A joint project between the U.S. National Aeronautics and Space Administration and Japan's Ministry of Economy, Trade, and Industry, ASTER has been acquiring data for 15 years, since March 2000. The archive now contains over 2.8 million scenes; for the majority of them, a stereo pair was collected using nadir and backward telescopes imaging in the NIR wavelength. The majority of users require only a few to a few dozen scenes for their work. Studies have ranged over numerous scientific disciplines, and many practical applications have benefited from ASTER's unique data. A few researchers have been able to mine the entire ASTER archive, that is now global in extent due to the long duration of the mission. Six examples of global products are described in this contribution: the ASTER Global Digital Elevation Model (GDEM), the most complete, highest resolution DEM available to all users; the ASTER Emissivity Database (ASTER GED), a global 5-band emissivity map of the land surface; the ASTER Global Urban Area Map (AGURAM), a 15-m resolution database of over 3500 cities; the ASTER Volcano Archive (AVA), an archive of over 1500 active volcanoes; ASTER Geoscience products of the continent of Australia; and the Global Ice Monitoring from Space (GLIMS) project.

  13. Tau Induces Cooperative Taxol Binding to Microtubules

    Science.gov (United States)

    Ross, Jennifer; Santangelo, Christian; Victoria, Makrides; Fygenson, Deborah

    2004-03-01

    Taxol and tau are two ligands which stabilize the microtubule (MT) lattice. Taxol is an anti-mitotic drug that binds β tubulin in the MT interior. Tau is a MT-associated protein that binds both α and β tubulin on the MT exterior. Both taxol and tau reduce MT dynamics and promote tubulin polymerization. Tau alone also acts as a buttress to bundle, stiffen, and space MTs. A structural study recently suggested that taxol and tau may interact by binding to the same site. Using fluorescence recovery after photobleaching, we find that tau induces taxol to bind MTs cooperatively depending on the tau concentration. We develop a model that correctly fits the data in the absence of tau and yields a measure of taxol cooperativity when tau is present.

  14. Microtubule as nanobioelectronic nonlinear circuit

    Directory of Open Access Journals (Sweden)

    Sekulić Dalibor L.

    2012-01-01

    Full Text Available In recent years, the use of biological molecules has offered exciting alternatives to conventional synthetic methods. Specific methods use various biological templates to direct the deposition and patterning of inorganic materials. Here we have established a new electrical model of microtubules as a biological nanoscale circuit based on polyelectrolyte features of cylindrical biopolymers. Our working hypothesis is that microtubules play an active role in sub-cellular computation and signaling via electronic and protonic conductivity and can thus be made useful in hybrid materials that offer novel electronic characteristics. We verify these hypotheses both computationally and analytically through a quantitative model based on the atomic resolution structures of the key functional proteins.

  15. Detailed Per-residue Energetic Analysis Explains the Driving Force for Microtubule Disassembly.

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    Ahmed T Ayoub

    2015-06-01

    Full Text Available Microtubules are long filamentous hollow cylinders whose surfaces form lattice structures of αβ-tubulin heterodimers. They perform multiple physiological roles in eukaryotic cells and are targets for therapeutic interventions. In our study, we carried out all-atom molecular dynamics simulations for arbitrarily long microtubules that have either GDP or GTP molecules in the E-site of β-tubulin. A detailed energy balance of the MM/GBSA inter-dimer interaction energy per residue contributing to the overall lateral and longitudinal structural stability was performed. The obtained results identified the key residues and tubulin domains according to their energetic contributions. They also identified the molecular forces that drive microtubule disassembly. At the tip of the plus end of the microtubule, the uneven distribution of longitudinal interaction energies within a protofilament generates a torque that bends tubulin outwardly with respect to the cylinder's axis causing disassembly. In the presence of GTP, this torque is opposed by lateral interactions that prevent outward curling, thus stabilizing the whole microtubule. Once GTP hydrolysis reaches the tip of the microtubule (lateral cap, lateral interactions become much weaker, allowing tubulin dimers to bend outwards, causing disassembly. The role of magnesium in the process of outward curling has also been demonstrated. This study also showed that the microtubule seam is the most energetically labile inter-dimer interface and could serve as a trigger point for disassembly. Based on a detailed balance of the energetic contributions per amino acid residue in the microtubule, numerous other analyses could be performed to give additional insights into the properties of microtubule dynamic instability.

  16. Microtubules in Dendritic Spine Development

    OpenAIRE

    2008-01-01

    It is generally believed that only the actin cytoskeleton resides in dendritic spines and controls spine morphology and plasticity. Here we report that microtubules (MTs) are present in spines and that shRNA knockdown of the MT-plus end binding protein EB3 significantly reduces spine formation. Furthermore, stabilization and inhibition of MTs by low doses of taxol and nocodazole enhance and impair spine formation elicited by BDNF, respectively. Therefore, MTs play an important role in the con...

  17. CYLD Regulates Noscapine Activity in Acute Lymphoblastic Leukemia via a Microtubule-Dependent Mechanism

    OpenAIRE

    Yang, Yunfan; Ran, Jie; Sun, Lei; Sun, Xiaodong; Luo, Youguang; Yan, Bing; Tala,; Liu, Min; Li, Dengwen; Zhang, Lei; Bao, Gang; Zhou, Jun

    2015-01-01

    Noscapine is an orally administrable drug used worldwide for cough suppression and has recently been demonstrated to disrupt microtubule dynamics and possess anticancer activity. However, the molecular mechanisms regulating noscapine activity remain poorly defined. Here we demonstrate that cylindromatosis (CYLD), a microtubule-associated tumor suppressor protein, modulates the activity of noscapine both in cell lines and in primary cells of acute lymphoblastic leukemia (ALL). Flow cytometry a...

  18. Detailed Per-residue Energetic Analysis Explains the Driving Force for Microtubule Disassembly

    OpenAIRE

    Ayoub, Ahmed T.; Mariusz Klobukowski; Tuszynski, Jack A

    2015-01-01

    Microtubules are long filamentous hollow cylinders whose surfaces form lattice structures of αβ-tubulin heterodimers. They perform multiple physiological roles in eukaryotic cells and are targets for therapeutic interventions. In our study, we carried out all-atom molecular dynamics simulations for arbitrarily long microtubules that have either GDP or GTP molecules in the E-site of β-tubulin. A detailed energy balance of the MM/GBSA inter-dimer interaction energy per residue contributing to t...

  19. Identificación de metabasaltos alterados mediante datos ASTER

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    Silvia Castro Godoy

    2010-06-01

    Full Text Available Los datos del sensor ASTER permitieron desarrollar una metodología sencilla, utilizando las bandas del infrarrojo de onda corta (subsistema SWIR, para prospectar cuerpos ígneos alterados que manifiestan diferencias espectrales con respecto a la roca de caja. A modo de ejemplo se detectó la alteración clorítica presente en metabasaltos intercalados en metasedimentitas de la Formación Puntilla de Uspallata en la Precordillera mendocina.

  20. Identificación de metabasaltos alterados mediante datos ASTER

    OpenAIRE

    Silvia Castro Godoy; Mariela Etcheverría; Marta Godeas

    2010-01-01

    Los datos del sensor ASTER permitieron desarrollar una metodología sencilla, utilizando las bandas del infrarrojo de onda corta (subsistema SWIR), para prospectar cuerpos ígneos alterados que manifiestan diferencias espectrales con respecto a la roca de caja. A modo de ejemplo se detectó la alteración clorítica presente en metabasaltos intercalados en metasedimentitas de la Formación Puntilla de Uspallata en la Precordillera mendocina.

  1. Prediction of winter wheat grain protein content by ASTER image

    Science.gov (United States)

    Huang, Wenjiang; Song, Xiaoyu; Wang, Jihua; Wang, Zhijie; Zhao, Chunjiang

    2008-10-01

    The Advanced technology in space-borne determination of grain crude protein content (CP) by remote sensing can help optimize the strategies for buyers in aiding purchasing decisions, and help farmers to maximize the grain output by adjusting field nitrogen (N) fertilizer inputs. We performed field experiments to study the relationship between grain quality indicators and foliar nitrogen concentration (FNC). FNC at anthesis stage was significantly correlated with CP, while spectral vegetation index was significantly correlated to FNC. Based on the relationships among nitrogen reflectance index (NRI), FNC and CP, a model for CP prediction was developed. NRI was able to evaluate FNC with a higher coefficient of determination of R2=0.7302. The method developed in this study could contribute towards developing optimal procedures for evaluating wheat grain quality by ASTER image at anthesis stage. The RMSE was 0.893 % for ASTER image model, and the R2 was 0.7194. It is thus feasible to forecast grain quality by NRI derived from ASTER image.

  2. Laulimalide induces dose-dependent modulation of microtubule behaviour in the C. elegans embryo.

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    Megha Bajaj

    Full Text Available Laulimalide is a microtubule-binding drug that was originally isolated from marine sponges. High concentrations of laulimalide stabilize microtubules and inhibit cell division similarly to paclitaxel; however, there are important differences with respect to the nature of the specific cellular defects between these two drugs and their binding sites on the microtubule. In this study, we used Caenorhabditis elegans embryos to investigate the acute effects of laulimalide on microtubules in vivo, with a direct comparison to paclitaxel. We observed surprising dose-dependent effects for laulimalide, whereby microtubules were stabilized at concentrations above 100 nM, but destabilized at concentrations between 50 and 100 nM. Despite this behaviour at low concentrations, laulimalide acted synergistically with paclitaxel to stabilize microtubules when both drugs were used at sub-effective concentrations, consistent with observations of synergistic interactions between these two drugs in other systems. Our results indicate that laulimalide induces a concentration-dependent, biphasic change in microtubule polymer dynamics in the C. elegans embryo.

  3. Long astral microtubules and RACK-1 stabilize polarity domains during maintenance phase in Caenorhabditis elegans embryos.

    Directory of Open Access Journals (Sweden)

    Erkang Ai

    Full Text Available Cell polarity is a very well conserved process important for cell differentiation, cell migration, and embryonic development. After the establishment of distinct cortical domains, polarity cues have to be stabilized and maintained within a fluid and dynamic membrane to achieve proper cell asymmetry. Microtubules have long been thought to deliver the signals required to polarize a cell. While previous studies suggest that microtubules play a key role in the establishment of polarity, the requirement of microtubules during maintenance phase remains unclear. In this study, we show that depletion of Caenorhabditis elegans RACK-1, which leads to short astral microtubules during prometaphase, specifically affects maintenance of cortical PAR domains and Dynamin localization. We then investigated the consequence of knocking down other factors that also abolish astral microtubule elongation during polarity maintenance phase. We found a correlation between short astral microtubules and the instability of PAR-6 and PAR-2 domains during maintenance phase. Our data support a necessary role for astral microtubules in the maintenance phase of cell polarity.

  4. Positioning of microtubule organizing centers by cortical pushing and pulling forces

    Science.gov (United States)

    Pavin, Nenad; Laan, Liedewij; Ma, Rui; Dogterom, Marileen; Jülicher, Frank

    2012-10-01

    Positioning of microtubule (MT) organizing centers with respect to the confining geometry of cells depends on pushing and/or pulling forces generated by MTs that interact with the cell cortex (Dogterom et al 2005 Curr. Opin. Cell Biol. 17 67-74). How, in living cells, these forces lead to proper positioning is still largely an open question. Recently, it was shown by in vitro experiments using artificial microchambers that in a square geometry, MT asters center more reliably by a combination of pulling and pushing forces than by pushing forces alone (Laan et al 2012a Cell 148 502-14). These findings were explained by a physical description of aster mechanics that includes slipping of pushing MT ends along chamber boundaries. In this paper, we extend that theoretical work by studying the influence of the shape of the confining geometry on the positioning process. We find that pushing and pulling forces can have centering or off-centering behavior in different geometries. Pushing forces center in a one-dimensional and a square geometry, but lead to off-centering in a circle if slipping is sufficiently pronounced. Pulling forces, however, do not center in a one-dimensional geometry, but improve centering in a circle and a square. In an elongated stadium geometry, positioning along the short axis depends mainly on pulling forces, while positioning along the long axis depends mainly on pushing forces. Our theoretical results suggest that different positioning strategies could be used by different cell types.

  5. Identificación de metabasaltos alterados mediante datos ASTER Identification of altered metabasalts from ASTER data

    Directory of Open Access Journals (Sweden)

    Silvia Castro Godoy

    2010-06-01

    Full Text Available Los datos del sensor ASTER permitieron desarrollar una metodología sencilla, utilizando las bandas del infrarrojo de onda corta (subsistema SWIR, para prospectar cuerpos ígneos alterados que manifiestan diferencias espectrales con respecto a la roca de caja. A modo de ejemplo se detectó la alteración clorítica presente en metabasaltos intercalados en metasedimentitas de la Formación Puntilla de Uspallata en la Precordillera mendocina.ASTER data allowed developing a simple methodology, using the Short Wave Infrared spectral bands (SWIR, in order to detect alteration minerals that affect igneous bodies and have spectral differences with the country rock. As an example chloritic alteration was detected in metabasalts interbedded in metasediments of Puntilla de Uspallata Formation, Precordillera Mendocina.

  6. The Drosophila microtubule-associated protein mars stabilizes mitotic spindles by crosslinking microtubules through its N-terminal region.

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    Gang Zhang

    Full Text Available Correct segregation of genetic material relies on proper assembly and maintenance of the mitotic spindle. How the highly dynamic microtubules (MTs are maintained in stable mitotic spindles is a key question to be answered. Motor and non-motor microtubule associated proteins (MAPs have been reported to stabilize the dynamic spindle through crosslinking adjacent MTs. Mars, a novel MAP, is essential for the early development of Drosophila embryos. Previous studies showed that Mars is required for maintaining an intact mitotic spindle but did not provide a molecular mechanism for this function. Here we show that Mars is able to stabilize the mitotic spindle in vivo. Both in vivo and in vitro data reveal that the N-terminal region of Mars functions in the stabilization of the mitotic spindle by crosslinking adjacent MTs.

  7. Anomalous motor mediated cargo transport in microtubule networks

    Science.gov (United States)

    Vandal, Steven; Macveigh-Fierro, Daniel; Shen, Zhiyuan; Lemoi, Kyle; Vidali, Luis; Ross, Jennifer; Tuzel, Erkan

    Cargo transport is an important biological mechanism by which cells locomote, self-organize, and actively transport organelles. This transport is mediated by the cytoskeletal network and molecular motors; however, it is not known how network self-organization and dynamics affect these transport processes. In order to develop a mechanistic understanding of cargo transport, we use a coarse-grained Brownian dynamics model that incorporates the dynamics of these networks, as well as experimentally determined motor properties. We will test these models with two experimental systems: (1) in vitro microtubule networks with kinesin-1 motors, and quantum dot cargos on recreated microtubule networks, and (2) an excellent model organism, the moss Physcomitrella patens, in which chloroplasts are transported via the microtubule network by means of kinesin-like proteins. Phenomenological network characterizations are made, both in vivo and in vitro, and cargo motility is characterized using Mean Squared Displacement (MSD) measurements. Our simulations shed light on the role of network density and motor properties on the observed transport behavior, and improve our understanding of cargo transport in cells.

  8. Dissecting the nanoscale distributions and functions of microtubule-end-binding proteins EB1 and ch-TOG in interphase HeLa cells.

    Directory of Open Access Journals (Sweden)

    Satoko Nakamura

    Full Text Available Recently, the EB1 and XMAP215/TOG families of microtubule binding proteins have been demonstrated to bind autonomously to the growing plus ends of microtubules and regulate their behaviour in in vitro systems. However, their functional redundancy or difference in cells remains obscure. Here, we compared the nanoscale distributions of EB1 and ch-TOG along microtubules using high-resolution microscopy techniques, and also their roles in microtubule organisation in interphase HeLa cells. The ch-TOG accumulation sites protruded ∼100 nm from the EB1 comets. Overexpression experiments showed that ch-TOG and EB1 did not interfere with each other's localisation, confirming that they recognise distinct regions at the ends of microtubules. While both EB1 and ch-TOG showed similar effects on microtubule plus end dynamics and additively increased microtubule dynamicity, only EB1 exhibited microtubule-cell cortex attachment activity. These observations indicate that EB1 and ch-TOG regulate microtubule organisation differently via distinct regions in the plus ends of microtubules.

  9. Joined at the hip: kinetochores, microtubules, and spindle assembly checkpoint signaling.

    Science.gov (United States)

    Sacristan, Carlos; Kops, Geert J P L

    2015-01-01

    Error-free chromosome segregation relies on stable connections between kinetochores and spindle microtubules. The spindle assembly checkpoint (SAC) monitors such connections and relays their absence to the cell cycle machinery to delay cell division. The molecular network at kinetochores that is responsible for microtubule binding is integrated with the core components of the SAC signaling system. Molecular-mechanistic understanding of how the SAC is coupled to the kinetochore-microtubule interface has advanced significantly in recent years. The latest insights not only provide a striking view of the dynamics and regulation of SAC signaling events at the outer kinetochore but also create a framework for understanding how that signaling may be terminated when kinetochores and microtubules connect.

  10. [The role of cortical microtubules in moss protonemal cells during dehydration/rehydration cycle].

    Science.gov (United States)

    Chen, Zhi-Ling; Ouyang, Hao-Miao; Liu, Xiang-Lin; Xia, Gui-Xian

    2003-05-01

    Plant cells response to water deficit through a variety of physiological processes. In this work, we studied the function of microtubule cytoskeleton during dehydration/rehydration cycle in moss (Atrichum undulatum) protonemal cells as a model system. The morphological and cytological change of protonemal cells during dehydration and rehydration cycle were first investigated. Under normal conditions, protonemal cells showed bright green colour and appeared wet and fresh. Numerous chloroplasts distributed regularly throughout the cytoplasm in each cell. After dehydration treatment, protonemal cells lost most of their chlorophylls and turned to look yellow and dry. In addition, dehydration caused plasmolysis in these cells. Upon rehydration, the cells could recover completely from the dehydrated state. These results indicated that moss had a remarkable intrinsic ability to survive from the extreme drought stress. Microtubule, an important component of cytoskeleton, is considered to play crucial roles in the responses to some environmental stresses such as cold and light. To see if it is also involved in the drought tolerance, dynamic organization of microtubules in protonemal cells of Atrichum undulatum subjected to drought and rehydration were examined by indirect immunofluorescence combined with confocal lasersharp scanning microscopy. The cortical microtubules were arranged into a fine structure with a predominant orientation parallel to the long axis of the cells in the control cells. After dehydration, the microtubule organization was remarkablly altered and the fine microtubule structure disappeared whereas some thicker cables formed. When the cells were grown under rehydration conditions, the fine microtubule arrays reappeared. These results provided a piece of evidence that microtubules play a role in the cellular responses to drought stress in moss. Furthermore, we analyzed the effects of the microtubule-disrupting agent colchicine on the morphology recovery

  11. Movement of chromosomes with severed kinetochore microtubules.

    Science.gov (United States)

    Forer, Arthur; Johansen, Kristen M; Johansen, Jørgen

    2015-05-01

    Experiments dating from 1966 and thereafter showed that anaphase chromosomes continued to move poleward after their kinetochore microtubules were severed by ultraviolet microbeam irradiation. These observations were initially met with scepticism as they contradicted the prevailing view that kinetochore fibre microtubules pulled chromosomes to the pole. However, recent experiments using visible light laser microbeam irradiations have corroborated these earlier experiments as anaphase chromosomes again were shown to move poleward after their kinetochore microtubules were severed. Thus, multiple independent studies using different techniques have shown that chromosomes can indeed move poleward without direct microtubule connections to the pole, with only a kinetochore 'stub' of microtubules. An issue not yet settled is: what propels the disconnected chromosome? There are two not necessarily mutually exclusive proposals in the literature: (1) chromosome movement is propelled by the kinetochore stub interacting with non-kinetochore microtubules and (2) chromosome movement is propelled by a spindle matrix acting on the stub. In this review, we summarise the data indicating that chromosomes can move with severed kinetochore microtubules and we discuss proposed mechanisms for chromosome movement with severed kinetochore microtubules.

  12. Assessment of Landscape Fragmentation Associated With Urban Centers Using ASTER Data

    Science.gov (United States)

    Stefanov, W. L.

    2002-12-01

    The role of humans as an integral part of the environment and ecosystem processes has only recently been accepted into mainstream ecological thought. The realization that virtually all ecosystems on Earth have experienced some degree of human alteration or impact has highlighted the need to incorporate humans (and their environmental effects) into ecosystem models. A logical starting point for investigation of human ecosystem dynamics is examination of the land cover characteristics of large urban centers. Land cover and land use changes associated with urbanization are important drivers of local geological, hydrological, ecological, and climatic change. Quantification and monitoring of urban land cover/land use change is part of the primary mission of the ASTER instrument on board the NASA Terra satellite, and comprises the fundamental research objective of the Urban Environmental Monitoring (UEM) Program at Arizona State University. The UEM program seeks to acquire day/night, visible through thermal infrared data twice per year for 100 global urban centers (with an emphasis on semi-arid cities) over the nominal six-year life of the Terra mission. Data have been acquired for the majority of the target urban centers and are used to compare landscape fragmentation patterns on the basis of land cover classifications. Land cover classifications of urban centers are obtained using visible through mid-infrared reflectance and emittance spectra together with calculated vegetation index and spatial variance texture information (all derived from raw ASTER data). This information is combined within a classification matrix, using an expert system framework, to obtain final pixel classifications. Landscape fragmentation is calculated using a pixel per unit area metric for comparison between 55 urban centers with varying geographic and climatic settings including North America, South America, Europe, central and eastern Asia, and Australia. Temporal variations in land cover

  13. Effects of 3-repeat tau on taxol mobility through microtubules

    Science.gov (United States)

    Park, Hyunjoo; Fygenson, Deborah; Kim, Mahn Won

    2005-03-01

    Both the anti-cancer drug taxol and the microtubule-associated protein tau suppress dynamics of microtubules (MT). We have observed taxol mobility with full-length 3-repeat tau, one of six tau isoforms, using fluorescence recovery after photobleaching (FRAP) on MTs and compare with earlier results on recombinant full-length adult 4-repeat tau. Taxol mobility becomes highly sensitive to taxol concentration in the presence of 3-repeat tau (up to 1:1 molar ratio) as it does in the presence of 4-repeat tau, but is 2 to 3 times faster at low taxol concentrations. Fitting to a mean-field binding reaction model [J.L. Ross et.al, PNAS 101:12910-5 (2004)] suggests that the presence of 3-repeat tau enhances taxol movement through pores in the MT walls.

  14. Mobility of Taxol in Microtubule Bundles

    Science.gov (United States)

    Ross, J.

    2003-06-01

    Mobility of taxol inside microtubules was investigated using fluorescence recovery after photobleaching (FRAP) on flow-aligned bundles. Bundles were made of microtubules with either GMPCPP or GTP at the exchangeable site on the tubulin dimer. Recovery times were sensitive to bundle thickness and packing, indicating that taxol molecules are able to move laterally through the bundle. The density of open binding sites along a microtubule was varied by controlling the concentration of taxol in solution for GMPCPP samples. With > 63% sites occupied, recovery times were independent of taxol concentration and, therefore, inversely proportional to the microscopic dissociation rate, k_{off}. It was found that 10*k_{off} (GMPCPP) ~ k_{off} (GTP), consistent with, but not fully accounting for, the difference in equilibrium constants for taxol on GMPCPP and GTP microtubules. With taxol along the microtubule interior is hindered by rebinding events when open sites are within ~7 nm of each other.

  15. Comparison of field emissivities with laboratory measurements and ASTER data

    Science.gov (United States)

    Mira, M.; Schmugge, T.; Valor, E.; Caselles, V.; Coll, C.

    2008-10-01

    Surface emissivity in the thermal infrared (TIR) region is an important parameter for determining the land surface temperature from remote sensing measurements. This work compares the emissivities measured by different field methods (the Box method and the Temperature and Emissivity Separation, TES, algorithm) as well as emissivity data from ASTER scenes and the spectra obtained from the ASTER Spectral Library. The study was performed with a field radiometer having TIR bands with central wavelengths at 11.3 μm, 10.6 μm, 9.1 μm, 8.7 μm and 8.4 μm, similar to the ASTER TIR bands. The measurements were made at two sites in southern New Mexico. The first was in the White Sands National Monument, and the second was an open shrub land in the Jornada Experimental Range, in the northern Chihuahuan Desert, New Mexico, USA. The measurements show that, in general, emissivities derived with the Box method agree within 3% with those derived with the TES method for the spectral bands centered at 10.6 μm and 11.3 μm. However, the emissivities for the shorter wavelength bands are higher when derived with the Box method than those with the TES algorithm (differences range from 2% to 7%). The field emissivities agree within 2% with the laboratory spectrum for the 8-13 μm, 11.3 μm and 10.6 μm bands. However, the field and laboratory measurements in general differ from 3% to 16% for the shorter wavelength bands, i.e., 9.1 μm, 8.6 μm and 8.4 μm. A good agreement between the experimental measurements and the ASTER TIR emissivity data is observed for White Sands, especially over the 9 - 12 μm range (agreement within 4%). The study showed an emissivity increase up to 17% in the 8 to 9 μm range and an increase of 8% in emissivity ratio of average channels (8.4 μm, 8.6 μm, 9.1 μm):(10.6 μm, 11.3 μm) for two gypsum samples with different water content.

  16. Assessing the land cover situation in Surkhang, Upper Mustang, Nepal, using an ASTER image

    NARCIS (Netherlands)

    Sharma, B.D.; Clevers, J.G.P.W.; Graaf, de N.R.; Chapagain, N.R.

    2003-01-01

    This paper describes the remote sensing technique used to prepare a land cover map of Surkhang, Upper Mustang Nepal. The latest ASTER image (October 2002) and an ASTER DEM were used for the land cover classification. The study was carried out in Surkhang Village Development Committee (area 799 km2)

  17. Laminin/β1 integrin signal triggers axon formation by promoting microtubule assembly and stabilization

    Institute of Scientific and Technical Information of China (English)

    Wen-Liang Lei; Shi-Ge Xing; Cai-Yun Deng; Xiang-Chun Ju; Xing-Yu Jiang; Zhen-Ge Luo

    2012-01-01

    Axon specification during neuronal polarization is closely associated with increased microtubule stabilization in one of the neurites of unpolarized neuron,but how this increased microtubule stability is achieved is unclear.Here,we show that extracellular matrix (ECM) component laminin promotes neuronal polarization via regulating directional microtubule assembly through β1 integrin (Itgb1).Contact with laminin coated on culture substrate or polystyrene beads was sufficient for axon specification of undifferentiated neurites in cultured hippocampal neurons and cortical slices.Active Itgb1 was found to be concentrated in laminin-contacting neurites.Axon formation was promoted and abolished by enhancing and attenuating Itgbl signaling,respectively.Interestingly,laminin contact promoted plus-end microtubule assembly in a manner that required Itgbl.Moreover,stabilizing microtubules partially prevented polarization defects caused by ltgbl downregulation.Finally,genetic ablation of ltgbl in dorsal telencephalic progenitors caused deficits in axon development of cortical pyramidal neurons.Thus,laminin/Itgb1 signaling plays an instructive role in axon initiation and growth,both in vitro and in vivo,through the regulation of microtubule assembly.This study has established a linkage between an extrinsic factor and intrinsic cytoskeletai dynamics during neuronal polarization.

  18. Symmetry Breaking in an Edgeless Epithelium by Fat2-Regulated Microtubule Polarity

    Directory of Open Access Journals (Sweden)

    Dong-Yuan Chen

    2016-05-01

    Full Text Available Planar cell polarity (PCP information is a critical determinant of organ morphogenesis. While PCP in bounded epithelial sheets is increasingly well understood, how PCP is organized in tubular and acinar tissues is not. Drosophila egg chambers (follicles are an acinus-like “edgeless epithelium” and exhibit a continuous, circumferential PCP that does not depend on pathways active in bounded epithelia; this follicle PCP directs formation of an ellipsoid rather than a spherical egg. Here, we apply an imaging algorithm to “unroll” the entire 3D tissue surface and comprehensively analyze PCP onset. This approach traces chiral symmetry breaking to plus-end polarity of microtubules in the germarium, well before follicles form and rotate. PCP germarial microtubules provide chiral information that predicts the direction of whole-tissue rotation as soon as independent follicles form. Concordant microtubule polarity, but not microtubule alignment, requires the atypical cadherin Fat2, which acts at an early stage to translate plus-end bias into coordinated actin-mediated collective cell migration. Because microtubules are not required for PCP or migration after follicle rotation initiates, while dynamic actin and extracellular matrix are, polarized microtubules lie at the beginning of a handoff mechanism that passes early chiral PCP of the cytoskeleton to a supracellular planar polarized extracellular matrix and elongates the organ.

  19. Early stages of spindle formation and independence of chromosome and microtubule cycles in Haemanthus endosperm.

    Science.gov (United States)

    Smirnova, E A; Bajer, A S

    1998-01-01

    We analyzed transformation of the interphase microtubular cytoskeleton into the prophase spindle and followed the pattern of spindle axis determination. Microtubules in endosperm of the higher plant Haemanthus (Scadoxus) were stained by the immunogold and immunogold silver-enhanced methods. Basic structural units involved in spindle morphogenesis were "microtubule converging centers." We emphasized the importance of relative independence of chromosomal and microtubular cycles, and the influence of these cycles on the progress of mitosis. Cells with moderately desynchronized cycles were functional, but extreme desynchronization led to aberrant mitosis. There were three distinct phases of spindle development. The first one comprised interphase and early to mid-prophase. During this phase, the interphase microtubule meshwork radiating from the nuclear surface into the cytoplasm rearranged and formed a dense microtubule cage around the nucleus. The second phase comprised mid to late prophase, and resulted in the formation of normal (bipolar) or transitory aberrant (apolar or multipolar) prophase spindles. The third phase comprised late prophase with prometaphase. The onset of prometaphase was accompanied by a rapid association of microtubule converging centers with kinetochores. In this stage aberrant spindles transformed invariably into bipolar ones. Lateral association of a few bipolar kinetochore fibers at early prometaphase established the core of the bipolar spindle and its alignment. We concluded that (1) spindle formation is a largely independent microtubular process modified by the chromosomal/kinetochore cycle; and (2) the initial polarity of the spindle is established by microtubule converging centers, which are a functional substitute of the centrosome/MTOC. We believe that the dynamics of microtubule converging centers is an expression of microtubule self-organization driven by motor proteins as proposed by Mitchison [1992: Philos. Trans. R. Soc. Lond. B

  20. Identification of a TPX2-like microtubule-associated protein in Drosophila.

    Directory of Open Access Journals (Sweden)

    Gohta Goshima

    Full Text Available Chromosome segregation during mitosis and meiosis relies on the spindle and the functions of numerous microtubule-associated proteins (MAPs. One of the best-studied spindle MAPs is the highly conserved TPX2, which has been reported to have characteristic intracellular dynamics and molecular activities, such as nuclear localisation in interphase, poleward movement in the metaphase spindle, microtubule nucleation, microtubule stabilisation, microtubule bundling, Aurora A kinase activation, kinesin-5 binding, and kinesin-12 recruitment. This protein has been shown to be essential for spindle formation in every cell type analysed so far. However, as yet, TPX2 homologues have not been found in the Drosophila genome. In this study, I found that the Drosophila protein Ssp1/Mei-38 has significant homology to TPX2. Sequence conservation was limited to the putative spindle microtubule-associated region of TPX2, and intriguingly, D-TPX2 (Ssp1/Mei-38 lacks Aurora A- and kinesin-5-binding domains, which are highly conserved in other animal and plant species, including many insects such as ants and bees. D-TPX2 uniformly localised to kinetochore microtubule-enriched regions of the metaphase spindle in the S2 cell line, and it had microtubule binding and bundling activities in vitro. In comparison with other systems, the contribution of D-TPX2 to cell division seems to be minor; live cell imaging of microtubules and chromosomes after RNAi knockdown identified significant delay in chromosome congression in only 18% of the cells. Thus, while this conserved spindle protein is present in Drosophila, other mechanisms may largely compensate for its spindle assembly and chromosome segregation functions.

  1. Decoherence Time of a Microtubule

    CERN Document Server

    Hiramatsu, T; Sakakibara, K; Hiramatsu, Takashi; Matsui, Tetsuo; Sakakibara, Kazuhiko

    2006-01-01

    We formulate and study a quantum field theory of a microtubule, a basic element of living cells. Following the quantum theory of consciousness by Hameroff and Penrose, we let the system to make self-reductions, and measure the decoherence time $\\tau_N$ (the mean interval between two successive reductions) of a cluster consisting of more than $N$ neighboring cells (tubulins). $\\tau_N$ is interpreted as an instance of the stream of consciousness. For a sufficiently small electron hopping amplitude, $\\tau_N$ obeys an exponential law, $\\tau_N \\sim \\exp(c' N)$, and may take realistic values $\\tau_N $ \\raisebox{-0.5ex} {$\\stackrel{>}{\\sim}$} $ 10^{-2}$ sec for $N \\raisebox{-0.5ex} {$\\stackrel{>}{\\sim}$} 1100$.

  2. Two-state mechanochemical model for microtubule growth

    CERN Document Server

    Zhang, Yunxin

    2011-01-01

    In this study, a two-state mechanochemical model is presented to describe the dynamic properties of microtubule (MT) growth in cells. The MT switches between two states, assembly state and disassembly state. In assembly state, the growth of microtubule includes two processes: GTP-tubulin binding to the tip of protofilament (PF) and conformational change of PF, during which the penultimate GTP is hydrolyzed and the first tubulin unit that curls out the MT surface is rearranged into MT surface using the energy released from GTP hydrolysis. In disassembly state, the shortening of microtubule is also described by two processes, the release of GDP-tibulin from the tip of PF and one new tubulin unit curls out from the MT surface. Switches between these two states, which are usually called rescue and catastrophe, happen stochastically with external force dependent rates. Using this two-state model with parameters obtained by fitting the recent experimental data, detailed properties of MT growth are obtained, we find...

  3. A mechanism for reorientation of cortical microtubule arrays driven by microtubule severing.

    Science.gov (United States)

    Lindeboom, Jelmer J; Nakamura, Masayoshi; Hibbel, Anneke; Shundyak, Kostya; Gutierrez, Ryan; Ketelaar, Tijs; Emons, Anne Mie C; Mulder, Bela M; Kirik, Viktor; Ehrhardt, David W

    2013-12-06

    Environmental and hormonal signals cause reorganization of microtubule arrays in higher plants, but the mechanisms driving these transitions have remained elusive. The organization of these arrays is required to direct morphogenesis. We discovered that microtubule severing by the protein katanin plays a crucial and unexpected role in the reorientation of cortical arrays, as triggered by blue light. Imaging and genetic experiments revealed that phototropin photoreceptors stimulate katanin-mediated severing specifically at microtubule intersections, leading to the generation of new microtubules at these locations. We show how this activity serves as the basis for a mechanism that amplifies microtubules orthogonal to the initial array, thereby driving array reorientation. Our observations show how severing is used constructively to build a new microtubule array.

  4. Structural comparison of the Caenorhabditis elegans and human Ndc80 complexes bound to microtubules reveals distinct binding behavior

    Science.gov (United States)

    Wilson-Kubalek, Elizabeth M.; Cheeseman, Iain M.; Milligan, Ronald A.

    2016-01-01

    During cell division, kinetochores must remain tethered to the plus ends of dynamic microtubule polymers. However, the molecular basis for robust kinetochore–microtubule interactions remains poorly understood. The conserved four-subunit Ndc80 complex plays an essential and direct role in generating dynamic kinetochore–microtubule attachments. Here we compare the binding of the Caenorhabditis elegans and human Ndc80 complexes to microtubules at high resolution using cryo–electron microscopy reconstructions. Despite the conserved roles of the Ndc80 complex in diverse organisms, we find that the attachment mode of these complexes for microtubules is distinct. The human Ndc80 complex binds every tubulin monomer along the microtubule protofilament, whereas the C. elegans Ndc80 complex binds more tightly to β-tubulin. In addition, the C. elegans Ndc80 complex tilts more toward the adjacent protofilament. These structural differences in the Ndc80 complex between different species may play significant roles in the nature of kinetochore–microtubule interactions. PMID:26941333

  5. Centriolar CPAP/SAS-4 Imparts Slow Processive Microtubule Growth.

    Science.gov (United States)

    Sharma, Ashwani; Aher, Amol; Dynes, Nicola J; Frey, Daniel; Katrukha, Eugene A; Jaussi, Rolf; Grigoriev, Ilya; Croisier, Marie; Kammerer, Richard A; Akhmanova, Anna; Gönczy, Pierre; Steinmetz, Michel O

    2016-05-23

    Centrioles are fundamental and evolutionarily conserved microtubule-based organelles whose assembly is characterized by microtubule growth rates that are orders of magnitude slower than those of cytoplasmic microtubules. Several centriolar proteins can interact with tubulin or microtubules, but how they ensure the exceptionally slow growth of centriolar microtubules has remained mysterious. Here, we bring together crystallographic, biophysical, and reconstitution assays to demonstrate that the human centriolar protein CPAP (SAS-4 in worms and flies) binds and "caps" microtubule plus ends by associating with a site of β-tubulin engaged in longitudinal tubulin-tubulin interactions. Strikingly, we uncover that CPAP activity dampens microtubule growth and stabilizes microtubules by inhibiting catastrophes and promoting rescues. We further establish that the capping function of CPAP is important to limit growth of centriolar microtubules in cells. Our results suggest that CPAP acts as a molecular lid that ensures slow assembly of centriolar microtubules and, thereby, contributes to organelle length control.

  6. The smallest active fragment of microtubule-associated protein 4 and its interaction with microtubules in phosphate buffer.

    Science.gov (United States)

    Hashi, Yurika; Nagase, Lisa; Matsushima, Kazuyuki; Kotani, Susumu

    2012-01-01

    To analyze the interaction between microtubule-associated protein (MAP) 4 and microtubules physicochemically, a MAP4 active site fragment was designed for nuclear magnetic resonance (NMR) use. The fragment was bacterially expressed and purified to homogeneity. The buffer conditions for NMR were optimized to support microtubule assembly. The fragment was found to bind to microtubules under the optimized buffer conditions.

  7. Applying Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) spectral indices for geological mapping and mineral identification on the Tibetan Plateau

    CERN Document Server

    Corrie, Robert; Aitchison, Jonathan

    2011-01-01

    The Tibetan Plateau holds clues to understanding the dynamics and mechanisms associated with continental growth. Part of the region is characterized by zones of ophiolitic melange believed to represent the remnants of ancient oceanic crust and underlying upper mantle emplaced during oceanic closures. However, due to the remoteness of the region and the inhospitable terrain many areas have not received detailed investigation. Increased spatial and spectral resolution of satellite sensors have made it possible to map in greater detail the mineralogy and lithology than in the past. Recent work by Yoshiki Ninomiya of the Geological Survey of Japan has pioneered the use of several spectral indices for the mapping of quartzose, carbonate, and silicate rocks using Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) thermal infrared (TIR) data. In this study, ASTER TIR indices have been applied to a region in western-central Tibet for the purposes of assessing their effectiveness for differentiatin...

  8. One-parameter nonrelativistic supersymmetry for microtubules

    CERN Document Server

    Rosu, H C

    2003-01-01

    The simple supersymmetric model of Caticha [PRA 51, 4264 (1995)], as used by Rosu [PRE 55, 2038 (1997)] for microtubules, is generalized to the case of Mielnik's one-parameter nonrelativistic susy [JMP 25, 3387 (1984)

  9. INTEGRATION OF PALSAR AND ASTER SATELLITE DATA FOR GEOLOGICAL MAPPING IN TROPICS

    Directory of Open Access Journals (Sweden)

    A. Beiranvand Pour

    2015-10-01

    Full Text Available This research investigates the integration of the Phased Array type L-band Synthetic Aperture Radar (PALSAR and the Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER satellite data for geological mapping applications in tropical environments. The eastern part of the central belt of peninsular Malaysia has been investigated to identify structural features and mineral mapping using PALSAR and ASTER data. Adaptive local sigma and directional filters were applied to PALSAR data for detecting geological structure elements in the study area. The vegetation, mineralogic and lithologic indices for ASTER bands were tested in tropical climate. Lineaments (fault and fractures and curvilinear (anticline or syncline were detected using PALSAR fused image of directional filters (N-S, NE-SW, and NW-SE.Vegetation index image map show vegetation cover by fusing ASTER VNIR bands. High concentration of clay minerals zone was detected using fused image map derived from ASTER SWIR bands. Fusion of ASTER TIR bands produced image map of the lithological units. Results indicate that data integration and data fusion from PALSAR and ASTER sources enhanced information extraction for geological mapping in tropical environments.

  10. The human kinesin-14 HSET tracks the tips of growing microtubules in vitro.

    Science.gov (United States)

    Braun, Marcus; Lansky, Zdenek; Bajer, Seweryn; Fink, Gero; Kasprzak, Andrzej A; Diez, Stefan

    2013-09-01

    Tip-tracking of kinesin-14 motor proteins is believed to be crucial for the assembly and maintenance of dynamic microtubule arrays. However, in contrast to other members of the kinesin-14 family, H. sapiens kinesin-14 HSET has so far never been observed to be prominently located at microtubule plus ends. Here, using an in vitro microtubule dynamics reconstitution assay we observe tip-tracking of GFP-HSET in the presence of H. sapiens EB1 (hsEB1). Tip-tracking depended on the SxIP-like motif in HSET as well as on the EB homology domain in hsEB1. D. melanogaster Ncd and S. pombe Klp2 tip-tracking reconstitution assays accompanied by kinesin-14 amino acid sequence comparisons suggest that SxIP-like motif mediated tip-tracking dependent on EB family proteins is conserved in the kinesin-14 family of molecular motors.

  11. Tsunami Inundation, North of Phuket, Thailand ASTER Images and SRTM Elevation Model

    Science.gov (United States)

    2005-01-01

    with critical information for surface mapping, and monitoring of dynamic conditions and temporal change. Example applications are: monitoring glacial advances and retreats; monitoring potentially active volcanoes; identifying crop stress; determining cloud morphology and physical properties; wetlands evaluation; thermal pollution monitoring; coral reef degradation; surface temperature mapping of soils and geology; and measuring surface heat balance. ASTER is one of five Earth-observing instruments launched December 18, 1999, on NASA's Terra satellite. The instrument was built by Japan's Ministry of Economy, Trade and Industry. A joint U.S./Japan science team is responsible for validation and calibration of the instrument and the data products. The U.S. science team is located at NASA's Jet Propulsion Laboratory, Pasadena, Calif. The Terra mission is part of NASA's Science Mission Directorate. Elevation data used in this image were acquired by the Shuttle Radar Topography Mission (SRTM) aboard the Space Shuttle Endeavour, launched on February 11, 2000. SRTM used the same radar instrument that comprised the Spaceborne Imaging Radar-C/X-Band Synthetic Aperture Radar (SIR-C/X-SAR) that flew twice on the Space Shuttle Endeavour in 1994. SRTM was designed to collect three-dimensional measurements of the Earth's surface. To collect the 3-D data, engineers added a 60-meter-long (200-foot) mast, installed additional C-band and X-band antennas, and improved tracking and navigation devices. The mission is a cooperative project between the National Aeronautics and Space Administration (NASA), the National Geospatial-Intelligence Agency (NGA) of the U.S. Department of Defense (DoD), and the German and Italian space agencies. It is managed by NASA's Jet Propulsion Laboratory, Pasadena, Calif., for NASA's Science Mission Directorate, Washington, D.C. Size: 9.75 x 27.6 kilometers (6.0 x 17.1 miles), Location: 8.6 degrees North latitude, 98.3 degrees East longitude Orientation: Top is

  12. Oxidative stress decreases microtubule growth and stability in ventricular myocytes.

    Science.gov (United States)

    Drum, Benjamin M L; Yuan, Can; Li, Lei; Liu, Qinghang; Wordeman, Linda; Santana, L Fernando

    2016-04-01

    Microtubules (MTs) have many roles in ventricular myocytes, including structural stability, morphological integrity, and protein trafficking. However, despite their functional importance, dynamic MTs had never been visualized in living adult myocytes. Using adeno-associated viral vectors expressing the MT-associated protein plus end binding protein 3 (EB3) tagged with EGFP, we were able to perform live imaging and thus capture and quantify MT dynamics in ventricular myocytes in real time under physiological conditions. Super-resolution nanoscopy revealed that EB1 associated in puncta along the length of MTs in ventricular myocytes. The vast (~80%) majority of MTs grew perpendicular to T-tubules at a rate of 0.06μm∗s(-1) and growth was preferentially (82%) confined to a single sarcomere. Microtubule catastrophe rate was lower near the Z-line than M-line. Hydrogen peroxide increased the rate of catastrophe of MTs ~7-fold, suggesting that oxidative stress destabilizes these structures in ventricular myocytes. We also quantified MT dynamics after myocardial infarction (MI), a pathological condition associated with increased production of reactive oxygen species (ROS). Our data indicate that the catastrophe rate of MTs increases following MI. This contributed to decreased transient outward K(+) currents by decreasing the surface expression of Kv4.2 and Kv4.3 channels after MI. On the basis of these data, we conclude that, under physiological conditions, MT growth is directionally biased and that increased ROS production during MI disrupts MT dynamics, decreasing K(+) channel trafficking.

  13. Motor function in interpolar microtubules during metaphase

    OpenAIRE

    Deutsch, J. M.; Lewis, Ian P.

