WorldWideScience

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. Shape-motion relationships of centering microtubule asters.

    Science.gov (United States)

    Tanimoto, Hirokazu; Kimura, Akatsuki; Minc, Nicolas

    2016-03-28

    Although mechanisms that contribute to microtubule (MT) aster positioning have been extensively studied, still little is known on how asters move inside cells to faithfully target a cellular location. Here, we study sperm aster centration in sea urchin eggs, as a stereotypical large-scale aster movement with extreme constraints on centering speed and precision. By tracking three-dimensional aster centration dynamics in eggs with manipulated shapes, we show that aster geometry resulting from MT growth and interaction with cell boundaries dictates aster instantaneous directionality, yielding cell shape-dependent centering trajectories. Aster laser surgery and modeling suggest that dynein-dependent MT cytoplasmic pulling forces that scale to MT length function to convert aster geometry into directionality. In contrast, aster speed remains largely independent of aster size, shape, or absolute dynein activity, which suggests it may be predominantly determined by aster growth rate rather than MT force amplitude. These studies begin to define the geometrical principles that control aster movements. PMID:27022090

  4. Collective behavior of minus-ended motors in mitotic microtubule asters gliding toward DNA

    International Nuclear Information System (INIS)

    Microtubules (MTs) nucleated by centrosomes form star-shaped structures referred to as asters. Aster motility and dynamics is vital for genome stability, cell division, polarization and differentiation. Asters move either toward the cell center or away from it. Here, we focus on the centering mechanism in a membrane independent system of Xenopus cytoplasmic egg extracts. Using live microscopy and single particle tracking, we find that asters move toward chromatinized DNA structures. The velocity and directionality profiles suggest a random-walk with drift directed toward DNA. We have developed a theoretical model that can explain this movement as a result of a gradient of MT length dynamics and MT gliding on immobilized dynein motors. In simulations, the antagonistic action of the motor species on the radial array of MTs leads to a tug-of-war purely due to geometric considerations and aster motility resembles a directed random-walk. Additionally, our model predicts that aster velocities do not change greatly with varying initial distance from DNA. The movement of asymmetric asters becomes increasingly super-diffusive with increasing motor density, but for symmetric asters it becomes less super-diffusive. The transition of symmetric asters from superdiffusive to diffusive mobility is the result of number fluctuations in bound motors in the tug-of-war. Overall, our model is in good agreement with experimental data in Xenopus cytoplasmic extracts and predicts novel features of the collective effects of motor-MT interactions. (paper)

  5. 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.

  6. 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.

  7. Regulation of microtubule dynamic instability

    NARCIS (Netherlands)

    B. van der Vaart (Babet); A.S. Akhmanova (Anna); A. Straube (Anne)

    2009-01-01

    textabstractProper regulation of MT (microtubule) dynamics is essential for various vital processes, including the segregation of chromosomes, directional cell migration and differentiation. MT assembly and disassembly is