    2013-01-01

    We analyze experimental observations of microtubules undergoing small fluctuations about a "balance point" when mixed in solution of two different kinesin motor proteins, KLP61F and Ncd. It has been proposed that the microtubule movement is due to stochastic variations in the densities of the two species of motor proteins. We test this hypothesis here by showing how it maps onto a one-dimensional random walk in a random environment. Our estimate of the amplitude of the fluctuations agrees wit...

  14. TIPsy tour guides: How microtubule plus-end tracking proteins (+TIPs facilitate axon guidance

    Directory of Open Access Journals (Sweden)

    Elizabeth A Bearce

    2015-06-01

    Full Text Available The growth cone is a dynamic cytoskeletal vehicle, which drives the end of a developing axon. It serves to interpret and navigate through the complex landscape and guidance cues of the early nervous system. The growth cone’s distinctive cytoskeletal organization offers a fascinating platform to study how extracellular cues can be translated into mechanical outgrowth and turning behaviors. While many studies of cell motility highlight the importance of actin networks in signaling, adhesion, and propulsion, both seminal and emerging works in the field have highlighted a unique and necessary role for microtubules in growth cone navigation. Here, we focus on the role of singular pioneer microtubules, which extend into the growth cone periphery and are regulated by a diverse family of microtubule plus-end tracking proteins (+TIPs. These +TIPs accumulate at the dynamic ends of microtubules, where they are well-positioned to encounter and respond to key signaling events downstream of guidance receptors, catalyzing immediate changes in microtubule stability and actin cross-talk, that facilitate both axonal outgrowth and turning events.

  15. Response of mycorrhizal periwinkle plants to aster yellows phytoplasma infection.

    Science.gov (United States)

    Kamińska, Maria; Klamkowski, Krzysztof; Berniak, Hanna; Sowik, Iwona

    2010-03-01

    The objective of our research was to assess if arbuscular mycorrhizal (AM) fungal colonization can modify the effect of infection by two aster yellows phytoplasma strains (AY1, AYSim) in Catharanthus roseus plants. Both phytoplasma strains had a negative effect on the root fresh weight, but they differed in symptoms appearance and in their influence on photosynthetic and transpiration rates of the periwinkle plants. AM plants showed significantly reduced shoot fresh weight, while the transpiration rate was significantly increased. AM fungal colonization significantly affected shoot height and fresh weight of the plants infected by each phytoplasma strains as well as the root system of plants infected with the more aggressive AYSim phytoplasma strain. Double inoculation did not reduce the negative effects induced with phytoplasma alone on the photosynthetic activity of phytoplasma-infected plants.

  16. Poly-Pattern Compressive Segmentation of ASTER Data for GIS

    Science.gov (United States)

    Myers, Wayne; Warner, Eric; Tutwiler, Richard

    2007-01-01

    Pattern-based segmentation of multi-band image data, such as ASTER, produces one-byte and two-byte approximate compressions. This is a dual segmentation consisting of nested coarser and finer level pattern mappings called poly-patterns. The coarser A-level version is structured for direct incorporation into geographic information systems in the manner of a raster map. GIs renderings of this A-level approximation are called pattern pictures which have the appearance of color enhanced images. The two-byte version consisting of thousands of B-level segments provides a capability for approximate restoration of the multi-band data in selected areas or entire scenes. Poly-patterns are especially useful for purposes of change detection and landscape analysis at multiple scales. The primary author has implemented the segmentation methodology in a public domain software suite.

  17. The role of microtubule-associated protein 1B in axonal growth and neuronal migration in the central nervous system

    Institute of Scientific and Technical Information of China (English)

    Maoguang Yang; Xiaoyu Yang; Minfei Wu; Peng Xia; Chunxin Wang; Peng Yan; Qi Gao; Jian Liu; Haitao Wang; Xingwei Duan

    2012-01-01

    In this review, we discuss the role of microtubule-associated protein 1B (MAP1B) and its phosphorylation in axonal development and regeneration in the central nervous system. MAP1B exhibits similar functions during axonal development and regeneration. MAP1B and phosphorylated MAP1B in neurons and axons maintain a dynamic balance between cytoskeletal components, and regulate the stability and interaction of microtubules and actin to promote axonal growth, neural connectivity and regeneration in the central nervous system.

  18. Pseudo-Spin Model for the Cytoskeletal Microtubule Surface

    Institute of Scientific and Technical Information of China (English)

    CHEN Ying; QIU Xi-Jun; LI Ru-Xin

    2004-01-01

    @@ Due to the inherent symmetry structures and the electric properties in the microtubule (MT), we treat the MT as a one-dimensional ferroelectric system and describe the nonlinear dynamics of dimer electric dipoles in one protofilament of the MT by virtue of the double-well potential. Consequently, the physical problem has been mapped onto the pseudo-spin system, and the mean-field approximation has been taken to obtain some physical results, including the expression for the phase transition temperature Tc and the estimated value of Tc (≈ 312 K).

  19. Thermal mapping of Hawaiian volcanoes with ASTER satellite data

    Science.gov (United States)

    Patrick, Matthew R.; Witzke, Coral-Nadine

    2011-01-01

    Thermal mapping of volcanoes is important to determine baseline thermal behavior in order to judge future thermal activity that may precede an eruption. We used cloud-free kinetic temperature images from the ASTER (Advanced Spaceborne Thermal Emission and Reflection Radiometer) sensor obtained between 2000 and 2010 to produce thermal maps for all five subaerial volcanoes in Hawai‘i that have had eruptions in the Holocene (Kīlauea, Mauna Loa, Hualālai, Mauna Kea, and Haleakalā). We stacked the images to provide time-averaged thermal maps, as well as to analyze temperature trends through time. Thermal areas are conspicuous at the summits and rift zones of Kīlauea and Mauna Loa, and the summit calderas of these volcanoes contain obvious arcuate, concentric linear thermal areas that probably result from channeling of rising gas along buried, historical intracaldera scarps. The only significant change in thermal activity noted in the study period is the opening of the Halema‘uma‘u vent at Kīlauea's summit in 2008. Several small thermal anomalies are coincident with pit craters on Hualālai. We suspect that these simply result from the sheltered nature of the depression, but closer inspection is warranted to determine if genuine thermal activity exists in the craters. Thermal areas were not detected on Haleakalā or Mauna Kea. The main limitation of the study is the large pixel size (90 m) of the ASTER images, which reduces our ability to detect subtle changes or to identify small, low-temperature thermal activity. This study, therefore, is meant to characterize the broad, large-scale thermal features on these volcanoes. Future work should study these thermal areas with thermal cameras and thermocouples, which have a greater ability to detect small, low-temperature thermal features.

  20. Role of Exchange Protein Activated by cAMP 1 in Regulating Rates of Microtubule Formation in Cystic Fibrosis Epithelial Cells.

    Science.gov (United States)

    Rymut, Sharon M; Ivy, Tracy; Corey, Deborah A; Cotton, Calvin U; Burgess, James D; Kelley, Thomas J

    2015-12-01

    The regulation of microtubule dynamics in cystic fibrosis (CF) epithelial cells and the consequences of reduced rates of microtubule polymerization on downstream CF cellular events, such as cholesterol accumulation, a marker of impaired intracellular transport, are explored here. It is identified that microtubules in both CF cell models and in primary CF nasal epithelial cells repolymerize at a slower rate compared with respective controls. Previous studies suggest a role for cAMP in modulating organelle transport in CF cells, implicating a role for exchange protein activated by cAMP (EPAC) 1, a regulator of microtubule elongation, as a potential mechanism. EPAC1 activity is reduced in CF cell models and in Cftr(-/-) mouse lung compared with respective non-CF controls. Stimulation of EPAC1 activity with the selective EPAC1 agonist, 8-cpt-2-O-Me-cAMP, stimulates microtubule repolymerization to wild-type rates in CF cells. EPAC1 activation also alleviates cholesterol accumulation in CF cells, suggesting a direct link between microtubule regulation and intracellular transport. To verify the relationship between transport and microtubule regulation, expression of the protein, tubulin polymerization-promoting protein, was knocked down in non-CF human tracheal (9/HTEo(-)) cells to mimic the microtubule dysregulation in CF cells. Transduced cells with short hairpin RNA targeting tubulin polymerization-promoting protein exhibit CF-like perinuclear cholesterol accumulation and other cellular manifestations of CF cells, thus supporting a role for microtubule regulation as a mechanism linking CFTR function to downstream cellular manifestation.

  1. Glacial Ice Velocity Determination and Correlation from Different Mountain Ranges Using Aster Imagery

    Science.gov (United States)

    Pleasants, M.; Haritashya, U. K.

    2013-12-01

    Mountain glaciers make good indicators of even slight changes in climatic conditions because of their sensitivity to temperature and other environmental changes. Due to the inaccessibility of most mountain glaciers, field based measurements of glacier dynamics, especially ice velocities, has proved to be difficult and unrealistic. Because of this, evaluation of satellite imagery has become useful in the determination of glacial ice velocities and production of ice flow models. The calculation and comparison of ice velocities from three glaciated regions (the Southern Alps of New Zealand, the North-east Nepali Himalaya, and the south-eastern Chugach Mountains of Alaska) is presented here. This study was completed to establish the accuracy and global applicability of the method of precise orthorectification, co-registration, and correlation using the software COSI-Corr and in-house filtering techniques. We chose glaciers from different mountain ranges that present different dynamics to establish a specific ice velocity method. Advanced Spacebourne Thermal Emission and Reflection Radiometer (ASTER) L1A images were chosen as close to annual pairs as possible from the available data at the USGS Global Visualization Viewer (GloVis). This study focuses on the Tasman glacier in New Zealand, the Khumbu glacier in the Himalayas, the previously unstudied Scott glacier in Alaska, and the comparison of these ice velocities to climate data for each region compiled from numerous sources in an attempt to draw conclusions of the behavior of these glaciers in response to current climatic changes.

  2. Rearrangements of microtubule cytoskeleton in stomatal closure of Arabidopsis induced by nitric oxide

    Institute of Scientific and Technical Information of China (English)

    ZHANG YongMei; WU ZhongYi; WANG XueChen; YU Rong

    2008-01-01

    NO (nitric oxide), known as a key signal molecule in plant, plays important roles in regulation of stomatal movement. In this study, microtubule dynamics and its possible mechanism in the NO signal pathway were investigated. The results were as follows: (ⅰ) In vivo stomatal aperture assays revealed that both vinblastine (microtubule-disrupting drug) and SNP (exogenous NO donor) prevented stomatal opening in the light, and vinblastine even could enhance the inhibitory effect of SNP, whereas taxol (a microtubule-stabilizing agent) was able to reduce this effect; (ⅱ) microtubules in the opening Arabi-dopsis guard cells expressing GFP:α-tubulin-6 (AtGFP:α-tubulin-6) were organized in parallel, straight and dense bundles, radiating from the ventral side to the dorsal side, and most of them were localized perpendicularly to the ventral wall; (ⅲ) under the same environmental conditions, treated with SNP for 30 min, the radial arrays of microtubules in guard cells began to break down, twisted partially and be-came oblique or exhibited a random pattern; (ⅳ) furthermore, the involvement of cytosolic Ca2+ in this event was tested. Stomatal aperture assays revealed that BAPTA-AM (a chelator of Ca2+) greatly sup-pressed the effect of NO on stomatal closure; however, it did not show the same function on stomatal closure induced by vinblastine. When BAPTA-AM was added to the SNP-pretreated solution, the SNP-induced disordered microtubulue cytoskeleton in guard cells underwent rearrangement in a time-dependent manner. After 30 min of treatment with BAPTA-AM, the cortical microtubules resumed the original radial distribution, almost the same as the control. All this indicates that NO may promote rearrangement of microtubule cytoskeleton via elevation of [Ca2+]cyt (free Ca2+ concentration in the cy-toplasm), finally leading to stomatal closure.

  3. Evidence on Possible Mycoplasma Etiology of Aster Yellows Disease II. Suppression of Aster Yellows in Insect Vectors.

    Science.gov (United States)

    Whitcomb, R F; Davis, R E

    1970-08-01

    Chlortetracycline or chloramphenicol (but not kanamycin, penicillin, or erythromycin), when administered in hydroponic solution to diseased aster, reduced the availability of the aster yellows (AY) agent to nymphs of Macrosteles fascifrons (Stål). Insects exposed to healthy plants whose roots were immersed in chlortetracycline were able to acquire AY agent from diseased plants the day after removal from the antibiotic-treated plants, but the latent period of the ensuing disease in the insects was prolonged. Chlortetracycline or tylosin tartrate blocked AY infection in nymphs injected with a mixture of antibiotic and the AY agent, but polymyxin, neomycin, vancomycin, penicillin, carbomycin, or chloramphenicol did not. All tetracyclines tested, methacycline, oxytetracycline, and chlortetracycline, produced a dramatic reduction in the ability of infected vectors to transmit AY agent. Tylosin tartrate also reduced transmission when injected into AY-transmitting vectors, but carbomycin, spectinomycin, cycloserine, penicillin, erythromycin, or kanamycin had no such effect. During the first 10 days after injection of tylosin tartrate or oxytetracycline into transmitting vectors, ability of the insects to transmit AY decayed rapidly. Transmission by insects injected with buffer alone, after decreasing the first day after injection, gradually returned to its normal level in less than 1 week. By 2 to 3 weeks after injection with tylosin or oxytetracycline, ability to transmit AY was regained by vectors. The results suggest that tetracycline antibiotics and tylosin tartrate inhibit multiplication of AY agent in the insect. The spectrum of antibiotic activity in the insect is consistent with the hypothesis that AY and other plant yellows diseases are caused by mycoplasma-like organisms.

  4. ASTER Global Emissivity Data Set Monthly 0.05 degree V041 - AG5KMMOH

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) Global Emissivity Dataset (GED) is a collection of monthly files (see known issues for gaps)...

  5. ASTER Global Emissivity Dataset Monthly 0.05 degree NetCDF4

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) Global Emissivity Dataset (GED) is a collection of monthly files (see known issues for gaps)...

  6. ASTER Global Emissivity Dataset 1-kilometer Binary V003 - AG1KMB

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — The Advanced Spaceborne Thermal Emission and Reflection radiometer Global Emissivity Database (ASTER GED) was developed by the National Aeronautics and Space...

  7. ASTER Global Emissivity Dataset 100-meter Binary V003 - AG100B

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — The Advanced Spaceborne Thermal Emission and Reflection radiometer Global Emissivity Database (ASTER GED) was developed by the National Aeronautics and Space...

  8. ASTER Global Emissivity Dataset 1-kilometer V003 - AG1KM

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — The Advanced Spaceborne Thermal Emission and Reflection radiometer Global Emissivity Database (ASTER GED) was developed by the National Aeronautics and Space...

  9. ASTER Global Emissivity Dataset Monthly 0.05 degree HDF5

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) Global Emissivity Dataset (GED) is a collection of monthly files (see known issues for gaps)...

  10. beta-Dystroglycan modulates the interplay between actin and microtubules in human-adhered platelets.

    Science.gov (United States)

    Cerecedo, Doris; Cisneros, Bulmaro; Suárez-Sánchez, Rocío; Hernández-González, Enrique; Galván, Iván

    2008-05-01

    To maintain the continuity of an injured blood vessel, platelets change shape, secrete granule contents, adhere, aggregate, and retract in a haemostatic plug. Ordered arrays of microtubules, microfilaments, and associated proteins are responsible for these platelet responses. In full-spread platelets, microfilament bundles in association with other cytoskeleton proteins are anchored in focal contacts. Recent studies in migrating cells suggest that co-ordination and direct physical interaction of microtubules and actin network modulate adhesion development. In platelets, we have proposed a feasible association between these two cytoskeletal systems, as well as the participation of the dystrophin-associated protein complex, as part of the focal adhesion complex. The present study analysed the participation of microtubules and actin during the platelet adhesion process. Confocal microscopy, fluorescence resonance transfer energy and immunoprecipitation assays were used to provide evidence of a cross-talk between these two cytoskeletal systems. Interestingly, beta-dystroglycan was found to act as an interplay protein between actin and microtubules and an additional communication between these two cytoskeleton networks was maintained through proteins of focal adhesion complex. Altogether our data are indicative of a dynamic co-participation of actin filaments and microtubules in modulating focal contacts to achieve platelet function.

  11. A study of microtubule dipole lattices

    Science.gov (United States)

    Nandi, Shubhendu

    Microtubules are cytoskeletal protein polymers orchestrating a host of important cellular functions including, but not limited to, cell support, cell division, cell motility and cell transport. In this thesis, we construct a toy-model of the microtubule lattice composed of vector Ising spins representing tubulin molecules, the building block of microtubules. Nearest-neighbor and next-to-nearest neighbor interactions are considered within an anisotropic dielectric medium. As a consequence of the helical topology, we observe that certain spin orientations render the lattice frustrated with nearest neighbor ferroelectric and next-to-nearest neighbor antiferroelectric bonds. Under these conditions, the lattice displays the remarkable property of stabilizing certain spin patterns that are robust to thermal fluctuations. We model this behavior in the framework of a generalized Ising model known as the J1 - J2 model and theoretically determine the set of stable patterns. Employing Monte-Carlo methods, we demonstrate the stability of such patterns in the microtubule lattice at human physiological temperatures. This suggests a novel biological mechanism for storing information in living organisms, whereby the tubulin spin (dipole moment) states become information bits and information gets stored in microtubules in a way that is robust to thermal fluctuations.

  12. On-Orbit Spatial Characterization of MODIS with ASTER Aboard the Terra Spacecraft

    Science.gov (United States)

    Xie, Yong; Xiong, Xiaoxiong

    2011-01-01

    This letter presents a novel approach for on-orbit characterization of MODerate resolution Imaging Spectroradiometer (MODIS) band-to-band registration (BBR) using Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) aboard the Terra spacecraft. The spatial resolution of ASTER spectral bands is much higher than that of MODIS, making it feasible to characterize MODIS on-orbit BBR using their simultaneous observations. The ground target selected for on-orbit MODIS BBR characterization in this letter is a water body, which is a uniform scene with high signal contrast relative to its neighbor areas. A key step of this approach is to accurately localize the measurements of each MODIS band in an ASTER measurement plane coordinate (AMPC). The ASTER measurements are first interpolated and aggregated to simulate the measurements of each MODIS band. The best measurement match between ASTER and each MODIS band is obtained when the measurement difference reaches its weighted minimum. The position of each MODIS band in the AMPC is then used to calculate the BBR. The results are compared with those derived from MODIS onboard Spectro-Radiometric Calibration Assembly. They are in good agreement, generally less than 0.1 MODIS pixel. This approach is useful for other sensors without onboard spatial characterization capability. Index Terms Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER), band-to-band registration (BBR), MODerate resolution Imaging Spectroradiometer (MODIS), spatial characterization.

  13. Novel insights into mammalian embryonic neural stem cell division: focus on microtubules.

    Science.gov (United States)

    Mora-Bermúdez, Felipe; Huttner, Wieland B

    2015-12-01

    During stem cell divisions, mitotic microtubules do more than just segregate the chromosomes. They also determine whether a cell divides virtually symmetrically or asymmetrically by establishing spindle orientation and the plane of cell division. This can be decisive for the fate of the stem cell progeny. Spindle defects have been linked to neurodevelopmental disorders, yet the role of spindle orientation for mammalian neurogenesis has remained controversial. Here we explore recent advances in understanding how the microtubule cytoskeleton influences mammalian neural stem cell division. Our focus is primarily on the role of spindle microtubules in the development of the cerebral cortex. We also highlight unique characteristics in the architecture and dynamics of cortical stem cells that are tightly linked to their mode of division. These features contribute to setting these cells apart as mitotic "rule breakers," control how asymmetric a division is, and, we argue, are sufficient to determine the fate of the neural stem cell progeny in mammals.

  14. Microtubule remodeling mediates the inhibition of store-operated calcium entry (SOCE) during mitosis in COS-7 cells.

    Science.gov (United States)

    Russa, Afadhali Denis; Ishikita, Naoyuki; Masu, Kazuki; Akutsu, Hitomi; Saino, Tomoyuki; Satoh, Yoh-ichi

    2008-12-01

    Regulation of the intracellular calcium ion concentration ([Ca(2+)](i)) is critical, because calcium signaling controls diverse and vital cellular processes such as secretion, proliferation, division, gene transcription, and apoptosis. Store-operated calcium entry (SOCE) is the main mechanism through which non-excitable cells replenish and thus maintain this delicate balance. There is limited evidence which indicates that SOCE may be inhibited during mitosis, and the mechanisms leading to the presumed inhibition has not been elucidated. In the present study, we examined and compared the [Ca(2+)](i) dynamics of COS-7 cells in mitotic and non-mitotic phases with special reference paid to SOCE. Laser scanning confocal microscopy to monitor [Ca(2+)](i) dynamics revealed that SOCE was progressively inhibited in mitosis and became virtually absent during the metaphase. We used various cytoskeletal modifying drugs and immunofluorescence to assess the contribution of microtubule and actin filaments in SOCE signaling. Nocodazole treatment caused microtubule reorganization and retraction from the cell periphery that mimicked the natural mitotic microtubule remodeling that was also accompanied by SOCE inhibition. Short exposure to paclitaxel, a microtubule-stabilizing drug, bolstered SOCE, whereas long exposure resulted in microtubule disruption and SOCE inhibition. Actin-modifying drugs did not affect SOCE. These findings indicate that mitotic microtubule remodeling plays a significant role in the inhibition of SOCE during mitosis.

  15. EOS Contract Report: The ASTER and MODIS Projects

    Science.gov (United States)

    Slater, P.; Thome, K. (Compiler)

    1997-01-01

    Three major tasks occupied the group's efforts during this six months. The first was measuring the bidirectional reflectance properties of four reflectance samples provided by NIST. S. Biggar and P. Spyak made these measurements in both the VNIR and SWIR. The second major task was the group's move to a new facility in March. This required that our calibration laboratory and blacklab be disassembled and reassembled in addition to moving offices and other equipment. The third task was the joint vicarious calibration that took place the latter half of June. This campaign included two weeks of laboratory measurements by the RSG and nine days in the field. Other work during the past six months consisted of Science Team support activities including the attendance at meetings related to MODIS and ASTER. In addition, K. Scott continued work on the cross-calibration software package by developing a graphical interface to 6S, an uncertainty analysis code, and an image registration module. M. Sicard used a trip to Cimel in France to change the Cimel TIR radiometer's field of view and then characterized this new field of view. Z. Rouf and Z. Murshalin processed radiance-based data from last summer's Lunar Lake campaign.

  16. Thermal remote sensing of ice-debris landforms using ASTER

    Science.gov (United States)

    Brenning, A.; Peña, M. A.; Long, S.; Soliman, A.

    2011-10-01

    Remote sensors face challenges in characterizing mountain permafrost and ground thermal conditions or mapping rock glaciers and debris-covered glaciers. We explore the potentials of thermal imaging and in particular thermal inertia mapping in mountain cryospheric research, focusing on the relationships between ground surface temperatures and the presence of ice-debris landforms on one side and land surface temperature (LST) and apparent thermal inertia (ATI) on the other. In our case study we utilize ASTER daytime and nighttime imagery and in-situ measurements of near-surface ground temperature (NSGT) in the Mediterranean Andes during a snow-free and dry observation period in late summer. Spatial patterns of LST and NSGT were mostly consistent with each other both at daytime and at nighttime. Daytime LST over ice-debris landforms was decreased and ATI consequently increased compared to other debris surfaces under otherwise equal conditions, but NSGT showed contradictory results, which underlines the complexity and possible scale dependence of ATI in heterogeneous substrates with the presence of a thermal mismatch and a heat sink at depth. While our results demonstrate the utility of thermal imaging and ATI mapping in a mountain cryospheric context, further research is needed for a better interpretation of ATI patterns in complex thermophysical conditions

  17. Evaluation of ASTER-Like Daily Land Surface Temperature by Fusing ASTER and MODIS Data during the HiWATER-MUSOEXE

    Directory of Open Access Journals (Sweden)

    Guijun Yang

    2016-01-01

    Full Text Available Land surface temperature (LST is an important parameter that is highly responsive to surface energy fluxes and has become valuable to many disciplines. However, it is difficult to acquire satellite LSTs with both high spatial and temporal resolutions due to tradeoffs between them. Thus, various algorithms/models have been developed to enhance the spatial or the temporal resolution of thermal infrared (TIR data or LST, but rarely both. The Spatial and Temporal Adaptive Reflectance Fusion Model (STARFM is the widely-used data fusion algorithm for Landsat and MODIS imagery to produce Landsat-like surface reflectance. In order to extend the STARFM application over heterogeneous areas, an enhanced STARFM (ESTARFM approach was proposed by introducing a conversion coefficient and the spectral unmixing theory. The aim of this study is to conduct a comprehensive evaluation of the ESTARFM algorithm for generating ASTER-like daily LST by three approaches: simulated data, ground measurements and remote sensing products, respectively. The datasets of LST ground measurements, MODIS, and ASTER images were collected in an arid region of Northwest China during the first thematic HiWATER-Multi-Scale Observation Experiment on Evapotranspiration (MUSOEXE over heterogeneous land surfaces in 2012 from May to September. Firstly, the results of the simulation test indicated that ESTARFM could accurately predict background with temperature variations, even coordinating with small ground objects and linear ground objects. Secondly, four temporal ASTER and MODIS data fusion LSTs (i.e., predicted ASTER-like LST products were highly consistent with ASTER LST products. Here, the four correlation coefficients were greater than 0.92, root mean square error (RMSE reached about 2 K and mean absolute error (MAE ranged from 1.32 K to 1.73 K. Finally, the results of the ground measurement validation indicated that the overall accuracy was high (R2 = 0.92, RMSE = 0.77 K, and the

  18. Biological Activity of 4-Substituted Methoxybenzoyl-Aryl-Thiazole (SMART): An Active Microtubule Inhibitor

    OpenAIRE

    2010-01-01

    Formation of microtubules is a dynamic process that involves polymerization and depolymerization of αβ-tubulin heterodimers. Drugs that enhance or inhibit tubulin polymerization can destroy this dynamic process, arresting cells in the G2/M phase of the cell cycle. Although drugs that target tubulin generally demonstrate cytotoxic potency in the sub-nanomolar range, resistance due to drug efflux is a common phenomenon among the antitubulin agents. We recently reported a class of 4-Substituted ...

  19. Microtubules guide root hair tip growth

    NARCIS (Netherlands)

    Sieberer, B.; Ketelaar, M.J.; Esseling, J.J.; Emons, A.M.C.

    2005-01-01

    The ability to establish cell polarity is crucial to form and function of an individual cell. Polarity underlies critical processes during cell development, such as cell growth, cell division, cell differentiation and cell signalling. Interphase cytoplasmic microtubules in tip-growing fission yeast

  20. Mechanical properties of organelles driven by microtubule-dependent molecular motors in living cells.

    Directory of Open Access Journals (Sweden)

    Luciana Bruno

    Full Text Available The organization of the cytoplasm is regulated by molecular motors which transport organelles and other cargoes along cytoskeleton tracks. Melanophores have pigment organelles or melanosomes that move along microtubules toward their minus and plus end by the action of cytoplasmic dynein and kinesin-2, respectively. In this work, we used single particle tracking to characterize the mechanical properties of motor-driven organelles during transport along microtubules. We tracked organelles with high temporal and spatial resolutions and characterized their dynamics perpendicular to the cytoskeleton track. The quantitative analysis of these data showed that the dynamics is due to a spring-like interaction between melanosomes and microtubules in a viscoelastic microenvironment. A model based on a generalized Langevin equation explained these observations and predicted that the stiffness measured for the motor complex acting as a linker between organelles and microtubules is ∼ one order smaller than that determined for motor proteins in vitro. This result suggests that other biomolecules involved in the interaction between motors and organelles contribute to the mechanical properties of the motor complex. We hypothesise that the high flexibility observed for the motor linker may be required to improve the efficiency of the transport driven by multiple copies of motor molecules.

  1. Discodermolide interferes with the binding of tau protein to microtubules.

    Science.gov (United States)

    Kar, Santwana; Florence, Gordon J; Paterson, Ian; Amos, Linda A

    2003-03-27

    We investigated whether discodermolide, a novel antimitotic agent, affects the binding to microtubules of tau protein repeat motifs. Like taxol, the new drug reduces the proportion of tau that pellets with microtubules. Despite their differing structures, discodermolide, taxol and tau repeats all bind to a site on beta-tubulin that lies within the microtubule lumen and is crucial in controlling microtubule assembly. Low concentrations of tau still bind strongly to the outer surfaces of preformed microtubules when the acidic C-terminal regions of at least six tubulin dimers are available for interaction with each tau molecule; otherwise binding is very weak.

  2. TCS1, a Microtubule-Binding Protein, Interacts with KCBP/ZWICHEL to Regulate Trichome Cell Shape in Arabidopsis thaliana.

    Directory of Open Access Journals (Sweden)

    Liangliang Chen

    2016-10-01

    Full Text Available How cell shape is controlled is a fundamental question in developmental biology, but the genetic and molecular mechanisms that determine cell shape are largely unknown. Arabidopsis trichomes have been used as a good model system to investigate cell shape at the single-cell level. Here we describe the trichome cell shape 1 (tcs1 mutants with the reduced trichome branch number in Arabidopsis. TCS1 encodes a coiled-coil domain-containing protein. Pharmacological analyses and observations of microtubule dynamics show that TCS1 influences the stability of microtubules. Biochemical analyses and live-cell imaging indicate that TCS1 binds to microtubules and promotes the assembly of microtubules. Further results reveal that TCS1 physically associates with KCBP/ZWICHEL, a microtubule motor involved in the regulation of trichome branch number. Genetic analyses indicate that kcbp/zwi is epistatic to tcs1 with respect to trichome branch number. Thus, our findings define a novel genetic and molecular mechanism by which TCS1 interacts with KCBP to regulate trichome cell shape by influencing the stability of microtubules.

  3. Evapotranspiration Estimation Using Multispectral Thermal Infrared Data from ASTER and MODIS

    Science.gov (United States)

    French, A. N.; Schmugge, T. J.; Kustas, W. P.; Prueger, J. H.

    2009-12-01

    Estimating evapotranspiration (ET) from space is important for monitoring water use at local and regional scales. Terra platform sensors ASTER and MODIS have been valuable for this goal because of their multispectral capabilities and high (90 m with ASTER) to moderate ( 1 km with MODIS) spatial resolutions. These capabilities have allowed discrimination of land cover conditions unobtainable from more conventional satellite imagery. In particular the multiple thermal channels provided by ASTER and MODIS have helped collect accurate observations of land surface temperature and emissivity which can be used to detect water stress and to distinguish between living and senescent vegetation. Each of these characteristics is important for modeling water fluxes. The frequent coverage by MODIS is also very important for this endeavor. To demonstrate how this can be accomplished, ASTER and Terra/MODIS data were modeled using images collected over the Jornada Experimental Range, a semi-arid research site in southern New Mexico. By combining 27 ASTER clear sky scenes with several hundred 1-km scale MODIS scenes between 2001 and 2003, it was feasible to estimate ET at weekly time steps and to also assess longer-term changes in vegetation distributions.

  4. Dual effect of procaine in sea urchin eggs. Inducer and inhibitor of microtubule assembly.

    Science.gov (United States)

    Coffe, G; Foucault, G; Raymond, M N; Pudles, J

    1985-01-01

    An increase in the amount of cytoplasmic filamentous structures (cytoplasmic matrix and aster) which were recovered after hexylene glycol/Triton X-100 treatment of sea urchin eggs (Paracentrotus lividus) activated by 0.2-2.5 mM procaine was observed. At higher activator concentrations, an opposite effect was observed and formation of these cytoplasmic structures was inhibited in the presence of 10 mM procaine. This inhibitory effect was reversed by diluting the drug in the incubation medium. DNase I inhibition assays on egg homogenates which were performed at different time points of the activation process, show that the same amount of actin was induced to polymerize in eggs activated either by 2.5 or 10 mM procaine. However, colchicine-binding assays on the 100 000 g particulate fractions of these homogenates show that in eggs activated by 10 mM procaine, in contrast to those activated by 2.5 mM, tubulin polymerization was inhibited and microtubules were disassembled. These results show that the dual effect of procaine in the organization of the egg cytoskeleton appears to be related to its effect on the state of tubulin.

  5. Doublecortin Is Excluded from Growing Microtubule Ends and Recognizes the GDP-Microtubule Lattice.

    Science.gov (United States)

    Ettinger, Andreas; van Haren, Jeffrey; Ribeiro, Susana A; Wittmann, Torsten

    2016-06-20

    Many microtubule (MT) functions are mediated by a diverse class of proteins (+TIPs) at growing MT plus ends that control intracellular MT interactions and dynamics and depend on end-binding proteins (EBs) [1]. Cryoelectron microscopy has recently identified the EB binding site as the interface of four tubulin dimers that undergoes a conformational change in response to β-tubulin GTP hydrolysis [2, 3]. Doublecortin (DCX), a MT-associated protein (MAP) required for neuronal migration during cortical development [4, 5], binds to the same site as EBs [6], and recent in vitro studies proposed DCX localization to growing MT ends independent of EBs [7]. Because this conflicts with observations in neurons [8, 9] and the molecular function of DCX is not well understood, we revisited intracellular DCX dynamics at low expression levels. Here, we report that DCX is not a +TIP in cells but, on the contrary, is excluded from the EB1 domain. In addition, we find that DCX-MT interactions are highly sensitive to MT geometry. In cells, DCX binding was greatly reduced at MT segments with high local curvature. Remarkably, this geometry-dependent binding to MTs was completely reversed in the presence of taxanes, which reconciles incompatible observations in cells [9] and in vitro [10]. We propose a model explaining DCX specificity for different MT geometries based on structural changes induced by GTP hydrolysis that decreases the spacing between adjacent tubulin dimers [11]. Our data are consistent with a unique mode of MT interaction in which DCX specifically recognizes this compacted GDP-like MT lattice.

  6. Microtubule-driven nuclear movements and linear elements as meiosis-specific characteristics of the fission yeasts Schizosaccharomyces versatilis and Schizosaccharomyces pombe.

    Science.gov (United States)

    Svoboda, A; Bähler, J; Kohli, J

    1995-11-01

    Meiotic prophase in Schizosaccharomyces pombe is characterized by striking nuclear movements and the formation of linear elements along chromosomes instead of tripartite synaptonemal complexes. We analysed the organization of nuclei and microtubules in cells of fission yeasts undergoing sexual differentiation. S. japonicus var. versatilis and S. pombe cells were studied in parallel, taking advantage of the better cytology in S. versatilis. During conjugation, microtubules were directed towards the mating projection. These microtubules seem to lead the haploid nuclei together in the zygote by interaction with the spindle pole bodies at the nuclear periphery. After karyogamy, arrays of microtubules emanating from the spindle pole body of the diploid nucleus extended to both cell poles. The same differentiated microtubule configuration was elaborated upon induction of azygotic meiosis in S. pombe. The cyclic movements of the elongated nuclei between the cell poles is reflected by a dynamic and coordinated shortening and lengthening of the two microtubule arrays. When the nucleus was at a cell end, one array was short while the other bridged the whole cell length. Experiments with inhibitors showed that microtubules are required for karyogamy and for the elongated shape and movement of nuclei during meiotic prophase. In both fission yeasts the SPBs and nucleoli are at the leading ends of the moving nuclei. Astral and cytoplasmic microtubules were also prominent during meiotic divisions and sporulation. We further show that in S. versatilis the linear elements formed during meiotic prophase are similar to those in S. pombe. Tripartite synaptonemal complexes were never detected. Taken together, these findings suggest that S. pombe and S. versatilis share basic characteristics in the organization of microtubules and the structure and behaviour of nuclei during their meiotic cell cycle. The prominent differentiations of microtubules and nuclei may be involved in the

  7. Recognition of a porphyry system using ASTER data in Bideghan - Qom province (central of Iran)

    Science.gov (United States)

    Feizi, F.; Mansouri, E.

    2014-07-01

    The Bideghan area is located south of the Qom province (central of Iran). The most impressive geological features in the studied area are the Eocene sequences which are intruded by volcanic rocks with basic compositions. Advanced Space borne Thermal Emission and Reflection Radiometer (ASTER) image processing have been used for hydrothermal alteration mapping and lineaments identification in the investigated area. In this research false color composite, band ratio, Principal Component Analysis (PCA), Least Square Fit (LS-Fit) and Spectral Angel Mapping (SAM) techniques were applied on ASTER data and argillic, phyllic, Iron oxide and propylitic alteration zones were separated. Lineaments were identified by aid of false color composite, high pass filters and hill-shade DEM techniques. The results of this study demonstrate the usefulness of remote sensing method and ASTER multi-spectral data for alteration and lineament mapping. Finally, the results were confirmed by field investigation.

  8. Recognition of a porphyry system using ASTER data in Bideghan – Qom province (central of Iran

    Directory of Open Access Journals (Sweden)

    F. Feizi

    2014-07-01

    Full Text Available The Bideghan area is located south of the Qom province (central of Iran. The most impressive geological features in the studied area are the Eocene sequences which are intruded by volcanic rocks with basic compositions. Advanced Space borne Thermal Emission and Reflection Radiometer (ASTER image processing have been used for hydrothermal alteration mapping and lineaments identification in the investigated area. In this research false color composite, band ratio, Principal Component Analysis (PCA, Least Square Fit (LS-Fit and Spectral Angel Mapping (SAM techniques were applied on ASTER data and argillic, phyllic, Iron oxide and propylitic alteration zones were separated. Lineaments were identified by aid of false color composite, high pass filters and hill-shade DEM techniques. The results of this study demonstrate the usefulness of remote sensing method and ASTER multi-spectral data for alteration and lineament mapping. Finally, the results were confirmed by field investigation.

  9. Mapping advanced argillic alteration zones with ASTER and Hyperion data in the Andes Mountains of Peru

    Science.gov (United States)

    Ramos, Yuddy; Goïta, Kalifa; Péloquin, Stéphane

    2016-04-01

    This study evaluates Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) and Hyperion hyperspectral sensor datasets to detect advanced argillic minerals. The spectral signatures of some alteration clay minerals, such as dickite and alunite, have similar absorption features; thus separating them using multispectral satellite images is a complex challenge. However, Hyperion with its fine spectral bands has potential for good separability of features. The Spectral Angle Mapper algorithm was used in this study to map three advanced argillic alteration minerals (alunite, kaolinite, and dickite) in a known alteration zone in the Peruvian Andes. The results from ASTER and Hyperion were analyzed, compared, and validated using a Portable Infrared Mineral Analyzer field spectrometer. The alterations corresponding to kaolinite and alunite were detected with both ASTER and Hyperion (80% to 84% accuracy). However, the dickite mineral was identified only with Hyperion (82% accuracy).

  10. Mapping temperature and radiant geothermal heat flux anomalies in the Yellowstone geothermal system using ASTER thermal infrared data

    Science.gov (United States)

    Vaughan, R. Greg; Lowenstern, Jacob B.; Keszthelyi, Laszlo P.; Jaworowski, Cheryl; Heasler, Henry

    2012-01-01

    The purpose of this work was to use satellite-based thermal infrared (TIR) remote sensing data to measure, map, and monitor geothermal activity within the Yellowstone geothermal area to help meet the missions of both the U.S. Geological Survey Yellowstone Volcano Observatory and the Yellowstone National Park Geology Program. Specifically, the goals were to: 1) address the challenges of remotely characterizing the spatially and temporally dynamic thermal features in Yellowstone by using nighttime TIR data from the Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) and 2) estimate the temperature, geothermal radiant emittance, and radiant geothermal heat flux (GHF) for Yellowstone’s thermal areas (both Park wide and for individual thermal areas). ASTER TIR data (90-m pixels) acquired at night during January and February, 2010, were used to estimate surface temperature, radiant emittance, and radiant GHF from all of Yellowstone’s thermal features, produce thermal anomaly maps, and update field-based maps of thermal areas. A background subtraction technique was used to isolate the geothermal component of TIR radiance from thermal radiance due to insolation. A lower limit for the Yellowstone’s total radiant GHF was established at ~2.0 GW, which is ~30-45% of the heat flux estimated through geochemical (Cl-flux) methods. Additionally, about 5 km2 was added to the geodatabase of mapped thermal areas. This work provides a framework for future satellite-based thermal monitoring at Yellowstone as well as exploration of other volcanic / geothermal systems on a global scale.

  11. The formin DIAPH1 (mDia1) regulates megakaryocyte proplatelet formation by remodeling the actin and microtubule cytoskeletons.

    Science.gov (United States)

    Pan, Jiajia; Lordier, Larissa; Meyran, Deborah; Rameau, Philippe; Lecluse, Yann; Kitchen-Goosen, Susan; Badirou, Idinath; Mokrani, Hayat; Narumiya, Shuh; Alberts, Arthur S; Vainchenker, William; Chang, Yunhua

    2014-12-18

    Megakaryocytes are highly specialized precursor cells that produce platelets via cytoplasmic extensions called proplatelets. Proplatelet formation (PPF) requires profound changes in microtubule and actin organization. In this work, we demonstrated that DIAPH1 (mDia1), a mammalian homolog of Drosophila diaphanous that works as an effector of the small GTPase Rho, negatively regulates PPF by controlling the dynamics of the actin and microtubule cytoskeletons. Moreover, we showed that inhibition of both DIAPH1 and the Rho-associated protein kinase (Rock)/myosin pathway increased PPF via coordination of both cytoskeletons. We provide evidence that 2 major effectors of the Rho GTPase pathway (DIAPH1 and Rock/myosin II) are involved not only in Rho-mediated stress fibers assembly, but also in the regulation of microtubule stability and dynamics during PPF.

  12. Reducing the Discrepancy Between ASTER and MODIS Land Surface Temperature Products

    Directory of Open Access Journals (Sweden)

    Changqing Ke

    2007-12-01

    Full Text Available Human-induced global warming has significantly increased the importance ofsatellite monitoring of land surface temperature (LST on a global scale. The MODerate-resolution Imaging Spectroradiometer (MODIS provides a 1-km resolution LST productwith almost daily coverage of the Earth, invaluable to both local and global change studies.The Advanced Spaceborne Thermal Emission Reflection Radiometer (ASTER provides aLST product with a high spatial resolution of 90-m and a 16-day recurrent cycle,simultaneously acquired at the same height and nadir view as MODIS. ASTER andMODIS are complementary in resolution, offering a unique opportunity for scale-relatedstudies. ASTER and MODIS LST have been widely used but the errors in LST were mostlydisregarded. Correction of ASTER-to-MODIS LST discrepancies is essential for studiesreliant upon the joint use of these sensors. In this study, we compared three correctionapproaches: the Wan et al.’s approach, the refined Wan et al.’s approach, and thegeneralized split window (GSW algorithm based approach. The Wan et al.’s approachcorrects the MODIS 1-km LST using MODIS 5-km LST. The refined approach modifiesthe Wan et al.’s approach through incorporating ASTER emissivity and MODIS 5-km data.The GSW algorithm approach does not use MODIS 5-km but only ASTER emissivity data. We examined the case over a semi-arid terrain area for the part of the Loess Plateau of China. All the approaches reduced the ASTER-to-MODIS LST discrepancy effectively. With terrain correction, the original ASTER-to-MODIS LST difference reduced from 2.7±1.28 K to -0.1±1.87 K for the Wan et al.’s approach, 0.2±1.57 K for the refined approach, and 0.1±1.33 K for the GSW algorithm based approach. Among all the approaches, the GSW algorithm based approach performed best in terms of mean, standard deviation, root mean square root, and correlation coefficient.

  13. How do Plants Organize Microtubules Without a Centrosome?

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    A microtubule nucleates from a γ-tubulin complex, which consists of γ-tubulin, proteins from the SPC97/SPC98 family, and the WD40 motif protein GCP-WD. We analyzed the phylogenetic relationships of the genes encoding these proteins and found that the components of this complex are widely conserved among land plants and other eukaryotes. By contrast,the interphase and mitotic arrays of microtubules in land plants differ from those in other eukaryotes. In the interphase cortical array, the majority of microtubules nucleate on existing microtubules in the absence of conspicuous microtubule organizing centers (MTOCs), such as a centrosome. During mitosis, the spindle also forms in the absence of conspicuous MTOCs. Both poles of the spindle are broad, and branched structures of microtubules called microtubule converging centers form at the poles. In this review, we hypothesize that the microtubule converging centers form via microtubuledependent microtubule nucleation, as in the case of the interphase arrays. The evolutionary insights arising from the molecular basis of the diversity in microtubule organization are discussed.

  14. Resolving bundled microtubules using anti-tubulin nanobodies.

    Science.gov (United States)

    Mikhaylova, Marina; Cloin, Bas M C; Finan, Kieran; van den Berg, Robert; Teeuw, Jalmar; Kijanka, Marta M; Sokolowski, Mikolaj; Katrukha, Eugene A; Maidorn, Manuel; Opazo, Felipe; Moutel, Sandrine; Vantard, Marylin; Perez, Frank; van Bergen en Henegouwen, Paul M P; Hoogenraad, Casper C; Ewers, Helge; Kapitein, Lukas C

    2015-08-11

    Microtubules are hollow biopolymers of 25-nm diameter and are key constituents of the cytoskeleton. In neurons, microtubules are organized differently between axons and dendrites, but their precise organization in different compartments is not completely understood. Super-resolution microscopy techniques can detect specific structures at an increased resolution, but the narrow spacing between neuronal microtubules poses challenges because most existing labelling strategies increase the effective microtubule diameter by 20-40 nm and will thereby blend neighbouring microtubules into one structure. Here we develop single-chain antibody fragments (nanobodies) against tubulin to achieve super-resolution imaging of microtubules with a decreased apparent diameter. To test the resolving power of these novel probes, we generate microtubule bundles with a known spacing of 50-70 nm and successfully resolve individual microtubules. Individual bundled microtubules can also be resolved in different mammalian cells, including hippocampal neurons, allowing novel insights into fundamental mechanisms of microtubule organization in cell- and neurobiology.

  15. Motor protein accumulation on antiparallel microtubule overlaps

    CERN Document Server

    Kuan, Hui-Shun

    2015-01-01

    Biopolymers serve as one-dimensional tracks on which motor proteins move to perform their biological roles. Motor protein phenomena have inspired theoretical models of one-dimensional transport, crowding, and jamming. Experiments studying the motion of Xklp1 motors on reconstituted antiparallel microtubule overlaps demonstrated that motors recruited to the overlap walk toward the plus end of individual microtubules and frequently switch between filaments. We study a model of this system that couples the totally asymmetric simple exclusion process (TASEP) for motor motion with switches between antiparallel filaments and binding kinetics. We determine steady-state motor density profiles for fixed-length overlaps using exact and approximate solutions of the continuum differential equations and compare to kinetic Monte Carlo simulations. The center region, far from the overlap ends, has a constant motor density as one would na\\"ively expect. However, rather than following a simple binding equilibrium, the center ...

  16. The actin-microtubule cross-linking activity of Drosophila Short stop is regulated by intramolecular inhibition.

    Science.gov (United States)

    Applewhite, Derek A; Grode, Kyle D; Duncan, Mara C; Rogers, Stephen L

    2013-09-01

    Actin and microtubule dynamics must be precisely coordinated during cell migration, mitosis, and morphogenesis--much of this coordination is mediated by proteins that physically bridge the two cytoskeletal networks. We have investigated the regulation of the Drosophila actin-microtubule cross-linker Short stop (Shot), a member of the spectraplakin family. Our data suggest that Shot's cytoskeletal cross-linking activity is regulated by an intramolecular inhibitory mechanism. In its inactive conformation, Shot adopts a "closed" conformation through interactions between its NH(2)-terminal actin-binding domain and COOH-terminal EF-hand-GAS2 domain. This inactive conformation is targeted to the growing microtubule plus end by EB1. On activation, Shot binds along the microtubule through its COOH-terminal GAS2 domain and binds to actin with its NH(2)-terminal tandem CH domains. We propose that this mechanism allows Shot to rapidly cross-link dynamic microtubules in response to localized activating signals at the cell cortex.

  17. Application Analysis of Global Public Data Resources ASTER GDEM%全球公众数据资源ASTER GDEM的应用研究

    Institute of Scientific and Technical Information of China (English)

    张涛

    2015-01-01

    Based on the global public data resources ASTER GDEM (space borne thermal emission and reflection ra⁃diometer global digital elevation model) data publicly announced by the USA space agency (NASA),this paper con⁃ducted practical processing and research, and carried out statistical validation on its accuracy.%本文利用美国太空总署(NASA)对外公布的全球公众数据资源ASTER GDEM(星载热发射和反射辐射仪全球数字高程模型)数据进行了实践性处理和研究,并对其精度进行统计验证。

  18. Microtubule-Targeting Therapy for Prostate Cancer

    Science.gov (United States)

    2007-02-01

    Cancer1828 Mol Cancer Ther 2005;4(12). December 2005 22. Sambrook J, Fritsch EF, Maniatis T. Molecular cloning : a laboratory manual. 2nd ed. Cold Spring...Harbor (NY): Cold Spring Harbor Laboratory; 1989. 23. Zhu XX, Kozarsky K, Strahler JR, et al. Molecular cloning of a novel human leukemia-associated...of Cancer Research, Abstract #4940, 2005. 3. Mistry, SJ, Atweh, GF. Microtubule targeting therapy: Anti-stathmin based molecular cancer

  19. MICROTUBULE ORGANIZATION 1 regulates structure and function of microtubule arrays during mitosis and cytokinesis in the Arabidopsis root.

    Science.gov (United States)

    Kawamura, Eiko; Himmelspach, Regina; Rashbrooke, Madeleine C; Whittington, Angela T; Gale, Kevin R; Collings, David A; Wasteneys, Geoffrey O

    2006-01-01

    MICROTUBULE ORGANIZATION 1 (MOR1) is a plant member of the highly conserved MAP215/Dis1 family of microtubule-associated proteins. Prior studies with the temperature-sensitive mor1 mutants of Arabidopsis (Arabidopsis thaliana), which harbor single amino acid substitutions in an N-terminal HEAT repeat, proved that MOR1 regulates cortical microtubule organization and function. Here we demonstrate by use of live cell imaging and immunolabeling that the mor1-1 mutation generates specific defects in the microtubule arrays of dividing vegetative cells. Unlike the universal cortical microtubule disorganization in elongating mor1-1 cells, disruption of mitotic and cytokinetic microtubule arrays was not detected in all dividing cells. Nevertheless, quantitative analysis identified distinct defects in preprophase bands (PPBs), spindles, and phragmoplasts. In nearly one-half of dividing cells at the restrictive temperature of 30 degrees C, PPBs were not detected prior to spindle formation, and those that did form were often disrupted. mor1-1 spindles and phragmoplasts were short and abnormally organized and persisted for longer times than in wild-type cells. The reduced length of these arrays predicts that the component microtubule lengths are also reduced, suggesting that microtubule length is a critical determinant of spindle and phragmoplast structure, orientation, and function. Microtubule organizational defects led to aberrant chromosomal arrangements, misaligned or incomplete cell plates, and multinucleate cells. Antiserum raised against an N-terminal MOR1 sequence labeled the full length of microtubules in interphase arrays, PPBs, spindles, and phragmoplasts. Continued immunolabeling of the disorganized and short microtubules of mor1-1 at the restrictive temperature demonstrated that the mutant mor1-1(L174F) protein loses function without dissociating from microtubules, providing important insight into the mechanism by which MOR1 may regulate microtubule length.

  20. Cellular Samurai: katanin and the severing of microtubules.

    Science.gov (United States)

    Quarmby, L

    2000-08-01

    Recent biochemical studies of the AAA ATPase, katanin, provide a foundation for understanding how microtubules might be severed along their length. These in vitro studies are complemented by a series of recent reports of direct in vivo observation of microtubule breakage, which indicate that the in vitro phenomenon of catalysed microtubule severing is likely to be physiological. There is also new evidence that microtubule severing by katanin is important for the production of non-centrosomal microtubules in cells such as neurons and epithelial cells. Although it has been difficult to establish the role of katanin in mitosis, new genetic evidence indicates that a katanin-like protein, MEI-1, plays an essential role in meiosis in C. elegans. Finally, new proteins involved in the severing of axonemal microtubules have been discovered in the deflagellation system of Chlamydomonas.

  1. Microtubule Associated Proteins in Plants and the Processes They Manage

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Microtubule associated proteins (MAPs) are proteins that physically bind to microtubules in eukaryotes. MAPs play important roles in regulating the polymerization and organization of microtubules and in using the ensuing microtubule arrays to carry out a variety of cellular functions. In plants, MAPs manage the construction, repositioning, and dismantling of four distinct microtubule arrays throughout the cell cycle. Three of these arrays, the cortical array, the preprophase band,and the phragmoplast, are prominent to plants and are responsible for facilitating cell wall deposition and modification,transducing signals, demarcating the plane of cell division, and forming the new cell plate during cytokinesis, This review highlights important aspects of how MAPs in plants establish and maintain microtubule arrays as well as regulate cell growth, cell division, and cellular responses to the environment.

  2. Cortical microtubule arrays are initiated from a nonrandom prepattern driven by atypical microtubule initiation.

    Science.gov (United States)

    Lindeboom, Jelmer J; Lioutas, Antonios; Deinum, Eva E; Tindemans, Simon H; Ehrhardt, David W; Emons, Anne Mie C; Vos, Jan W; Mulder, Bela M

    2013-03-01

    The ordered arrangement of cortical microtubules in growing plant cells is essential for anisotropic cell expansion and, hence, for plant morphogenesis. These arrays are dismantled when the microtubule cytoskeleton is rearranged during mitosis and reassembled following completion of cytokinesis. The reassembly of the cortical array has often been considered as initiating from a state of randomness, from which order arises at least partly through self-organizing mechanisms. However, some studies have shown evidence for ordering at early stages of array assembly. To investigate how cortical arrays are initiated in higher plant cells, we performed live-cell imaging studies of cortical array assembly in tobacco (Nicotiana tabacum) Bright Yellow-2 cells after cytokinesis and drug-induced disassembly. We found that cortical arrays in both cases did not initiate randomly but with a significant overrepresentation of microtubules at diagonal angles with respect to the cell axis, which coincides with the predominant orientation of the microtubules before their disappearance from the cell cortex in preprophase. In Arabidopsis (Arabidopsis thaliana) root cells, recovery from drug-induced disassembly was also nonrandom and correlated with the organization of the previous array, although no diagonal bias was observed in these cells. Surprisingly, during initiation, only about one-half of the new microtubules were nucleated from locations marked by green fluorescent protein-γ-tubulin complex protein2-tagged γ-nucleation complexes (γ-tubulin ring complex), therefore indicating that a large proportion of early polymers was initiated by a noncanonical mechanism not involving γ-tubulin ring complex. Simulation studies indicate that the high rate of noncanonical initiation of new microtubules has the potential to accelerate the rate of array repopulation.

  3. Drosophila Stathmin: A Microtubule-destabilizing Factor Involved in Nervous System Formation

    OpenAIRE

    Ozon, Sylvie; Guichet, Antoine; Gavet, Olivier; Roth, Siegfried; Sobel, André

    2002-01-01

    Stathmin is a ubiquitous regulatory phosphoprotein, the generic element of a family of neural phosphoproteins in vertebrates that possess the capacity to bind tubulin and interfere with microtubule dynamics. Although stathmin and the other proteins of the family have been associated with numerous cell regulations, their biological roles remain elusive, as in particular inactivation of the stathmin gene in the mouse resulted in no clear deleterious phenotype. We identified stathmin phosphoprot...

  4. First report of Alternaria alternata causing leaf spot on Ruth's golden aster (Pityopsis ruthii) in Tennessee

    Science.gov (United States)

    Ruth’s golden aster, Pityopsis ruthii (Small), is an endangered, herbaceous perennial plant that is only endemic to small sections of the Hiwassee and Ocoee Rivers, in Polk County, Tennessee. In July 2015, a greenhouse grown plant exhibited symptoms of disease that included elongated brown lesions o...

  5. Improvement of dem Generation from Aster Images Using Satellite Jitter Estimation and Open Source Implementation

    Science.gov (United States)

    Girod, L.; Nuth, C.; Kääb, A.

    2015-12-01

    The Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) system embarked on the Terra (EOS AM-1) satellite has been a source of stereoscopic images covering the whole globe at a 15m resolution at a consistent quality for over 15 years. The potential of this data in terms of geomorphological analysis and change detection in three dimensions is unrivaled and needs to be exploited. However, the quality of the DEMs and ortho-images currently delivered by NASA (ASTER DMO products) is often of insufficient quality for a number of applications such as mountain glacier mass balance. For this study, the use of Ground Control Points (GCPs) or of other ground truth was rejected due to the global "big data" type of processing that we hope to perform on the ASTER archive. We have therefore developed a tool to compute Rational Polynomial Coefficient (RPC) models from the ASTER metadata and a method improving the quality of the matching by identifying and correcting jitter induced cross-track parallax errors. Our method outputs more accurate DEMs with less unmatched areas and reduced overall noise. The algorithms were implemented in the open source photogrammetric library and software suite MicMac.

  6. Cross-Calibration of Earth Observing System Terra Satellite Sensors MODIS and ASTER

    Science.gov (United States)

    McCorkel, J.

    2014-01-01

    The Advanced Spaceborne Thermal Emissive and Reflection Radiometer (ASTER) and Moderate Resolution Imaging Spectrometer (MODIS) are two of the five sensors onboard the Earth Observing System's Terra satellite. These sensors share many similar spectral channels while having much different spatial and operational parameters. ASTER is a tasked sensor and sometimes referred to a zoom camera of the MODIS that collects a full-earth image every one to two days. It is important that these sensors have a consistent characterization and calibration for continued development and use of their data products. This work uses a variety of test sites to retrieve and validate intercalibration results. The refined calibration of Collection 6 of the Terra MODIS data set is leveraged to provide the up-to-date reference for trending and validation of ASTER. Special attention is given to spatially matching radiance measurements using prelaunch spatial response characterization of MODIS. Despite differences in spectral band properties and spatial scales, ASTER-MODIS is an ideal case for intercomparison since the sensors have nearly identical views and acquisitions times and therefore can be used as a baseline of intercalibration performance of other satellite sensor pairs.

  7. Bewaren van onbeworteld stek : bewaring van onbeworteld stek van aster en solidago

    NARCIS (Netherlands)

    Eveleens, B.; Krijger, D.; Kouwenhoven, D.; Berg, van den T.; Telgen, van H.J.

    2002-01-01

    In de praktijk worden geoogste stekken van Aster ericoides en Solidago maximaal een week bewaard. Bij langere bewaring (of transport) treden vaak problemen op als uitval ten gevolge van rot en/of slechte beworteling. Een langere bewaarduur is gewenst, zowel voor planning van werkzaamheden als behoud

  8. Calculation of the Electromagnetic Field Around a Microtubule

    Directory of Open Access Journals (Sweden)

    D. Havelka

    2009-01-01

    Full Text Available Microtubules are important structures in the cytoskeleton which organizes the cell. A single microtubule is composed of electrically polar structures, tubulin heterodimers, which have a strong electric dipole moment. Vibrations are expected to be generated in microtubules, thus tubulin heterodimers oscillate as electric dipoles. This gives rise to an electromagnetic field which is detected around the cells. We calculate here the electromagnetic field of microtubules if they are excited at 1 GHz. This paper includes work done for the bachelor thesis of the first author. 

  9. Glacier Volume Change Estimation Using Time Series of Improved Aster Dems

    Science.gov (United States)

    Girod, Luc; Nuth, Christopher; Kääb, Andreas

    2016-06-01

    Volume change data is critical to the understanding of glacier response to climate change. The Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) system embarked on the Terra (EOS AM-1) satellite has been a unique source of systematic stereoscopic images covering the whole globe at 15m resolution and at a consistent quality for over 15 years. While satellite stereo sensors with significantly improved radiometric and spatial resolution are available to date, the potential of ASTER data lies in its long consistent time series that is unrivaled, though not fully exploited for change analysis due to lack of data accuracy and precision. Here, we developed an improved method for ASTER DEM generation and implemented it in the open source photogrammetric library and software suite MicMac. The method relies on the computation of a rational polynomial coefficients (RPC) model and the detection and correction of cross-track sensor jitter in order to compute DEMs. ASTER data are strongly affected by attitude jitter, mainly of approximately 4 km and 30 km wavelength, and improving the generation of ASTER DEMs requires removal of this effect. Our sensor modeling does not require ground control points and allows thus potentially for the automatic processing of large data volumes. As a proof of concept, we chose a set of glaciers with reference DEMs available to assess the quality of our measurements. We use time series of ASTER scenes from which we extracted DEMs with a ground sampling distance of 15m. Our method directly measures and accounts for the cross-track component of jitter so that the resulting DEMs are not contaminated by this process. Since the along-track component of jitter has the same direction as the stereo parallaxes, the two cannot be separated and the elevations extracted are thus contaminated by along-track jitter. Initial tests reveal no clear relation between the cross-track and along-track components so that the latter seems not to be

  10. Human SAS-6 C-Terminus Nucleates and Promotes Microtubule Assembly in Vitro by Binding to Microtubules.

    Science.gov (United States)

    Gupta, Hindol; Badarudeen, Binshad; George, Athira; Thomas, Geethu Emily; Gireesh, K K; Manna, Tapas K

    2015-10-20

    Centrioles are essential components of the animal centrosome and play crucial roles in the formation of cilia and flagella. They are cylindrical structures composed of nine triplet microtubules organized around a central cartwheel. Recent studies have identified spindle assembly abnormal protein SAS-6 as a critical component necessary for formation of the cartwheel. However, the molecular details of how the cartwheel participates in centriolar microtubule assembly have not been clearly understood. In this report, we show that the C-terminal tail (residues 470-657) of human SAS-6, HsSAS-6 C, the region that has been shown to extend toward the centriolar wall where the microtubule triplets are organized, nucleated and induced microtubule polymerization in vitro. The N-terminus (residues 1-166) of HsSAS-6, the domain known to be involved in formation of the central hub of the cartwheel, did not, however, exert any effect on microtubule polymerization. HsSAS-6 C bound to the microtubules and localized along the lengths of the microtubules in vitro. Microtubule pull-down and coimmunoprecipitation (Co-IP) experiments with S-phase synchronized HeLa cell lysates showed that the endogenous HsSAS-6 coprecipitated with the microtubules, and it mediated interaction with tubulin. Isothermal calorimetry titration and size exclusion chromatography showed that HsSAS-6 C bound to the αβ-tubulin dimer in vitro. The results demonstrate that HsSAS-6 possesses an intrinsic microtubule assembly promoting activity and further implicate that its outer exposed C-terminal tail may play critical roles in microtubule assembly and stabilizing microtubule attachment with the centriolar cartwheel.

  11. Interplay between kinesin-1 and cortical dynein during axonal outgrowth and microtubule organization in Drosophila neurons.

    Science.gov (United States)

    del Castillo, Urko; Winding, Michael; Lu, Wen; Gelfand, Vladimir I

    2015-12-28

    In this study, we investigated how microtubule motors organize microtubules in Drosophila neurons. We showed that, during the initial stages of axon outgrowth, microtubules display mixed polarity and minus-end-out microtubules push the tip of the axon, consistent with kinesin-1 driving outgrowth by sliding antiparallel microtubules. At later stages, the microtubule orientation in the axon switches from mixed to uniform polarity with plus-end-out. Dynein knockdown prevents this rearrangement and results in microtubules of mixed orientation in axons and accumulation of microtubule minus-ends at axon tips. Microtubule reorganization requires recruitment of dynein to the actin cortex, as actin depolymerization phenocopies dynein depletion, and direct recruitment of dynein to the membrane bypasses the actin requirement. Our results show that cortical dynein slides 'minus-end-out' microtubules from the axon, generating uniform microtubule arrays. We speculate that differences in microtubule orientation between axons and dendrites could be dictated by differential activity of cortical dynein.

  12. ASTER, ALI and Hyperion sensors data for lithological mapping and ore minerals exploration.

    Science.gov (United States)

    Beiranvand Pour, Amin; Hashim, Mazlan

    2014-01-01

    This paper provides a review of the Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER), Advanced Land Imager (ALI), and Hyperion data and applications of the data as a tool for ore minerals exploration, lithological and structural mapping. Spectral information extraction from ASTER, ALI, and Hyperion data has great ability to assist geologists in all disciplines to map the distribution and detect the rock units exposed at the earth's surface. The near coincidence of Earth Observing System (EOS)/Terra and Earth Observing One (EO-1) platforms allows acquiring ASTER, ALI, and Hyperion imagery of the same ground areas, resulting accurate information for geological mapping applications especially in the reconnaissance stages of hydrothermal copper and gold exploration, chromite, magnetite, massive sulfide and uranium ore deposits, mineral components of soils and structural interpretation at both regional and district scales. Shortwave length infrared and thermal infrared bands of ASTER have sufficient spectral resolution to map fundamental absorptions of hydroxyl mineral groups and silica and carbonate minerals for regional mapping purposes. Ferric-iron bearing minerals can be discriminated using six unique wavelength bands of ALI spanning the visible and near infrared. Hyperion visible and near infrared bands (0.4 to 1.0 μm) and shortwave infrared bands (0.9 to 2.5 μm) allowed to produce image maps of iron oxide minerals, hydroxyl-bearing minerals, sulfates and carbonates in association with hydrothermal alteration assemblages, respectively. The techniques and achievements reviewed in the present paper can further introduce the efficacy of ASTER, ALI, and Hyperion data for future mineral and lithological mapping and exploration of the porphyry copper, epithermal gold, chromite, magnetite, massive sulfide and uranium ore deposits especially in arid and semi-arid territory.

  13. Inaccessible Biodiversity on Limestone Cliffs: Aster tianmenshanensis (Asteraceae, a New Critically Endangered Species from China.

    Directory of Open Access Journals (Sweden)

    Guo-Jin Zhang

    Full Text Available Aster tianmenshanensis G. J. Zhang & T. G. Gao, a new species of Asteraceae from southern China is described and illustrated based on evidence from morphology, micromorphology and molecular phylogeny. The new species is superficially similar to Aster salwinensis Onno in having rosettes of spatulate leaves and a solitary, terminal capitulum, but it differs by its glabrous leaf margins, unequal disc floret lobes and 1-seriate pappus. The molecular phylogenetic analysis, based on nuclear sequences ITS, ETS and chloroplast sequence trnL-F, showed that the new species was nested within the genus Aster and formed a well supported clade with Aster verticillatus (Reinw. Brouillet et al. The new species differs from the latter in having unbranched stems, much larger capitula, unequal disc floret lobes, beakless achenes and persistent pappus. In particular, A. tianmenshanensis has very short stigmatic lines, only ca. 0.18 mm long and less than 1/3 of the length of sterile style tip appendages, remarkably different from its congeners. This type of stigmatic line, as far as we know, has not been found in any other species of Aster. The very short stigmatic lines plus the unequal disc floret lobes imply that the new species may have a very specialized pollination system, which may be a consequence of habitat specialization. The new species grows only on the limestone cliffs of Mt. Tianmen, Hunan Province, at the elevation of 1400 m. It could only be accessed when a plank walkway was built across the cliffs for tourists. As it is known only from an area estimated at less than 10 km2 and a walkway passes through this location, its habitat could be easily disturbed. This species should best be treated as Critically Endangered based on the International Union for Conservation of Nature Red List Categories and Criteria B2a.

  14. Lithological Discrimination of the Mafic-Ultramafic Complex, Huitongshan, Beishan, China:Using ASTER Data

    Institute of Scientific and Technical Information of China (English)

    Lei Liu; Jun Zhou; Dong Jiang; Dafang Zhuang; Lamin R Mansaray

    2014-01-01

    The Beishan area has more than seventy mafic-ultramafic complexes sparsely distributed in the area and is of a big potential in mineral resources related to mafic-ultramafic intrusions. Many mafic-ultramafic intrusions which are mostly in small sizes have been omitted by previous works. This research takes Huitongshan as the study area, which is a major district for mafic-ultramafic occur-rences in Beishan. Advanced spaceborne thermal emission and reflection radiometer (ASTER) data have been processed and interpreted for mapping the mafic-ultramafic complex. ASTER data were processed by different techniques that were selected based on image reflectance and laboratory emis-sivity spectra. The visible near-infrared (VNIR) and short wave infrared (SWIR) data were trans-formed using band ratios and minimum noise fraction (MNF), while the thermal infrared (TIR) data were processed using mafic index (MI) and principal components analysis (PCA). ASTER band ratios (6/8, 5/4, 2/1) in RGB image and MNF (1, 2, 4) in RGB image were powerful in distinguishing the subtle differences between the various rock units. PCA applied to all five bands of ASTER TIR imagery high-lighted marked differences among the mafic rock units and was more effective than the MI in differen-tiating mafic-ultramafic rocks. Our results were consistent with information derived from local geolog-ical maps. Based on the remote sensing results and field inspection, eleven gabbroic intrusions and a pyroxenite occurrence were recognized for the first time. A new geologic map of the Huitongshan area was created by integrating the results of remote sensing, previous geological maps and field inspection. It is concluded that the workflow of ASTER image processing, interpretation and ground inspection has great potential for mafic-ultramafic rocks identifying and relevant mineral targeting in the sparsely vegetated arid region of northwestern China.

  15. Tubers from patients with tuberous sclerosis complex are characterized by changes in microtubule biology through ROCK2 signalling.

    Science.gov (United States)

    Ferrer, Isidre; Mohan, Pooja; Chen, Helen; Castellsague, Joan; Gómez-Baldó, Laia; Carmona, Marga; García, Nadia; Aguilar, Helena; Jiang, Jihong; Skowron, Margaretha; Nellist, Mark; Ampuero, Israel; Russi, Antonio; Lázaro, Conxi; Maxwell, Christopher A; Pujana, Miguel Angel

    2014-07-01

    Most patients with tuberous sclerosis complex (TSC) develop cortical tubers that cause severe neurological disabilities. It has been suggested that defects in neuronal differentiation and/or migration underlie the appearance of tubers. However, the precise molecular alterations remain largely unknown. Here, by combining cytological and immunohistochemical analyses of tubers from nine TSC patients (four of them diagnosed with TSC2 germline mutations), we show that alteration of microtubule biology through ROCK2 signalling contributes to TSC neuropathology. All tubers showed a larger number of binucleated neurons than expected relative to control cortex. An excess of normal and altered cytokinetic figures was also commonly observed. Analysis of centrosomal markers suggested increased microtubule nucleation capacity, which was supported by the analysis of an expression dataset from cortical tubers and control cortex, and subsequently linked to under-expression of Rho-associated coiled-coil containing kinase 2 (ROCK2). Thus, augmented microtubule nucleation capacity was observed in mouse embryonic fibroblasts and human fibroblasts deficient in the Tsc2/TSC2 gene product, tuberin. Consistent with ROCK2 under-expression, microtubule acetylation was found to be increased with tuberin deficiency; this alteration was abrogated by rapamycin treatment and mimicked by HDAC6 inhibition. Together, the results of this study support the hypothesis that loss of TSC2 expression can alter microtubule organization and dynamics, which, in turn, deregulate cell division and potentially impair neuronal differentiation.

  16. Microtubule binding by the formin Cappuccino and its implications for Drosophila oogenesis

    OpenAIRE

    Roth-Johnson, Elizabeth Anne

    2014-01-01

    Coordination of actin and microtubule cytoskeletal networks is required for a number of fundamental cellular processes. Formin family actin nucleators are emerging coordinators of the actin and microtubule cytoskeletons, as they can both nucleate actin filaments and bind microtubules in vitro. To gain a more detailed mechanistic understanding of formin-microtubule interactions and formin-mediated actin-microtubule crosstalk, we studied microtubule binding by Cappuccino (Capu), a formin involv...

  17. In vivo FRET imaging revealed a regulatory role of RanGTP in kinetochore-microtubule attachments via Aurora B kinase.

    Directory of Open Access Journals (Sweden)

    Yoke-Peng Lee

    Full Text Available Under the fluctuating circumstances provided by the innate dynamics of microtubules and opposing tensions resulted from microtubule-associated motors, it is vital to ensure stable kinetochore-microtubule attachments for accurate segregation. However, a comprehensive understanding of how this regulation is mechanistically achieved remains elusive. Using our newly designed live cell FRET time-lapse imaging, we found that post-metaphase RanGTP is crucial in the maintenance of stable kinetochore-microtubule attachments by regulating Aurora B kinase via the NES-bearing Mst1. More importantly, our study demonstrates that by ensuring stable alignment of metaphase chromosomes prior to segregation, RanGTP is indispensible in governing the genomic integrity and the fidelity of cell cycle progression. Our findings suggest an additional role of RanGTP beyond its known function in mitotic spindle assembly during the prometaphase-metaphase transition.

  18. Molecular Pathway of Microtubule Organization at the Golgi Apparatus

    NARCIS (Netherlands)

    Wu, Jingchao; de Heus, Cecilia; Liu, Qingyang; Bouchet, Benjamin P; Noordstra, Ivar; Jiang, Kai; Hua, Shasha; Martin, Maud; Yang, Chao; Grigoriev, Ilya; Katrukha, Eugene A; Altelaar, A F Maarten; Hoogenraad, Casper C; Qi, Robert Z; Klumperman, Judith; Akhmanova, Anna

    2016-01-01

    The Golgi apparatus controls the formation of non-centrosomal microtubule arrays important for Golgi organization, polarized transport, cell motility, and cell differentiation. Here, we show that CAMSAP2 stabilizes and attaches microtubule minus ends to the Golgi through a complex of AKAP450 and myo

  19. Resolving bundled microtubules using anti-tubulin nanobodies

    NARCIS (Netherlands)

    Mikhaylova, Marina; Cloin, Bas M C; Finan, Kieran; van den Berg, Robert; Teeuw, Jalmar; Kijanka, Marta M; Sokolowski, Mikolaj; Katrukha, Eugene A; Maidorn, Manuel; Opazo, Felipe; Moutel, Sandrine; Vantard, Marylin; Perez, Frank; van Bergen en Henegouwen, Paul M P; Hoogenraad, Casper C; Ewers, Helge; Kapitein, Lukas C

    2015-01-01

    Microtubules are hollow biopolymers of 25-nm diameter and are key constituents of the cytoskeleton. In neurons, microtubules are organized differently between axons and dendrites, but their precise organization in different compartments is not completely understood. Super-resolution microscopy techn

  20. Interconnections between cell wall polymers, wall mechanics, and cortical microtubules: Teasing out causes and consequences.

    Science.gov (United States)

    Xiao, Chaowen; Anderson, Charles T

    2016-09-01

    In plants, cell wall components including cellulose, hemicelluloses, and pectins interact with each other to form complex extracellular network structures that control cell growth and maintain cell shape. However, it is still not clear exactly how different wall polymers interact, how the conformations and interactions of cell wall polymers relate to wall mechanics, and how these factors impinge on intracellular structures such as the cortical microtubule cytoskeleton. Here, based on studies of Arabidopsis thaliana xxt1 xxt2 mutants, which lack detectable xyloglucan in their walls and display aberrant wall mechanics, altered cellulose patterning and biosynthesis, and reduced cortical microtubule stability, we discuss the potential relationships between cell wall biosynthesis, wall mechanics, and cytoskeletal dynamics in an effort to better understand their roles in controlling plant growth and morphogenesis.

  1. Pironetin reacts covalently with cysteine-316 of α-tubulin to destabilize microtubule

    Science.gov (United States)

    Yang, Jianhong; Wang, Yuxi; Wang, Taijing; Jiang, Jian; Botting, Catherine H.; Liu, Huanting; Chen, Qiang; Yang, Jinliang; Naismith, James H.; Zhu, Xiaofeng; Chen, Lijuan

    2016-06-01

    Molecules that alter the normal dynamics of microtubule assembly and disassembly include many anticancer drugs in clinical use. So far all such therapeutics target β-tubulin, and structural biology has explained the basis of their action and permitted design of new drugs. However, by shifting the profile of β-tubulin isoforms, cancer cells become resistant to treatment. Compounds that bind to α-tubulin are less well characterized and unexploited. The natural product pironetin is known to bind to α-tubulin and is a potent inhibitor of microtubule polymerization. Previous reports had identified that pironetin reacts with lysine-352 residue however analogues designed on this model had much lower potency, which was difficult to explain, hindering further development. We report crystallographic and mass spectrometric data that reveal that pironetin forms a covalent bond to cysteine-316 in α-tubulin via a Michael addition reaction. These data provide a basis for the rational design of α-tubulin targeting chemotherapeutics.

  2. Asymptotic Analysis of Microtubule-Based Transport by Multiple Identical Molecular Motors

    CERN Document Server

    McKinley, Scott A; Fricks, John; Kramer, Peter R

    2011-01-01

    We describe a system of stochastic differential equations (SDEs) which model the interaction between processive molecular motors, such as kinesin and dynein, and the biomolecular cargo they tow as part of microtubule-based intracellular transport. We show that the classical experimental environment fits within a parameter regime which is qualitatively distinct from conditions one expects to find in living cells. Through an asymptotic analysis of our system of SDEs, we develop a means for applying in vitro observations of the nonlinear response by motors to forces induced on the attached cargo to make analytical predictions for two parameter regimes that have thus far eluded direct experimental observation: 1) highly viscous in vivo transport and 2) dynamics when multiple identical motors are attached to the cargo and microtubule.

  3. Microtubules in Plant Cells: Strategies and Methods for Immunofluorescence, Transmission Electron Microscopy and Live Cell Imaging

    Science.gov (United States)

    Celler, Katherine; Fujita, Miki; Kawamura, Eiko; Ambrose, Chris; Herburger, Klaus; Wasteneys, Geoffrey O.

    2016-01-01

    Microtubules are required throughout plant development for a wide variety of processes, and different strategies have evolved to visualize and analyze them. This chapter provides specific methods that can be used to analyze microtubule organization and dynamic properties in plant systems and summarizes the advantages and limitations for each technique. We outline basic methods for preparing samples for immunofluorescence labelling, including an enzyme-based permeabilization method, and a freeze-shattering method, which generates microfractures in the cell wall to provide antibodies access to cells in cuticle-laden aerial organs such as leaves. We discuss current options for live cell imaging of MTs with fluorescently tagged proteins (FPs), and provide chemical fixation, high pressure freezing/freeze substitution, and post-fixation staining protocols for preserving MTs for transmission electron microscopy and tomography. PMID:26498784

  4. Role of the Number of Microtubules in Chromosome Segregation during Cell Division

    CERN Document Server

    Bertalan, Zsolt; La Porta, Caterina A M; Zapperi, Stefano

    2015-01-01

    Faithful segregation of genetic material during cell division requires alignment of chromosomes between two spindle poles and attachment of their kinetochores to each of the poles. Failure of these complex dynamical processes leads to chromosomal instability (CIN), a characteristic feature of several diseases including cancer. While a multitude of biological factors regulating chromosome congression and bi-orientation have been identified, it is still unclear how they are integrated so that coherent chromosome motion emerges from a large collection of random and deterministic processes. Here we address this issue by a three dimensional computational model of motor-driven chromosome congression and bi-orientation during mitosis. Our model reveals that successful cell division requires control of the total number of microtubules: if this number is too small bi-orientation fails, while if it is too large not all the chromosomes are able to congress. The optimal number of microtubules predicted by our model compa...

  5. HILI destabilizes microtubules by suppressing phosphorylation and Gigaxonin-mediated degradation of TBCB

    Science.gov (United States)

    Tan, Hao; Liao, Hua; Zhao, Lianfang; Lu, Yilu; Jiang, Siyuan; Tao, Dachang; Liu, Yunqiang; Ma, Yongxin

    2017-01-01

    Human PIWIL2, aka HILI, is a member of PIWI protein family and overexpresses in various tumors. However, the underlying mechanisms of HILI in tumorigenesis remain largely unknown. TBCB has a critical role in regulating microtubule dynamics and is overexpressed in many cancers. Here we report that HILI inhibits Gigaxonin-mediated TBCB ubiquitination and degradation by interacting with TBCB, promoting the binding between HSP90 and TBCB, and suppressing the interaction between Gigaxonin and TBCB. Meanwhile, HILI can also reduce phosphorylation level of TBCB induced by PAK1. Our results showed that HILI suppresses microtubule polymerization and promotes cell proliferation, migration and invasion via TBCB for the first time, revealing a novel mechanism for HILI in tumorigenesis. PMID:28393858

  6. Multimodal microtubule binding by the Ndc80 kinetochore complex.

    Science.gov (United States)

    Alushin, Gregory M; Musinipally, Vivek; Matson, Daniel; Tooley, John; Stukenberg, P Todd; Nogales, Eva

    2012-11-01

    The Ndc80 complex is a key site of kinetochore-microtubule attachment during cell division. The human complex engages microtubules with a globular 'head' formed by tandem calponin-homology domains and an 80-amino-acid unstructured 'tail' that contains sites of phosphoregulation by the Aurora B kinase. Using biochemical, cell biological and electron microscopy analyses, we dissected the roles of the tail in binding of microtubules and mediation of cooperative interactions between Ndc80 complexes. Two segments of the tail that contain Aurora B phosphorylation sites become ordered at interfaces; one with tubulin and the second with an adjacent Ndc80 head on the microtubule surface, forming interactions that are disrupted by phosphorylation. We propose a model in which Ndc80's interaction with either growing or shrinking microtubule ends can be tuned by the phosphorylation state of its tail.

  7. Tensile stress stimulates microtubule outgrowth in living cells

    Science.gov (United States)

    Kaverina, Irina; Krylyshkina, Olga; Beningo, Karen; Anderson, Kurt; Wang, Yu-Li; Small, J. Victor

    2002-01-01

    Cell motility is driven by the sum of asymmetric traction forces exerted on the substrate through adhesion foci that interface with the actin cytoskeleton. Establishment of this asymmetry involves microtubules, which exert a destabilising effect on adhesion foci via targeting events. Here, we demonstrate the existence of a mechano-sensing mechanism that signals microtubule polymerisation and guidance of the microtubules towards adhesion sites under increased stress. Stress was applied either by manipulating the body of cells moving on glass with a microneedle or by stretching a flexible substrate that cells were migrating on. We propose a model for this mechano-sensing phenomenon whereby microtubule polymerisation is stimulated and guided through the interaction of a microtubule tip complex with actin filaments under tension.

  8. Monitoring and predicting eutrophication of Sri Lankan inland waters using ASTER satellite data

    Science.gov (United States)

    Dahanayaka, D. D. G. L.; Wijeyaratne, M. J. S.; Tonooka, H.; Minato, A.; Ozawa, S.; Perera, B. D. C.

    2014-10-01

    This study focused on determining the past changes and predicting the future trends in eutrophication of the Bolgoda North lake, Sri Lanka using in situ Chlorophyll-a (Chl-a) measurements and Advanced Spaceborne Thermal Emission and Reflectance Radiometer (ASTER) satellite data. This Lake is located in a mixed land use area with industries, some agricultural lands, middle income and high income housing, tourist hotels and low income housing. From March to October 2013, water samples from five sampling sites were collected once a month parallel to ASTER overpass and Chl-a, nitrate and phosphate contents of each sample were measured using standard laboratory methods. Cloud-free ASTER scenes over the lake during the 2000-2013 periods were acquired for Chl-a estimation and trend analysis. All ASTER images were atmospherically corrected using FLAASH software and in-situ Chl-a data were regressed with atmospherically corrected three ASTER VNIR band ratios of the same date. The regression equation of the band ratio and Chl-a content with the highest correlation, which was the green/red band ratio was used to develop algorithm for generation of 15-m resolution Chl-a distribution maps. According to the ASTER based Chl-a distribution maps it was evident that eutrophication of this lake has gradually increased from 2008-2011. Results also indicated that there had been significantly high eutrophic conditions throughout the year 2013 in several regions, especially in water stagnant areas and adjacent to freshwater outlets. Field observations showed that this lake is receiving various discharges from factories. Unplanned urbanization and inadequacy of proper facilities in the nearby industries for waste management have resulted in the eutrophication of the water body. If the present trends of waste disposal and unplanned urbanization continue, enormous environmental problems would be resulted in future. Results of the present study showed that information from satellite remote

  9. Efficient event-driven simulations shed new light on microtubule organization in the plant cortical array

    Science.gov (United States)

    Tindemans, Simon H.; Deinum, Eva E.; Lindeboom, Jelmer J.; Mulder, Bela M.

    2014-04-01

    The dynamics of the plant microtubule cytoskeleton is a paradigmatic example of the complex spatiotemporal processes characterising life at the cellular scale. This system is composed of large numbers of spatially extended particles, each endowed with its own intrinsic stochastic dynamics, and is capable of non-equilibrium self-organisation through collisional interactions of these particles. To elucidate the behaviour of such a complex system requires not only conceptual advances, but also the development of appropriate computational tools to simulate it. As the number of parameters involved is large and the behaviour is stochastic, it is essential that these simulations be fast enough to allow for an exploration of the phase space and the gathering of sufficient statistics to accurately pin down the average behaviour as well as the magnitude of fluctuations around it. Here we describe a simulation approach that meets this requirement by adopting an event-driven methodology that encompasses both the spontaneous stochastic changes in microtubule state as well as the deterministic collisions. In contrast with finite time step simulations this technique is intrinsically exact, as well as several orders of magnitude faster, which enables ordinary PC hardware to simulate systems of ˜ 10^3 microtubules on a time scale ˜ 10^{3} faster than real time. In addition we present new tools for the analysis of microtubule trajectories on curved surfaces. We illustrate the use of these methods by addressing a number of outstanding issues regarding the importance of various parameters on the transition from an isotropic to an aligned and oriented state.

  10. Efficient event-driven simulations shed new light on microtubule organisation in the plant cortical array

    Directory of Open Access Journals (Sweden)

    Simon H. Tindemans

    2014-04-01

    Full Text Available The dynamics of the plant microtubule cytoskeleton is a paradigmatic example of the complex spatiotemporal processes characterising life at the cellular scale. This system is composed of large numbers of spatially extended particles, each endowed with its own intrinsic stochastic dynamics, and is capable of non-equilibrium self-organisation through collisional interactions of these particles. To elucidate the behaviour of such a complex system requires not only conceptual advances, but also the development of appropriate computational tools to simulate it. As the number of parameters involved is large and the behaviour is stochastic, it is essential that these simulations be fast enough to allow for an exploration of the phase space and the gathering of sufficient statistics to accurately pin down the average behaviour as well as the magnitude of fluctuations around it. Here we describe a simulation approach that meets this requirement by adopting an event-driven methodology that encompasses both the spontaneous stochastic changes in microtubule state as well as the deterministic collisions. In contrast with finite time step simulations this technique is intrinsically exact, as well as several orders of magnitude faster, which enables ordinary PC hardware to simulate systems of $sim 10^3$ microtubules on a time scale $sim 10^{3}$ faster than real time. In addition we present new tools for the analysis of microtubule trajectories on curved surfaces. We illustrate the use of these methods by addressing a number of outstanding issues regarding the importance of various parameters on the transition from an isotropic to an aligned and oriented state.

  11. Examination of actin and microtubule dependent APC localisations in living mammalian cells

    Directory of Open Access Journals (Sweden)

    Adams Matthew

    2006-01-01

    Full Text Available Abstract Background The trafficking of the adenomatous polyposis coli (APC tumour suppressor protein in mammalian cells is a perennially controversial topic. Immunostaining evidence for an actin-associated APC localisation at intercellular junctions has been previously presented, though live imaging of mammalian junctional APC has not been documented. Results Using live imaging of transfected COS-7 cells we observed intercellular junction-associated pools of GFP-APC in addition to previously documented microtubule-associated GFP-APC and a variety of minor localisations. Although both microtubule and junction-associated populations could co-exist within individual cells, they differed in their subcellular location, dynamic behaviour and sensitivity to cytoskeletal poisons. GFP-APC deletion mutant analysis indicated that a protein truncated immediately after the APC armadillo repeat domain retained the ability to localise to adhesive membranes in transfected cells. Supporting this, we also observed junctional APC immunostaining in cultures of human colorectal cancer cell line that express truncated forms of APC. Conclusion Our data indicate that APC can be found in two spatially separate populations at the cell periphery and these populations can co-exist in the same cell. The first localisation is highly dynamic and associated with microtubules near free edges and in cell vertices, while the second is comparatively static and is closely associated with actin at sites of cell-cell contact. Our imaging confirms that human GFP-APC possesses many of the localisations and behaviours previously seen by live imaging of Xenopus GFP-APC. However, we report the novel finding that GFP-APC puncta can remain associated with the ends of shrinking microtubules. Deletion analysis indicated that the N-terminal region of the APC protein mediated its junctional localisation, consistent with our observation that truncated APC proteins in colon cancer cell lines are

  12. KIF7 Controls the Proliferation of Cells of the Respiratory Airway through Distinct Microtubule Dependent Mechanisms.

    Directory of Open Access Journals (Sweden)

    Garry L Coles

    2015-10-01

    Full Text Available The cell cycle must be tightly coordinated for proper control of embryonic development and for the long-term maintenance of organs such as the lung. There is emerging evidence that Kinesin family member 7 (Kif7 promotes Hedgehog (Hh signaling during embryonic development, and its misregulation contributes to diseases such as ciliopathies and cancer. Kif7 encodes a microtubule interacting protein that controls Hh signaling through regulation of microtubule dynamics within the primary cilium. However, whether Kif7 has a function in nonciliated cells remains largely unknown. The role Kif7 plays in basic cell biological processes like cell proliferation or cell cycle progression also remains to be elucidated. Here, we show that Kif7 is required for coordination of the cell cycle, and inactivation of this gene leads to increased cell proliferation in vivo and in vitro. Immunostaining and transmission electron microscopy experiments show that Kif7dda/dda mutant lungs are hyperproliferative and exhibit reduced alveolar epithelial cell differentiation. KIF7 depleted C3H10T1/2 fibroblasts and Kif7dda/dda mutant mouse embryonic fibroblasts have increased growth rates at high cellular densities, suggesting that Kif7 may function as a general regulator of cellular proliferation. We ascertained that in G1, Kif7 and microtubule dynamics regulate the expression and activity of several components of the cell cycle machinery known to control entry into S phase. Our data suggest that Kif7 may function to regulate the maintenance of the respiratory airway architecture by controlling cellular density, cell proliferation, and cycle exit through its role as a microtubule associated protein.

  13. Recent variation of the Las Vacas Glacier Mt. Aconcagua region, Central Andes, Argentina, based on ASTER stereoscopic images

    Science.gov (United States)

    Lenzano, M. G.; Leiva, J. C.; Lenzano, L.

    2010-01-01

    This work presents the results of the ASTER stereoscopic image processing to calculate the volume changes of Las Vacas Glacier. The processing of medium resolution satellite images (ASTER level 1A - pixel 15 m) from February 2001 and 2007 was performed applying the satellite digital photogrammetry method (Kääb, 2005). The comparison of the two generated DTM returns results that are acceptable within the parameters and precisions that can be obtained with this kind of sensor and the processing methodology.

  14. Marine boundary layer cloud property retrievals from high-resolution ASTER observations: case studies and comparison with Terra MODIS

    Science.gov (United States)

    Werner, Frank; Wind, Galina; Zhang, Zhibo; Platnick, Steven; Di Girolamo, Larry; Zhao, Guangyu; Amarasinghe, Nandana; Meyer, Kerry

    2016-12-01

    A research-level retrieval algorithm for cloud optical and microphysical properties is developed for the Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) aboard the Terra satellite. It is based on the operational MODIS algorithm. This paper documents the technical details of this algorithm and evaluates the retrievals for selected marine boundary layer cloud scenes through comparisons with the operational MODIS Data Collection 6 (C6) cloud product. The newly developed, ASTER-specific cloud masking algorithm is evaluated through comparison with an independent algorithm reported in [Zhao and Di Girolamo(2006)]. To validate and evaluate the cloud optical thickness (τ) and cloud effective radius (reff) from ASTER, the high-spatial-resolution ASTER observations are first aggregated to the same 1000 m resolution as MODIS. Subsequently, τaA and reff, aA retrieved from the aggregated ASTER radiances are compared with the collocated MODIS retrievals. For overcast pixels, the two data sets agree very well with Pearson's product-moment correlation coefficients of R > 0.970. However, for partially cloudy pixels there are significant differences between reff, aA and the MODIS results which can exceed 10 µm. Moreover, it is shown that the numerous delicate cloud structures in the example marine boundary layer scenes, resolved by the high-resolution ASTER retrievals, are smoothed by the MODIS observations. The overall good agreement between the research-level ASTER results and the operational MODIS C6 products proves the feasibility of MODIS-like retrievals from ASTER reflectance measurements and provides the basis for future studies concerning the scale dependency of satellite observations and three-dimensional radiative effects.

  15. Effects of spermine binding on Taxol-stabilized microtubules

    Science.gov (United States)

    Cheng, Shengfeng; Regmi, Chola

    Previous studies have shown that polyamines such as spermine present in cells at physiological concentrations can facilitate the polymerization of tubulins into microtubules (MTs). A recent experiment demonstrates that in the presence of high-concentration spermine, Taxol-stabilized MTs undergo a shape transformation into inverted tubulin tubules (ITTs), the outside surface of which corresponds to the inside surface of a regular MT. However, the molecular mechanism underlying the shape transformation of MTs into ITTs is unclear. We perform all atom molecular dynamics simulations on Taxol-stabilized MT sheets containing two protofilaments surrounded by spermine ions. The spermine concentration is varied from 0 to 25mM to match the range probed experimentally. We identify important spermine binding regions on the MT surface and the influence of the spermine binding on the structure and dynamics of MTs. In contrast to Taxol, our results show that spermine binding seems to decrease the flexibility of tubulin proteins, resulting in weaker tubulin-tubulin contacts and promoting the bending of protofilaments into curved protofilaments, inverted rings, and eventually inverted tubules.

  16. A pseudo-spin model for the wall of cytoskeletal microtubule

    Institute of Scientific and Technical Information of China (English)

    Chen Ying; Qiu Xi-Jun; Li Ru-Xin

    2005-01-01

    A pseudo-spin model is intended to describe the physical dynamics of unbound electrons in the wall of cytoskeletal microtubule (MT). Due to the inherent symmetry of the structure and the electric properties in the MT, one may treat it as a one-dimensional ferroelectric system, and describe the nonlinear dynamics of dimer electric dipoles in one protofilament of the MT by virtue of the double-well potential. Consequently, the physical problem has been mapped onto the pseudo-spin system, and the mean-field approximation has been taken to get some physical results.

  17. Visual interpretation of ASTER satellite data, Part II: Land use mapping in Mpumalanga,South Africa

    Directory of Open Access Journals (Sweden)

    Elna van Niekerk

    2007-09-01

    Full Text Available Since the initiation in 1960 of the era of satellite remote sensing to detect the different characteristics of the earth, a powerful tool was created to aid researchers. Many land-use studies were undertaken using Landsat MSS, Landsat TM and ETM, as well as SPOT satellite data. The application of these data to the mapping of land use and land cover at smaller scales was constrained by the limited spectral and/or spatial resolution of the data provided by these satellite sensors. In view of the relatively high cost of SPOT data, and uncertainty regarding the future continuation of the Landsat series, alternative data sources need to be investigated. In the absence of published previous research on this issue in South Africa, the purpose of this article is to investigate the value of visual interpretation of ASTER satellite images for the identification and mapping of land-use in an area in South Africa. The study area is situated in Mpumalanga, in the area of Witbank, around the Witbank and Doorndraai dams. This area is characterised by a variety of urban, rural and industrial land uses. Digital image processing of one Landsat 5 TM, one Landsat 7 ETM and one ASTER satellite image was undertaken, including atmospheric correction and georeferencing, natural colour composites, photo infrared colour composites (or false colour satellite images, band ratios, Normalised Difference Indices, as well as the Brightness, Greenness and Wetness Indices. The efficacy with which land use could be identified through the visual interpretation of the processed Landsat 5 TM, Landsat 7 TM and ASTER satellite images was compared. The published 1:50 000 topographical maps of the area were used for the purpose of initial verification. Findings of the visual interpretation process were verified by field visits to the study area. The study found that the ASTER satellite data produced clearer results and therefore have a higher mapping ability and capacity than the

  18. Effect of colour and size grading of China aster (Callistephus chinensis Nees seeds on their germination

    Directory of Open Access Journals (Sweden)

    Agnieszka Rosińska

    2012-12-01

    Full Text Available Seeds of 3 commercial China aster (Callistephus chinensis Nees lots were divided by hand into 3 grades with different colours: dark brown, brown and light brown, and 2 grades with a different size: length below 3.9 mm (small and above 3.9 mm (large. The colour grading was done based on the Royal Horticultural Society Colour Chart and size grading was done by hand for each seed. Then, seeds were routinely germinated based on the International Seed Testing Association (ISTA rules. The size of seeds had no effect on their germination. The dark brown seeds germinated better than the light brown ones. Removing light brown seeds from the China aster seed lot improved their germination.

  19. Estimation of Tree Size Diversity Using Object Oriented Texture Analysis and Aster Imagery.

    Science.gov (United States)

    Ozdemir, Ibrahim; Norton, David A; Ozkan, Ulas Yunus; Mert, Ahmet; Senturk, Ozdemir

    2008-08-11

    This study investigates the potential of object-based texture parameters extracted from 15m spatial resolution ASTER imagery for estimating tree size diversity in a Mediterranean forested landscape in Turkey. Tree size diversity based on tree basal area was determined using the Shannon index and Gini Coefficient at the sampling plot level. Image texture parameters were calculated based on the grey level co-occurrence matrix (GLCM) for various image segmentation levels. Analyses of relationships between tree size diversity and texture parameters found that relationships between the Gini Coefficient and the GLCM values were the most statistically significant, with the highest correlation (r=0.69) being with GLCM Homogeneity values. In contrast, Shannon Index values were weakly correlated with image derived texture parameters. The results suggest that 15m resolution Aster imagery has considerable potential in estimating tree size diversity based on the Gini Coefficient for heterogeneous Mediterranean forests.

  20. Digital Mapping of Soil Properties Using Multivariate Statistical Analysis and ASTER Data in an Arid Region

    Directory of Open Access Journals (Sweden)

    Said Nawar

    2015-01-01

    Full Text Available Modeling and mapping of soil properties has been identified as key for effective land degradation management and mitigation. The ability to model and map soil properties at sufficient accuracy for a large agriculture area is demonstrated using Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER imagery. Soil samples were collected in the El-Tina Plain, Sinai, Egypt, concurrently with the acquisition of ASTER imagery, and measured for soil electrical conductivity (ECe, clay content and soil organic matter (OM. An ASTER image covering the study area was preprocessed, and two predictive models, multivariate adaptive regression splines (MARS and the partial least squares regression (PLSR, were constructed based on the ASTER spectra. For all three soil properties, the results of MARS models were better than those of the respective PLSR models, with cross-validation estimated R2 of 0.85 and 0.80 for ECe, 0.94 and 0.90 for clay content and 0.79 and 0.73 for OM. Independent validation of ECe, clay content and OM maps with 32 soil samples showed the better performance of the MARS models, with R2 = 0.81, 0.89 and 0.73, respectively, compared to R2 = 0.78, 0.87 and 0.71 for the PLSR models. The results indicated that MARS is a more suitable and superior modeling technique than PLSR for the estimation and mapping of soil salinity (ECe, clay content and OM. The method developed in this paper was found to be reliable and accurate for digital soil mapping in arid and semi-arid environments.

  1. The microtubule plus-end tracking proteins SPR1 and EB1b interact to maintain polar cell elongation and directional organ growth in Arabidopsis.

    Science.gov (United States)

    Galva, Charitha; Kirik, Viktor; Lindeboom, Jelmer J; Kaloriti, Despoina; Rancour, David M; Hussey, Patrick J; Bednarek, Sebastian Y; Ehrhardt, David W; Sedbrook, John C

    2014-11-01

    The microtubule plus-end tracking proteins (+TIPs) END BINDING1b (EB1b) and SPIRAL1 (SPR1) are required for normal cell expansion and organ growth. EB proteins are viewed as central regulators of +TIPs and cell polarity in animals; SPR1 homologs are specific to plants. To explore if EB1b and SPR1 fundamentally function together, we combined genetic, biochemical, and cell imaging approaches in Arabidopsis thaliana. We found that eb1b-2 spr1-6 double mutant roots exhibit substantially more severe polar expansion defects than either single mutant, undergoing right-looping growth and severe axial twisting instead of waving on tilted hard-agar surfaces. Protein interaction assays revealed that EB1b and SPR1 bind each other and tubulin heterodimers, which is suggestive of a microtubule loading mechanism. EB1b and SPR1 show antagonistic association with microtubules in vitro. Surprisingly, our combined analyses revealed that SPR1 can load onto microtubules and function independently of EB1 proteins, setting SPR1 apart from most studied +TIPs in animals and fungi. Moreover, we found that the severity of defects in microtubule dynamics in spr1 eb1b mutant hypocotyl cells correlated well with the severity of growth defects. These data indicate that SPR1 and EB1b have complex interactions as they load onto microtubule plus ends and direct polar cell expansion and organ growth in response to directional cues.

  2. The Microtubule Plus-End Tracking Proteins SPR1 and EB1b Interact to Maintain Polar Cell Elongation and Directional Organ Growth in Arabidopsis[W

    Science.gov (United States)

    Galva, Charitha; Kirik, Viktor; Lindeboom, Jelmer J.; Kaloriti, Despoina; Rancour, David M.; Hussey, Patrick J.; Bednarek, Sebastian Y.; Ehrhardt, David W.; Sedbrook, John C.

    2014-01-01

    The microtubule plus-end tracking proteins (+TIPs) END BINDING1b (EB1b) and SPIRAL1 (SPR1) are required for normal cell expansion and organ growth. EB proteins are viewed as central regulators of +TIPs and cell polarity in animals; SPR1 homologs are specific to plants. To explore if EB1b and SPR1 fundamentally function together, we combined genetic, biochemical, and cell imaging approaches in Arabidopsis thaliana. We found that eb1b-2 spr1-6 double mutant roots exhibit substantially more severe polar expansion defects than either single mutant, undergoing right-looping growth and severe axial twisting instead of waving on tilted hard-agar surfaces. Protein interaction assays revealed that EB1b and SPR1 bind each other and tubulin heterodimers, which is suggestive of a microtubule loading mechanism. EB1b and SPR1 show antagonistic association with microtubules in vitro. Surprisingly, our combined analyses revealed that SPR1 can load onto microtubules and function independently of EB1 proteins, setting SPR1 apart from most studied +TIPs in animals and fungi. Moreover, we found that the severity of defects in microtubule dynamics in spr1 eb1b mutant hypocotyl cells correlated well with the severity of growth defects. These data indicate that SPR1 and EB1b have complex interactions as they load onto microtubule plus ends and direct polar cell expansion and organ growth in response to directional cues. PMID:25415978

  3. Microtubules in the Cerebral Cortex: Role in Memory and Consciousness

    Science.gov (United States)

    Woolf, Nancy J.

    This chapter raises the question whether synaptic connections in the cerebral cortex are adequate in accounting for higher cognition, especially cognition involving multimodal processing. A recent and novel approach to brain mechanics is outlined, one that involves microtubules and microtubule-associated protein-2 (MAP2). In addition to effects on the neuronal membrane, neurotransmitters exert actions on microtubules. These neurotransmitter effects alter the MAP2 phosphorylation state and rates of microtubule polymerization and transport. It is argued that these processes are important to the physical basis of memory and consciousness. In support of this argument, MAP2 is degraded with learning in discrete cortical modules. How this relates to synaptic change related to learning is unknown. The specific proposal is advanced that learning alters microtubules in the subsynaptic zone lying beneath the synapse, and that this forms the physical basis of long-term memory storage because microtubule networks determine the synapse strength by directing contacts with actin filaments and transport of synaptic proteins. It is argued that this is more probable than memory-related physical storage in the synapse itself. Comparisons to consciousness are made and it is concluded that there is a link between microtubules, memory and consciousness.

  4. An improved quantitative analysis method for plant cortical microtubules.

    Science.gov (United States)

    Lu, Yi; Huang, Chenyang; Wang, Jia; Shang, Peng

    2014-01-01

    The arrangement of plant cortical microtubules can reflect the physiological state of cells. However, little attention has been paid to the image quantitative analysis of plant cortical microtubules so far. In this paper, Bidimensional Empirical Mode Decomposition (BEMD) algorithm was applied in the image preprocessing of the original microtubule image. And then Intrinsic Mode Function 1 (IMF1) image obtained by decomposition was selected to do the texture analysis based on Grey-Level Cooccurrence Matrix (GLCM) algorithm. Meanwhile, in order to further verify its reliability, the proposed texture analysis method was utilized to distinguish different images of Arabidopsis microtubules. The results showed that the effect of BEMD algorithm on edge preserving accompanied with noise reduction was positive, and the geometrical characteristic of the texture was obvious. Four texture parameters extracted by GLCM perfectly reflected the different arrangements between the two images of cortical microtubules. In summary, the results indicate that this method is feasible and effective for the image quantitative analysis of plant cortical microtubules. It not only provides a new quantitative approach for the comprehensive study of the role played by microtubules in cell life activities but also supplies references for other similar studies.

  5. An Improved Quantitative Analysis Method for Plant Cortical Microtubules

    Directory of Open Access Journals (Sweden)

    Yi Lu

    2014-01-01

    Full Text Available The arrangement of plant cortical microtubules can reflect the physiological state of cells. However, little attention has been paid to the image quantitative analysis of plant cortical microtubules so far. In this paper, Bidimensional Empirical Mode Decomposition (BEMD algorithm was applied in the image preprocessing of the original microtubule image. And then Intrinsic Mode Function 1 (IMF1 image obtained by decomposition was selected to do the texture analysis based on Grey-Level Cooccurrence Matrix (GLCM algorithm. Meanwhile, in order to further verify its reliability, the proposed texture analysis method was utilized to distinguish different images of Arabidopsis microtubules. The results showed that the effect of BEMD algorithm on edge preserving accompanied with noise reduction was positive, and the geometrical characteristic of the texture was obvious. Four texture parameters extracted by GLCM perfectly reflected the different arrangements between the two images of cortical microtubules. In summary, the results indicate that this method is feasible and effective for the image quantitative analysis of plant cortical microtubules. It not only provides a new quantitative approach for the comprehensive study of the role played by microtubules in cell life activities but also supplies references for other similar studies.

  6. Oscillatory fluid flow influences primary cilia and microtubule mechanics.

    Science.gov (United States)

    Espinha, Lina C; Hoey, David A; Fernandes, Paulo R; Rodrigues, Hélder C; Jacobs, Christopher R

    2014-07-01

    Many tissues are sensitive to mechanical stimuli; however, the mechanotransduction mechanism used by cells remains unknown in many cases. The primary cilium is a solitary, immotile microtubule-based extension present on nearly every mammalian cell which extends from the basal body. The cilium is a mechanosensitive organelle and has been shown to transduce fluid flow-induced shear stress in tissues, such as the kidney and bone. The majority of microtubules assemble from the mother centriole (basal body), contributing significantly to the anchoring of the primary cilium. Several studies have attempted to quantify the number of microtubules emanating from the basal body and the results vary depending on the cell type. It has also been shown that cellular response to shear stress depends on microtubular integrity. This study hypothesizes that changing the microtubule attachment of primary cilia in response to a mechanical stimulus could change primary cilia mechanics and, possibly, mechanosensitivity. Oscillatory fluid flow was applied to two different cell types and the microtubule attachment to the ciliary base was quantified. For the first time, an increase in microtubules around primary cilia both with time and shear rate in response to oscillatory fluid flow stimulation was demonstrated. Moreover, it is presented that the primary cilium is required for this loading-induced cellular response. This study has demonstrated a new role for the cilium in regulating alterations in the cytoplasmic microtubule network in response to mechanical stimulation, and therefore provides a new insight into how cilia may regulate its mechanics and thus the cells mechanosensitivity.

  7. Roles for microtubule and microfilament cytoskeletons in animal cell cytokinesis

    Institute of Scientific and Technical Information of China (English)

    CHEN Zhongcai; CAI Shang; JIANG Qing; ZHANG Chuanmao; TANG Xiaowei

    2005-01-01

    Microtubule and microfilament cytoskeletons play key roles in the whole process of cytokinesis. Although a number of hypotheses have been proposed to elucidate the mechanism of cytokinesis by microtubule and actin filament cytoskeletons, many reports are conflicting. In our study, combining the cytoskeletons drug treatments with the time-lapse video technology, we retested the key roles of microtubule and actin filament in cytokinesis. The results showed that depolymerization of microtubules by Nocodazole after the initiation of furrowing would not inhibit the furrow ingression, but obviously decrease the stiffness of daughter cells. Depolymerizing actin filaments by Cytochalasin B before metaphase would inhibit the initiation of furrowing but not chromosome segregation, resulting in the formation of binucleate cells; however, depolymerizing actin filaments during anaphase would prevent furrowing and lead to the regress of established furrow, also resulting in the formation of binucleate cells. Further, depolymerizing microtubules and actin filaments simultaneously after metaphase would cause the quick regress of the furrow and the formation of binucleate cells. From these results we propose that a successful cytokinesis requires functions and coordination of both the microtubule and actin filament cytoskeletons. Microtubule cytoskeleton may function in the positioning and initiation of cleavage furrow, and the actin filament cytoskeleton may play key roles in the initiation and ingression of the furrow.

  8. PEMETAAN SUHU PERMUKAAN LAUT (SPL MENGGUNAKAN CITRA SATELIT ASTER DI PERAIRAN LAUT JAWA BAGIAN BARAT MADURA

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    Dyah Ayu Sulistyo Rini

    2010-10-01

    Full Text Available Oceanographical temperature in Java Sea is very important to be considered. This research was combines in-site observation technique, Geographical Information System (GLS and remote sensing in order to get accurate, present and updateable data. The aim of this research is to determine the distribution of sea-surface temperature and accuration-test value in Java Sea especially on western coast of Madura using ASTER satellite imagery. This research were used software of ENVI 4.5, ILWIS 3.3, and ArcGIS 9.3 and also changed the radian value until °C. Result showed that using ASTER satellite imagery within band 10 range between 32 "C-35 "C. Band 11,between 24.9"C 25,2"C. Band 12 between 16,7"C to 17"C. Band while band 13 abd 14 between 30.7, band 28. Band 11 is more accurate compared to Band 10, 12, 13. 14, the RMS Error on band 11 showed lower value compared to the other band.Keywords: Sea-surface Temperature. ASTER satellite imagery. Java Sea, Western coast of Madura

  9. Remote sensing of thermal state of volcanoes in Turkey and neighbouring countries using ASTER nighttime images

    Science.gov (United States)

    Ulusoy, İnan; Diker, Caner

    2016-04-01

    Ongoing studies are increasingly revealing that Holocene and historical activity has been reported for many of the Anatolian volcanoes. So far, hydrothermal activity have been observed on Nemrut, Tendürek, Aǧrı (Ararat), Hasan daǧ and Kula. Fumaroles, steam vents, steam/gas emission and zones of hot grounds have been reported. Thermal state of Anatolian volcanoes have been investigated using ASTER nighttime satellite imagery. We have analyzed the nighttime thermal images of Aǧrı, Akça, Çandarlı, Erciyes, Gölcük, Göllüdaǧ, Hasandaǧ, Kula, Meydan, Nemrut, Süphan and Tendürek volcanoes in Turkey and Demavand and Nisyros volcanoes in the neighboring countries. In order to quantify the current thermal state of the volcanos studied, we have used ASTER Thermal Infrared spectra. Several ASTER nighttime images have been used to calculate land surface temperature, surface thermal anomaly and relative radiative heat flux on the volcanoes. Following the atmospheric correction of thermal images, temperature and emissivity have been separated and then land surface temperature have been calculated from 5 thermal bands. Surface temperature images have been topographically corrected. Relative radiative heat flux have been calculated using corrected surface temperature data, emissivity, vapor pressure and height-dependent air temperature values. These values have been correlated with ongoing activity observed on active Indonesian volcanoes Sinabung, Semeru and Bromo Tengger. (This study have been financially supported by TUBITAK project no: 113Y032).

  10. Detection of Alteration Induced by Onshore Gas Seeps from ASTER and WorldView-2 Data

    Directory of Open Access Journals (Sweden)

    Sanaz Salati

    2014-04-01

    Full Text Available Hydrocarbon seeps cause chemical and mineralogical changes at the surface, which can be detected by remote sensing. This paper aims at the detection of mineral alteration induced by gas seeps in a marly limestone formation, SW Iran. For this purpose, the multispectral Advance Spaceborne Thermal Emission and Reflection Radiometer (ASTER and the high spatial resolution WorldView-2 (WV-2 data were utilized for mapping surficial rock alteration. In addition, the potential of Visible Near Infrared (VNIR bands of the WV-2 and its high spatial resolution for mapping alterations was determined. Band ratioing, principal component analysis (PCA, data fusion and the boosted regression trees (BRT were applied to enhance and classify the altered and unaltered marly limestone formation. The alteration zones were identified and mapped by remote sensing analyses. Integrating the WV-2 into the ASTER data improved the spatial accuracy of the BRT classifications. The results showed that the BRT classification of the multiple band imagery (created from ASTER and WV-2 using regions of interest (ROIs around field data provides the best discrimination between altered and unaltered areas. It is suggested that the WV-2 dataset can provide a potential tool along higher spectral resolution data for mapping alteration minerals related to hydrocarbon seeps in arid and semi-arid areas.

  11. Structural insights into microtubule doublet interactions inaxonemes

    Energy Technology Data Exchange (ETDEWEB)

    Downing, Kenneth H.; Sui, Haixin

    2007-06-06

    Coordinated sliding of microtubule doublets, driven by dynein motors, produces periodic beating of the axoneme. Recent structural studies of the axoneme have used cryo-electron tomography to reveal new details of the interactions among some of the multitude of proteins that form the axoneme and regulate its movement. Connections among the several sets of dyneins, in particular, suggest ways in which their actions may be coordinated. Study of the molecular architecture of isolated doublets has provided a structural basis for understanding the doublet's mechanical properties that are related to the bending of the axoneme, and has also offered insight into its potential role in the mechanism of dynein activity regulation.

  12. Centrosome and microtubule instability in aging Drosophila cells

    Science.gov (United States)

    Schatten, H.; Chakrabarti, A.; Hedrick, J.

    1999-01-01

    Several cytoskeletal changes are associated with aging which includes alterations in muscle structure leading to muscular atrophy, and weakening of the microtubule network which affects cellular secretion and maintenance of cell shape. Weakening of the microtubule network during meiosis in aging oocytes can result in aneuploidy or trisomic zygotes with increasing maternal age. Imbalances of cytoskeletal organization can lead to disease such as Alzheimer's, muscular disorders, and cancer. Because many cytoskeletal diseases are related to age we investigated the effects of aging on microtubule organization in cell cultures of the Drosophila cell model system (Schneider S-1 and Kc23 cell lines). This cell model is increasingly being used as an alternative system to mammalian cell cultures. Drosophila cells are amenable to genetic manipulations and can be used to identify and manipulate genes which are involved in the aging processes. Immunofluorescence, scanning, and transmission electron microscopy were employed for the analysis of microtubule organizing centers (centrosomes) and microtubules at various times after subculturing cells in fresh medium. Our results reveal that centrosomes and the microtubule network becomes significantly affected in aging cells after 5 days of subculture. At 5-14 days of subculture, 1% abnormal out of 3% mitoses were noted which were clearly distinguishable from freshly subcultured control cells in which 3% of cells undergo normal mitosis with bipolar configurations. Microtubules are also affected in the midbody during cell division. The midbody in aging cells becomes up to 10 times longer when compared with midbodies in freshly subcultured cells. During interphase, microtubules are often disrupted and disorganized, which may indicate improper function related to transport of cell organelles along microtubules. These results are likely to help explain some cytoskeletal disorders and diseases related to aging.

  13. Assembly of bipolar microtubule structures by passive cross-linkers and molecular motors

    Science.gov (United States)

    Johann, D.; Goswami, D.; Kruse, K.

    2016-06-01

    During cell division, sister chromatids are segregated by the mitotic spindle, a bipolar assembly of interdigitating antiparallel polar filaments called microtubules. The spindle contains the midzone, a stable region of overlapping antiparallel microtubules, that is essential for maintaining bipolarity. Although a lot is known about the molecular players involved, the mechanism underlying midzone formation and maintenance is still poorly understood. We study the interaction of polar filaments that are cross-linked by molecular motors moving directionally and by passive cross-linkers diffusing along microtubules. Using a particle-based stochastic model, we find that the interplay of motors and passive cross-linkers can generate a stable finite overlap between a pair of antiparallel polar filaments. We develop a mean-field theory to study this mechanism in detail and investigate the influence of steric interactions between motors and passive cross-linkers on the overlap dynamics. In the presence of interspecies steric interactions, passive cross-linkers mimic the behavior of molecular motors and stable finite overlaps are generated even for non-cross-linking motors. Finally, we develop a mean-field theory for a bundle of aligned polar filaments and show that they can self-organize into a spindlelike pattern. Our work suggests possible ways as to how cells can generate spindle midzones and control their extensions.

  14. Microtubule-associated proteins and tubulin interaction by isothermal titration calorimetry.

    Science.gov (United States)

    Tsvetkov, P O; Barbier, P; Breuzard, G; Peyrot, V; Devred, F

    2013-01-01

    Microtubules play an important role in a number of vital cell processes such as cell division, intracellular transport, and cell architecture. The highly dynamic structure of microtubules is tightly regulated by a number of stabilizing and destabilizing microtubule-associated proteins (MAPs), such as tau and stathmin. Because of their importance, tubulin-MAPs interactions have been extensively studied using various methods that provide researchers with complementary but sometimes contradictory thermodynamic data. Isothermal titration calorimetry (ITC) is the only direct thermodynamic method that enables a full thermodynamic characterization (stoichiometry, enthalpy, entropy of binding, and association constant) of the interaction after a single titration experiment. This method has been recently applied to study tubulin-MAPs interactions in order to bring new insights into molecular mechanisms of tubulin regulation. In this chapter, we review the technical specificity of this method and then focus on the use of ITC in the investigation of tubulin-MAPs binding. We describe technical issues which could arise during planning and carrying out the ITC experiments, in particular with fragile proteins such as tubulin. Using examples of stathmin and tau, we demonstrate how ITC can be used to gain major insights into tubulin-MAP interaction.

  15. 3D reconstruction of cortical microtubules using multi-angle total internal reflection fluorescence microscopy

    Science.gov (United States)

    Jin, Luhong; Xiu, Peng; Zhou, Xiaoxu; Fan, Jiannan; Kuang, Cuifang; Liu, Xu; Xu, Yingke

    2017-01-01

    Total internal reflection fluorescence microscopy (TIRFM) has been widely used in biomedical research to visualize cellular processes near the cell surface. In this study, a novel multi-angle ring-illuminated TIRFM system, equipped with two galvo mirrors that are on conjugate plan of a 4f optical system was developed. Multi-angle TIRFM generates images with different penetration depths through the controlled variation of the incident angle of illuminating laser. We presented a method to perform three-dimensional (3-D) reconstruction of microtubules from multi-angle TIRFM images. The performance of our method was validated in simulated microtubules with variable signal-to-noise ratios (SNR) and the axial resolution and accuracy of reconstruction were evaluated in selecting different numbers of illumination angles or in different SNR conditions. In U373 cells, we reconstructed the 3-D localization of microtubules near the cell surface with high resolution using over a hundred different illumination angles. Theoretically, the presented TIRFM setup and 3-D reconstruction method can achieve 40 nm axial resolution in experimental conditions where SNR is as low as 2, with 35 different illumination angles. Moreover, our system and reconstruction method have the potential to be used in live cells to track membrane dynamics in 3-D.

  16. Structural microtubule cap: Stability, catastrophe, rescue, and third state

    DEFF Research Database (Denmark)

    Flyvbjerg, H.; Chretien, D.; Janosi, I.M.

    2002-01-01

    Microtubules polymerize from GTP-liganded tubulin dinners, but are essentially made of GDP-liganded tubulin. We investigate the tug-of-war resulting from the fact that GDP-liganded tubulin favors a curved configuration, but is forced to remain in a straight one when part of a microtubule. We point...... of two well-established facts: protofilaments made of GDP-liganded tubulin have intrinsic curvature, and microtubules are elastic, made from material that can yield to forces, in casu its own intrinsic forces. We explore possible properties of this structural cap, and demonstrate 1) how it allows both...

  17. Regulation of localization and activity of the microtubule depolymerase MCAK.

    Science.gov (United States)

    Tanenbaum, Marvin E; Medema, René H; Akhmanova, Anna

    2011-03-01

    Mitotic Centromere Associated Kinesin (MCAK) is a potent microtubule depolymerizing and catastrophe-inducing factor, which uses the energy of ATP hydrolysis to destabilize microtubule ends. MCAK is localized to inner centromeres, kinetochores and spindle poles of mitotic cells, and is also present in the cytoplasm. Both in interphase and in mitosis, MCAK can specifically accumulate at the growing microtubule ends. Here we discuss the mechanisms, which modulate subcellular localization and activity of MCAK through the interaction with the End Binding (EB) proteins and phosphorylation.

  18. Mitochondrial inheritance is mediated by microtubules in mammalian cell division.

    Science.gov (United States)

    Lawrence, Elizabeth; Mandato, Craig

    2013-11-01

    The mitochondrial network fragments and becomes uniformly dispersed within the cytoplasm when mammalian cells enter mitosis. Such morphology and distribution of mitochondria was previously thought to facilitate the stochastic inheritance of mitochondria by daughter cells. In contrast, we recently reported that mitochondria in dividing mammalian cells are inherited by an ordered mechanism of inheritance mediated by microtubules. We showed that mitochondria are progressively enriched at the cell equator and depleted at the poles throughout division. Furthermore, the mitochondrial distribution during division is dependent on microtubules, indicating an ordered inheritance strategy. The microtubule-mediated positioning of mitochondria in dividing mammalian cells may have functional consequences for cell division and/or mitochondrial inheritance.

  19. Visualizing and Analyzing Branching Microtubule Nucleation Using Meiotic Xenopus Egg Extracts and TIRF Microscopy

    Science.gov (United States)

    King, Matthew; Petry, Sabine

    2016-01-01

    Mitotic and meiotic spindles consist primarily of microtubules, which originate from centrosomes and within the vicinity of chromatin. Indirect evidence suggested that microtubules also originate throughout the spindle, but the high microtubule density within the spindle precludes the direct observation of this phenomenon. By using meiotic Xenopus laevis egg extract and employing total internal reflection (TIRF) microscopy, microtubule nucleation from preexisting microtubules could be demonstrated and analyzed. Branching microtubule nucleation is an ideal mechanism to assemble and maintain a mitotic spindle, because microtubule numbers are amplified while preserving their polarity. Here, we describe the assays that made these findings possible and the experiments that helped identify the key molecular players involved. PMID:27193844

  20. Opposing microtubule motors control motility, morphology and cargo segregation during ER-to-Golgi transport

    Directory of Open Access Journals (Sweden)

    Anna K. Brown

    2014-04-01

    Full Text Available We recently demonstrated that dynein and kinesin motors drive multiple aspects of endosomal function in mammalian cells. These functions include driving motility, maintaining morphology (notably through providing longitudinal tension to support vesicle fission, and driving cargo sorting. Microtubule motors drive bidirectional motility during traffic between the endoplasmic reticulum (ER and Golgi. Here, we have examined the role of microtubule motors in transport carrier motility, morphology, and domain organization during ER-to-Golgi transport. We show that, consistent with our findings for endosomal dynamics, microtubule motor function during ER-to-Golgi transport of secretory cargo is required for motility, morphology, and cargo sorting within vesicular tubular carriers en route to the Golgi. Our data are consistent with previous findings that defined roles for dynein-1, kinesin-1 (KIF5B and kinesin-2 in this trafficking step. Our high resolution tracking data identify some intriguing aspects. Depletion of kinesin-1 reduces the number of motile structures seen, which is in line with other findings relating to the role of kinesin-1 in ER export. However, those transport carriers that were produced had a much greater run length suggesting that this motor can act as a brake on anterograde motility. Kinesin-2 depletion did not significantly reduce the number of motile transport carriers but did cause a similar increase in run length. These data suggest that kinesins act as negative regulators of ER-to-Golgi transport. Depletion of dynein not only reduced the number of motile carriers formed but also caused tubulation of carriers similar to that seen for sorting nexin-coated early endosomes. Our data indicated that the previously observed anterograde–retrograde polarity of transport carriers in transit to the Golgi from the ER is maintained by microtubule motor function.

  1. Identification of interphase functions for the NIMA kinase involving microtubules and the ESCRT pathway.

    Directory of Open Access Journals (Sweden)

    Meera Govindaraghavan

    2014-03-01

    Full Text Available The Never in Mitosis A (NIMA kinase (the founding member of the Nek family of kinases has been considered a mitotic specific kinase with nuclear restricted roles in the model fungus Aspergillus nidulans. By extending to A. nidulans the results of a synthetic lethal screen performed in Saccharomyces cerevisiae using the NIMA ortholog KIN3, we identified a conserved genetic interaction between nimA and genes encoding proteins of the Endosomal Sorting Complex Required for Transport (ESCRT pathway. Absence of ESCRT pathway functions in combination with partial NIMA function causes enhanced cell growth defects, including an inability to maintain a single polarized dominant cell tip. These genetic insights suggest NIMA potentially has interphase functions in addition to its established mitotic functions at nuclei. We therefore generated endogenously GFP-tagged NIMA (NIMA-GFP which was fully functional to follow its interphase locations using live cell spinning disc 4D confocal microscopy. During interphase some NIMA-GFP locates to the tips of rapidly growing cells and, when expressed ectopically, also locates to the tips of cytoplasmic microtubules, suggestive of non-nuclear interphase functions. In support of this, perturbation of NIMA function either by ectopic overexpression or through partial inactivation results in marked cell tip growth defects with excess NIMA-GFP promoting multiple growing cell tips. Ectopic NIMA-GFP was found to locate to the plus ends of microtubules in an EB1 dependent manner, while impairing NIMA function altered the dynamic localization of EB1 and the cytoplasmic microtubule network. Together, our genetic and cell biological analyses reveal novel non-nuclear interphase functions for NIMA involving microtubules and the ESCRT pathway for normal polarized fungal cell tip growth. These insights extend the roles of NIMA both spatially and temporally and indicate that this conserved protein kinase could help integrate cell

  2. Estimating surface fluxes over the north Tibetan Plateau area with ASTER imagery

    Directory of Open Access Journals (Sweden)

    Weiqiang Ma

    2009-01-01

    Full Text Available Surface fluxes are important boundary conditions for climatological modeling and Asian monsoon system. The recent availability of high-resolution, multi-band imagery from the ASTER (Advanced Space-borne Thermal Emission and Reflection radiometer sensor has enabled us to estimate surface fluxes to bridge the gap between local scale flux measurements using micrometeorological instruments and regional scale land-atmosphere exchanges of water and heat fluxes that are fundamental for the understanding of the water cycle in the Asian monsoon system. A parameterization method based on ASTER data and field observations has been proposed and tested for deriving surface albedo, surface temperature, Normalized Difference Vegetation Index (NDVI, Modified Soil Adjusted Vegetation Index (MSAVI, vegetation coverage, Leaf Area Index (LAI, net radiation flux, soil heat flux, sensible heat flux and latent heat flux over heterogeneous land surface in this paper. As a case study, the methodology was applied to the experimental area of the Coordinated Enhanced Observing Period (CEOP Asia-Australia Monsoon Project (CAMP on the Tibetan Plateau (CAMP/Tibet, located at the north Tibetan Plateau. The ASTER data of 24 July 2001, 29 November 2001 and 12 March 2002 was used in this paper for the case of summer, winter and spring. To validate the proposed methodology, the ground-measured surface variables (surface albedo and surface temperature and land surface heat fluxes (net radiation flux, soil heat flux, sensible heat flux and latent heat flux were compared to the ASTER derived values. The results show that the derived surface variables and land surface heat fluxes in three different months over the study area are in good accordance with the land surface status. Also, the estimated land surface variables and land surface heat fluxes are in good accordance with ground measurements, and all their absolute percentage difference (APD is less than 10% in the validation sites

  3. Effects of the KIF2C neck peptide on microtubules: lateral disintegration of microtubules and β-structure formation.

    Science.gov (United States)

    Shimizu, Youské; Shimizu, Takashi; Nara, Masayuki; Kikumoto, Mahito; Kojima, Hiroaki; Morii, Hisayuki

    2013-04-01

    Members of the kinesin-13 sub-family, including KIF2C, depolymerize microtubules. The positive charge-rich 'neck' region extending from the N-terminus of the catalytic head is considered to be important in the depolymerization activity. Chemically synthesized peptides, covering the basic region (A182-E200), induced a sigmoidal increase in the turbidity of a microtubule suspension. The increase was suppressed by salt addition or by reduction of basicity by amino acid substitutions. Electron microscopic observations revealed ring structures surrounding the microtubules at high peptide concentrations. Using the peptide A182-D218, we also detected free thin straight filaments, probably protofilaments disintegrated from microtubules. Therefore, the neck region, even without the catalytic head domain, may induce lateral disintegration of microtubules. With microtubules lacking anion-rich C-termini as a result of subtilisin treatment, addition of the peptide induced only a moderate increase in turbidity, and rings and protofilaments were rarely detected, while aggregations, also thought to be caused by lateral disintegration, were often observed in electron micrographs. Thus, the C-termini are not crucial for the action of the peptides in lateral disintegration but contribute to structural stabilization of the protofilaments. Previous structural studies indicated that the neck region of KIF2C is flexible, but our IR analysis suggests that the cation-rich region (K190-A204) forms β-structure in the presence of microtubules, which may be of significance with regard to the action of the neck region. Therefore, the neck region of KIF2C is sufficient to cause disintegration of microtubules into protofilaments, and this may contribute to the ability of KIF2C to cause depolymerization of microtubules.

  4. Enantioselective effects of (+)- and (-)-citronellal on animal and plant microtubules.

    Science.gov (United States)

    Altshuler, Osnat; Abu-Abied, Mohamad; Chaimovitsh, David; Shechter, Alona; Frucht, Hilla; Dudai, Nativ; Sadot, Einat

    2013-09-27

    Citronellal is a major component of Corymbia citriodora and Cymbopogon nardus essential oils. Herein it is shown that whereas (+)-citronellal (1) is an effective microtubule (MT)-disrupting compound, (-)-citronellal (2) is not. Quantitative image analysis of fibroblast cells treated with 1 showed total fluorescence associated with fibers resembling that in cells treated with the MT-disrupting agents colchicine and vinblastine; in the presence of 2, the fluorescence more closely resembled that in control cells. The distribution of tubulin in soluble and insoluble fractions in the presence of 1 also resembled that in the presence of colchicine, whereas similar tubulin distribution was obtained in the presence of 2 and in control cells. In vitro polymerization of MTs was inhibited by 1 but not 2. Measurements of MT dynamics in plant cells showed similar MT elongation and shortening rates in control and 2-treated cells, whereas in the presence of 1, much fewer and shorter MTs were observed and no elongation or shrinkage was detected. Taken together, the MT system is suggested to be able to discriminate between different enantiomers of the same compound. In addition, the activity of essential oils rich in citronellal is affected by the relative content of the two enantiomers of this monoterpenoid.

  5. Feedback Mechanism for Microtubule Length Regulation by Stathmin Gradients

    Science.gov (United States)

    Zeitz, Maria; Kierfeld, Jan

    2014-12-01

    We formulate and analyze a theoretical model for the regulation of microtubule (MT) polymerization dynamics by the signaling proteins Rac1 and stathmin. In cells, the MT growth rate is inhibited by cytosolic stathmin, which, in turn, is inactivated by Rac1. Growing MTs activate Rac1 at the cell edge, which closes a positive feedback loop. We investigate both tubulin sequestering and catastrophe promotion as mechanisms for MT growth inhibition by stathmin. For a homogeneous stathmin concentration in the absence of Rac1, we find a switch-like regulation of the MT mean length by stathmin. For constitutively active Rac1 at the cell edge, stathmin is deactivated locally, which establishes a spatial gradient of active stathmin. In this gradient, we find a stationary bimodal MT length distributions for both mechanisms of MT growth inhibition by stathmin. One subpopulation of the bimodal length distribution can be identified with fast growing and long pioneering MTs in the region near the cell edge, which have been observed experimentally. The feedback loop is closed through Rac1 activation by MTs. For tubulin sequestering by stathmin, this establishes a bistable switch with two stable states: one stable state corresponds to upregulated MT mean length and bimodal MT length distributions, i.e., pioneering MTs; the other stable state corresponds to an interrupted feedback with short MTs. Stochastic effects as well as external perturbations can trigger switching events. For catastrophe promoting stathmin we do not find bistability.

  6. Microtubule as a Transmission Line for Ionic Currents

    Institute of Scientific and Technical Information of China (English)

    ILI(C) D.I.; SATARI(C) M.V.; RALEVI(C) N.

    2009-01-01

    We establish a new model for ionic waves along microtubules based on polyelectrolyte features of cylindrical biopolymers. The nonlinear transmission line described by a nonlinear differential equation is obtained with stable kink solution pertinent to the shape of the front of accompanying potential. The localized ionic wave could be used to explain the behavior of microtubules as biomolecular transistors capable of amplifying electrical information in neurons.

  7. Transport properties of melanosomes along microtubules interpreted by a tug-of-war model with loose mechanical coupling.

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    Sebastián Bouzat

    Full Text Available In this work, we explored theoretically the transport of organelles driven along microtubules by molecular motors of opposed polarities using a stochastic model that considers a Langevin dynamics for the cargo, independent cargo-motor linkers and stepping motion for the motors. It has been recently proposed that the stiffness of the motor plays an important role when multiple motors collectively transport a cargo. Therefore, we considered in our model the recently reported values for the stiffness of the cargo-motor linker determined in living cells (∼0.01 pN/nm, which is significantly lower than the motor stiffness obtained in in vitro assays and used in previous studies. Our model could reproduce the multimodal velocity distributions and typical trajectory characteristics including the properties of the reversions in the overall direction of motion observed during melanosome transport along microtubules in Xenopus laevis melanophores. Moreover, we explored the contribution of the different motility states of the cargo-motor system to the different modes of the velocity distributions and could identify the microscopic mechanisms of transport leading to trajectories compatible with those observed in living cells. Finally, by changing the attachment and detachment rates, the model could reproduce the different velocity distributions observed during melanosome transport along microtubules in Xenopus laevis melanophores stimulated for aggregation and dispersion. Our analysis suggests that active tug-of-war processes with loose mechanical coupling can account for several aspects of cargo transport along microtubules in living cells.

  8. Dynein prevents erroneous kinetochore-microtubule attachments in mitosis.

    Science.gov (United States)

    Barisic, Marin; Maiato, Helder

    2015-01-01

    Equal distribution of the genetic material during cell division relies on efficient congression of chromosomes to the metaphase plate. Prior to their alignment, the Dynein motor recruited to kinetochores transports a fraction of laterally-attached chromosomes along microtubules toward the spindle poles. By doing that, Dynein not only contributes to chromosome movements, but also prevents premature stabilization of end-on kinetochore-microtubule attachments. This is achieved by 2 parallel mechanisms: 1) Dynein-mediated poleward movement of chromosomes counteracts opposite polar-ejection forces (PEFs) on chromosome arms by the microtubule plus-end-directed motors chromokinesins. Otherwise, they could stabilize erroneous syntelic kinetochore-microtubule attachments and lead to the random ejection of chromosomes away from the spindle poles; and 2) By transporting chromosomes to the spindle poles, Dynein brings the former to the zone of highest Aurora A kinase activity, further destabilizing kinetochore-microtubule attachments. Thus, Dynein plays an important role in keeping chromosome segregation error-free by preventing premature stabilization of kinetochore-microtubule attachments near the spindle poles.

  9. Cortical microtubules are responsible for gravity resistance in plants.

    Science.gov (United States)

    Hoson, Takayuki; Matsumoto, Shouhei; Soga, Kouichi; Wakabayashi, Kazuyuki

    2010-06-01

    Mechanical resistance to the gravitational force is a principal gravity response in plants distinct from gravitropism. In the final step of gravity resistance, plants increase the rigidity of their cell walls. Here we discuss the role of cortical microtubules, which sustain the function of the cell wall, in gravity resistance. Hypocotyls of Arabidopsis tubulin mutants were shorter and thicker than the wild-type, and showed either left-handed or right-handed helical growth at 1 g. The degree of twisting phenotype was intensified under hypergravity conditions. Hypergravity also induces reorientation of cortical microtubules from transverse to longitudinal directions in epidermal cells. In tubulin mutants, the percentage of cells with longitudinal microtubules was high even at 1 g, and it was further increased by hypergravity. The left-handed helical growth mutants had right-handed microtubule arrays, whereas the right-handed mutant had left-handed arrays. Moreover, blockers of mechanoreceptors suppressed both the twisting phenotype and reorientation of microtubules in tubulin mutants. These results support the hypothesis that cortical microtubules play an essential role in maintenance of normal growth phenotype against the gravitational force, and suggest that mechanoreceptors are involved in signal perception in gravity resistance. Space experiments will confirm whether this view is applicable to plant resistance to 1 g gravity, as to the resistance to hypergravity.

  10. Regional mapping methods using ASTER data to map minerals in the U.S. Basin and Range

    Science.gov (United States)

    Mars, J. C.

    2011-12-01

    The U.S. Geological Survey is currently mapping minerals associated with hydrothermal alteration in a study area that covers most of the U.S. Basin and Range. The study area was mapped using AST_L1B radiance data from the Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER). ASTER measures reflected radiation in three bands in the 0.52-0.86 μm wavelength region (VNIR); six bands in the 1.6-2.43 μm wavelength region (SWIR); and five bands of emitted radiation in the 8.125-11.65 μm wavelength region (TIR) with 15 m, 30 m, and 90 m resolution, respectively. ASTER VNIR and SWIR radiance data were calibrated to reflectance data using ACORN atmospheric correction software. TIR radiance data were calibrated to emissivity data using atmospheric removal and emissivity normalization algorithms in ENVI. Rocks containing hydrothermal silica, calcite-dolomite, epidote-chlorite, alunite-kaolinite, and sericite were mapped using Interactive Data Language (IDL) logical operators. IDL logical operators string together band thresholds and band ratios to map spectral features of minerals. ASTER SWIR band ratios are used in IDL logical operators to map Al-O-H spectral absorption features of alunite, kaolinite and sericite. The ASTER SWIR 4/5 ratio maps the 2.165 μm spectral absorption feature in alunite and kaolinite, and the ASTER SWIR 4/6 and 7/6 ratios map the 2.2 μm spectral absorption feature exhibited in alunite, kaolinite and sericite. Hydrothermal silica was mapped using ASTER SWIR and ASTER TIR band ratios. The ASTER SWIR band ratio 4/7 is typically higher for hydrothermal silica-rich rocks which have lower overall SWIR reflectance in the 2.0 to 2.4 μm region than non-hydrothermal silica-rich rocks due to residual molecular water or an O-H absorption feature spanning 2.26 to 2.4 μm. The ASTER TIR band ratio 13/12 maps the 9.09 μm quartz restralen absorption feature. Thus, silica-rich rocks were mapped using the TIR emissivity data and hydrothermal

  11. Adjacent effect and cross talk of land surfaces on coastal water in the Aster VNIR and SWIR

    Science.gov (United States)

    Takashima, Tsutomu; Masuda, Kazuhiko; Sato, Isao; Tsuchida, Satoshi

    2002-12-01

    The adjacency effect is discussed at coastal areas of main land and peninsula using VNIR and SWIR on ASTER sensor, although the cross-talk phenomenon is apparently noted on some SWIR. The purpose of the analysis is to derive optical characteristics of atmospheric aerosol. The aerosol model is in accordance to the dust-like model. This model is adopted to ASTER and MISR on Terra satellite. Data is the Atsumi Peninsula near Nagoya (34° 40'N, 134° 00'E) GMT1.55 on July 10,2000. The ASTER SWIR(1.65μm-2.395μm) cross-talk phenomenon is noted in the data. This is known as a result of a structure of ASTER sensor. It is relatively large (5-6 DN counts and 100 lines or 3km length). On the other hands, when ASTER observe heterogeneous surface of coastal water, the adjacency effect due to the scattering by atmosphere might partly be contaminated to the above effect. In the SWIR region of spectrum, molecular scattering is practically neglected. However, some aerosol model indicates strong scattering effect at SWIR wavelengths. The main results are (1) The Japan Main land indicates 6~20 times more effect than the peninsula on adjacent radiance from ocean water. (2) SWIR & VNIR exhibit similar adjacent effect which might indicate aerosol or large particles.

  12. Live Cell Imaging Reveals Structural Associations between the Actin and Microtubule Cytoskeleton in Arabidopsis [W] [OA

    Science.gov (United States)

    Sampathkumar, Arun; Lindeboom, Jelmer J.; Debolt, Seth; Gutierrez, Ryan; Ehrhardt, David W.; Ketelaar, Tijs; Persson, Staffan

    2011-01-01

    In eukaryotic cells, the actin and microtubule (MT) cytoskeletal networks are dynamic structures that organize intracellular processes and facilitate their rapid reorganization. In plant cells, actin filaments (AFs) and MTs are essential for cell growth and morphogenesis. However, dynamic interactions between these two essential components in live cells have not been explored. Here, we use spinning-disc confocal microscopy to dissect interaction and cooperation between cortical AFs and MTs in Arabidopsis thaliana, utilizing fluorescent reporter constructs for both components. Quantitative analyses revealed altered AF dynamics associated with the positions and orientations of cortical MTs. Reorganization and reassembly of the AF array was dependent on the MTs following drug-induced depolymerization, whereby short AFs initially appeared colocalized with MTs, and displayed motility along MTs. We also observed that light-induced reorganization of MTs occurred in concert with changes in AF behavior. Our results indicate dynamic interaction between the cortical actin and MT cytoskeletons in interphase plant cells. PMID:21693695

  13. Microtubules and associated molecular motors in Neurospora crassa.

    Science.gov (United States)

    Mouriño-Pérez, Rosa Reyna; Riquelme, Meritxell; Callejas-Negrete, Olga Alicia; Galván-Mendoza, José Iván

    2016-01-01

    The cytoskeleton provides structure, shape and movement to various cells. Microtubules (MTs) are tubular structures made of α and β-tubulin heterodimers organized in 13 protofilaments, forming a hollow cylinder. A vast group of MT-associated proteins determines the function, behavior and interaction of the MTs with other cellular components. Among these proteins, molecular motors such as the dynein-dynactin complex and kinesin superfamily play roles in MT organization and organelle transport. This article focuses on the MT cytoskeleton and associated molecular motors in the filamentous fungus Neurospora crassa In addition to reviewing current available information for this fungus and contrasting it with knowledge of other fungal species, we present new experimental results that support the role of dynein, dynactin and conventional kinesin in MT organization, dynamics and transport of subcellular structures (nuclei and secretory vesicles). In wild type hyphae of N. crassa, cytoplasmic MTs are arranged longitudinally along hyphae and display a helical curvature. They interlace with one another to form a network throughout the cytoplasm. N. crassa dynein and dynactin mutants have a scant and disorganized MT cytoskeleton, an erratic and reduced Spitzenkörper (Spk) and distorted hyphal morphology. In contrast, hyphae of mutants with defective conventional kinesin exhibit only minor disruptions in MT and Spk organization. Although nuclear positioning is affected in all mutants, the MT-associated motor proteins are not major contributors to nuclear movement during hyphal growth. Cytoplasmic bulk flow is the vehicle for nuclear displacement in growing hyphal regions of N. crassa Motors are involved in nuclei saltatory movements in both retrograde or anterograde direction. In the dynein and kinesin mutants, micro and macrovesicles can reach the Spk, although growth is slightly impaired and the Spk displays an erratic path. Hyphal growth requires MTs, and their associated

  14. The Katanin Microtubule Severing Protein in Plants

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Katanin is a heterodimeric microtubule (MT) severing protein that uses energy from ATP hydrolysis to generate internal breaks along MTs. Katanin p60, one of the two subunits, possesses ATPase and MT-bindinglsevering activities, and the p80 subunit is responsible for targeting of katanin to certain subcellular locations. In animals, katanin plays an important role in the release of MTs from their nucleation sites in the centrosome. It is also involved in severing MTs into smaller fragments which can serve as templates for further polymerization to increase MT number during meiotic and mitotic spindle assembly. Katanin homologs are present in a wide variety of plant species. The Arabidopsis katanin homolog has been shown to possess ATP-dependent MT severing activity in vitro and exhibit a punctate localization pattern at the cell cortex and the perinuclear region. Disruption of katanin functions by genetic mutations causes a delay in the disappearance of the perinuclear MT array and results in an aberrant organization of cortical MTs in elongating cells. Consequently, katanin mutations lead to defects in cell elongation, cellulose microfibril deposition, and hormonal responses. Studies of katanin in plants provide new insights into our understanding of its roles in cellular functions.

  15. Microtubules, polarity and vertebrate neural tube morphogenesis.

    Science.gov (United States)

    Cearns, Michael D; Escuin, Sarah; Alexandre, Paula; Greene, Nicholas D E; Copp, Andrew J

    2016-07-01

    Microtubules (MTs) are key cellular components, long known to participate in morphogenetic events that shape the developing embryo. However, the links between the cellular functions of MTs, their effects on cell shape and polarity, and their role in large-scale morphogenesis remain poorly understood. Here, these relationships were examined with respect to two strategies for generating the vertebrate neural tube: bending and closure of the mammalian neural plate; and cavitation of the teleost neural rod. The latter process has been compared with 'secondary' neurulation that generates the caudal spinal cord in mammals. MTs align along the apico-basal axis of the mammalian neuroepithelium early in neural tube closure, participating functionally in interkinetic nuclear migration, which indirectly impacts on cell shape. Whether MTs play other functional roles in mammalian neurulation remains unclear. In the zebrafish, MTs are important for defining the neural rod midline prior to its cavitation, both by localizing apical proteins at the tissue midline and by orienting cell division through a mirror-symmetric MT apparatus that helps to further define the medial localization of apical polarity proteins. Par proteins have been implicated in centrosome positioning in neuroepithelia as well as in the control of polarized morphogenetic movements in the neural rod. Understanding of MT functions during early nervous system development has so far been limited, partly by techniques that fail to distinguish 'cause' from 'effect'. Future developments will likely rely on novel ways to selectively impair MT function in order to investigate the roles they play.

  16. Tau neurofibrillary pathology and microtubule stability.

    Science.gov (United States)

    Michaelis, Mary L; Dobrowsky, Rick T; Li, Guibin

    2002-12-01

    We previously reported that nonomolar concentrations of Taxol and several structurally diverse microtubule (MT)-stabilizing agents significantly enhanced the survival of neurons in the presence of fibrils of amyloid beta peptide (Abeta). Pretreatment of neurons with MT-stabilizing drugs also blocked Abeta-induced activation of tau hyperphosphorylation. Although tau is a substrate for several kinases, we initially focused on cdk5, as this tau kinase has been shown to be activated in Abeta-treated neurons and Alzheimer's disease (AD) brain. In an in vitro kinase assay, Taxol inhibited activation of cdk5 by Abeta. In addition, the proposed cellular cascade in which calpain activation leads to cleavage of the cdk5 regulator, p35, to the strong kinase activator p25 was also prevented. Taxol did not directly inhibit the activity of either cdk5 or calpain, indicating that other cellular components are required for the effect of the drug on Abeta activation of tau phosphorylation. Our results suggest that drugs that interact with MTs can alter signaling events in neurons, possibly because some MTs play a role in organizing protein complexes involved in responses to Abeta. Thus the cytoskeletal network may serve as a biosensor of cellular well-being.

  17. On the significance of microtubule flexural behavior in cytoskeletal mechanics.

    Directory of Open Access Journals (Sweden)

    Mehrdad Mehrbod

    Full Text Available Quantitative description of cell mechanics has challenged biological scientists for the past two decades. Various structural models have been attempted to analyze the structure of the cytoskeleton. One important aspect that has been largely ignored in all these modeling approaches is related to the flexural and buckling behavior of microtubular filaments. The objective of this paper is to explore the influence of this flexural and buckling behavior in cytoskeletal mechanics.In vitro the microtubules are observed to buckle in the first mode, reminiscent of a free, simply-supported beam. In vivo images of microtubules, however, indicate that the buckling mostly occurs in higher modes. This buckling mode switch takes place mostly because of the lateral support of microtubules via their connections to actin and intermediate filaments. These lateral loads are exerted throughout the microtubule length and yield a considerable bending behavior that, unless properly accounted for, would produce erroneous results in the modeling and analysis of the cytoskeletal mechanics.One of the promising attempts towards mechanical modeling of the cytoskeleton is the tensegrity model, which simplifies the complex network of cytoskeletal filaments into a combination merely of tension-bearing actin filaments and compression-bearing microtubules. Interestingly, this discrete model can qualitatively explain many experimental observations in cell mechanics. However, evidence suggests that the simplicity of this model may undermine the accuracy of its predictions, given the model's underlying assumption that "every single member bears solely either tensile or compressive behavior," i.e. neglecting the flexural behavior of the microtubule filaments. We invoke an anisotropic continuum model for microtubules and compare the bending energy stored in a single microtubule with its axial strain energy at the verge of buckling. Our results suggest that the bending energy can

  18. Gene organization, evolution and expression of the microtubule-associated protein ASAP (MAP9

    Directory of Open Access Journals (Sweden)

    Giorgi Dominique

    2008-09-01

    tissues rich in microtubules. ASAP associates to the mitotic spindle and cytoplasmic microtubules, and represents a key factor of mitosis with possible involvement in other cell cycle processes. It may have a role in spermatogenesis and also represents a potential new target for antitumoral drugs. Possible involvement in neuron dynamics also highlights ASAP as a candidate target in neurodegenerative diseases.

  19. Association between microtubules and Golgi vesicles isolated from rat parotid glands.

    Science.gov (United States)

    Coffe, G; Raymond, M N

    1990-01-01

    We report an isolation procedure of trans-Golgi vesicles (GVs) from rat parotid glands. Various organelle markers were used, particularly galactosyl transferase as a trans-Golgi marker, to test the purity of the GV fraction. A quantitative in vitro binding assay between microtubules and GVs is described. The vesicles were incubated with taxol-induced microtubules, layered between 50% and 43% sucrose cushions and subjected to centrifugation. Unlike free microtubules which were sedimented, the GV-bound microtubules co-migrated upward with GVs. Quantification of these bound microtubules was carried out by densitometric scanning of Coomassie blue-stained gels. The association between microtubules and GVs followed a saturation curve, with a plateau value of 20 micrograms of microtubule protein bound to 500 micrograms of GV fraction. The half-saturation of the GV sites was obtained with a microtubule concentration of 20 micrograms/ml. Electron microscopy of negatively stained re-floated material showed numerous microtubule-vesicle complexes. Coating of microtubules with an excess of brain microtubule-associated proteins (MAPs) abolished binding. In the absence of exogenous microtubules, we showed that the GV fraction was already interacting with a class of endogenous rat parotid microtubules. This class of colcemid and cold-stable microtubules represents 10-20% of the total tubulin content of the parotid cell.

  20. LKB1 destabilizes microtubules in myoblasts and contributes to myoblast differentiation.

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    Isma Mian

    Full Text Available BACKGROUND: Skeletal muscle myoblast differentiation and fusion into multinucleate myotubes is associated with dramatic cytoskeletal changes. We find that microtubules in differentiated myotubes are highly stabilized, but premature microtubule stabilization blocks differentiation. Factors responsible for microtubule destabilization in myoblasts have not been identified. FINDINGS: We find that a transient decrease in microtubule stabilization early during myoblast differentiation precedes the ultimate microtubule stabilization seen in differentiated myotubes. We report a role for the serine-threonine kinase LKB1 in both microtubule destabilization and myoblast differentiation. LKB1 overexpression reduced microtubule elongation in a Nocodazole washout assay, and LKB1 RNAi increased it, showing LKB1 destabilizes microtubule assembly in myoblasts. LKB1 levels and activity increased during myoblast differentiation, along with activation of the known LKB1 substrates AMP-activated protein kinase (AMPK and microtubule affinity regulating kinases (MARKs. LKB1 overexpression accelerated differentiation, whereas RNAi impaired it. CONCLUSIONS: Reduced microtubule stability precedes myoblast differentiation and the associated ultimate microtubule stabilization seen in myotubes. LKB1 plays a positive role in microtubule destabilization in myoblasts and in myoblast differentiation. This work suggests a model by which LKB1-induced microtubule destabilization facilitates the cytoskeletal changes required for differentiation. Transient destabilization of microtubules might be a useful strategy for enhancing and/or synchronizing myoblast differentiation.

  1. According to Regression Combine with Interpolation to Mend ASTER GDEM Data%回归与内插法处理ASTER GDEM数据异常值的研究

    Institute of Scientific and Technical Information of China (English)

    惠珊; 李远华

    2012-01-01

    How to solve the common no data or data abnormality problem in ASTER GDEM is mainly studied. At first, detecting the section which no data or data abnormality in ASTER GDEM slope analysis together with density divide method isused. Then SRTM DEM is used according to regression fusing combine with interpolation to simulate and mend the ASTER GDEM data . At Last, the results through using the mended ASTER GDEM data to set up 3D model and Hydrological Model are evaluated. The study results indicate that regression fusing and interpolation method can produce a less error margin and advanced precision data . The mended data is primly matched with remote sensing data and the hydrological model is reasonable. Above all, this method mendes no data and abnormality section in ASTER GDEM improving the data integrality and applicability.%为了解决ASTER GDEM数据中普遍存在的数据空洞及异常问题,在利用坡度分析与密度分割等方法,提取ASTERGDEM数据中存在的空洞及异常值后,借助SRTM DEM数据,利用回归与内插法对空洞及异常区域的高程进行模拟修复.并通过三维建模和实例应用评估其模拟质量.研究结果表明:回归与内插法误差较小、精度较高,所得DEM三维模型良好,能有效提高ASTER GDEM数据的完整性和可用性.

  2. Comparative mineral mapping in the Colorado Mineral Belt using AVIRIS and ASTER remote sensing data

    Science.gov (United States)

    Rockwell, Barnaby W.

    2013-01-01

    This report presents results of interpretation of spectral remote sensing data covering the eastern Colorado Mineral Belt in central Colorado, USA, acquired by the Airborne Visible/Infrared Imaging Spectrometer (AVIRIS) and Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) sensors. This study was part of a multidisciplinary mapping and data integration project at the U.S. Geological Survey that focused on long-term resource planning by land-managing entities in Colorado. The map products were designed primarily for the regional mapping and characterization of exposed surface mineralogy, including that related to hydrothermal alteration and supergene weathering of pyritic rocks. Alteration type was modeled from identified minerals based on standard definitions of alteration mineral assemblages. Vegetation was identified using the ASTER data and subdivided based on per-pixel chlorophyll content (depth of 0.68 micrometer absorption band) and dryness (fit and depth of leaf biochemical absorptions in the shortwave infrared spectral region). The vegetation results can be used to estimate the abundance of fire fuels at the time of data acquisition (2002 and 2003). The AVIRIS- and ASTER-derived mineral mapping results can be readily compared using the toggleable layers in the GeoPDF file, and by using the provided GIS-ready raster datasets. The results relating to mineral occurrence and distribution were an important source of data for studies documenting the effects of mining and un-mined, altered rocks on aquatic ecosystems at the watershed level. These studies demonstrated a high correlation between metal concentrations in streams and the presence of hydrothermal alteration and (or) pyritic mine waste as determined by analysis of the map products presented herein. The mineral mapping results were also used to delineate permissive areas for various mineral deposit types.

  3. Self-organizing feature map neural network classification of the ASTER data based on wavelet fusion

    Institute of Scientific and Technical Information of China (English)

    HASI Bagan; MA Jianwen; LI Qiqing; HAN Xiuzhen; LIU Zhili

    2004-01-01

    Most methods for classification of remote sensing data are based on the statistical parameter evaluation with the assumption that the samples obey the normal distribution. However, more accurate classification results can be obtained with the neural network method through getting knowledge from environments and adjusting the parameter (or weight) step by step by a specific measurement. This paper focuses on the double-layer structured Kohonen self-organizing feature map (SOFM), for which all neurons within the two layers are linked one another and those of the competition layers are linked as well along the sides. Therefore, the self-adapting learning ability is improved due to the effective competition and suppression in this method. The SOFM has become a hot topic in the research area of remote sensing data classification. The Advanced Spaceborne Thermal Emission and Reflectance Radiometer (ASTER) is a new satellite-borne remote sensing instrument with three 15-m resolution bands and three 30-m resolution bands at the near infrared. The ASTER data of Dagang district, Tianjin Municipality is used as the test data in this study. At first, the wavelet fusion is carried out to make the spatial resolutions of the ASTER data identical; then, the SOFM method is applied to classifying the land cover types. The classification results are compared with those of the maximum likelihood method (MLH). As a consequence, the classification accuracy of SOFM increases about by 7% in general and, in particular, it is almost as twice as that of the MLH method in the town.

  4. EXTRACTION OF FOREST STANDS PARAMETERS FROM ASTER DATA IN OPEN FOREST

    Directory of Open Access Journals (Sweden)

    M. Abbasi

    2012-07-01

    Full Text Available Tree crown size (CS and stem number per hectare (SN has become increasingly important for forest management and ecosystem monitoring. Tree crown size is also strongly related to other canopy structural parameters, such as diameter at breast height, tree height and biomass. For both issues, remote sensing data are attractive for their large-area and up-to-date mapping capacities. The QuickBird and ASTER imagery used in this study was acquired over Zagros Forests in southern Zagros region, Fars province of Iran on 1 August 2005 and 1 July 2005, respectively. For the forest site investigated in this study, we concentrated on stands of Quercus Persica which is the dominant species in Zagros region. This study was conducted to investigate the capabilities of ASTER-L1B data to estimate some of forest parameters at individual tree and stand level in dry area. The forest stand parameters are crown area, crown density, average crown area. Obtaining the accuracy of classification the ground truth map was prepared by tree crown delineation using the panchromatic band of QuickBird data. Individual tree crowns were automatically delineated by color segmentation of QuickBird imagery. Simple linear regression procedure was used to show the relationships between spectral variables and the individual trees and forest stand parameters. With decreasing the crown density the effects of background will increase. Our results indicated that crown size could be accurately extracted from panchromatic band of QuickBird images especially for open forest stands. This paper demonstrates that using high-resolution satellite imagery in the open forest offers a unique opportunity for deriving single tree attributes and allowing reliable ground truth map to estimate stand structure. ASTER data and its indices showed good capability to estimate crown area in this study.

  5. ASTER GDEM validation using LiDAR data over coastal regions of Greenland

    DEFF Research Database (Denmark)

    Hvidegaard, Sine Munk; Sørensen, Louise Sandberg; Forsberg, René

    2011-01-01

    Elevation data from airborne Light Detection and Ranging (LiDAR) campaigns are used in an attempt to evaluate the accuracy of the Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) global digital elevation model (GDEM) in Greenland. The LiDAR elevation data set is characterized...... by a high spatial resolution of about 1 m and elevation accuracy of 20–30 cm root mean square error (RMSE). The LiDAR data sets used were acquired during ice-monitoring campaigns carried out from 2003 to 2008. The study areas include ice-free regions, local ice caps and the ice sheet margin. A linear error...

  6. An application of the Self Organizing Map Algorithm to computer aided classification of ASTER multispectral data

    Directory of Open Access Journals (Sweden)

    Ferdinando Giacco

    2008-01-01

    Full Text Available In this paper we employ the Kohonen’s Self Organizing Map (SOM as a strategy for an unsupervised analysis of ASTER multispectral (MS images. In order to obtain an accurate clusterization we introduce as input for the network, in addition to spectral data, some texture measures extracted from IKONOS images, which gives a contribution to the classification of manmade structures. After clustering of SOM outcomes, we associated each cluster with a major land cover and compared them with prior knowledge of the scene analyzed.

  7. Technoscientific Diplomacy: The Practice of International Politics in the ASTER Collaboration

    Science.gov (United States)

    Plafcan, Dan

    Most chapters in this volume focus on the scientific and technical aspects of the design, performance, operations, and applications of the MODIS and Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) instruments. In contrast, this final chapter focuses on politics - specifically, the politics of technical decision making and scientific judgment. When scientific objectives, engineering design decisions, and familiar forms of scientific and technical authority are uncertain or otherwise unsettled, how do they become certain and settled? What facilitates collective judgment and the exercise of power in efforts to advance and achieve common scientific goals, especially in the international arena?

  8. Direct Cytoplasmic Delivery and Nuclear Targeting Delivery of HPMA-MT Conjugates in a Microtubules Dependent Fashion.

    Science.gov (United States)

    Zhong, Jiaju; Zhu, Xi; Luo, Kui; Li, Lian; Tang, Manlin; Liu, Yanxi; Zhou, Zhou; Huang, Yuan

    2016-09-06

    As the hearts of tumor cells, the nucleus is the ultimate target of many chemotherapeutic agents and genes. However, nuclear drug delivery is always hampered by multiple intracellular obstacles, such as low efficiency of lysosome escape and insufficient nuclear trafficking. Herein, an N-(2-hydroxypropyl) methacrylamide (HPMA) polymer-based drug delivery system was designed, which could achieve direct cytoplasmic delivery by a nonendocytic pathway and transport into the nucleus in a microtubules dependent fashion. A special targeting peptide (MT), derived from an endogenic parathyroid hormone-related protein, was conjugated to the polymer backbone, which could accumulate into the nucleus a by microtubule-mediated pathway. The in vitro studies found that low temperature and NaN3 could not influence the cell internalization of the conjugates. Besides, no obvious overlay of the conjugates with lysosome demonstrated that the polymer conjugates could enter the tumor cell cytoplasm by a nonendocytic pathway, thus avoiding the drug degradation in the lysosome. Furthermore, after suppression of the microtubule dynamics with microtubule stabilizing docetaxel (DTX) and destabilizing nocodazole (Noc), the nuclear accumulation of polymeric conjugates was significantly inhibited. Living cells fluorescence recovery after photobleaching study found that the nuclear import rate of conjugates was 2-fold faster compared with the DTX and Noc treated groups. These results demonstrated that the conjugates transported into the nucleus in a microtubules dependent way. Therefore, in addition to direct cytoplasmic delivery, our peptide conjugated polymeric platform could simultaneously mediate nuclear drug accumulation, which may open a new path for further intracellular genes/peptides delivery.

  9. Regional Lithological Mapping Using ASTER-TIR Data: Case Study for the Tibetan Plateau and the Surrounding Area

    Directory of Open Access Journals (Sweden)

    Yoshiki Ninomiya

    2016-09-01

    Full Text Available The mineralogical indices the Quartz Index (QI, Carbonate Index (CI and Mafic Index (MI for ASTER multispectral thermal infrared (TIR data were applied to various geological materials for regional lithological mapping on the Tibetan Plateau. Many lithological and structural features are not currently well understood in the central Tibetan Plateau, including the distribution of mafic-ultramafic rocks related to the suture zones, the quartzose and carbonate sedimentary rocks accreted to the Eurasian continent, and sulfate layers related to the Tethys and neo-Tethys geological setting. These rock types can now be mapped with the interpretation of the processed ASTER TIR images described in this paper. A methodology is described for the processing of ASTER TIR data applied to a very wide region of the Tibetan Plateau. The geometrical and radiometric performance of the processed images is discussed, and the advantages of using ortho-rectified data are shown. The challenges of using ASTER data with a small footprint in addition to selecting an appropriate subset of scenes are also examined. ASTER scenes possess a narrow swath width when compared to LANDSAT data (60 km vs. 185 km, respectively. Furthermore, the ASTER data archive is vast, consisting of approximately three million images. These details can present an added level of complexity during an image processing workflow. Finally, geological interpretations made on the maps of the indices are compared with prior geological field studies. The results from the investigations suggest that the indices perform well in the classification of quartzose rocks based on the carbonate and mafic mineral content, in addition to the granitic rocks based on the feldspar content.

  10. Microtubule dynamics. II. Kinetics of self-assembly

    DEFF Research Database (Denmark)

    Flyvbjerg, H.; Jobs, E.

    1997-01-01

    Inverse scattering theory describes the conditions necessary and sufficient to determine an unknown potential from known scattering data. No similar theory exists for when and how one may deduce the kinetics of an unknown chemical reaction from quantitative information about its final state and its...... various initial concentrations of monomers. Distinct mathematical properties of the kinetics of the unknown reaction pathway are found. These properties are shown to restrict the kinetics to a single model that generalizes Oosawa's classical nucleation-polymerization model. The methods used here...

  11. Thermodynamic and structural analysis of microtubule assembly: the role of GTP hydrolysis.

    Science.gov (United States)

    Vulevic, B; Correia, J J

    1997-03-01

    Different models have been proposed that link the tubulin heterodimer nucleotide content and the role of GTP hydrolysis with microtubule assembly and dynamics. Here we compare the thermodynamics of microtubule assembly as a function of nucleotide content by van't Hoff analysis. The thermodynamic parameters of tubulin assembly in 30-100 mM piperazine-N,N'-bis(2-ethanesulfonic acid), 1 mM MgSO4, 2 mM EGTA, pH 6.9, in the presence of a weakly hydrolyzable analog, GMPCPP, the dinucleotide analog GMPCP plus 2 M glycerol, and GTP plus 2 M glycerol were obtained together with data for taxol-GTP/GDP tubulin assembly (GMPCPP and GMPCP are the GTP and GDP nucleotide analogs where the alpha beta oxygen has been replaced by a methylene, -CH2-). All of the processes studied are characterized by a positive enthalpy, a positive entropy, and a large, negative heat capacity change. GMPCP-induced assembly has the largest negative heat capacity change and GMPCPP has the second largest, whereas GTP/2 M glycerol- and taxol-induced assembly have more positive values, respectively. A large, negative heat capacity is most consistent with the burial of water-accessible hydrophobic surface area, which gives rise to the release of bound water. The heat capacity changes observed with GTP/2 M glycerol-induced and with taxol-induced assembly are very similar, -790 +/- 190 cal/mol/k, and correspond to the burial of 3330 +/- 820 A2 of nonpolar surface area. This value is shown to be very similar to an estimate of the buried nonpolar surface in a reconstructed microtubule lattice. Polymerization data from GMPCP- and GMPCPP-induced assembly are consistent with buried nonpolar surface areas that are 3 and 6 times larger. A linear enthalpy-entropy and enthalpy-free energy plot for tubulin polymerization reactions verifies that enthalpy-entropy compensation for this system is based upon true biochemical correlation, most likely corresponding to a dominant hydrophobic effect. Entropy analysis suggests

  12. Intracellular spatial localization regulated by the microtubule network.

    Directory of Open Access Journals (Sweden)

    Jing Chen

    Full Text Available The commonly recognized mechanisms for spatial regulation inside the cell are membrane-bounded compartmentalization and biochemical association with subcellular organelles. We use computational modeling to investigate another spatial regulation mechanism mediated by the microtubule network in the cell. Our results demonstrate that the mitotic spindle can impose strong sequestration and concentration effects on molecules with binding affinity for microtubules, especially dynein-directed cargoes. The model can recapitulate the essence of three experimental observations on distinct microtubule network morphologies: the sequestration of germ plasm components by the mitotic spindles in the Drosophila syncytial embryo, the asymmetric cell division initiated by the time delay in centrosome maturation in the Drosophila neuroblast, and the diffusional block between neighboring energids in the Drosophila syncytial embryo. Our model thus suggests that the cell cycle-dependent changes in the microtubule network are critical for achieving different spatial regulation effects. The microtubule network provides a spatially extensive docking platform for molecules and gives rise to a "structured cytoplasm", in contrast to a free and fluid environment.

  13. A Quantitative Method for Microtubule Analysis in Fluorescence Images.

    Science.gov (United States)

    Lan, Xiaodong; Li, Lingfei; Hu, Jiongyu; Zhang, Qiong; Dang, Yongming; Huang, Yuesheng

    2015-12-01

    Microtubule analysis is of significant value for a better understanding of normal and pathological cellular processes. Although immunofluorescence microscopic techniques have proven useful in the study of microtubules, comparative results commonly rely on a descriptive and subjective visual analysis. We developed an objective and quantitative method based on image processing and analysis of fluorescently labeled microtubular patterns in cultured cells. We used a multi-parameter approach by analyzing four quantifiable characteristics to compose our quantitative feature set. Then we interpreted specific changes in the parameters and revealed the contribution of each feature set using principal component analysis. In addition, we verified that different treatment groups could be clearly discriminated using principal components of the multi-parameter model. High predictive accuracy of four commonly used multi-classification methods confirmed our method. These results demonstrated the effectiveness and efficiency of our method in the analysis of microtubules in fluorescence images. Application of the analytical methods presented here provides information concerning the organization and modification of microtubules, and could aid in the further understanding of structural and functional aspects of microtubules under normal and pathological conditions.

  14. Microtubules contribute to maintain nucleus shape in epithelial cell monolayer

    Science.gov (United States)

    Tremblay, Dominique; Andrzejewski, Lukasz; Pelling, Andrew

    2013-03-01

    INTRODUCTION: Tissue strains can result in significant nuclear deformations and may regulate gene expression. However, the precise role of the cytoskeleton in regulating nuclear mechanics remains poorly understood. Here, we investigate the nuclear deformability of Madin-Darky canine kidney cells (MDCK) under various stretching conditions to clarify the role of the microtubules and actin network on the mechanical behavior of the nucleus. METHODS: A custom-built cell-stretching device allowing for real time imaging of MDCK nuclei was used. Cells were seeded on a silicone membrane coated with rat-tail collagen I. A nuclear stain, Hoechst-33342, was used to image nuclei during stretching. We exposed cells to a compressive and non-compressive stretching strain field of 25%. Nocodazole and cytochalasin-D were used to depolymerize the microtubules and actin network. RESULTS: Nuclei in control cells stretched more along their minor axis than major axis with a deformation of 5% and 2% respectively. This anisotropy vanished completely in microtubule-deprived cells and these cells showed a very high nuclear deformability along the minor axis when exposed to a compressive stretching strain field. CONCLUSIONS: The microtubules drive the anisotropic deformability of MDCK nuclei in a monolayer and maintain nuclear shape when exposed to compressive strain. Such intrinsic mechanical behavior indicates that microtubules are essential to maintain nuclear shape and may prevent down regulation of gene expression.

  15. The feasibility of coherent energy transfer in microtubules.

    Science.gov (United States)

    Craddock, Travis John Adrian; Friesen, Douglas; Mane, Jonathan; Hameroff, Stuart; Tuszynski, Jack A

    2014-11-06

    It was once purported that biological systems were far too 'warm and wet' to support quantum phenomena mainly owing to thermal effects disrupting quantum coherence. However, recent experimental results and theoretical analyses have shown that thermal energy may assist, rather than disrupt, quantum coherent transport, especially in the 'dry' hydrophobic interiors of biomolecules. Specifically, evidence has been accumulating for the necessary involvement of quantum coherent energy transfer between uniquely arranged chromophores in light harvesting photosynthetic complexes. The 'tubulin' subunit proteins, which comprise microtubules, also possess a distinct architecture of chromophores, namely aromatic amino acids, including tryptophan. The geometry and dipolar properties of these aromatics are similar to those found in photosynthetic units indicating that tubulin may support coherent energy transfer. Tubulin aggregated into microtubule geometric lattices may support such energy transfer, which could be important for biological signalling and communication essential to living processes. Here, we perform a computational investigation of energy transfer between chromophoric amino acids in tubulin via dipole excitations coupled to the surrounding thermal environment. We present the spatial structure and energetic properties of the tryptophan residues in the microtubule constituent protein tubulin. Plausibility arguments for the conditions favouring a quantum mechanism of signal propagation along a microtubule are provided. Overall, we find that coherent energy transfer in tubulin and microtubules is biologically feasible.

  16. Interaction between microtubules and the Drosophila formin Cappuccino and its effect on actin assembly.

    Science.gov (United States)

    Roth-Johnson, Elizabeth A; Vizcarra, Christina L; Bois, Justin S; Quinlan, Margot E

    2014-02-14

    Formin family actin nucleators are potential coordinators of the actin and microtubule cytoskeletons, as they can both nucleate actin filaments and bind microtubules in vitro. To gain a more detailed mechanistic understanding of formin-microtubule interactions and formin-mediated actin-microtubule cross-talk, we studied microtubule binding by Cappuccino (Capu), a formin involved in regulating actin and microtubule organization during Drosophila oogenesis. We found that two distinct domains within Capu, FH2 and tail, work together to promote high-affinity microtubule binding. The tail domain appears to bind microtubules through nonspecific charge-based interactions. In contrast, distinct residues within the FH2 domain are important for microtubule binding. We also report the first visualization of a formin polymerizing actin filaments in the presence of microtubules. Interestingly, microtubules are potent inhibitors of the actin nucleation activity of Capu but appear to have little effect on Capu once it is bound to the barbed end of an elongating filament. Because Capu does not simultaneously bind microtubules and assemble actin filaments in vitro, its actin assembly and microtubule binding activities likely require spatial and/or temporal regulation within the Drosophila oocyte.

  17. Interaction between Microtubules and the Drosophila Formin Cappuccino and Its Effect on Actin Assembly*

    Science.gov (United States)

    Roth-Johnson, Elizabeth A.; Vizcarra, Christina L.; Bois, Justin S.; Quinlan, Margot E.

    2014-01-01

    Formin family actin nucleators are potential coordinators of the actin and microtubule cytoskeletons, as they can both nucleate actin filaments and bind microtubules in vitro. To gain a more detailed mechanistic understanding of formin-microtubule interactions and formin-mediated actin-microtubule cross-talk, we studied microtubule binding by Cappuccino (Capu), a formin involved in regulating actin and microtubule organization during Drosophila oogenesis. We found that two distinct domains within Capu, FH2 and tail, work together to promote high-affinity microtubule binding. The tail domain appears to bind microtubules through nonspecific charge-based interactions. In contrast, distinct residues within the FH2 domain are important for microtubule binding. We also report the first visualization of a formin polymerizing actin filaments in the presence of microtubules. Interestingly, microtubules are potent inhibitors of the actin nucleation activity of Capu but appear to have little effect on Capu once it is bound to the barbed end of an elongating filament. Because Capu does not simultaneously bind microtubules and assemble actin filaments in vitro, its actin assembly and microtubule binding activities likely require spatial and/or temporal regulation within the Drosophila oocyte. PMID:24362037

  18. Inter-Band Radiometric Comparison and Calibration of ASTER Visible and Near-Infrared Bands

    Directory of Open Access Journals (Sweden)

    Kenta Obata

    2015-11-01

    Full Text Available The present study evaluates inter-band radiometric consistency across the Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER visible and near-infrared (VNIR bands and develops an inter-band calibration algorithm to improve radiometric consistency. Inter-band radiometric comparison of current ASTER data shows a root mean square error (RMSE of 3.8%–5.7% among radiance outputs of spectral bands due primarily to differences between calibration strategies of the NIR band for nadir-looking (Band 3N and the other two bands (green and red bands, corresponding to Bands 1 and 2. An algorithm for radiometric calibration of Bands 2 and 3N with reference to Band 1 is developed based on the band translation technique and is used to obtain new radiometric calibration coefficients (RCCs for sensor sensitivity degradation. The systematic errors between radiance outputs are decreased by applying the derived RCCs, which result in reducing the RMSE from 3.8%–5.7% to 2.2%–2.9%. The remaining errors are approximately equal to or smaller than the intrinsic uncertainties of inter-band calibration derived by sensitivity analysis. Improvement of the radiometric consistency would increase the accuracy of band algebra (e.g., vegetation indices and its application. The algorithm can be used to evaluate inter-band radiometric consistency, as well as for the calibration of other sensors.

  19. Morphometric analysis of sub-watershed in parts of Western Ghats, South India using ASTER DEM

    Directory of Open Access Journals (Sweden)

    Evangelin Ramani Sujatha

    2015-05-01

    Full Text Available Morphometric analysis is a key to understand the hydrological process and hence is a prerequisite for the assessment of hydrological characteristics of surface water basin. Morphometric analysis to determine the drainage characteristics of Palar sub-watershed, a part of Shanmukha watershed in the Amaravati sub-catchment is done using Advanced Space-borne Thermal Emission and Reflection Global Digital Elevation Model (ASTER GDEM data, and is supplemented with topographical maps in geographical information systems platform. This study uses ASTER GDEM data to extract morphometric features of a mountain stream at micro-watershed level. The sub-watershed is divided into six micro-watersheds. The sub-watershed includes a sixth-order stream. Lower stream orders, in particular first-order streams, dominate the sub-watershed. Development of stream segments is controlled by slope and local relief. Drainage pattern of the sub-watershed and micro-watersheds is dendritic in general. The mean bifurcation ratio of the sub-watershed is 3.69 but its variation between the various stream orders suggests structural control in the development of stream network. The shape factors reveal the elongation of the sub-watershed and micro-watersheds.The relief ratio reveals the high discharge capability of the sub-watershed and meagre groundwater potential. This study is a useful tool for planning strategies in control of soil erosion and soil conservation.

  20. Estimating Evapotranspiration from an Improved Two-Source Energy Balance Model Using ASTER Satellite Imagery

    Directory of Open Access Journals (Sweden)

    Qifeng Zhuang

    2015-11-01

    Full Text Available Reliably estimating the turbulent fluxes of latent and sensible heat at the Earth’s surface by remote sensing is important for research on the terrestrial hydrological cycle. This paper presents a practical approach for mapping surface energy fluxes using Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER images from an improved two-source energy balance (TSEB model. The original TSEB approach may overestimate latent heat flux under vegetative stress conditions, as has also been reported in recent research. We replaced the Priestley-Taylor equation used in the original TSEB model with one that uses plant moisture and temperature constraints based on the PT-JPL model to obtain a more accurate canopy latent heat flux for model solving. The collected ASTER data and field observations employed in this study are over corn fields in arid regions of the Heihe Watershed Allied Telemetry Experimental Research (HiWATER area, China. The results were validated by measurements from eddy covariance (EC systems, and the surface energy flux estimates of the improved TSEB model are similar to the ground truth. A comparison of the results from the original and improved TSEB models indicates that the improved method more accurately estimates the sensible and latent heat fluxes, generating more precise daily evapotranspiration (ET estimate under vegetative stress conditions.

  1. Surface energy balance and resolution effects using ASTER imagery over Jornada, New Mexico

    Science.gov (United States)

    French, A.; Schmugge, T.; Kustas, W.; Prueger, J.; Jacob, F.

    2003-04-01

    Knowledge of the relationship between the accuracy of surface energy balance estimates and remote sensing resolution is important for the integration of instantaneous satellite observations with regional and global scale hydrological models. Higher resolution thermal infrared imagery, such as from the ASTER sensor (90 m), provide the needed observations to distinguish between hydrological end-member conditions and are expected to be more accurate than imagery with resolutions >1 km. But higher resolution observations are instantaneous and usually not repeatable at time intervals less than 2 to 4 weeks. On the other hand, lower resolution imagery from sensors such as GOES, in combination with hydrological assimilation models, can estimate the diurnal variations in surface energy balance. To assess the relative merits of resolution vs. imaging frequency, we have collected a series of ASTER observations, between May and October 2002 over the Jornada Experimental Range in New Mexico, and modeled the surface energy balance. Estimates of the energy components are made from the original resolution (90 m) up to an aggregated resolution of 4 km. These estimates are compared with ground based flux observations, and then compared with each other. Loss in range of data and expected biases due to resolution effects will be discussed.

  2. A Forest Fire Risk Assessment Using ASTER Images in Peninsular Malaysia

    Institute of Scientific and Technical Information of China (English)

    PENG Guang-xiong; LI Jing; CHEN Yun-hao; NORIZAN Abdul-patah

    2007-01-01

    Based on the physical concept of heat energy of pre-ignition, a new fire susceptibility index (FSI) is used to estimate forest fire risk. This physical basis allows calculation of ignition probabilities and comparisons of fire risk across eco-regions. The computation of the index requires inputs of fuel temperature and fuel moisture content (FMC), both of which can be estimated using remote sensing data. While ASTER data for land surface temperatures (LST) was used as proxys for fuel temperatures, fuel moisture content is estimated by regression technique utilizing the ratio NDVI/LST of ASTER data. FSIs are computed in peninsular Malaysia for nine days before the fires of 2004 and 2005 and validated with fire occurrence data. Results show that the FSI increases as the day approaches the fire day. This trend can be observed clearly about four days before the day of fire. It suggests that FSI can be a good estimator of fire risk. The physical basis provides a more meaningful FSI, allows calculation of ignition probabilities and facilitates the development of a future class of fire risk models. FSI can be used to compare fire risk across different eco-regions and time periods. FSI retains the flexibility to be localized to a vegetation type or eco-regions for improved performance.

  3. ASTER L1B satellite data applied to geothermal in Cuba

    Directory of Open Access Journals (Sweden)

    V. González-Acosta

    2015-12-01

    Full Text Available The 83 ASTER L1B thermal channels of Cuban territorial scenes, from 2000 to 2008 years, selected and processed with geothermal aims showed almost 50% of cloudy coverage. The vortex coordinated as well as other data from such metadata facilitated completing the designed database. From a preliminary mosaic with the images existent these were subsequently processed in order to obtain temperature images. Such images were then integrated into another mosaic with a suitable reclassification resulting in 11 classes with 3°C each. This allowed delimitating those anomalous zones where the greater distribution of pixels oscillated from 25°C to 37°C, and the cloudy coverage temperature aroused up to 20°C approximately. In the resulting temperature map, 69 polygons were a priori delimitated and categorized, as for their perspective and the temperature value above 40°C. These polygons were later overlapped to Google Earth images with the aim to identify those from anthropogenic origins. Finally it was obtained an estimation of the temperature value of the surface coverage of the national territory as well as the understanding of that the eastern zone is the most perspective. This is an experimental application, using satellite images ASTER L1B with geothermic purpose.

  4. AUTOMATED CONSTRUCTION OF COVERAGE CATALOGUES OF ASTER SATELLITE IMAGE FOR URBAN AREAS OF THE WORLD

    Directory of Open Access Journals (Sweden)

    H. Miyazaki

    2012-07-01

    Full Text Available We developed an algorithm to determine a combination of satellite images according to observation extent and image quality. The algorithm was for testing necessity for completing coverage of the search extent. The tests excluded unnecessary images with low quality and preserve necessary images with good quality. The search conditions of the satellite images could be extended, indicating the catalogue could be constructed with specified periods required for time series analysis. We applied the method to a database of metadata of ASTER satellite images archived in GEO Grid of National Institute of Advanced Industrial Science and Technology (AIST, Japan. As indexes of populated places with geographical coordinates, we used a database of 3372 populated place of more than 0.1 million populations retrieved from GRUMP Settlement Points, a global gazetteer of cities, which has geographical names of populated places associated with geographical coordinates and population data. From the coordinates of populated places, 3372 extents were generated with radiuses of 30 km, a half of swath of ASTER satellite images. By merging extents overlapping each other, they were assembled into 2214 extents. As a result, we acquired combinations of good quality for 1244 extents, those of low quality for 96 extents, incomplete combinations for 611 extents. Further improvements would be expected by introducing pixel-based cloud assessment and pixel value correction over seasonal variations.

  5. ASTER Global Digital Elevation Model Version 2 - summary of validation results

    Science.gov (United States)

    Tachikawa, Tetushi; Kaku, Manabu; Iwasaki, Akira; Gesch, Dean B.; Oimoen, Michael J.; Zhang, Z.; Danielson, Jeffrey J.; Krieger, Tabatha; Curtis, Bill; Haase, Jeff; Abrams, Michael; Carabajal, C.; Meyer, Dave

    2011-01-01

    On June 29, 2009, NASA and the Ministry of Economy, Trade and Industry (METI) of Japan released a Global Digital Elevation Model (GDEM) to users worldwide at no charge as a contribution to the Global Earth Observing System of Systems (GEOSS). This “version 1” ASTER GDEM (GDEM1) was compiled from over 1.2 million scenebased DEMs covering land surfaces between 83°N and 83°S latitudes. A joint U.S.-Japan validation team assessed the accuracy of the GDEM1, augmented by a team of 20 cooperators. The GDEM1 was found to have an overall accuracy of around 20 meters at the 95% confidence level. The team also noted several artifacts associated with poor stereo coverage at high latitudes, cloud contamination, water masking issues and the stacking process used to produce the GDEM1 from individual scene-based DEMs (ASTER GDEM Validation Team, 2009). Two independent horizontal resolution studies estimated the effective spatial resolution of the GDEM1 to be on the order of 120 meters.

  6. Quantum Computation in Brain Microtubules? Decoherence and Biological Feasibility

    CERN Document Server

    Hagan, S; Tuszynski, J A

    2000-01-01

    The Penrose-Hameroff (`Orch OR') model of quantum computation in brain microtubules has been criticized as regards the issue of environmental decoherence. A recent report by Tegmark finds that microtubules can maintain quantum coherence for only $10^{-13}$ s, far too short to be neurophysiologically relevant. Here, we critically examine the assumptions behind Tegmark's calculation and find that: 1) Tegmark's commentary is not aimed at an existing model in the literature but rather at a hybrid that replaces the superposed protein conformations of the `Orch OR' theory with a soliton in superposition along the microtubule, 2) Tegmark predicts decreasing decoherence times at lower temperature, in direct contradiction of the observed behavior of quantum states, 3) recalculation after correcting Tegmark's equation for differences between his model and the `Orch OR' model (superposition separation, charge vs. dipole, dielectric constant) lengthens the decoherence time to $10^{-5} - 10^{-4}$ s and invalidates a criti...

  7. Disruption of microtubule integrity initiates mitosis during CNS repair.

    Science.gov (United States)

    Bossing, Torsten; Barros, Claudia S; Fischer, Bettina; Russell, Steven; Shepherd, David

    2012-08-14

    Mechanisms of CNS repair have vital medical implications. We show that traumatic injury to the ventral midline of the embryonic Drosophila CNS activates cell divisions to replace lost cells. A pilot screen analyzing transcriptomes of single cells during repair pointed to downregulation of the microtubule-stabilizing GTPase mitochondrial Rho (Miro) and upregulation of the Jun transcription factor Jun-related antigen (Jra). Ectopic Miro expression can prevent midline divisions after damage, whereas Miro depletion destabilizes cortical β-tubulin and increases divisions. Disruption of cortical microtubules, either by chemical depolymerization or by overexpression of monomeric tubulin, triggers ectopic mitosis in the midline and induces Jra expression. Conversely, loss of Jra renders midline cells unable to replace damaged siblings. Our data indicate that upon injury, the integrity of the microtubule cytoskeleton controls cell division in the CNS midline, triggering extra mitosis to replace lost cells. The conservation of the identified molecules suggests that similar mechanisms may operate in vertebrates.

  8. Septins localize to microtubules during nutritional limitation in Saccharomyces cerevisiae

    Directory of Open Access Journals (Sweden)

    Vázquez de Aldana Carlos R

    2008-10-01

    Full Text Available Abstract Background In Saccharomyces cerevisiae, nutrient limitation stimulates diploid cells to undergo DNA replication and meiosis, followed by the formation of four haploid spores. Septins are a family of proteins that assemble a ring structure at the mother-daughter neck during vegetative growth, where they control cytokinesis. In sporulating cells, the septin ring disassembles and septins relocalize to the prospore membrane. Results Here, we demonstrate that nutrient limitation triggers a change in the localization of at least two vegetative septins (Cdc10 and Cdc11 from the bud neck to the microtubules. The association of Cdc10 and Cdc11 with microtubules persists into meiosis, and they are found associated with the meiotic spindle until the end of meiosis II. In addition, the meiosis-specific septin Spr28 displays similar behavior, suggesting that this is a common feature of septins. Septin association to microtubules is a consequence of the nutrient limitation signal, since it is also observed when haploid cells are incubated in sporulation medium and when haploid or diploid cells are grown in medium containing non-fermentable carbon sources. Moreover, during meiosis II, when the nascent prospore membrane is formed, septins moved from the microtubules to this membrane. Proper organization of the septins on the membrane requires the sporulation-specific septins Spr3 and Spr28. Conclusion Nutrient limitation in S. cerevisiae triggers the sporulation process, but it also induces the disassembly of the septin bud neck ring and relocalization of the septin subunits to the nucleus. Septins remain associated with microtubules during the meiotic divisions and later, during spore morphogenesis, they are detected associated to the nascent prospore membranes surrounding each nuclear lobe. Septin association to microtubules also occurs during growth in non-fermentable carbon sources.

  9. Three-dimensional reconstruction of tubulin sheets and re-investigation of microtubule surface lattice.

    Science.gov (United States)

    Schultheiss, R; Mandelkow, E

    1983-10-25

    Sheets are incomplete microtubule walls observed as polymorphic variants of microtubule assembly. Their substructure is similar to that of microtubules, as shown by two-dimensional optical and computer reconstruction. We have extended earlier studies by computing a three-dimensional reconstruction. From a re-investigation of the surface lattice it appears that the three-start helix of microtubules is right-handed rather than left-handed.

  10. In vivo and in vitro effects of the mitochondrial uncoupler FCCP on microtubules.

    OpenAIRE

    Maro, B.; Marty, M C; Bornens, M

    1982-01-01

    FCCP (carbonylcyanide-p-trifluoromethoxyphenylhydrazone), a potent uncoupler of oxidative phosphorylation, induces the complete disruption of cellular microtubules. A further analysis of this effect on BHK21 cells has shown that a decrease in the number of microtubules can be observed 15 min after adding FCCP and there is complete disruption after 60 min. Regrowth of microtubules was initiated 30 min after removal of FCCP, in marked contrast with the rapid reversion observed when microtubules...

  11. Kinesin-3 in the basidiomycete Ustilago maydis transports organelles along the entire microtubule array.

    Science.gov (United States)

    Steinberg, Gero

    2015-01-01

    The molecular motor kinesin-3 transports early endosomes along microtubules in filamentous fungi. It was reported that kinesin-3 from the ascomycete fungi Aspergillus nidulans and Neurospora crassa use a subset of post-translationally modified and more stable microtubules. Here, I show that kinesin-3 from the basidiomycete Ustilago maydis moves along all hyphal microtubules. This difference is likely due to variation in cell cycle control and associated organization of the microtubule array.

  12. Sliding of microtubules by a team of dynein motors: Understanding the effect of spatial distribution of motor tails and mutual exclusion of motor heads on microtubules

    Science.gov (United States)

    Singh, Hanumant Pratap; Takshak, Anjneya; Mall, Utkarsh; Kunwar, Ambarish

    2016-06-01

    Molecular motors are natural nanomachines that use the free energy released from ATP hydrolysis to generate mechanical forces. Cytoplasmic dynein motors often work collectively as a team to drive important processes such as axonal growth, proplatelet formation and mitosis, as forces generated by single motors are insufficient. A large team of dynein motors is used to slide cytoskeletal microtubules with respect to one another during the process of proplatelet formation and axonal growth. These motors attach to a cargo microtubule via their tail domains, undergo the process of detachment and reattachment of their head domains on another track microtubule, while sliding the cargo microtubule along the track. Traditional continuum/mean-field approaches used in the past are not ideal for studying the sliding mechanism of microtubules, as they ignore spatial and temporal fluctuations due to different possible distributions of motor tails on cargo filament, as well as binding/unbinding of motors from their track. Therefore, these models cannot be used to address important questions such as how the distribution of motor tails on microtubules, or how the mutual exclusion of motor heads on microtubule tracks affects the sliding velocity of cargo microtubule. To answer these, here we use a computational stochastic model where we model each dynein motor explicitly. In our model, we use both random as well as uniform distributions of dynein motors on cargo microtubule, as well as mutual exclusion of motors on microtubule tracks. We find that sliding velocities are least affected by the distribution of motor tails on microtubules, whereas they are greatly affected by mutual exclusion of motor heads on microtubule tracks. We also find that sliding velocity depends on the length of cargo microtubule if mutual exclusion among motor heads is considered.

  13. High-quality seamless DEM generation blending SRTM-1, ASTER GDEM v2 and ICESat/GLAS observations

    Science.gov (United States)

    Yue, Linwei; Shen, Huanfeng; Zhang, Liangpei; Zheng, Xianwei; Zhang, Fan; Yuan, Qiangqiang

    2017-01-01

    The absence of a high-quality seamless global digital elevation model (DEM) dataset has been a challenge for the Earth-related research fields. Recently, the 1-arc-second Shuttle Radar Topography Mission (SRTM-1) data have been released globally, covering over 80% of the Earth's land surface (60°N-56°S). However, voids and anomalies still exist in some tiles, which has prevented the SRTM-1 dataset from being directly used without further processing. In this paper, we propose a method to generate a seamless DEM dataset blending SRTM-1, ASTER GDEM v2, and ICESat laser altimetry data. The ASTER GDEM v2 data are used as the elevation source for the SRTM void filling. To get a reliable filling source, ICESat GLAS points are incorporated to enhance the accuracy of the ASTER data within the void regions, using an artificial neural network (ANN) model. After correction, the voids in the SRTM-1 data are filled with the corrected ASTER GDEM values. The triangular irregular network based delta surface fill (DSF) method is then employed to eliminate the vertical bias between them. Finally, an adaptive outlier filter is applied to all the data tiles. The final result is a seamless global DEM dataset. ICESat points collected from 2003 to 2009 were used to validate the effectiveness of the proposed method, and to assess the vertical accuracy of the global DEM products in China. Furthermore, channel networks in the Yangtze River Basin were also extracted for the data assessment.

  14. Phospholipase D activation correlates with microtubule reorganization in living plant cells

    NARCIS (Netherlands)

    P.B. Dhonukshe; A.M. Laxalt; J. Goedhart; Th.W.J. Gadella; T. Munnik

    2003-01-01

    A phospholipase D (PLD) was shown recently to decorate microtubules in plant cells. Therefore, we used tobacco BY-2 cells expressing the microtubule reporter GFP-MAP4 to test whether PLD activation affects the organization of plant microtubules. Within 30 min of adding n-butanol, a potent activator

  15. Cortical Microtubule Arrays Are Initiated from a Nonrandom Prepattern Driven by Atypical Microtubule Initiation1[W][OA

    Science.gov (United States)

    Lindeboom, Jelmer J.; Lioutas, Antonios; Deinum, Eva E.; Tindemans, Simon H.; Ehrhardt, David W.; Emons, Anne Mie C.; Vos, Jan W.; Mulder, Bela M.

    2013-01-01

    The ordered arrangement of cortical microtubules in growing plant cells is essential for anisotropic cell expansion and, hence, for plant morphogenesis. These arrays are dismantled when the microtubule cytoskeleton is rearranged during mitosis and reassembled following completion of cytokinesis. The reassembly of the cortical array has often been considered as initiating from a state of randomness, from which order arises at least partly through self-organizing mechanisms. However, some studies have shown evidence for ordering at early stages of array assembly. To investigate how cortical arrays are initiated in higher plant cells, we performed live-cell imaging studies of cortical array assembly in tobacco (Nicotiana tabacum) Bright Yellow-2 cells after cytokinesis and drug-induced disassembly. We found that cortical arrays in both cases did not initiate randomly but with a significant overrepresentation of microtubules at diagonal angles with respect to the cell axis, which coincides with the predominant orientation of the microtubules before their disappearance from the cell cortex in preprophase. In Arabidopsis (Arabidopsis thaliana) root cells, recovery from drug-induced disassembly was also nonrandom and correlated with the organization of the previous array, although no diagonal bias was observed in these cells. Surprisingly, during initiation, only about one-half of the new microtubules were nucleated from locations marked by green fluorescent protein-γ-tubulin complex protein2-tagged γ-nucleation complexes (γ-tubulin ring complex), therefore indicating that a large proportion of early polymers was initiated by a noncanonical mechanism not involving γ-tubulin ring complex. Simulation studies indicate that the high rate of noncanonical initiation of new microtubules has the potential to accelerate the rate of array repopulation. PMID:23300168

  16. Paleohydrologic Analysis of Debris-Flow Inundation at Mount Rainier, Washington Using ASTER and SRTM Derived Topography

    Science.gov (United States)

    Hubbard, B. E.; Crowley, J. K.; Mars, J.

    2002-05-01

    Methods used in extracting digital topography from remote sensing data include photogrammetry, interferometry, altimetry, and photoclinometry. Two recent spaceborne missions: ASTER (Advanced Spaceborne Thermal Emission and Reflection Radiometer) and SRTM (Shuttle Radar Topography Mission) use some of these methods for generating global DEM coverages at horizontal resolutions less than 100 m per pixel. This study evaluates the utility of such data for estimating inundation levels of past debris-flows based on the geomorphometric characteristics of terraces preserved along river valleys draining Mount Rainier. Deposits representing debris-flow volumes spanning several orders of magnitude are used as case examples. ASTER DEMs are derived photogrammetrically by measuring the parallax between a stereo pair of images acquired simultaneously by nadir- and aft-viewing instruments. The two channels used (3N and 3B) have a near-infrared bandwidth of 0.76-0.86 microns and a base/height ratio of 0.6 for the stereo pair. SRTM DEMs are generated interferometrically from C- (5.6 cm wavelength) and X-band (3 cm wavelength) synthetic aperture radar (SAR) by measuring the phase differences between SAR images acquired by two antennas spaced 60 m apart. Terrace deposits of the Osceola mudflow, Electron mudflow, National lahar, and Tahoma lahar were all resolved to varying degrees in cross-sectional profiles extracted from the ASTER and SRTM DEMs. Profiles from these two datasets were compared to profiles extracted from a level 2 USGS DEM. The latter dataset was derived by interpolation of USGS digital contour plots, which have been stereoscopically corrected for errors in elevation caused by canopy height variations. Both ASTER and SRTM DEMs were spatially registered to the USGS DEM, which has a spatial resolution of 30 m and vertical accuracy within +15 m. The vertical accuracies of ASTER and STRM DEMs vary within +30 m and +16 m, respectively. The extent of canopy cover throughout

  17. Estimating surface fluxes over middle and upper streams of the Heihe River Basin with ASTER imagery

    Directory of Open Access Journals (Sweden)

    W. Ma

    2011-05-01

    Full Text Available Land surface heat fluxes are essential measures of the strengths of land-atmosphere interactions involving energy, heat and water. Correct parameterization of these fluxes in climate models is critical. Despite their importance, state-of-the-art observation techniques cannot provide representative areal averages of these fluxes comparable to the model grid. Alternative methods of estimation are thus required. These alternative approaches use (satellite observables of the land surface conditions. In this study, the Surface Energy Balance System (SEBS algorithm was evaluated in a cold and arid environment, using land surface parameters derived from Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER data. Field observations and estimates from SEBS were compared in terms of net radiation flux (Rn, soil heat flux (G0, sensible heat flux (H and latent heat flux (λE over a heterogeneous land surface. As a case study, this methodology was applied to the experimental area of the Watershed Allied Telemetry Experimental Research (WATER project, located on the mid-to-upstream sections of the Heihe River in northwest China. ASTER data acquired between 3 May and 4 June 2008, under clear-sky conditions were used to determine the surface fluxes. Ground-based measurements of land surface heat fluxes were compared with values derived from the ASTER data. The results show that the derived surface variables and the land surface heat fluxes furnished by SEBS in different months over the study area are in good agreement with the observed land surface status under the limited cases (some cases looks poor results. So SEBS can be used to estimate turbulent heat fluxes with acceptable accuracy in areas where there is partial vegetation cover in exceptive conditions. It is very important to perform calculations using ground-based observational data for parameterization in SEBS in the future

  18. Decadal region-wide and glacier-wide mass balances derived from multi-temporal ASTER satellite digital elevation models. Validation over the Mont-Blanc area

    Science.gov (United States)

    Berthier, Etienne; Cabot, Vincent; Vincent, Christian; Six, Delphine

    2016-06-01

    Since 2000, a vast archive of stereo-images has been built by the Advanced Spaceborne Thermal Emission and Reflection (ASTER) satellite. Several studies already extracted glacier mass balances from multi-temporal ASTER digital elevation models (DEMs) but they lacked accurate independent data for validation. Here, we apply a linear regression to a time series of 3D-coregistered ASTER DEMs to estimate the rate of surface elevation changes (dh/dtASTER) and geodetic mass balances of Mont-Blanc glaciers (155 km²) between 2000 and 2014. Validation using field and spaceborne geodetic measurements reveals large errors at the individual pixel level (> 1 m a-1) and an accuracy of 0.2-0.3 m a-1 for dh/dtASTER averaged over areas larger than 1 km². For all Mont-Blanc glaciers, the ASTER region-wide mass balance (-1.05±0.37 m water equivalent (w.e.) a-1) agrees remarkably with the one measured using Spot5 and Pléiades DEMs (-1.06±0.23 m w.e. a-1) over their common 2003-2012 period. This multi-temporal ASTER DEM strategy leads to smaller errors than the simple differencing of two ASTER DEMs. By extrapolating dh/dtASTER to mid-February 2000, we infer a mean penetration depth of about 9±3 m for the C-band Shuttle Radar Topographic Mission (SRTM) radar signal, with a strong altitudinal dependency (range 0-12 m). This methodology thus reveals the regional pattern of glacier surface elevation changes and improves our knowledge of the penetration of the radar signal into snow and ice.

  19. Microtubules accelerate the kinase activity of Aurora-B by a reduction in dimensionality.

    Science.gov (United States)

    Noujaim, Michael; Bechstedt, Susanne; Wieczorek, Michal; Brouhard, Gary J

    2014-01-01

    Aurora-B is the kinase subunit of the Chromosome Passenger Complex (CPC), a key regulator of mitotic progression that corrects improper kinetochore attachments and establishes the spindle midzone. Recent work has demonstrated that the CPC is a microtubule-associated protein complex and that microtubules are able to activate the CPC by contributing to Aurora-B auto-phosphorylation in trans. Aurora-B activation is thought to occur when the local concentration of Aurora-B is high, as occurs when Aurora-B is enriched at centromeres. It is not clear, however, whether distributed binding to large structures such as microtubules would increase the local concentration of Aurora-B. Here we show that microtubules accelerate the kinase activity of Aurora-B by a "reduction in dimensionality." We find that microtubules increase the kinase activity of Aurora-B toward microtubule-associated substrates while reducing the phosphorylation levels of substrates not associated to microtubules. Using the single molecule assay for microtubule-associated proteins, we show that a minimal CPC construct binds to microtubules and diffuses in a one-dimensional (1D) random walk. The binding of Aurora-B to microtubules is salt-dependent and requires the C-terminal tails of tubulin, indicating that the interaction is electrostatic. We show that the rate of Aurora-B auto-activation is faster with increasing concentrations of microtubules. Finally, we demonstrate that microtubules lose their ability to stimulate Aurora-B when their C-terminal tails are removed by proteolysis. We propose a model in which microtubules act as scaffolds for the enzymatic activity of Aurora-B. The scaffolding activity of microtubules enables rapid Aurora-B activation and efficient phosphorylation of microtubule-associated substrates.

  20. Microtubule organization : from the centrosome to the Golgi apparatus

    NARCIS (Netherlands)

    Wu, J.

    2017-01-01

    Similar to the skeleton of a human body, every cell possesses the so-called cytoskeleton, a system of filaments that support cell shape and enable cells to divide and move. One of the major types of cytoskeletal fibers are microtubules, microscopic tubes that cells use as rails to transport their co

  1. Structure of the microtubule-binding domain of flagellar dynein.

    Science.gov (United States)

    Kato, Yusuke S; Yagi, Toshiki; Harris, Sarah A; Ohki, Shin-ya; Yura, Kei; Shimizu, Youské; Honda, Shinya; Kamiya, Ritsu; Burgess, Stan A; Tanokura, Masaru

    2014-11-04

    Flagellar dyneins are essential microtubule motors in eukaryotes, as they drive the beating motions of cilia and flagella. Unlike myosin and kinesin motors, the track binding mechanism of dyneins and the regulation between the strong and weak binding states remain obscure. Here we report the solution structure of the microtubule-binding domain of flagellar dynein-c/DHC9 (dynein-c MTBD). The structure reveals a similar overall helix-rich fold to that of the MTBD of cytoplasmic dynein (cytoplasmic MTBD), but dynein-c MTBD has an additional flap, consisting of an antiparallel b sheet. The flap is positively charged and highly flexible. Despite the structural similarity to cytoplasmic MTBD, dynein-c MTBD shows only a small change in the microtubule- binding affinity depending on the registry change of coiled coil-sliding, whereby lacks the apparent strong binding state. The surface charge distribution of dynein-c MTBD also differs from that of cytoplasmic MTBD, which suggests a difference in the microtubule-binding mechanism.

  2. Fission yeast Scp3 potentially maintains microtubule orientation through bundling.

    Directory of Open Access Journals (Sweden)

    Kanako Ozaki

    Full Text Available Microtubules play important roles in organelle transport, the maintenance of cell polarity and chromosome segregation and generally form bundles during these processes. The fission yeast gene scp3+ was identified as a multicopy suppressor of the cps3-81 mutant, which is hypersensitive to isopropyl N-3-chlorophenylcarbamate (CIPC, a poison that induces abnormal multipolar spindle formation in higher eukaryotes. In this study, we investigated the function of Scp3 along with the effect of CIPC in the fission yeast Schizosaccharomyces pombe. Microscopic observation revealed that treatment with CIPC, cps3-81 mutation and scp3+ gene deletion disturbed the orientation of microtubules in interphase cells. Overexpression of scp3+ suppressed the abnormal orientation of microtubules by promoting bundling. Functional analysis suggested that Scp3 functions independently from Ase1, a protein largely required for the bundling of the mitotic spindle. A strain lacking the ase1+ gene was more sensitive to CIPC, with the drug affecting the integrity of the mitotic spindle, indicating that CIPC has a mitotic target that has a role redundant with Ase1. These results suggested that multiple systems are independently involved to ensure microtubule orientation by bundling in fission yeast.

  3. Neuronal microtubule organization: from minus end to plus end

    NARCIS (Netherlands)

    Yau, K.W.

    2016-01-01

    Neurons are highly polarized cells consisting of a dendritic part and axonal part. Dendrites receive signals from other cells while axons transmit signals to other cells. In this thesis, mostly hippocampal neurons from rat embryos are used to study fundamental aspects of the microtubule organization

  4. Total synthesis of the potent microtubule-stabilizing agent (+)-discodermolide.

    Science.gov (United States)

    Harried, Scott S; Lee, Christopher P; Yang, Ge; Lee, Tony I H; Myles, David C

    2003-08-22

    The total synthesis of the potent microtubule-stabilizing, antimitotic agent (+)-discodermolide is described. The convergent synthetic strategy takes advantage of the diastereoselective alkylation of a ketone enolate to establish the key C15-C16 bond. The synthesis is amenable to preparation of gram-scale quantities of (+)-discodermolide and analogues.

  5. Direct Modulation of Microtubule Stability Contributes to Anthracene General Anesthesia

    Science.gov (United States)

    Emerson, Daniel J.; Weiser, Brian P.; Psonis, John; Liao, Zhengzheng; Taratula, Olena; Fiamengo, Ashley; Wang, Xiaozhao; Sugasawa, Keizo; Smith, Amos B.; Eckenhoff, Roderic G; Dmochowski, Ivan J.

    2013-01-01

    Recently, we identified 1-aminoanthracene as a fluorescent general anesthetic. To investigate the mechanism of action, a photoactive analogue, 1-azidoanthracene, was synthesized. Administration of 1-azidoanthracene to albino stage 40–47 tadpoles was found to immobilize animals upon near-UV irradiation of the forebrain region. The immobilization was often reversible, but it was characterized by a longer duration consistent with covalent attachment of the ligand to functionally important targets. IEF/SDS-PAGE examination of irradiated tadpole brain homogenate revealed labeled protein, identified by mass spectrometry as β-tubulin. In vitro assays with aminoanthracene-cross-linked tubulin indicated inhibition of microtubule polymerization, similar to colchicine. Tandem mass spectrometry confirmed anthracene binding near the colchicine site. Stage 40–47 tadpoles were also incubated 1 h with microtubule stabilizing agents, epothilone D or discodermolide, followed by dosing with 1-aminoanthracene. The effective concentration of 1-aminoanthracene required to immobilize the tadpoles was significantly increased in the presence of either microtubule stabilizing agent. Epothilone D similarly mitigated the effects of a clinical neurosteroid general anesthetic, allopregnanolone, believed to occupy the colchicine site in tubulin. We conclude that neuronal microtubules are “on-pathway” targets for anthracene general anesthetics and may also represent functional targets for some neurosteroid general anesthetics. PMID:23484901

  6. BMP signaling and microtubule organization regulate synaptic strength.

    Science.gov (United States)

    Ball, R W; Peled, E S; Guerrero, G; Isacoff, E Y

    2015-04-16

    The strength of synaptic transmission between a neuron and multiple postsynaptic partners can vary considerably. We have studied synaptic heterogeneity using the glutamatergic Drosophila neuromuscular junction (NMJ), which contains multiple synaptic connections of varying strengths between a motor axon and muscle fiber. In larval NMJs, there is a gradient of synaptic transmission from weak proximal to strong distal boutons. We imaged synaptic transmission with the postsynaptically targeted fluorescent calcium sensor SynapCam, to investigate the molecular pathways that determine synaptic strength and set up this gradient. We discovered that mutations in the Bone Morphogenetic Protein (BMP) signaling pathway disrupt production of strong distal boutons. We find that strong connections contain unbundled microtubules in the boutons, suggesting a role for microtubule organization in transmission strength. The spastin mutation, which disorganizes microtubules, disrupted the transmission gradient, supporting this interpretation. We propose that the BMP pathway, shown previously to function in the homeostatic regulation of synaptic growth, also boosts synaptic transmission in a spatially selective manner that depends on the microtubule system.

  7. ASTERICS : Addressing Cross-Cutting Synergies and Common Challenges for the Next Decade Astronomy Facilities

    CERN Document Server

    Pasian, Fabio; Genova, Francoise; Lamanna, Giovanni; Serjeant, Stephen; Szomoru, Arpad; van der Meer, Rob

    2016-01-01

    The large infrastructure projects for the next decade will allow a new quantum leap in terms of new possible science. ESFRI, the European Strategy Forum on Research Infrastructures, a strategic initiative to develop the scientific integration of Europe, has identified four facilities (SKA, CTA, KM3Net and E-ELT) deserving priority in support. The ASTERICS project aims to address the cross-cutting synergies and common challenges shared by the various Astronomy ESFRI and other world-class facilities. The project (22 partners across Europe) is funded by the EU Horizon 2020 programme with 15 MEuro in 4 years. It brings together for the first time the astronomy, astrophysics and particle astrophysics communities, in addition to other related research infrastructures.

  8. Aster albescens Rust Caused by Aecidium asterum and Its Hyperparasitic Fungus Tuberculina persicina

    Institute of Scientific and Technical Information of China (English)

    HUANG Yun; YE Hua-zhi; CHEN Guang-yan; LIU Zi-ying; DONG Bao-chen

    2005-01-01

    Aster albescens is a medicinal plant. The rust caused by Aecidium asterum is a new record in China and an important disease of A. albescens in Feng Tong-zhai national reservation area. The percentages of diseased plant and the diseased leaf reach 100 and 28% respectively when the disease is serious. The disease results in leaf spot and leaf cast. Tuberculina persicina is a natural hyperparasite of A. asterum and is firstly reported in the world. The hyperparasite attacks aecium of A. asterum and hinderes the release of aeciospores. The symptom of the rust and the morphological characteristics of A.asterum is reported. The hyperparasitism of A. asterum and the morphological characteristics of the Tuberculina persicina are also reported.

  9. VALIDATION OF THE ASTER GLOBAL DIGITAL ELEVATION MODEL VERSION 2 OVER THE CONTERMINOUS UNITED STATES

    Directory of Open Access Journals (Sweden)

    D. Gesch

    2012-07-01

    Full Text Available The ASTER Global Digital Elevation Model Version 2 (GDEM v2 was evaluated over the conterminous United States in a manner similar to the validation conducted for the original GDEM Version 1 (v1 in 2009. The absolute vertical accuracy of GDEM v2 was calculated by comparison with more than 18,000 independent reference geodetic ground control points from the National Geodetic Survey. The root mean square error (RMSE measured for GDEM v2 is 8.68 meters. This compares with the RMSE of 9.34 meters for GDEM v1. Another important descriptor of vertical accuracy is the mean error, or bias, which indicates if a DEM has an overall vertical offset from true ground level. The GDEM v2 mean error of –0.20 meters is a significant improvement over the GDEM v1 mean error of –3.69 meters. The absolute vertical accuracy assessment results, both mean error and RMSE, were segmented by land cover to examine the effects of cover types on measured errors. The GDEM v2 mean errors by land cover class verify that the presence of aboveground features (tree canopies and built structures cause a positive elevation bias, as would be expected for an imaging system like ASTER. In open ground classes (little or no vegetation with significant aboveground height, GDEM v2 exhibits a negative bias on the order of 1 meter. GDEM v2 was also evaluated by differencing with the Shuttle Radar Topography Mission (SRTM dataset. In many forested areas, GDEM v2 has elevations that are higher in the canopy than SRTM.

  10. Mycorrhizal symbiosis and local adaptation in Aster amellus: a field transplant experiment.

    Directory of Open Access Journals (Sweden)

    Hana Pánková

    Full Text Available Many plant populations have adapted to local soil conditions. However, the role of arbuscular mycorrhizal fungi is often overlooked in this context. Only a few studies have used reciprocal transplant experiments to study the relationships between soil conditions, mycorrhizal colonisation and plant growth. Furthermore, most of the studies were conducted under controlled greenhouse conditions. However, long-term field experiments can provide more realistic insights into this issue. We conducted a five-year field reciprocal transplant experiment to study the relationships between soil conditions, arbuscular mycorrhizal fungi and plant growth in the obligate mycotrophic herb Aster amellus. We conducted this study in two regions in the Czech Republic that differ significantly in their soil nutrient content, namely Czech Karst (region K and Ceske Stredohori (region S. Plants that originated from region S had significantly higher mycorrhizal colonisation than plants from region K, indicating that the percentage of mycorrhizal colonisation has a genetic basis. We found no evidence of local adaptation in Aster amellus. Instead, plants from region S outperformed the plants from region K in both target regions. Similarly, plants from region S showed more mycorrhizal colonisation in all cases, which was likely driven by the lower nutrient content in the soil from that region. Thus, plant aboveground biomass and mycorrhizal colonisation exhibited corresponding differences between the two target regions and regions of origin. Higher mycorrhizal colonisation in the plants from region with lower soil nutrient content (region S in both target regions indicates that mycorrhizal colonisation is an adaptive trait. However, lower aboveground biomass in the plants with lower mycorrhizal colonisation suggests that the plants from region K are in fact maladapted by their low inherent mycorrhizal colonization. We conclude that including mycorrhizal symbiosis in local

  11. Fidgetin-Like 2: A Microtubule-Based Regulator of Wound Healing.

    Science.gov (United States)

    Charafeddine, Rabab A; Makdisi, Joy; Schairer, David; O'Rourke, Brian P; Diaz-Valencia, Juan D; Chouake, Jason; Kutner, Allison; Krausz, Aimee; Adler, Brandon; Nacharaju, Parimala; Liang, Hongying; Mukherjee, Suranjana; Friedman, Joel M; Friedman, Adam; Nosanchuk, Joshua D; Sharp, David J

    2015-09-01

    Wound healing is a complex process driven largely by the migration of a variety of distinct cell types from the wound margin into the wound zone. In this study, we identify the previously uncharacterized microtubule-severing enzyme, Fidgetin-like 2 (FL2), as a fundamental regulator of cell migration that can be targeted in vivo using nanoparticle-encapsulated small interfering RNA (siRNA) to promote wound closure and regeneration. In vitro, depletion of FL2 from mammalian tissue culture cells results in a more than twofold increase in the rate of cell movement, in part due to a significant increase in directional motility. Immunofluorescence analyses indicate that FL2 normally localizes to the cell edge, importantly to the leading edge of polarized cells, where it regulates the organization and dynamics of the microtubule cytoskeleton. To clinically translate these findings, we utilized a nanoparticle-based siRNA delivery platform to locally deplete FL2 in both murine full-thickness excisional and burn wounds. Topical application of FL2 siRNA nanoparticles to either wound type results in a significant enhancement in the rate and quality of wound closure both clinically and histologically relative to controls. Taken together, these results identify FL2 as a promising therapeutic target to promote the regeneration and repair of cutaneous wounds.

  12. Statistical mechanics provides novel insights into microtubule stability and mechanism of shrinkage.

    Science.gov (United States)

    Jain, Ishutesh; Inamdar, Mandar M; Padinhateeri, Ranjith

    2015-02-01

    Microtubules are nano-machines that grow and shrink stochastically, making use of the coupling between chemical kinetics and mechanics of its constituent protofilaments (PFs). We investigate the stability and shrinkage of microtubules taking into account inter-protofilament interactions and bending interactions of intrinsically curved PFs. Computing the free energy as a function of PF tip position, we show that the competition between curvature energy, inter-PF interaction energy and entropy leads to a rich landscape with a series of minima that repeat over a length-scale determined by the intrinsic curvature. Computing Langevin dynamics of the tip through the landscape and accounting for depolymerization, we calculate the average unzippering and shrinkage velocities of GDP protofilaments and compare them with the experimentally known results. Our analysis predicts that the strength of the inter-PF interaction (E(s)(m)) has to be comparable to the strength of the curvature energy (E(b)(m)) such that E(s)(m) - E(b)(m) ≈ 1kBT, and questions the prevalent notion that unzippering results from the domination of bending energy of curved GDP PFs. Our work demonstrates how the shape of the free energy landscape is crucial in explaining the mechanism of MT shrinkage where the unzippered PFs will fluctuate in a set of partially peeled off states and subunit dissociation will reduce the length.

  13. The microtubule stabilizing agent discodermolide is a potent inducer of accelerated cell senescence.

    Science.gov (United States)

    Klein, Laura E; Freeze, B Scott; Smith, Amos B; Horwitz, Susan Band

    2005-03-01

    Discodermolide is a microtubule stabilizing agent that suppresses dynamic instability and blocks cells in mitosis. Selection of A549 nonsmall cell lung carcinoma cells with increasing concentrations of discodermolide yielded a clone that proliferated in 8 nM. When these cells were exposed to any concentration greater than 8 nM, replication ceased and the cells developed a flattened, enlarged, granular morphology. Accelerated senescence was demonstrated by a functional beta-galactosidase activity at pH 6. When parental A549 cells were treated with IC50-concentrations of doxorubicin, Taxol or discodermolide, the latter two drugs quickly produced aberrant mitosis. However, discodermolide, but not Taxol, also produced a large increase in senescence-associated beta-galactosidase activity and altered levels of known senescence markers. Although some of these differences between Taxol and discodermolide were dose dependent, only discodermolide produced a doxorubicin-like induction of a senescence phenotype, including a senescence-associated beta-galactosidase activity, up-regulation of PAI-1 and p66Shc, and a strong, sustained, Erk1/2 activation. This research provides insights into the mechanism of action of discodermolide and provides the first demonstration of a microtubule stabilizing agent that inhibits tumor cell growth with a powerful induction of accelerated senescence.

  14. 53BP1 and the LINC Complex Promote Microtubule-Dependent DSB Mobility and DNA Repair.

    Science.gov (United States)

    Lottersberger, Francisca; Karssemeijer, Roos Anna; Dimitrova, Nadya; de Lange, Titia

    2015-11-05

    Increased mobility of chromatin surrounding double-strand breaks (DSBs) has been noted in yeast and mammalian cells but the underlying mechanism and its contribution to DSB repair remain unclear. Here, we use a telomere-based system to track DNA damage foci with high resolution in living cells. We find that the greater mobility of damaged chromatin requires 53BP1, SUN1/2 in the linker of the nucleoskeleton, and cytoskeleton (LINC) complex and dynamic microtubules. The data further demonstrate that the excursions promote non-homologous end joining of dysfunctional telomeres and implicated Nesprin-4 and kinesins in telomere fusion. 53BP1/LINC/microtubule-dependent mobility is also evident at irradiation-induced DSBs and contributes to the mis-rejoining of drug-induced DSBs in BRCA1-deficient cells showing that DSB mobility can be detrimental in cells with numerous DSBs. In contrast, under physiological conditions where cells have only one or a few lesions, DSB mobility is proposed to prevent errors in DNA repair.

  15. Pironetin reacts covalently with cysteine-316 of α-tubulin to destabilize microtubule

    Science.gov (United States)

    Yang, Jianhong; Wang, Yuxi; Wang, Taijing; Jiang, Jian; Botting, Catherine H.; Liu, Huanting; Chen, Qiang; Yang, Jinliang; Naismith, James H.; Zhu, Xiaofeng; Chen, Lijuan

    2016-01-01

    Molecules that alter the normal dynamics of microtubule assembly and disassembly include many anticancer drugs in clinical use. So far all such therapeutics target β-tubulin, and structural biology has explained the basis of their action and permitted design of new drugs. However, by shifting the profile of β-tubulin isoforms, cancer cells become resistant to treatment. Compounds that bind to α-tubulin are less well characterized and unexploited. The natural product pironetin is known to bind to α-tubulin and is a potent inhibitor of microtubule polymerization. Previous reports had identified that pironetin reacts with lysine-352 residue however analogues designed on this model had much lower potency, which was difficult to explain, hindering further development. We report crystallographic and mass spectrometric data that reveal that pironetin forms a covalent bond to cysteine-316 in α-tubulin via a Michael addition reaction. These data provide a basis for the rational design of α-tubulin targeting chemotherapeutics. PMID:27357539

  16. Statistical mechanics provides novel insights into microtubule stability and mechanism of shrinkage.

    Directory of Open Access Journals (Sweden)

    Ishutesh Jain

    2015-02-01

    Full Text Available Microtubules are nano-machines that grow and shrink stochastically, making use of the coupling between chemical kinetics and mechanics of its constituent protofilaments (PFs. We investigate the stability and shrinkage of microtubules taking into account inter-protofilament interactions and bending interactions of intrinsically curved PFs. Computing the free energy as a function of PF tip position, we show that the competition between curvature energy, inter-PF interaction energy and entropy leads to a rich landscape with a series of minima that repeat over a length-scale determined by the intrinsic curvature. Computing Langevin dynamics of the tip through the landscape and accounting for depolymerization, we calculate the average unzippering and shrinkage velocities of GDP protofilaments and compare them with the experimentally known results. Our analysis predicts that the strength of the inter-PF interaction (E(s(m has to be comparable to the strength of the curvature energy (E(b(m such that E(s(m - E(b(m ≈ 1kBT, and questions the prevalent notion that unzippering results from the domination of bending energy of curved GDP PFs. Our work demonstrates how the shape of the free energy landscape is crucial in explaining the mechanism of MT shrinkage where the unzippered PFs will fluctuate in a set of partially peeled off states and subunit dissociation will reduce the length.

  17. Microtubule-dependent modulation of adhesion complex composition.

    Science.gov (United States)

    Ng, Daniel H J; Humphries, Jonathan D; Byron, Adam; Millon-Frémillon, Angélique; Humphries, Martin J

    2014-01-01

    The microtubule network regulates the turnover of integrin-containing adhesion complexes to stimulate cell migration. Disruption of the microtubule network results in an enlargement of adhesion complex size due to increased RhoA-stimulated actomyosin contractility, and inhibition of adhesion complex turnover; however, the microtubule-dependent changes in adhesion complex composition have not been studied in a global, unbiased manner. Here we used label-free quantitative mass spectrometry-based proteomics to determine adhesion complex changes that occur upon microtubule disruption with nocodazole. Nocodazole-treated cells displayed an increased abundance of the majority of known adhesion complex components, but no change in the levels of the fibronectin-binding α5β1 integrin. Immunofluorescence analyses confirmed these findings, but revealed a change in localisation of adhesion complex components. Specifically, in untreated cells, α5-integrin co-localised with vinculin at peripherally located focal adhesions and with tensin at centrally located fibrillar adhesions. In nocodazole-treated cells, however, α5-integrin was found in both peripherally located and centrally located adhesion complexes that contained both vinculin and tensin, suggesting a switch in the maturation state of adhesion complexes to favour focal adhesions. Moreover, the switch to focal adhesions was confirmed to be force-dependent as inhibition of cell contractility with the Rho-associated protein kinase inhibitor, Y-27632, prevented the nocodazole-induced conversion. These results highlight a complex interplay between the microtubule cytoskeleton, adhesion complex maturation state and intracellular contractile force, and provide a resource for future adhesion signaling studies. The proteomics data have been deposited in the ProteomeXchange with identifier PXD001183.

  18. Cell edges accumulate gamma tubulin complex components and nucleate microtubules following cytokinesis in Arabidopsis thaliana.

    Directory of Open Access Journals (Sweden)

    Chris Ambrose

    Full Text Available Microtubules emanate from distinct organizing centers in fungal and animal cells. In plant cells, by contrast, microtubules initiate from dispersed sites in the cell cortex, where they then self-organize into parallel arrays. Previous ultrastructural evidence suggested that cell edges participate in microtubule nucleation but so far there has been no direct evidence for this. Here we use live imaging to show that components of the gamma tubulin nucleation complex (GCP2 and GCP3 localize at distinct sites along the outer periclinal edge of newly formed crosswalls, and that microtubules grow predominantly away from these edges. These data confirm a role for cell edges in microtubule nucleation, and suggest that an asymmetric distribution of microtubule nucleation factors contributes to cortical microtubule organization in plants, in a manner more similar to other kingdoms than previously thought.

  19. Dissecting the molecular mechanism underlying the intimate relationship between cellulose microfibrils and cortical microtubules

    Directory of Open Access Journals (Sweden)

    Lei eLei

    2014-03-01

    Full Text Available A central question in plant cell development is how the cell wall determines directional cell expansion and therefore the final shape of the cell. As the major load-bearing component of the cell wall, cellulose microfibrils are laid down transversely to the axis of elongation, thus forming a spring-like structure that reinforces the cell laterally and while favoring longitudinal expansion in most growing cells. Mounting evidence suggests that cortical microtubules organize the deposition of cellulose microfibrils, but the precise molecular mechanisms linking microtubules to cellulose organization have remained unclear until the recent discovery of CSI1, a linker protein between the cortical microtubules and the cellulose biosynthesizing machinery. In this review, we will focus on the intimate relationship between cellulose microfibrils and cortical microtubules, in particular, we will discuss microtubule arrangement and cell wall architecture, the linkage between cellulose synthase complexes and microtubules, and the feedback mechanisms between cell wall and microtubules.

  20. Sulfo-SMCC Prevents Annealing of Taxol-Stabilized Microtubules In Vitro

    CERN Document Server

    Prabhune, Meenakshi; Schmidt, Christoph F

    2015-01-01

    Microtubule structure and functions have been widely studied in vitro and in cells. Research has shown that cysteines on tubulin play a crucial role in the polymerization of microtubules. Here, we show that blocking sulfhydryl groups of cysteines in taxol-stabilized polymerized microtubules with a commonly used chemical crosslinker prevents temporal end-to-end annealing of microtubules in vitro. This can dramatically affect the length distribution of the microtubules. The crosslinker sulfosuccinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate, sulfo-SMCC, consists of a maleimide and a N-hydroxysuccinimide ester group to bind to sulfhydryl groups and primary amines, respectively. Interestingly, addition of a maleimide dye alone does not show the same prevention of annealing in stabilized microtubules. This study shows that the sulfhydryl groups of cysteines of tubulin that are vital for the polymerization are also important for the subsequent annealing of microtubules.

  1. Siva1 suppresses epithelial-mesenchymal transition and metastasis of tumor cells by inhibiting stathmin and stabilizing microtubules.

    Science.gov (United States)

    Li, Nan; Jiang, Peng; Du, Wenjing; Wu, Zhengsheng; Li, Cong; Qiao, Mengran; Yang, Xiaolu; Wu, Mian

    2011-08-02

    Epithelial-mesenchymal transition (EMT) enables epithelial cells to acquire motility and invasiveness that are characteristic of mesenchymal cells. It plays an important role in development and tumor cell metastasis. However, the mechanisms of EMT and their dysfunction in cancer cells are still not well understood. Here we report that Siva1 interacts with stathmin, a microtubule destabilizer. Siva1 inhibits stathmin's activity directly as well as indirectly through Ca(2+)/calmodulin-dependent protein kinase II-mediated phosphorylation of stathmin at Ser16. Via the inhibition of stathmin, Siva1 enhances the formation of microtubules and impedes focal adhesion assembly, cell migration, and EMT. Low levels of Siva1 and Ser16-phosphorylated stathmin correlate with high metastatic states of human breast cancer cells. In mouse models, knockdown of Siva1 promotes cancer dissemination, whereas overexpression of Siva1 inhibits it. These results suggest that microtubule dynamics are critical for EMT. Furthermore, they reveal an important role for Siva1 in suppressing cell migration and EMT and indicate that down-regulation of Siva1 may contribute to tumor cell metastasis.

  2. Chemical genetic approach identifies microtubule affinity-regulating kinase 1 as a leucine-rich repeat kinase 2 substrate.

    Science.gov (United States)

    Krumova, Petranka; Reyniers, Lauran; Meyer, Marc; Lobbestael, Evy; Stauffer, Daniela; Gerrits, Bertran; Muller, Lionel; Hoving, Sjouke; Kaupmann, Klemens; Voshol, Johannes; Fabbro, Doriano; Bauer, Andreas; Rovelli, Giorgio; Taymans, Jean-Marc; Bouwmeester, Tewis; Baekelandt, Veerle

    2015-07-01

    Mutations in leucine-rich repeat kinase 2 (LRRK2) are the most common cause of autosomal-dominant forms of Parkinson's disease. LRRK2 is a modular, multidomain protein containing 2 enzymatic domains, including a kinase domain, as well as several protein-protein interaction domains, pointing to a role in cellular signaling. Although enormous efforts have been made, the exact pathophysiologic mechanisms of LRRK2 are still not completely known. In this study, we used a chemical genetics approach to identify LRRK2 substrates from mouse brain. This approach allows the identification of substrates of 1 particular kinase in a complex cellular environment. Several of the identified peptides are involved in the regulation of microtubule (MT) dynamics, including microtubule-associating protein (MAP)/microtubule affinity-regulating kinase 1 (MARK1). MARK1 is a serine/threonine kinase known to phosphorylate MT-binding proteins such as Tau, MAP2, and MAP4 at KXGS motifs leading to MT destabilization. In vitro kinase assays and metabolic-labeling experiments in living cells confirmed MARK1 as an LRRK2 substrate. Moreover, we also showed that LRRK2 and MARK1 are interacting in eukaryotic cells. Our findings contribute to the identification of physiologic LRRK2 substrates and point to a potential mechanism explaining the reported effects of LRRK2 on neurite morphology.

  3. Heuristic consequences of a load of oxygen in microtubules.

    Science.gov (United States)

    Denis, Pierre A

    2014-04-01

    The current cell oxygen paradigm shows some major gaps that have not yet been resolved. Something seems to be lacking for the comprehensive statement of the oxygen distribution in the cell, especially the low cytoplasmic oxygen level. The entrapment of oxygen in microtubules (MTs) resolves the latter observation, as well as the occurrence of an extensive cytoplasmic foam formation. It leads to a novel oxygen paradigm for cells. During the steady-state treadmilling, the mobile cavity would absorb oxygenated cytoplasm forward, entrap gas nuclei and concentrate them. A fluorescence method is described to confirm the in vitro load of oxygen in MTs during their periodic growths and shrinkages. The latter operating mechanism is called the gas dynamic instability (GDI) of MTs. Several known biosystems could rest on the GDI. (1) The GTP-cap is linked with the gas meniscus encountered in a tube filled with gas. The GTP hydrolysis is linked to the conformational change of the GTPase domain according to the bubble pressure, and to the shaking of protofilaments with gas particles (soliton-like waves). (2) The GDI provides a free energy water pump because water molecules have to escape from MT pores when foam concentrates within the MT. Beside ATP hydrolysis in motor proteins, the GDI provides an additional driving force in intracellular transport of cargo. The water streams flowing from the MT through slits organize themselves as water layers between the cargo and the MT surface, and break ionic bridges. It makes the cargo glide over a water rail. (3) The GDI provides a universal motor for chromosome segregation because the depolymerization of kinetochorial MTs is expected to generate a strong cytoplasmic foam. Chromosomes are sucked up according to the pressure difference (or density difference) applied to opposite sides of the kinetochore, which is in agreement with Archimedes' principle of buoyancy. Non-kinetochorial MTs reabsorb foam during GDI. Last, the mitotic spindle

  4. Modulation of microtubule assembly by the HIV-1 Tat protein is strongly dependent on zinc binding to Tat

    Directory of Open Access Journals (Sweden)

    Muller Sylviane

    2008-07-01

    Full Text Available Abstract Background During HIV-1 infection, the Tat protein plays a key role by transactivating the transcription of the HIV-1 proviral DNA. In addition, Tat induces apoptosis of non-infected T lymphocytes, leading to a massive loss of immune competence. This apoptosis is notably mediated by the interaction of Tat with microtubules, which are dynamic components essential for cell structure and division. Tat binds two Zn2+ ions through its conserved cysteine-rich region in vitro, but the role of zinc in the structure and properties of Tat is still controversial. Results To investigate the role of zinc, we first characterized Tat apo- and holo-forms by fluorescence correlation spectroscopy and time-resolved fluorescence spectroscopy. Both of the Tat forms are monomeric and poorly folded but differ by local conformational changes in the vicinity of the cysteine-rich region. The interaction of the two Tat forms with tubulin dimers and microtubules was monitored by analytical ultracentrifugation, turbidity measurements and electron microscopy. At 20°C, both of the Tat forms bind tubulin dimers, but only the holo-Tat was found to form discrete complexes. At 37°C, both forms promoted the nucleation and increased the elongation rates of tubulin assembly. However, only the holo-Tat increased the amount of microtubules, decreased the tubulin critical concentration, and stabilized the microtubules. In contrast, apo-Tat induced a large amount of tubulin aggregates. Conclusion Our data suggest that holo-Tat corresponds to the active form, responsible for the Tat-mediated apoptosis.

  5. TCTEX1D4, a novel protein phosphatase 1 interactor: connecting the phosphatase to the microtubule network

    Directory of Open Access Journals (Sweden)

    Luís Korrodi-Gregório

    2013-03-01

    Reversible phosphorylation plays an important role as a mechanism of intracellular control in eukaryotes. PPP1, a major eukaryotic Ser/Thr-protein phosphatase, acquires its specificity by interacting with different protein regulators, also known as PPP1 interacting proteins (PIPs. In the present work we characterized a physiologically relevant PIP in testis. Using a yeast two-hybrid screen with a human testis cDNA library, we identified a novel PIP of PPP1CC2 isoform, the T-complex testis expressed protein 1 domain containing 4 (TCTEX1D4 that has recently been described as a Tctex1 dynein light chain family member. The overlay assays confirm that TCTEX1D4 interacts with the different spliced isoforms of PPP1CC. Also, the binding domain occurs in the N-terminus, where a consensus PPP1 binding motif (PPP1BM RVSF is present. The distribution of TCTEX1D4 in testis suggests its involvement in distinct functions, such as TGFβ signaling at the blood–testis barrier and acrosome cap formation. Immunofluorescence in human ejaculated sperm shows that TCTEX1D4 is present in the flagellum and in the acrosome region of the head. Moreover, TCTEX1D4 and PPP1 co-localize in the microtubule organizing center (MTOC and microtubules in cell cultures. Importantly, the TCTEX1D4 PPP1BM seems to be relevant for complex formation, for PPP1 retention in the MTOC and movement along microtubules. These novel results open new avenues to possible roles of this dynein, together with PPP1. In essence TCTEX1D4/PPP1C complex appears to be involved in microtubule dynamics, sperm motility, acrosome reaction and in the regulation of the blood–testis barrier.

  6. Hydrothermal Alteration Maps of the Central and Southern Basin and Range Province of the United States Compiled From Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) Data

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) data and Interactive Data Language (IDL) logical operator algorithms were used to map...

  7. The nucleation of microtubules in Aspergillus nidulans germlings

    Directory of Open Access Journals (Sweden)

    Cristina de Andrade-Monteiro

    1999-09-01

    Full Text Available Microtubules are filaments composed of dimers of alpha- and beta-tubulins, which have a variety of functions in living cells. In fungi, the spindle pole bodies usually have been considered to be microtubule-organizing centers. We used the antimicrotubule drug Benomyl in block/release experiments to depolymerize and repolymerize microtubules in Aspergillus nidulans germlings to learn more about the microtubule nucleation process in this filamentous fungus. Twenty seconds after release from Benomyl short microtubules were formed from several bright (immunofluorescent dots distributed along the germlings, suggesting that microtubule nucleation is randomly distributed in A. nidulans germlings. Since nuclear movement is dependent on microtubules in A. nidulans we analyzed whether mutants defective in nuclear distribution along the growing hyphae (nud mutants have some obvious microtubule defect. Cytoplasmic, astral and spindle microtubules were present and appeared to be normal in all nud mutants. However, significant changes in the percentage of short versus long mitotic spindles were observed in nud mutants. This suggests that some of the nuclei of nud mutants do not reach the late stage of cell division at normal temperatures.Microtúbulos são filamentos compostos por dímeros das tubulinas a e b e têm uma variedade de funções nas células vivas. Em fungos, os corpúsculos polares dos fusos são geralmente considerados os centros organizadores dos microtúbulos. Com o objetivo de contribuir para uma melhor compreensão dos processos de nucleação dos microtúbulos no fungo filamentoso A. nidulans, nós utilizamos a droga antimicrotúbulo Benomil em experimentos de bloqueio e liberação para depolimerizar e repolimerizar os microtúbulos. Após 20 segundos de reincubação em meio sem Benomil, pequenos microtúbulos foram formados a partir de pontos distribuídos pela célula, sugerindo que os pontos de nucleação de microtúbulos s

  8. Analysis of Topographic Feature with SRTM DEM and ASTER GDEM Data and Railway Alignment%基于SRTM DEM,ASTER GDEM 地貌特征分析与铁路选线

    Institute of Scientific and Technical Information of China (English)

    高山

    2012-01-01

    研究目的:SRTM DEM,ASTER GDEM 数据具有全球范围、免费获取、精度较高的优点,结合复杂山区铁路工程,评价其数据精度,研究线路地貌特征,辅助铁路选线.研究结论:SRTM DEM,ASTER GDEM 数据由于各自存在较大的系统误差,通过采用高程精度评价、DEM融合、高程基准偏移、DEM重采样方法提高数字高程精度,进而构建大范围、高精度、可视化的三维数字地貌模型,有助于分析构造地貌特征、岩溶水文地质条件、地质灾害发育规律,提高铁路选线质量和效率.%Research purposes: The SRTM DEM and ASTER GDEM data have the advantages of the global scope, free access and high precision. The data were appliied in building railways in mountain area with complex geological condition to evaluate the data accuracy and study the topographic feature for assisting the railway alignment. Research conclusions:As the SRTM DEM and ASTER GDEM data have its own system error, the measures, such as the digital elevation accuracy evaluation, DEM fusion, elevation datum offset and DEM re - sampling, should be taken for improving the DEM accuracy and building a large range, high accuracy and visual digital topographic feature model to assist the analysis of the structural topographic feature, the geological condition of karst hydrology and the development regulation of geological disaster in order to enhance the quality and efficiency of railway alignment.

  9. Mineral mapping on the Chilean-Bolivian Altiplano using co-orbital ALI, ASTER and Hyperion imagery: Data dimensionality issues and solutions

    Science.gov (United States)

    Hubbard, B.E.; Crowley, J.K.

    2005-01-01

    Hyperspectral data coverage from the EO-1 Hyperion sensor was useful for calibrating Advanced Land Imager (ALI) and Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) images of a volcanic terrane area of the Chilean-Bolivian Altiplano. Following calibration, the ALI and ASTER datasets were co-registered and joined to produce a 13-channel reflectance cube spanning the Visible to Short Wave Infrared (0.4-2.4 ??m). Eigen analysis and comparison of the Hyperion data with the ALI + ASTER reflectance data, as well as mapping results using various ALI+ASTER data subsets, provided insights into the information dimensionality of all the data. In particular, high spectral resolution, low signal-to-noise Hyperion data were only marginally better for mineral mapping than the merged 13-channel, low spectral resolution, high signal-to-noise ALI + ASTER dataset. Neither the Hyperion nor the combined ALI + ASTER datasets had sufficient information dimensionality for mapping the diverse range of surface materials exposed on the Altiplano. However, it is possible to optimize the use of the multispectral data for mineral-mapping purposes by careful data subsetting, and by employing other appropriate image-processing strategies.

  10. Changes in microtubule stability and density in myelin-deficient shiverer mouse CNS axons

    Science.gov (United States)

    Kirkpatrick, L. L.; Witt, A. S.; Payne, H. R.; Shine, H. D.; Brady, S. T.

    2001-01-01

    Altered axon-Schwann cell interactions in PNS myelin-deficient Trembler mice result in changed axonal transport rates, neurofilament and microtubule-associated protein phosphorylation, neurofilament density, and microtubule stability. To determine whether PNS and CNS myelination have equivalent effects on axons, neurofilaments, and microtubules in CNS, myelin-deficient shiverer axons were examined. The genetic defect in shiverer is a deletion in the myelin basic protein (MBP) gene, an essential component of CNS myelin. As a result, shiverer mice have little or no compact CNS myelin. Slow axonal transport rates in shiverer CNS axons were significantly increased, in contrast to the slowing in demyelinated PNS nerves. Even more striking were substantial changes in the composition and properties of microtubules in shiverer CNS axons. The density of axonal microtubules is increased, reflecting increased expression of tubulin in shiverer, and the stability of microtubules is drastically reduced in shiverer axons. Shiverer transgenic mice with two copies of a wild-type myelin basic protein transgene have an intermediate level of compact myelin, making it possible to determine whether the actual level of compact myelin is an important regulator of axonal microtubules. Both increased microtubule density and reduced microtubule stability were still observed in transgenic mouse nerves, indicating that signals beyond synaptogenesis and the mere presence of compact myelin are required for normal regulation of the axonal microtubule cytoskeleton.

  11. The Ndc80 complex uses a tripartite attachment point to couple microtubule depolymerization to chromosome movement.

    Science.gov (United States)

    Tooley, John G; Miller, Stephanie A; Stukenberg, P Todd

    2011-04-15

    In kinetochores, the Ndc80 complex couples the energy in a depolymerizing microtubule to perform the work of moving chromosomes. The complex directly binds microtubules using an unstructured, positively charged N-terminal tail located on Hec1/Ndc80. Hec1/Ndc80 also contains a calponin homology domain (CHD) that increases its affinity for microtubules in vitro, yet whether it is required in cells and how the tail and CHD work together are critical unanswered questions. Human kinetochores containing Hec1/Ndc80 with point mutations in the CHD fail to align chromosomes or form productive microtubule attachments. Kinetochore architecture and spindle checkpoint protein recruitment are unaffected in these mutants, and the loss of CHD function cannot be rescued by removing Aurora B sites from the tail. The interaction between the Hec1/Ndc80 CHD and a microtubule is facilitated by positively charged amino acids on two separate regions of the CHD, and both are required for kinetochores to make stable attachments to microtubules. Chromosome congression in cells also requires positive charge on the Hec1 tail to facilitate microtubule contact. In vitro binding data suggest that charge on the tail regulates attachment by directly increasing microtubule affinity as well as driving cooperative binding of the CHD. These data argue that in vertebrates there is a tripartite attachment point facilitating the interaction between Hec1/Ndc80 and microtubules. We discuss how such a complex microtubule-binding interface may facilitate the coupling of depolymerization to chromosome movement.

  12. Cyclin G-associated kinase promotes microtubule outgrowth from chromosomes during spindle assembly.

    Science.gov (United States)

    Tanenbaum, Marvin E; Vallenius, Tea; Geers, Erica F; Greene, Lois; Mäkelä, Tomi P; Medema, Rene H

    2010-08-01

    During mitosis, all chromosomes must attach to microtubules of the mitotic spindle to ensure correct chromosome segregation. Microtubule attachment occurs at specialized structures at the centromeric region of chromosomes, called kinetochores. These kinetochores can generate microtubule attachments through capture of centrosome-derived microtubules, but in addition, they can generate microtubules themselves, which are subsequently integrated with centrosome-derived microtubules to form the mitotic spindle. Here, we have performed a large scale RNAi screen and identify cyclin G-associated kinase (GAK) as a novel regulator of microtubule generation at kinetochores/chromatin. This function of GAK requires its C-terminal J-domain, which is essential for clathrin recycling from endocytic vesicles. Consistently, cells lacking GAK show strongly reduced levels of clathrin on the mitotic spindle, and reduction of clathrin levels also inhibits microtubule generation at kinetochores/chromosomes. Finally, we present evidence that association of clathrin with the spindle is promoted by a signal coming from the chromosomes. These results identify a role for GAK and clathrin in microtubule outgrowth from kinetochores/chromosomes and suggest that GAK acts through clathrin to control microtubule outgrowth around chromosomes.

  13. The non-catalytic domains of Drosophila katanin regulate its abundance and microtubule-disassembly activity.

    Directory of Open Access Journals (Sweden)

    Kyle D Grode

    Full Text Available Microtubule severing is a biochemical reaction that generates an internal break in a microtubule and regulation of microtubule severing is critical for cellular processes such as ciliogenesis, morphogenesis, and meiosis and mitosis. Katanin is a conserved heterodimeric ATPase that severs and disassembles microtubules, but the molecular determinants for regulation of microtubule severing by katanin remain poorly defined. Here we show that the non-catalytic domains of Drosophila katanin regulate its abundance and activity in living cells. Our data indicate that the microtubule-interacting and trafficking (MIT domain and adjacent linker region of the Drosophila katanin catalytic subunit Kat60 cooperate to regulate microtubule severing in two distinct ways. First, the MIT domain and linker region of Kat60 decrease its abundance by enhancing its proteasome-dependent degradation. The Drosophila katanin regulatory subunit Kat80, which is required to stabilize Kat60 in cells, conversely reduces the proteasome-dependent degradation of Kat60. Second, the MIT domain and linker region of Kat60 augment its microtubule-disassembly activity by enhancing its association with microtubules. On the basis of our data, we propose that the non-catalytic domains of Drosophila katanin serve as the principal sites of integration of regulatory inputs, thereby controlling its ability to sever and disassemble microtubules.

  14. In vivo and in vitro effects of the mitochondrial uncoupler FCCP on microtubules.

    Science.gov (United States)

    Maro, B; Marty, M C; Bornens, M

    1982-01-01

    FCCP (carbonylcyanide-p-trifluoromethoxyphenylhydrazone), a potent uncoupler of oxidative phosphorylation, induces the complete disruption of cellular microtubules. A further analysis of this effect on BHK21 cells has shown that a decrease in the number of microtubules can be observed 15 min after adding FCCP and there is complete disruption after 60 min. Regrowth of microtubules was initiated 30 min after removal of FCCP, in marked contrast with the rapid reversion observed when microtubules are disrupted by nocodazole. A similar delay was required for the recovery of mitochondrial function as assessed by rhodamine 123 labelling. The effect of FCCP on microtubules was partially inhibited by preincubation of the cells with NaN3, suggesting that FCCP acts on microtubules through mitochondria. FCCP did not depolymerize microtubules of cells permeabilized with Triton X-100. In vitro polymerisation of microtubule protein was only slightly diminished by concentrations of FCCP which provoke complete disassembly in vivo. SDS-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of the microtubules polymerized in vitro in the presence of FCCP showed a reduced amount of high mol. wt. proteins, mainly MAP 2, associated with them. In an attempt to reproduce the mitochondrial effects of FCCP in vitro, we checked the effects of alkaline pH and calcium on microtubule protein polymerization in the presence of FCCP. FCCP did not influence the calcium inhibitory effect but did significantly increase the inhibitory effect of alkaline pH. We conclude that FCCP could depolymerise microtubules in vivo through a dual operation: increasing the intracellular pH by the disruption of the mitochondrial H+ gradient and decreasing the stability of microtubules by impairing the binding of microtubule-associated proteins.

  15. FRET and FRAP imaging: approaches to characterise tau and stathmin interactions with microtubules in cells.

    Science.gov (United States)

    Nouar, Roqiya; Devred, François; Breuzard, Gilles; Peyrot, Vincent

    2013-04-01

    Microtubules (MTs) are involved in many crucial processes such as cell morphogenesis, mitosis and motility. These dynamic structures resulting from the complex assembly of tubulin are tightly regulated by stabilising MT-associated proteins (MAPs) such as tau and destabilising proteins, notably stathmin. Because of their key role, these MAPs and their interactions have been extensively studied using biochemical and biophysical approaches, particularly in vitro. Nevertheless, numerous questions remain unanswered and the mechanisms of interaction between MT and these proteins are still unclear in cells. Techniques coupling cell imaging and fluorescence methods, such as Förster resonance energy transfer and fluorescence recovery after photobleaching, are excellent tools to study these interactions in situ. After describing these methods, we will present emblematic data from the literature and unpublished experimental results from our laboratory concerning the interactions between MTs, tau and stathmin in cells.

  16. Localized Aurora B activity spatially controls non-kinetochore microtubules during spindle assembly.

    Science.gov (United States)

    Tanenbaum, Marvin E; Medema, René H

    2011-12-01

    Efficient spindle assembly involves the generation of spatial cues around chromosomes that locally stabilize microtubule (MT) plus-ends. In addition to the small GTPase Ran, there is evidence that Aurora B kinase might also generate a spatial cue around chromosomes but direct proof for this is still lacking. Here, we find that the Aurora B substrate MCAK localizes to MT plus-ends throughout the mitotic spindle, but its accumulation is strongly reduced on MT plus-ends near chromatin, suggesting that a signal emanating from chromosomes negatively regulates MCAK plus-end binding. Indeed, we show that Aurora B is the kinase responsible for producing this chromosome-derived signal. These results are the first to visualize spatially restricted Aurora B kinase activity around chromosomes on an endogenous substrate and explain how Aurora B could spatially control the dynamics of non-kinetochore MTs during spindle assembly.

  17. Surface Heat Balance Analysis of Tainan City on March 6, 2001 Using ASTER and Formosat-2 Data

    Directory of Open Access Journals (Sweden)

    Chen-Yi Sun

    2008-09-01

    Full Text Available The urban heat island phenomenon occurs as a mixed result of anthropogenic heat discharge, decreased vegetation, and increased artificial impervious surfaces. To clarify the contribution of each factor to the urban heat island, it is necessary to evaluate the surface heat balance. Satellite remote sensing data of Tainan City, Taiwan, obtained from Terra ASTER and Formosat-2 were used to estimate surface heat balance in this study. ASTER data is suitable for analyzing heat balance because of the wide spectral range. We used Formosat-2 multispectral data to classify the land surface, which was used to interpolate some surface parameters for estimating heat fluxes. Because of the high spatial resolution of the Formosat-2 image, more roads, open spaces and small vegetation areas could be distinguished from buildings in urban areas; however, misclassifications of land cover in such areas using ASTER data would overestimate the sensible heat flux. On the other hand, the small vegetated areas detected from the Formosat-2 image slightly increased the estimation of latent heat flux. As a result, the storage heat flux derived from Formosat-2 is higher than that derived from ASTER data in most areas. From these results, we can conclude that the higher resolution land coverage map increases accuracy of the heat balance analysis. Storage heat flux occupies about 60 to 80% of the net radiation in most of the artificial surface areas in spite of their usages. Because of the homogeneity of the building roof materials, there is no contrast between the storage heat flux in business and residential areas. In sparsely vegetated urban areas, more heat is stored and latent heat is smaller than that in the forested suburbs. This result implies that density of vegetation has a significant influence in decreasing temperatures.

  18. The Feasibility of Coherent Energy Transfer in Microtubules

    CERN Document Server

    Craddock, Travis John Adrian; Mane, Jonathan; Hameroff, Stuart; Tuszynski, Jack A

    2014-01-01

    It was once purported that biological systems were far too warm and wet to support quantum phenomena mainly due to thermal effects disrupting quantum coherence. However recent experimental results and theoretical analyses have shown that thermal energy may assist, rather than disrupt, quantum coherence, especially in the dry hydrophobic interiors of biomolecules. Specifically, evidence has been accumulating for the necessary involvement of quantum coherence and entanglement between uniquely arranged chromophores in light harvesting photosynthetic complexes. Amazingly, the tubulin subunit proteins, which comprise microtubules, also possess a distinct architecture of chromophores, namely aromatic amino acids including tryptophan. The geometry and dipolar properties of these aromatics are similar to those found in photosynthetic units indicating that tubulin may support coherent energy transfer. Tubulin aggregated into microtubule geometric lattices may support such energy transfer, which could be of import for ...

  19. The Oligomeric Outer Dynein Arm Assembly Factor CCDC103 Is Tightly Integrated within the Ciliary Axoneme and Exhibits Periodic Binding to Microtubules*

    Science.gov (United States)

    King, Stephen M.; Patel-King, Ramila S.

    2015-01-01

    CCDC103 is an ∼29-kDa protein consisting of a central RPAP3_C domain flanked by N- and C-terminal coiled coils. Defects in CCDC103 lead to primary ciliary dyskinesia caused by the loss of outer dynein arms. This protein is present along the entire length of the ciliary axoneme and does not require other dynein or docking complex components for its integration. Unlike other known dynein assembly factors within the axoneme, CCDC103 is not solubilized by 0.6 m NaCl and requires more chaotropic conditions, such as 0.5 m KI. Alternatively, it can be extracted using 0.3% sarkosyl. CCDC103 forms stable dimers and other oligomers in solution through interactions involving the central domain. The smallest particle observed by dynamic light scattering has a hydrodynamic diameter of ∼25 nm. Furthermore, CCDC103 binds microtubules directly, forming ∼9-nm diameter particles that exhibit a 12-nm spacing on the microtubule lattice, suggesting that there may be two CCDC103 units per outer arm dynein repeat. Although the outer dynein arm docking complex is necessary to form arrays of dyneins along microtubules, it is not sufficient to set up a single array in a precise location on each axonemal doublet. We propose that CCDC103 helps generate a high-affinity site on the doublets for outer arm assembly, either through direct interactions or indirectly, perhaps by modifying the underlying microtubule lattice. PMID:25572396

  20. Kinetochore microtubule establishment is defective in oocytes from aged mice

    OpenAIRE

    Shomper, Maria; Lappa, Christina; FitzHarris, Greg

    2014-01-01

    Errors in chromosome segregation in mammalian oocytes increase in number with advancing maternal age, and are a major cause of pregnancy loss. Why chromosome segregation errors are more common in oocytes from older females remains poorly understood. In mitosis, accurate chromosome segregation is enabled by attachment of kinetochores to microtubules from appropriate spindle poles, and erroneous attachments increase the likelihood of mis-segregation. Whether attachment errors are responsible fo...

  1. The role of microtubules in contractile ring function

    Science.gov (United States)

    Conrad, A. H.; Paulsen, A. Q.; Conrad, G. W.; Spooner, B. S. (Principal Investigator)

    1992-01-01

    During cytokinesis, a cortical contractile ring forms around a cell, constricts to a stable tight neck and terminates in separation of the daughter cells. At first cleavage, Ilyanassa obsoleta embryos form two contractile rings simultaneously. The cleavage furrow (CF), in the animal hemisphere between the spindle poles, constricts to a stable tight neck and separates the daughter cells. The third polar lobe constriction (PLC-3), in the vegetal hemisphere below the spindle, constricts to a transient tight neck, but then relaxes, allowing the polar lobe cytoplasm to merge with one daughter cell. Eggs exposed to taxol, a drug that stabilizes microtubules, before the CF or the PLC-3 develop, fail to form CFs, but form stabilized tight PLCs. Eggs exposed to taxol at the time of PLC-3 formation develop varied numbers of constriction rings in their animal hemispheres and one PLC in their vegetal hemisphere, none of which relax. Eggs exposed to taxol after PLC-3 initiation form stabilized tight CFs and PLCs. At maximum constriction, control embryos display immunolocalization of nonextractable alpha-tubulin in their CFs, but not in their PLCs, and reveal, via electron microscopy, many microtubules extending through their CFs, but not through their PLCs. Embryos which form stabilized tightly constricted CFs and PLCs in the presence of taxol display immunolocalization of nonextractable alpha-tubulin in both constrictions and show many polymerized microtubules extending through both CFs and PLCs. These results suggest that the extension of microtubules through a tight contractile ring may be important for stabilizing that constriction and facilitating subsequent cytokinesis.

  2. The Microtubule Minus-End-Binding Protein Patronin/PTRN-1 Is Required for Axon Regeneration in C. elegans

    Directory of Open Access Journals (Sweden)

    Marian Chuang

    2014-11-01

    Full Text Available Precise regulation of microtubule (MT dynamics is increasingly recognized as a critical determinant of axon regeneration. In contrast to developing neurons, mature axons exhibit noncentrosomal microtubule nucleation. The factors regulating noncentrosomal MT architecture in axon regeneration remain poorly understood. We report that PTRN-1, the C. elegans member of the Patronin/Nezha/calmodulin- and spectrin-associated protein (CAMSAP family of microtubule minus-end-binding proteins, is critical for efficient axon regeneration in vivo. ptrn-1-null mutants display generally normal developmental axon outgrowth but significantly impaired regenerative regrowth after laser axotomy. Unexpectedly, mature axons in ptrn-1 mutants display elevated numbers of dynamic axonal MTs before and after injury, suggesting that PTRN-1 inhibits MT dynamics. The CKK domain of PTRN-1 is necessary and sufficient for its functions in axon regeneration and MT dynamics and appears to stabilize MTs independent of minus-end localization. Whereas in developing neurons, PTRN-1 inhibits activity of the DLK-1 mitogen-activated protein kinase (MAPK cascade, we find that, in regeneration, PTRN-1 and DLK-1 function together to promote axonal regrowth.

  3. Using ASTER Imagery in Land Use/cover Classification of Eastern Mediterranean Landscapes According to CORINE Land Cover Project

    Directory of Open Access Journals (Sweden)

    Recep Gundogan

    2008-02-01

    Full Text Available The satellite imagery has been effectively utilized for classifying land covertypes and detecting land cover conditions. The Advanced Spaceborne Thermal Emissionand Reflection Radiometer (ASTER sensor imagery has been widely used in classificationprocess of land cover. However, atmospheric corrections have to be made by preprocessingsatellite sensor imagery since the electromagnetic radiation signals received by the satellitesensors can be scattered and absorbed by the atmospheric gases and aerosols. In this study,an ASTER sensor imagery, which was converted into top-of-atmosphere reflectance(TOA, was used to classify the land use/cover types, according to COoRdination ofINformation on the Environment (CORINE land cover nomenclature, for an arearepresenting the heterogonous characteristics of eastern Mediterranean regions inKahramanmaras, Turkey. The results indicated that using the surface reflectance data ofASTER sensor imagery can provide accurate (i.e. overall accuracy and kappa values of83.2% and 0.79, respectively and low-cost cover mapping as a part of inventory forCORINE Land Cover Project.

  4. Long-Term Volcanic Activity at Shiveluch Volcano: Nine Years of ASTER Spaceborne Thermal Infrared Observations  

    Directory of Open Access Journals (Sweden)

    Adam Carter

    2010-11-01

    Full Text Available Shiveluch (Kamchatka, Russia is the most active andesitic volcano of the Kuril-Kamchatka arc, typically exhibiting near-continual high-temperature fumarolic activity and periods of exogenous lava dome emplacement punctuated by discrete large explosive eruptions. These eruptions can produce large pyroclastic flow (PF deposits, which are common on the southern flank of the volcano. Since 2000, six explosive eruptions have occurred that generated ash fall and PF deposits. Over this same time period, the Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER instrument has been acquiring image-based visible/near infrared (VNIR, short wave infrared (SWIR and thermal infrared (TIR data globally, with a particular emphasis on active volcanoes. Shiveluch was selected as an ASTER target of interest early in the mission because of its frequent activity and potential impact to northern Pacific air transportation. The north Pacific ASTER archive was queried for Shiveluch data and we present results from 2000 to 2009 that documents three large PF deposits emplaced on 19 May 2001, 9 May 2004, and 28 February 2005. The long-term archive of infrared data provides an excellent record on the changing activity and eruption state of the volcano.

  5. Using aster multispectral imagery for mapping woody invasive species in pico da vara natural reserve (Azores Islands, Portugal

    Directory of Open Access Journals (Sweden)

    Artur Gil

    2014-06-01

    Full Text Available This paper aims to assess the effectiveness of ASTER imagery to support the mapping of Pittosporum undulatum, an invasive woody species, in Pico da Vara Natural Reserve (S. Miguel Island, Archipelago of the Azores, Portugal. This assessment was done by applying K-Nearest Neighbor (KNN, Support Vector Machine (SVM and Maximum Likelihood (MLC pixel-based supervised classifications to 4 different geographic and remote sensing datasets constituted by the Visible, Near-Infrared (VNIR and Short Wave Infrared (SWIR of the ASTER sensor and by digital cartography associated to orography (altitude and "distance to water streams" of which the spatial distribution of Pittosporum undulatum directly depends. Overall, most performed classifications showed a strong agreement and high accuracy. At targeted species level, the two higher classification accuracies were obtained when applying MLC and KNN to the VNIR bands coupled with auxiliary geographic information use. Results improved significantly by including ecology and occurrence information of species (altitude and distance to water streams in the classification scheme. These results show that the use of ASTER sensor VNIR spectral bands, when coupled to relevant ancillary GIS data, can constitute an effective and low cost approach for the evaluation and continuous assessment of Pittosporum undulatum woodland propagation and distribution within Protected Areas of the Azores Islands.

  6. Cross-satellite comparison of operational land surface temperature products derived from MODIS and ASTER data over bare soil surfaces

    Science.gov (United States)

    Duan, Si-Bo; Li, Zhao-Liang; Cheng, Jie; Leng, Pei

    2017-04-01

    The collection 6 (C6) MODIS land surface temperature (LST) product is publicly available for the user community. Compared to the collection 5 (C5) MODIS LST product, the C6 MODIS LST product has been refined over bare soil pixels. Assessing the accuracy of the C6 MODIS LST product will help to facilitate the use of the LST product in various applications. In this study, we present a cross-satellite comparison to evaluate the accuracy of the C6 MODIS LST product (MOD11_L2) over bare soil surfaces under various atmospheric and surface conditions using the ASTER LST product as a reference. For comparison, the C5 MODIS LST product was also used in the analysis. The absolute biases (0.2-1.5 K) of the differences between the C6 MODIS LST and ASTER LST over bare soil surfaces are approximately two times less than those (0.6-3.8 K) of the differences between the C5 MODIS LST and ASTER LST. Furthermore, the RMSEs (0.7-2.3 K) over bare soil surfaces for the C6 MODIS LST are significantly smaller than those (0.9-4.2 K) for the C5 MODIS LST. These results indicate that the accuracy of the C6 MODIS LST product is much better than that of the C5 MODIS LST product. We recommend that the user community employs the C6 MODIS LST product in their applications.

  7. Validation of the ASTER Global Digital Elevation Model version 3 over the conterminous United States

    Science.gov (United States)

    Gesch, Dean B.; Oimoen, Michael J.; Danielson, Jeffrey J.; Meyer, David

    2016-01-01

    The ASTER Global Digital Elevation Model Version 3 (GDEM v3) was evaluated over the conterminous United States in a manner similar to the validation conducted for the original GDEM Version 1 (v1) in 2009 and GDEM Version 2 (v2) in 2011. The absolute vertical accuracy of GDEM v3 was calculated by comparison with more than 23,000 independent reference geodetic ground control points from the U.S. National Geodetic Survey. The root mean square error (RMSE) measured for GDEM v3 is 8.52 meters. This compares with the RMSE of 8.68 meters for GDEM v2. Another important descriptor of vertical accuracy is the mean error, or bias, which indicates if a DEM has an overall vertical offset from true ground level. The GDEM v3 mean error of −1.20 meters reflects an overall negative bias in GDEM v3. The absolute vertical accuracy assessment results, both mean error and RMSE, were segmented by land cover type to provide insight into how GDEM v3 performs in various land surface conditions. While the RMSE varies little across cover types (6.92 to 9.25 meters), the mean error (bias) does appear to be affected by land cover type, ranging from −2.99 to +4.16 meters across 14 land cover classes. These results indicate that in areas where built or natural aboveground features are present, GDEM v3 is measuring elevations above the ground level, a condition noted in assessments of previous GDEM versions (v1 and v2) and an expected condition given the type of stereo-optical image data collected by ASTER. GDEM v3 was also evaluated by differencing with the Shuttle Radar Topography Mission (SRTM) dataset. In many forested areas, GDEM v3 has elevations that are higher in the canopy than SRTM. The overall validation effort also included an evaluation of the GDEM v3 water mask. In general, the number of distinct water polygons in GDEM v3 is much lower than the number in a reference land cover dataset, but the total areas compare much more closely.

  8. ASTER/AVHRR Data Hybridization to determine Pyroclastic Flow cooling curves

    Science.gov (United States)

    Reath, K. A.; Wright, R.; Ramsey, M. S.

    2014-12-01

    Shiveluch Volcano (Kamchatka, Russia) has been in a consistent state of eruption for the past 15 years. During this period different eruption styles have been documented including: sub-plinian events, dome growth and collapse, and subsequent debris flow deposits. For example, on June 25-26, 2009 a pyroclastic debris flow was emplaced and the eruption onset that produced it was recorded by a series of seismic events spanning several hours. However, due to cloud cover, visual confirmation of the exact emplacement time was obscured. Orbital remote sensing was able to image the deposit repeatedly over the subsequent months. ASTER is a high spatial resolution (90m), low temporal resolution (2 - 4 days at the poles, 16 days at the equator) thermal infrared (TIR) sensor on the NASA Terra satellite. AVHRR is a high temporal resolution (minutes to several hours), low spatial resolution (1km) spaceborne TIR sensor on a series of NOAA satellites. Combined, these sensors provide a unique opportunity to fuse high-spatial and high-temporal resolution data to better observe changes on the surface of the deposit over time. For example, ASTER data were used to determine the flow area and to provide several data points for average temperature while AVHRR data were used to increase the amount of data points. Through this method an accurate average cooling rate over a three month period was determined. This cooling curve was then examined to derive several features about the deposit that were previously unknown. The time of emplacement and period of time needed for negligible thermal output were first determined by extrapolating the cooling curve in time. The total amount of heat output and total flow volume of the deposit were also calculated. This volume was then compared to the volume of the dome to calculate the percentage of collapse. This method can be repeated for other flow deposits to determine if there is a consistent correlation between the dome growth rate, the average

  9. Developing New Coastal Forest Restoration Products Based on Landsat, ASTER, and MODIS Data

    Science.gov (United States)

    Spruce, Joseph P.; Graham, William; Smoot, James

    2009-01-01

    This paper discusses an ongoing effort to develop new geospatial information products for aiding coastal forest restoration and conservation efforts in coastal Louisiana and Mississippi. This project employs Landsat, Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER), and Moderate Resolution Imaging Spectroradiometer (MODIS) satellite data in conjunction with airborne elevation data to compute coastal forest cover type maps and change detection products. Improved forest mapping products are needed to aid coastal forest restoration and management efforts of State and Federal agencies in the Northern Gulf of Mexico (NGOM) region. In particular, such products may aid coastal forest land acquisition and conservation easement procurements. This region's forests are often disturbed and subjected to multiple biotic and abiotic threats, including subsidence, salt water intrusion, hurricanes, sea-level rise, insect-induced defoliation and mortality, altered hydrology, wildfire, and conversion to non-forest land use. In some cases, such forest disturbance has led to forest loss or loss of regeneration capacity. In response, a case study was conducted to assess and demonstrate the potential of satellite remote sensing products for improving forest type maps and for assessing forest change over the last 25 years. Change detection products are needed for assessing risks for specific priority coastal forest types, such as live oak and baldcypress-dominated forest. Preliminary results indicate Landsat time series data are capable of generating the needed forest type and change detection products. Useful classifications were obtained using 2 strategies: 1) general forest classification based on use of 3 seasons of Landsat data from the same year; and 2) classification of specific forest types of concern using a single date of Landsat data in which a given targeted type is spectrally distinct compared to adjacent forested cover. When available, ASTER data was

  10. Validation of the Aster Global Digital Elevation Model Version 3 Over the Conterminous United States

    Science.gov (United States)

    Gesch, D.; Oimoen, M.; Danielson, J.; Meyer, D.

    2016-06-01

    The ASTER Global Digital Elevation Model Version 3 (GDEM v3) was evaluated over the conterminous United States in a manner similar to the validation conducted for the original GDEM Version 1 (v1) in 2009 and GDEM Version 2 (v2) in 2011. The absolute vertical accuracy of GDEM v3 was calculated by comparison with more than 23,000 independent reference geodetic ground control points from the U.S. National Geodetic Survey. The root mean square error (RMSE) measured for GDEM v3 is 8.52 meters. This compares with the RMSE of 8.68 meters for GDEM v2. Another important descriptor of vertical accuracy is the mean error, or bias, which indicates if a DEM has an overall vertical offset from true ground level. The GDEM v3 mean error of -1.20 meters reflects an overall negative bias in GDEM v3. The absolute vertical accuracy assessment results, both mean error and RMSE, were segmented by land cover type to provide insight into how GDEM v3 performs in various land surface conditions. While the RMSE varies little across cover types (6.92 to 9.25 meters), the mean error (bias) does appear to be affected by land cover type, ranging from -2.99 to +4.16 meters across 14 land cover classes. These results indicate that in areas where built or natural aboveground features are present, GDEM v3 is measuring elevations above the ground level, a condition noted in assessments of previous GDEM versions (v1 and v2) and an expected condition given the type of stereo-optical image data collected by ASTER. GDEM v3 was also evaluated by differencing with the Shuttle Radar Topography Mission (SRTM) dataset. In many forested areas, GDEM v3 has elevations that are higher in the canopy than SRTM. The overall validation effort also included an evaluation of the GDEM v3 water mask. In general, the number of distinct water polygons in GDEM v3 is much lower than the number in a reference land cover dataset, but the total areas compare much more closely.

  11. Microtubule-Actin Cross-Linking Factor 1: Domains, Interaction Partners, and Tissue-Specific Functions.

    Science.gov (United States)

    Goryunov, Dmitry; Liem, Ronald K H

    2016-01-01

    The cytoskeleton of most eukaryotic cells is composed of three principal filamentous components: actin filaments, microtubules (MTs), and intermediate filaments. It is a highly dynamic system that plays crucial roles in a wide range of cellular processes, including migration, adhesion, cytokinesis, morphogenesis, intracellular traffic and signaling, and structural flexibility. Among the large number of cytoskeleton-associated proteins characterized to date, microtubule-actin cross-linking factor 1 (MACF1) is arguably the most versatile integrator and modulator of cytoskeleton-related processes. MACF1 belongs to the plakin family of proteins, and within it, to the spectraplakin subfamily. These proteins are characterized by the ability to bridge MT and actin cytoskeletal networks in a dynamic fashion, which underlies their involvement in the regulation of cell migration, axonal extension, and vesicular traffic. Studying MACF1 functions has provided insights not only into the regulation of the cytoskeleton but also into molecular mechanisms of both normal cellular physiology and cellular pathology. Multiple MACF1 isoforms exist, composed of a large variety of alternatively spliced domains. Each of these domains mediates a specific set of interactions and functions. These functions are manifested in tissue and cell-specific phenotypes observed in conditional MACF1 knockout mice. The conditional models described to date reveal critical roles of MACF1 in mammalian skin, nervous system, heart muscle, and intestinal epithelia. Complete elimination of MACF1 is early embryonic lethal, indicating an essential role for MACF1 in early development. Further studies of MACF1 domains and their interactions will likely reveal multiple new roles of this protein in various tissues.

  12. Molecular Quantum Robotics: Particle and Wave Solutions, illustrated by "Leg-over-Leg" Walking along Microtubules

    Directory of Open Access Journals (Sweden)

    Paul eLevi

    2015-05-01

    Full Text Available Remarkable biological examples of molecular robots are the proteins kinesin-1 and dynein, which move and transport cargo down microtubule highways, e.g. of the axon, to final nerve nodes or along dendrites. They convert the energy of ATP hydrolysis into mechanical forces and can thereby push them forwards or backwards step by step. Such mechano-chemical cycles that generate conformal changes are essential for transport on all different types of substrate lanes. The step length of an individual molecular robot is a matter of nanometers but the dynamics of each individual step cannot be predicted with certainty (as it is a random process. Hence, our proposal is to involve the methods of quantum field theory (QFT to describe an overall reliable, multi–robot system that is composed of a huge set of unreliable, local elements. The methods of QFT deliver techniques that are also computationally demanding to synchronize the motion of these molecular robots on one substrate lane as well as across lanes.Three different challenging types of solutions are elaborated. The impact solution reflects the particle point of view; the two remaining solutions are wave based. The second solution outlines coherent robot motions on different lanes. The third solution describes running waves. Experimental investigations are needed to clarify under which biological conditions such different solutions occur.Moreover, such a nano-chemical system can be stimulated by external signals, and this opens a new, hybrid approach to analyze and control the combined system of robots and microtubules externally. Such a method offers the chance to detect mal-functions of the biological system. In our framework, such defects can be characterized by the distortion of typical features of dynamic systems like attractive fixed points, limit cycles, etc. However, such additional details would overload this presentation and obscure the essentials that we wish to point out.

  13. GIT1 enhances neurite outgrowth by stimulating microtubule assembly

    Institute of Scientific and Technical Information of China (English)

    Yi-sheng Li; Li-xia Qin; Jie Liu; Wei-liang Xia; Jian-ping Li; Hai-lian Shen; Wei-Qiang Gao

    2016-01-01

    GIT1, a G-protein-coupled receptor kinase interacting protein, has been reported to be involved in neurite outgrowth. However, the neu-robiological functions of the protein remain unclear. In this study, we found that GIT1 was highly expressed in the nervous system, and its expression was maintained throughout all stages of neuritogenesis in the brain. In primary cultured mouse hippocampal neurons from GIT1 knockout mice, there was a signiifcant reduction in total neurite length per neuron, as well as in the average length of axon-like struc-tures, which could not be prevented by nerve growth factor treatment. Overexpression of GIT1 signiifcantly promoted axon growth and fully rescued the axon outgrowth defect in the primary hippocampal neuron cultures from GIT1 knockout mice. The GIT1 N terminal region, including the ADP ribosylation factor-GTPase activating protein domain, the ankyrin domains and the Spa2 homology domain, were sufifcient to enhance axonal extension. Importantly, GIT1 bound to many tubulin proteins and microtubule-associated proteins, and it accelerated microtubule assemblyin vitro. Collectively, our ifndings suggest that GIT1 promotes neurite outgrowth, at least partially by stimulating microtubule assembly. This study provides new insight into the cellular and molecular pathogenesis of GIT1-associated neuro-logical diseases.

  14. GIT1 enhances neurite outgrowth by stimulating microtubule assembly

    Directory of Open Access Journals (Sweden)

    Yi-sheng Li

    2016-01-01

    Full Text Available GIT1, a G-protein-coupled receptor kinase interacting protein, has been reported to be involved in neurite outgrowth. However, the neurobiological functions of the protein remain unclear. In this study, we found that GIT1 was highly expressed in the nervous system, and its expression was maintained throughout all stages of neuritogenesis in the brain. In primary cultured mouse hippocampal neurons from GIT1 knockout mice, there was a significant reduction in total neurite length per neuron, as well as in the average length of axon-like structures, which could not be prevented by nerve growth factor treatment. Overexpression of GIT1 significantly promoted axon growth and fully rescued the axon outgrowth defect in the primary hippocampal neuron cultures from GIT1 knockout mice. The GIT1 N terminal region, including the ADP ribosylation factor-GTPase activating protein domain, the ankyrin domains and the Spa2 homology domain, were sufficient to enhance axonal extension. Importantly, GIT1 bound to many tubulin proteins and microtubule-associated proteins, and it accelerated microtubule assembly in vitro. Collectively, our findings suggest that GIT1 promotes neurite outgrowth, at least partially by stimulating microtubule assembly. This study provides new insight into the cellular and molecular pathogenesis of GIT1-associated neurological diseases.

  15. Site-specific phosphorylation of Tau protein is associated with deacetylation of microtubules in mouse spermatogenic cells during meiosis.

    Science.gov (United States)

    Inoue, Hiroki; Hiradate, Yuuki; Shirakata, Yoshiki; Kanai, Kenta; Kosaka, Keita; Gotoh, Aina; Fukuda, Yasuhiro; Nakai, Yutaka; Uchida, Takafumi; Sato, Eimei; Tanemura, Kentaro

    2014-05-29

    Tau is one of the microtubule-associated proteins and a major component of paired helical filaments, a hallmark of Alzheimer's disease. Its expression has also been indicated in the testis. However, its function and modification in the testis have not been established. Here, we analyzed the dynamics of phosphorylation patterns during spermatogenesis. The expression of Tau protein and its phosphorylation were shown in the mouse testis. Immunohistochemistry revealed that the phosphorylation was strongly detected during meiosis. Correspondingly, the expression of acetylated tubulin was inversely weakened during meiosis. These results suggest that phosphorylation of Tau protein contributes to spermatogenesis, especially in meiosis.

  16. Quantitative comparison of Landsat 7 and ASTER multispectral measurements for the NDBI and IBI%Landsat-7 ETM+与ASTER建筑指数的定量比较

    Institute of Scientific and Technical Information of China (English)

    徐涵秋; 张铁军; 黄绍霖

    2013-01-01

    Accompanying with accelerated urbanization processes,urban built-up lands have expanded dramatically and caused negative impacts on environments and natural resources of the world.Thus,there is an increasing need to monitor the dynamics of the urban built-up lands.Of the medium-scale earth observation systems,Landsat and ASTER systems are most suitable for this monitoring task.Nevertheless,the relationship between the built-up indices of the two sensors has not yet been revealed so far.To quantitatively investigate this relationship,this study crossly compares the NDBI and IBI built-up indices of Landsat-7 ETM+ and ASTER sensors based on three tandem image pairs.Regression analysis was used to examine the quantitative relationship through the scatter plots of the built-up index values of the two sensors.This yielded the models for the conversion between two sensors' built-up indices.Results indicate that there are differences in the compared built-up indices between two sensors in spite of a high degree of correlation between them.The values of the built-up indices of ASTER are greater than those of ETM+ by 48.6% in NDBI or 37.5% in IBI.Accordingly,the built-up index information derived from ASTER data cannot be compared directly with those derived from ETM+ data in the applications.A conversion between them using the conversion models obtained in this study is recommended.Detailed spectral analysis reveals that the differences in band wavelengths and spectral response functions of the near infrared and shortwave infrared bands between the ETM + and ASTER sensors contribute to the built-up index difference between the two sensors.This is because the NDBI and IBI are composed merely or mostly by these two bands.%城市化进程导致的城市建筑用地急剧扩张已对全球的环境资源产生了负面影响,因此亟需对其进行监测.在中尺度卫星对地观测系统中,Landsat和ASTER数据最适合于监测城市建筑用地变化.但是迄今为

  17. Kinesin-1 Translocation along Human Breast Cancer Cell Microtubules in Vitro

    Science.gov (United States)

    Shojania Feizabadi, Mitra; Jun, Yonggun

    2015-03-01

    A principle approach to better understand intra-cellular microtubule based transport is to study such it in vitro. Such in vitro examinations have predominantly used microtubules polymerized from bovine brain tubulin, but motor function can also in principle be affected by the specific tubulin isotypes present in different cells. The human breast cancer cells carry different beta tubulin isotype distribution. However, it is entirely unknown whether transport along the microtubules is different in these cells. In this work we have characterized, for the first time, the translocation specifications of kinesin-1 along human breast cancer cell microtubules polymerized in vitro. We found that as compared with the translocation along bovine brain microtubules, kinesin-1 shows a fifty percent shorter processive run length and slightly slower velocity under similar experimental conditions. These first time results support the regulatory role of tubulin isotypes in regards to motor protein translocations, and quantify the translocation specifications of kinesin-1 along microtubules of human breast cancer cells.

  18. Selective extraction of isolated mitotic apparatus. Evidence that typical microtubule protein is extracted by organic mercurial.

    Science.gov (United States)

    Bibring, T; Baxandall, J

    1971-02-01

    Mitotic apparatus isolated from sea urchin eggs has been treated with meralluride sodium under conditions otherwise resembling those of its isolation. The treatment causes a selective morphological disappearance of microtubules while extracting a major protein fraction, probably consisting of two closely related proteins, which constitutes about 10% of mitotic apparatus protein. Extraction of other cell particulates under similar conditions yields much less of this protein. The extracted protein closely resembles outer doublet microtubule protein from sea urchin sperm tail in properties considered typical of microtubule proteins: precipitation by calcium ion and vinblastine, electrophoretic mobility in both acid and basic polyacrylamide gels, sedimentation coefficient, molecular weight, and, according to a preliminary determination, amino acid composition. An antiserum against a preparation of sperm tail outer doublet microtubules cross-reacts with the extract from mitotic apparatus. On the basis of these findings it appears that microtubule protein is selectively extracted from isolated mitotic apparatus by treatment with meralluride, and is a typical microtubule protein.

  19. Regulation of developmental and environmental signaling by interaction between microtubules and membranes in plant cells

    Directory of Open Access Journals (Sweden)

    Qun Zhang

    2015-12-01

    Full Text Available ABSTRACT Cell division and expansion require the ordered arrangement of microtubules, which are subject to spatial and temporal modifications by developmental and environmental factors. Understanding how signals translate to changes in cortical microtubule organization is of fundamental importance. A defining feature of the cortical microtubule array is its association with the plasma membrane; modules of the plasma membrane are thought to play important roles in the mediation of microtubule organization. In this review, we highlight advances in research on the regulation of cortical microtubule organization by membrane-associated and membrane-tethered proteins and lipids in response to phytohormones and stress. The transmembrane kinase receptor Rho-like guanosine triphosphatase, phospholipase D, phosphatidic acid, and phosphoinositides are discussed with a focus on their roles in microtubule organization.

  20. Relative velocity of sliding of microtubules by the action of Kinesin-5

    CERN Document Server

    Roy, Sthitadhi

    2011-01-01

    Kinesin-5, also known as Eg5 in vertebrates is a processive motor with 4 heads, which moves on filamentous tracks called microtubules. The basic function of Kinesin-5 is to slide apart two anti-parallel microtubules by simultaneously walking on both the microtubules. We develop an analytical expression for the steady-state relative velocity of this sliding in terms of the rates of attachments and detachments of motor heads with the ATPase sites on the microtubules. We first analyse the motion of one pair of motor heads on one microtubule and then couple it to the motion of the other pair of motor heads of the same motor on the second microtubule to get the relative velocity of sliding.

  1. High rectifying efficiencies of microtubule motility on kinesin-coated gold nanostructures.

    Science.gov (United States)

    van den Heuvel, Martin G L; Butcher, Christopher T; Smeets, Ralph M M; Diez, Stefan; Dekker, Cees

    2005-06-01

    We demonstrate highly efficient rectification of microtubule motility on gold nanofabricated structures. First, we present a novel nanofabrication process for the creation of gold tracks for microtubule motility recessed in silicon oxide. This approach is particularly useful because it enables the use of the well-understood PEG-silane chemistry on SiO2 for the blocking of kinesin, whereas the gold tracks allow possible electrical control. We demonstrate excellent confinement of microtubule motility to the gold nanostructures and that microtubules move on the gold with speeds comparable to that on glass. Second, we present designs of three advanced rectifier geometries. We analyze the microtubule pathways through the geometries, and we demonstrate highly efficient rectification with up to 92% efficiency. As a result, we find that up to 97% of the microtubules move unidirectionally.

  2. Mitosis and microtubule organizational changes in rice root-tip cells

    Institute of Scientific and Technical Information of China (English)

    XUSHIXIONG(SYZEE); CHUNGUILI; CHENGZHU

    1993-01-01

    The pattern of change of the microtubule cytoskeleton of the root-tip cells of rice during mitosis was studied using immunofluorescence technic and confocal laser scanning microscopy. All the major stages of ceil division including preprophase, prophase, metaphase, anaphase and telophase were observed. The most significant finding was that in the preprophase cells microtubules radiating from the nuclear surface to the cortex were frequently seen. During development these microtubules became closely associated with the preprophase band and prophase spindie indicating that the microtubules radiating from the nuclear surface, the preprophase band and the prophazc spindle were structurally and functionally closely related to each other. Granule-like anchorage sites for the radiating microtubules at the muclear surface were often seen and the possibility that these gramle-like anchorage sites might represent the microtubule organizing centres was discussed.

  3. Polo-like kinase 1 regulates Nlp, a centrosome protein involved in microtubule nucleation.

    Science.gov (United States)

    Casenghi, Martina; Meraldi, Patrick; Weinhart, Ulrike; Duncan, Peter I; Körner, Roman; Nigg, Erich A

    2003-07-01

    In animal cells, most microtubules are nucleated at centrosomes. At the onset of mitosis, centrosomes undergo a structural reorganization, termed maturation, which leads to increased microtubule nucleation activity. Centrosome maturation is regulated by several kinases, including Polo-like kinase 1 (Plk1). Here, we identify a centrosomal Plk1 substrate, termed Nlp (ninein-like protein), whose properties suggest an important role in microtubule organization. Nlp interacts with two components of the gamma-tubulin ring complex and stimulates microtubule nucleation. Plk1 phosphorylates Nlp and disrupts both its centrosome association and its gamma-tubulin interaction. Overexpression of an Nlp mutant lacking Plk1 phosphorylation sites severely disturbs mitotic spindle formation. We propose that Nlp plays an important role in microtubule organization during interphase, and that the activation of Plk1 at the onset of mitosis triggers the displacement of Nlp from the centrosome, allowing the establishment of a mitotic scaffold with enhanced microtubule nucleation activity.

  4. Tracking of plus-ends reveals microtubule functional diversity in different cell types

    Science.gov (United States)

    Shaebani, M. Reza; Pasula, Aravind; Ott, Albrecht; Santen, Ludger

    2016-07-01

    Many cellular processes are tightly connected to the dynamics of microtubules (MTs). While in neuronal axons MTs mainly regulate intracellular trafficking, they participate in cytoskeleton reorganization in many other eukaryotic cells, enabling the cell to efficiently adapt to changes in the environment. We show that the functional differences of MTs in different cell types and regions is reflected in the dynamic properties of MT tips. Using plus-end tracking proteins EB1 to monitor growing MT plus-ends, we show that MT dynamics and life cycle in axons of human neurons significantly differ from that of fibroblast cells. The density of plus-ends, as well as the rescue and catastrophe frequencies increase while the growth rate decreases toward the fibroblast cell margin. This results in a rather stable filamentous network structure and maintains the connection between nucleus and membrane. In contrast, plus-ends are uniformly distributed along the axons and exhibit diverse polymerization run times and spatially homogeneous rescue and catastrophe frequencies, leading to MT segments of various lengths. The probability distributions of the excursion length of polymerization and the MT length both follow nearly exponential tails, in agreement with the analytical predictions of a two-state model of MT dynamics.

  5. 用Asterisk和AsterCRM部署心理咨询呼叫中心

    Institute of Scientific and Technical Information of China (English)

    李正鹏; 王迪

    2010-01-01

    随着社会快速发展与变革,人们对心理咨询的需求日益增多,如何提高客户服务质量和效率是心理咨询机构成功的关键因素.由于呼叫中心(CallCenter)能提供便利、规范和高效的客户服务,因此在心理咨询客户诉聊中具有很好的应用价值,Asterisk是目前使用最广泛的开源IP-PBX系统软件,AsterCRM是一款基于web的针对asterisk开发的开源呼叫中心软件,文章详细介绍了Asterisk软交换平台和AstetCRM呼叫中心平台,并设计实现了心理咨询呼叫中心.

  6. Accuracy Improvement of ASTER Stereo Satellite Generated DEM Using Texture Filter

    Institute of Scientific and Technical Information of China (English)

    Mandla V. Ravibabu; Kamal Jain; Surendra Pal Singh; Naga Jyothi Meeniga

    2010-01-01

    The grid DEM (digital elevation model) generation can be from any of a number of sources: for instance, analogue to digital conversion of contour maps followed by application of the TIN model, or direct elevation point modelling via digital photogrammetry applied to airborne images or satellite images. Currently, apart from the deployment of point-clouds from LiDAR data acquisition, the generally favoured approach refers to applications of digital photogrammetry. One of the most important steps in such deployment is the stereo matching process for conjugation point (pixel) establishment: very difficult in modelling any homogenous areas like water cover or forest canopied areas due to the lack of distinct spatial features. As a result, application of automated procedures is sure to generate erroneous elevation values. In this paper, we present and apply a method for improving the quality of stereo DEMs generated via utilization of an entropy texture filter. The filter was applied for extraction of homogenous areas before stereo matching so that a statistical texture filter could then be applied for removing anomalous evaluation values prior to interpolation and accuracy assessment via deployment of a spatial correlation technique. For exemplification, we used a stereo pair of ASTER 1B images.

  7. Inversion of Land Surface Temperature (LST Using Terra ASTER Data: A Comparison of Three Algorithms

    Directory of Open Access Journals (Sweden)

    Milton Isaya Ndossi

    2016-12-01

    Full Text Available Land Surface Temperature (LST is an important measurement in studies related to the Earth surface’s processes. The Advanced Space-borne Thermal Emission and Reflection Radiometer (ASTER instrument onboard the Terra spacecraft is the currently available Thermal Infrared (TIR imaging sensor with the highest spatial resolution. This study involves the comparison of LSTs inverted from the sensor using the Split Window Algorithm (SWA, the Single Channel Algorithm (SCA and the Planck function. This study has used the National Oceanic and Atmospheric Administration’s (NOAA data to model and compare the results from the three algorithms. The data from the sensor have been processed by the Python programming language in a free and open source software package (QGIS to enable users to make use of the algorithms. The study revealed that the three algorithms are suitable for LST inversion, whereby the Planck function showed the highest level of accuracy, the SWA had moderate level of accuracy and the SCA had the least accuracy. The algorithms produced results with Root Mean Square Errors (RMSE of 2.29 K, 3.77 K and 2.88 K for the Planck function, the SCA and SWA respectively.

  8. Per-Field Irrigated Crop Classification in Arid Central Asia Using SPOT and ASTER Data

    Directory of Open Access Journals (Sweden)

    Christopher Conrad

    2010-04-01

    Full Text Available The overarching goal of this research was to explore accurate methods of mapping irrigated crops, where digital cadastre information is unavailable: (a Boundary separation by object-oriented image segmentation using very high spatial resolution (2.5–5 m data was followed by (b identification of crops and crop rotations by means of phenology, tasselled cap, and rule-based classification using high resolution (15–30 m bi-temporal data. The extensive irrigated cotton production system of the Khorezm province in Uzbekistan, Central Asia, was selected as a study region. Image segmentation was carried out on pan-sharpened SPOT data. Varying combinations of segmentation parameters (shape, compactness, and color were tested for optimized boundary separation. The resulting geometry was validated against polygons digitized from the data and cadastre maps, analysing similarity (size, shape and congruence. The parameters shape and compactness were decisive for segmentation accuracy. Differences between crop phenologies were analyzed at field level using bi-temporal ASTER data. A rule set based on the tasselled cap indices greenness and brightness allowed for classifying crop rotations of cotton, winter-wheat and rice, resulting in an overall accuracy of 80 %. The proposed field-based crop classification method can be an important tool for use in water demand estimations, crop yield simulations, or economic models in agricultural systems similar to Khorezm.

  9. Spatial distributions of the leafminer Ophiomyia maura (Diptera: Agromyzidae) in host plant Aster ageratoides

    Institute of Scientific and Technical Information of China (English)

    Yoshiko Ayabe; Ei'ichi Shibata

    2008-01-01

    The seasonal occurrence and among-plant and within-plant spatial distribution of the multivoltine leafminer Ophiomyia maura Meigen (Diptera: Agromyzidae) on the herbaceous plant Aster ageratoides Turcz. subsp, ovatus (Asteraceae) were investigated in the field. O. maura has at least four generations a year and mines per leaf fluctuate with a mean of 0.007 throughout the occurrence period. Seasonal occurrence is associated with abundance of new host leaves, suggesting O. maura females prefer to oviposit in newly emerged leaves. The among-plant distribution of O. maura is described by a Poisson distribution early in the season but tends to be weakly clumped later. The within-plant vertical distribution of larval mines increased from middle to upper leaves during plantdevelopment, because mined leaves in the middle position early in the season move downward with the emergence of new leaves, shifting mined leaves from the position where O. maura oviposits eggs. Later in the season, mined leaves remain where they are deposited because few new leaves emerge. The spatial distribution of O. rnaura, resource utilization patterns, and host plant characteristics are discussed.

  10. Thermal remote sensing of ice-debris landforms using ASTER: an example from the Chilean Andes

    Science.gov (United States)

    Brenning, A.; Peña, M. A.; Long, S.; Soliman, A.

    2012-03-01

    Remote sensors face challenges in characterizing mountain permafrost and ground thermal conditions or mapping rock glaciers and debris-covered glaciers. We explore the potential of thermal imaging and in particular thermal inertia mapping in mountain cryospheric research, focusing on the relationships between ground surface temperatures and the presence of ice-debris landforms on one side and land surface temperature (LST) and apparent thermal inertia (ATI) on the other. In our case study we utilize ASTER daytime and nighttime imagery and in-situ measurements of near-surface ground temperature (NSGT) in the Mediterranean Andes during a snow-free and dry observation period in late summer. Spatial patterns of LST and NSGT were mostly consistent with each other both at daytime and at nighttime. Daytime LST over ice-debris landforms was decreased and ATI consequently increased compared to other debris surfaces under otherwise equal conditions, but NSGT showed contradictory results, which underlines the complexity and possible scale dependence of ATI in heterogeneous substrates with the presence of a thermal mismatch and a heat sink at depth. While our results demonstrate the utility of thermal imaging and ATI mapping in a mountain cryospheric context, further research is needed for a better interpretation of ATI patterns in complex thermophysical conditions.

  11. Alteration, slope-classified alteration, and potential lahar inundation maps of volcanoes for the Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) Volcano Archive

    Science.gov (United States)

    Mars, John C.; Hubbard, Bernard E.; Pieri, David; Linick, Justin

    2015-01-01

    This study identifies areas prone to lahars from hydrothermally altered volcanic edifices on a global scale, using visible and near infrared (VNIR) and short wavelength infrared (SWIR) reflectance data from the Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) and digital elevation data from the ASTER Global Digital Elevation Model (GDEM) dataset. This is the first study to create a global database of hydrothermally altered volcanoes showing quantitatively compiled alteration maps and potentially affected drainages, as well as drainage-specific maps illustrating modeled lahars and their potential inundation zones. We (1) identified and prioritized 720 volcanoes based on population density surrounding the volcanoes using the Smithsonian Institution Global Volcanism Program database (GVP) and LandScan™ digital population dataset; (2) validated ASTER hydrothermal alteration mapping techniques using Airborne Visible and Infrared Imaging Spectrometer (AVIRIS) and ASTER data for Mount Shasta, California, and Pico de Orizaba (Citlaltépetl), Mexico; (3) mapped and slope-classified hydrothermal alteration using ASTER VNIR-SWIR reflectance data on 100 of the most densely populated volcanoes; (4) delineated drainages using ASTER GDEM data that show potential flow paths of possible lahars for the 100 mapped volcanoes; (5) produced potential alteration-related lahar inundation maps using the LAHARZ GIS code for Iztaccíhuatl, Mexico, and Mount Hood and Mount Shasta in the United States that illustrate areas likely to be affected based on DEM-derived volume estimates of hydrothermally altered rocks and the ~2x uncertainty factor inherent within a statistically-based lahar model; and (6) saved all image and vector data for 3D and 2D display in Google Earth™, ArcGIS® and other graphics display programs. In addition, these data are available from the ASTER Volcano Archive (AVA) for distribution (available at http://ava.jpl.nasa.gov/recent_alteration_zones.php).

  12. Myomegalin is necessary for the formation of centrosomal and Golgi-derived microtubules

    Directory of Open Access Journals (Sweden)

    Régine Roubin

    2012-12-01

    The generation of cellular microtubules is initiated at specific sites such as the centrosome and the Golgi apparatus that contain nucleation complexes rich in γ-tubulin. The microtubule growing plus-ends are stabilized by plus-end tracking proteins (+TIPs, mainly EB1 and associated proteins. Myomegalin was identified as a centrosome/Golgi protein associated with cyclic nucleotide phosphodiesterase. We show here that Myomegalin exists as several isoforms. We characterize two of them. One isoform, CM-MMG, harbors a conserved domain (CM1, recently described as a nucleation activator, and is related to a family of γ-tubulin binding proteins, which includes Drosophila centrosomin. It localizes at the centrosome and at the cis-Golgi in an AKAP450-dependent manner. It recruits γ-tubulin nucleating complexes and promotes microtubule nucleation. The second isoform, EB-MMG, is devoid of CM1 domain and has a unique N-terminus with potential EB1-binding sites. It localizes at the cis-Golgi and can localize to microtubule plus-ends. EB-MMG binds EB1 and affects its loading on microtubules and microtubule growth. Depletion of Myomegalin by small interfering RNA delays microtubule growth from the centrosome and Golgi apparatus, and decreases directional migration of RPE1 cells. In conclusion, the Myomegalin gene encodes different isoforms that regulate microtubules. At least two of these have different roles, demonstrating a previously unknown mechanism to control microtubules in vertebrate cells.

  13. Inhibition of kinesin-5 improves regeneration of injured axons by a novel microtubule-based mechanism

    Institute of Scientific and Technical Information of China (English)

    Peter W. Baas; Andrew J. Matamoros

    2015-01-01

    Microtubules have been identiifed as a powerful target for augmenting regeneration of injured adult axons in the central nervous system. Drugs that stabilize microtubules have shown some promise, but there are concerns that abnormally stabilizing microtubules may have only limited beneifts for regeneration, while at the same time may be detrimental to the normal work that microtubules perform for the axon. Kinesin-5 (also called kif11 or Eg5), a molecular motor protein best known for its crucial role in mitosis, acts as a brake on microtubule movements by other motor proteins in the axon. Drugs that inhibit kinesin-5, originally developed to treat cancer, result in greater mobility of microtubules in the axon and an overall shift in the forces on the microtubule array. As a result, the axon grows faster, retracts less, and more readily enters environments that are inhibitory to axonal regeneration. Thus, drugs that inhibit kinesin-5 offer a novel microtubule-based means to boost axonal regeneration without the concerns that ac-company abnormal stabilization of the microtubule array. Even so, inhibiting kinesin-5 is not without its own caveats, such as potential problems with navigation of the regenerating axon to its target, as well as morphological effects on dendrites that could affect learning and memory if the drugs reach the brain.

  14. Microtubule polarity and the direction of pigment transport reverse simultaneously in surgically severed melanophore arms.

    Science.gov (United States)

    McNiven, M A; Wang, M; Porter, K R

    1984-07-01

    The transport of pigment through the long cytoplasmic extensions (arms) of teleost melanophores is a microtubule-dependent event. We have severed the arms from melanophores to test whether microtubules isolated from the centrosome maintain their original polarity and disposition. In addition, we have tested whether arms containing microtubules of mixed polarities alter the direction of pigment transport. We find that microtubules within severed arms eventually change their polarity and reorganize from the arm center as if to form a new minicell. Concomitant with this change is a reversal in the direction of pigment transport.

  15. Measurement of Breaking Force of Fluorescence Labelled Microtubules with Optical Tweezers

    Institute of Scientific and Technical Information of China (English)

    LIU Chun-Xiang; GUO Hong-Lian; XU Chun-Hua; YUAN Ming; LI Znao-Lin; CHENG Bing-Ying; ZHANG Dao-Zhong

    2005-01-01

    @@ Under illumination of excitation light, the force that can make fluorescent dye-labelled microtubules break up is measured by using dual-beam optical tweezers. It is found that this force is about several piconewtons, which is two orders of magnitude smaller than that without fluorescence label. Microtubules can be elongated about 20% and the increase of the tensile force is nonlinear with the microtubule elongation. Some qualitative explanations are given for the mechanisms about the breakup and elongation of microtubules exposed to excitation light.

  16. Combing and self-assembly phenomena in dry films of Taxol-stabilized microtubules

    Directory of Open Access Journals (Sweden)

    Rose Franck

    2007-01-01

    Full Text Available AbstractMicrotubules are filamentous proteins that act as a substrate for the translocation of motor proteins. As such, they may be envisioned as a scaffold for the self-assembly of functional materials and devices. Physisorption, self-assembly and combing are here investigated as a potential prelude to microtubule-templated self-assembly. Dense films of self-assembled microtubules were successfully produced, as well as patterns of both dendritic and non-dendritic bundles of microtubules. They are presented in the present paper and the mechanism of their formation is discussed.

  17. Inhibition of kinesin-5 improves regeneration of injured axons by a novel microtubule-based mechanism

    Directory of Open Access Journals (Sweden)

    Peter W Baas

    2015-01-01

    Full Text Available Microtubules have been identified as a powerful target for augmenting regeneration of injured adult axons in the central nervous system. Drugs that stabilize microtubules have shown some promise, but there are concerns that abnormally stabilizing microtubules may have only limited benefits for regeneration, while at the same time may be detrimental to the normal work that microtubules perform for the axon. Kinesin-5 (also called kif11 or Eg5, a molecular motor protein best known for its crucial role in mitosis, acts as a brake on microtubule movements by other motor proteins in the axon. Drugs that inhibit kinesin-5, originally developed to treat cancer, result in greater mobility of microtubules in the axon and an overall shift in the forces on the microtubule array. As a result, the axon grows faster, retracts less, and more readily enters environments that are inhibitory to axonal regeneration. Thus, drugs that inhibit kinesin-5 offer a novel microtubule-based means to boost axonal regeneration without the concerns that accompany abnormal stabilization of the microtubule array. Even so, inhibiting kinesin-5 is not without its own caveats, such as potential problems with navigation of the regenerating axon to its target, as well as morphological effects on dendrites that could affect learning and memory if the drugs reach the brain.

  18. cAMP-dependent protein kinase and c-Jun N-terminal kinase mediate stathmin phosphorylation for the maintenance of interphase microtubules during osmotic stress.

    Science.gov (United States)

    Yip, Yan Y; Yeap, Yvonne Y C; Bogoyevitch, Marie A; Ng, Dominic C H

    2014-01-24

    Dynamic microtubule changes after a cell stress challenge are required for cell survival and adaptation. Stathmin (STMN), a cytoplasmic microtubule-destabilizing phosphoprotein, regulates interphase microtubules during cell stress, but the signaling mechanisms involved are poorly defined. In this study ectopic expression of single alanine-substituted phospho-resistant mutants demonstrated that STMN Ser-38 and Ser-63 phosphorylation were specifically required to maintain interphase microtubules during hyperosmotic stress. STMN was phosphorylated on Ser-38 and Ser-63 in response to hyperosmolarity, heat shock, and arsenite treatment but rapidly dephosphorylated after oxidative stress treatment. Two-dimensional PAGE and Phos-tag gel analysis of stress-stimulated STMN phospho-isoforms revealed rapid STMN Ser-38 phosphorylation followed by subsequent Ser-25 and Ser-63 phosphorylation. Previously, we delineated stress-stimulated JNK targeting of STMN. Here, we identified cAMP-dependent protein kinase (PKA) signaling as responsible for stress-induced STMN Ser-63 phosphorylation. Increased cAMP levels induced by cholera toxin triggered potent STMN Ser-63 phosphorylation. Osmotic stress stimulated an increase in PKA activity and elevated STMN Ser-63 and CREB (cAMP-response element-binding protein) Ser-133 phosphorylation that was substantially attenuated by pretreatment with H-89, a PKA inhibitor. Interestingly, PKA activity and subsequent phosphorylation of STMN were augmented in the absence of JNK activation, indicating JNK and PKA pathway cross-talk during stress regulation of STMN. Taken together our study indicates that JNK- and PKA-mediated STMN Ser-38 and Ser-63 phosphorylation are required to preserve interphase microtubules in response to hyperosmotic stress.

  19. TRESK background K(+ channel is inhibited by PAR-1/MARK microtubule affinity-regulating kinases in Xenopus oocytes.

    Directory of Open Access Journals (Sweden)

    Gabriella Braun

    Full Text Available TRESK (TWIK-related spinal cord K(+ channel, KCNK18 is a major background K(+ channel of sensory neurons. Dominant-negative mutation of TRESK is linked to familial migraine. This important two-pore domain K(+ channel is uniquely activated by calcineurin. The calcium/calmodulin-dependent protein phosphatase directly binds to the channel and activates TRESK current several-fold in Xenopus oocytes and HEK293 cells. We have recently shown that the kinase, which is responsible for the basal inhibition of the K(+ current, is sensitive to the adaptor protein 14-3-3. Therefore we have examined the effect of the 14-3-3-inhibited PAR-1/MARK, microtubule-associated-protein/microtubule affinity-regulating kinase on TRESK in the Xenopus oocyte expression system. MARK1, MARK2 and MARK3 accelerated the return of TRESK current to the resting state after the calcium-dependent activation. Several other serine-threonine kinase types, generally involved in the modulation of other ion channels, failed to influence TRESK current recovery. MARK2 phosphorylated the primary determinant of regulation, the cluster of three adjacent serine residues (S274, 276 and 279 in the intracellular loop of mouse TRESK. In contrast, serine 264, the 14-3-3-binding site of TRESK, was not phosphorylated by the kinase. Thus MARK2 selectively inhibits TRESK activity via the S274/276/279 cluster, but does not affect the direct recruitment of 14-3-3 to the channel. TRESK is the first example of an ion channel phosphorylated by the dynamically membrane-localized MARK kinases, also known as general determinants of cellular polarity. These results raise the possibility that microtubule dynamics is coupled to the regulation of excitability in the neurons, which express TRESK background potassium channel.

  20. Conjugation in S. pombe: identification of a microtubule-organising centre, a requirement for microtubules and a role for Mad2.

    Science.gov (United States)

    Petersen, J; Heitz, M J; Hagan, I M

    1998-08-27

    During the G1 phase of the cell cycle, cells of the fission yeast Schizosaccharomyces pombe can be induced to mate by nitrogen starvation and the presence of mating pheromones. Polarised growth towards cells of the opposite mating type (P or M) leads to the formation of a projection tip and, upon contact, localised cell wall degradation results in conjugation and cell fusion [1]. Here, we have investigated the role of microtubules in this process. We describe a previously unidentified microtubule-organising centre (MTOC) that forms at projection tips upon cell-to-cell contact, before cells fuse. Treatment of mating cells with the microtubule-destabilising drug thiabendazole (TBZ) showed that microtubule integrity was required for mating at two distinct stages: during projection tip formation and cell fusion. Projection tip formation requires filamentous (F) actin function [2] and microtubules are required for the localisation of F actin to the projection tip. We also identify a role during mating for Mad2--a mitotic checkpoint protein that is required in all eukaryotes to maintain the mitotic state in response to microtubule depolymerisation [3]. S. pombe mad2 mutant cells were compromised in their ability to mate upon removal of TBZ, indicating that in fission yeast, in the absence of microtubules, Mad2 is also required to maintain mating competence.

  1. Dependency of microtubule-associated proteins (MAPs) for tubulin stability and assembly; use of estramustine phosphate in the study of microtubules.

    Science.gov (United States)

    Fridén, B; Wallin, M

    1991-07-10

    Microtubule-associated proteins (MAPs) were separated from tubulin with several different methods. The ability of the isolated MAPs to reinduce assembly of phosphocellulose purified tubulin differed markedly between the different methods. MAPs isolated by addition of 0.35 M NaCl to taxol-stabilized microtubules stimulated tubulin assembly most effectively, while addition of 0.6 M NaCl produced MAPs with a substantially lower ability to stimulate tubulin assembly. The second best preparation was achieved with phosphocellulose chromatographic separation of MAPs with 0.6 M NaCl elution. The addition of estramustine phosphate to microtubules reconstituted of MAPs prepared by 0.35 M NaCl or phosphocellulose chromatography, induced less disassembly than for microtubules assembled from unseparated proteins, and was almost without effect on microtubules reconstituted from MAPs prepared by taxol and 0.6 M NaCl. Estramustine phosphate binds to the tubulin binding part of the MAPs, and the results do therefore indicate that the MAPs are altered by the separation methods. Since the MAPs are regarded as highly stable molecules, one probable alteration could be aggregation of the MAPs, as also indicated by the results. The purified tubulin itself seemed not to be affected by the phosphocellulose purification, since the microtubule proteins were unchanged by the low buffer strenght used during the cromatography. However, the assembly competence after a prolonged incubation of the microtubule proteins at 4 degrees C was dependent on intact bindings between the tubulin and MAPs.

  2. Molecular quantum robotics: particle and wave solutions, illustrated by "leg-over-leg" walking along microtubules.

    Science.gov (United States)

    Levi, Paul

    2015-01-01

    Remarkable biological examples of molecular robots are the proteins kinesin-1 and dynein, which move and transport cargo down microtubule "highways," e.g., of the axon, to final nerve nodes or along dendrites. They convert the energy of ATP hydrolysis into mechanical forces and can thereby push them forwards or backwards step by step. Such mechano-chemical cycles that generate conformal changes are essential for transport on all different types of substrate lanes. The step length of an individual molecular robot is a matter of nanometers but the dynamics of each individual step cannot be predicted with certainty (as it is a random process). Hence, our proposal is to involve the methods of quantum field theory (QFT) to describe an overall reliable, multi-robot system that is composed of a huge set of unreliable, local elements. The methods of QFT deliver techniques that are also computationally demanding to synchronize the motion of these molecular robots on one substrate lane as well as across lanes. Three different challenging types of solutions are elaborated. The impact solution reflects the particle point of view; the two remaining solutions are wave based. The second solution outlines coherent robot motions on different lanes. The third solution describes running waves. Experimental investigations are needed to clarify under which biological conditions such different solutions occur. Moreover, such a nano-chemical system can be stimulated by external signals, and this opens a new, hybrid approach to analyze and control the combined system of robots and microtubules externally. Such a method offers the chance to detect mal-functions of the biological system.

  3. Structure and Dynamics of Single-isoform Recombinant Neuronal Human Tubulin.

    Science.gov (United States)

    Vemu, Annapurna; Atherton, Joseph; Spector, Jeffrey O; Szyk, Agnieszka; Moores, Carolyn A; Roll-Mecak, Antonina

    2016-06-17

    Microtubules are polymers that cycle stochastically between polymerization and depolymerization, i.e. they exhibit "dynamic instability." This behavior is crucial for cell division, motility, and differentiation. Although studies in the last decade have made fundamental breakthroughs in our understanding of how cellular effectors modulate microtubule dynamics, analysis of the relationship between tubulin sequence, structure, and dynamics has been held back by a lack of dynamics measurements with and structural characterization of homogeneous isotypically pure engineered tubulin. Here, we report for the first time the cryo-EM structure and in vitro dynamics parameters of recombinant isotypically pure human tubulin. α1A/βIII is a purely neuronal tubulin isoform. The 4.2-Å structure of post-translationally unmodified human α1A/βIII microtubules shows overall similarity to that of heterogeneous brain microtubules, but it is distinguished by subtle differences at polymerization interfaces, which are hot spots for sequence divergence between tubulin isoforms. In vitro dynamics assays show that, like mosaic brain microtubules, recombinant homogeneous microtubules undergo dynamic instability, but they polymerize slower and have fewer catastrophes. Interestingly, we find that epitaxial growth of α1A/βIII microtubules from heterogeneous brain seeds is inefficient but can be fully rescued by incorporating as little as 5% of brain tubulin into the homogeneous α1A/βIII lattice. Our study establishes a system to examine the structure and dynamics of mammalian microtubules with well defined tubulin species and is a first and necessary step toward uncovering how tubulin genetic and chemical diversity is exploited to modulate intrinsic microtubule dynamics.

  4. ErbB2-dependent chemotaxis requires microtubule capture and stabilization coordinated by distinct signaling pathways.

    Directory of Open Access Journals (Sweden)

    Khedidja Benseddik

    Full Text Available Activation of the ErbB2 receptor tyrosine kinase stimulates breast cancer cell migration. Cell migration is a complex process that requires the synchronized reorganization of numerous subcellular structures including cell-to-matrix adhesions, the actin cytoskeleton and microtubules. How the multiple signaling pathways triggered by ErbB2 coordinate, in time and space, the various processes involved in cell motility, is poorly defined. We investigated the mechanism whereby ErbB2 controls microtubules and chemotaxis. We report that activation of ErbB2 increased both cell velocity and directed migration. Impairment of the Cdc42 and RhoA GTPases, but not of Rac1, prevented the chemotactic response. RhoA is a key component of the Memo/ACF7 pathway whereby ErbB2 controls microtubule capture at the leading edge. Upon Memo or ACF7 depletion, microtubules failed to reach the leading edge and cells lost their ability to follow the chemotactic gradient. Constitutive ACF7 targeting to the membrane in Memo-depleted cells reestablished directed migration. ErbB2-mediated activation of phospholipase C gamma (PLCγ also contributed to cell guidance. We further showed that PLCγ signaling, via classical protein kinases C, and Memo signaling converged towards a single pathway controlling the microtubule capture complex. Finally, inhibiting the PI3K/Akt pathway did not affect microtubule capture, but disturbed microtubule stability, which also resulted in defective chemotaxis. PI3K/Akt-dependent stabilization of microtubules involved repression of GSK3 activity on the one hand and inhibition of the microtubule destabilizing protein, Stathmin, on the other hand. Thus, ErbB2 triggers distinct and complementary pathways that tightly coordinate microtubule capture and microtubule stability to control chemotaxis.

  5. Buckling of microtubules: An insight by molecular and continuum mechanics

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Jin; Meguid, S. A., E-mail: meguid@mie.utoronto.ca [Mechanics and Aerospace Design Laboratory, Department of Mechanical and Industrial Engineering, University of Toronto, Toronto, Ontario M5S 3G8 (Canada)

    2014-10-27

    The molecular structural mechanics method has been extended to investigate the buckling of microtubules (MTs) with various configurations. The results indicate that for relative short MTs the shear deformation effect, rather than the nonlocal effect, is mainly responsible for the limitation of their widely used Euler beam description and the observed length-dependence of their bending stiffness. In addition, the configuration effect of MTs is also studied and considered as an explanation for the large scattering of the critical buckling force and bending stiffness observed in existing experiments. This configuration effect is also found to mainly originate from the geometry of the MTs and is mainly determined by the protofilament number.

  6. Connections between microtubules and endoplasmic reticulum in mitotic spindle

    Directory of Open Access Journals (Sweden)

    J. A. Tarkowska

    2015-01-01

    Full Text Available Dividing endosperm cells of Haemanthus katherinae Bak. were treated with an 0.025 per cent aqueous solution of an oleander glycosides mixture which produces severe disturtaances in the mitotic spindle and high hypertrophy of the endoplasmic reticulum (ER in the whole cells. There appear between the kinetochore microtubules (MTs numerous elongated and narrow ER cisterns, particularly well visible when the number of kinetochore MTs is reduced. Both these structures (MTs and ER are frequently connected by cross-bridges. The presumable role of these connections is discused.

  7. Measurement of in vitro microtubule polymerization by turbidity and fluorescence.

    Science.gov (United States)

    Mirigian, Matthew; Mukherjee, Kamalika; Bane, Susan L; Sackett, Dan L

    2013-01-01

    Tubulin polymerization may be conveniently monitored by the increase in turbidity (optical density, or OD) or by the increase in fluorescence intensity of diamidino-phenylindole. The resulting data can be a quantitative measure of microtubule (MT) assembly, but some care is needed in interpretation, especially of OD data. Buffer formulations used for the assembly reaction significantly influence the polymerization, both by altering the critical concentration for polymerization and by altering the exact polymer produced-for example, by increasing the production of sheet polymers in addition to MT. Both the turbidity and the fluorescence methods are useful for demonstrating the effect of MT-stabilizing or -destabilizing additives.

  8. Biochemical characterization of tektins from sperm flagellar doublet microtubules

    OpenAIRE

    1987-01-01

    Tektins, protein components of stable protofilaments from sea urchin sperm flagellar outer doublet microtubules (Linck, R. W., and G. L. Langevin, 1982, J. Cell Sci., 58:1-22), are separable by preparative SDS PAGE into 47-, 51-, and 55-kD equimolar components. High resolution two-dimensional tryptic peptide mapping reveals 63-67% coincidence among peptides of the 51-kD tektin chain and its 47- and 55-kD counterparts, greater than 70% coincidence between the 47- and 55-kD tektins, but little ...

  9. Fluid mechanics in dentinal microtubules provides mechanistic insights into the difference between hot and cold dental pain.

    Science.gov (United States)

    Lin, Min; Luo, Zheng Yuan; Bai, Bo Feng; Xu, Feng; Lu, Tian Jian

    2011-03-23

    Dental thermal pain is a significant health problem in daily life and dentistry. There is a long-standing question regarding the phenomenon that cold stimulation evokes sharper and more shooting pain sensations than hot stimulation. This phenomenon, however, outlives the well-known hydrodynamic theory used to explain dental thermal pain mechanism. Here, we present a mathematical model based on the hypothesis that hot or cold stimulation-induced different directions of dentinal fluid flow and the corresponding odontoblast movements in dentinal microtubules contribute to different dental pain responses. We coupled a computational fluid dynamics model, describing the fluid mechanics in dentinal microtubules, with a modified Hodgkin-Huxley model, describing the discharge behavior of intradental neuron. The simulated results agreed well with existing experimental measurements. We thence demonstrated theoretically that intradental mechano-sensitive nociceptors are not "equally sensitive" to inward (into the pulp) and outward (away from the pulp) fluid flows, providing mechanistic insights into the difference between hot and cold dental pain. The model developed here could enable better diagnosis in endodontics which requires an understanding of pulpal histology, neurology and physiology, as well as their dynamic response to the thermal stimulation used in dental practices.

  10. Avaliação de fluxos de calor e evapotranspiração pelo modelo SEBAL com uso de dados do sensor ASTER Evaluation of heat fluxes and evapotranspiration using SEBAL model with data from ASTER sensor

    Directory of Open Access Journals (Sweden)

    Thiago Veloso dos Santos

    2010-05-01

    Full Text Available O objetivo deste trabalho foi avaliar a eficiência da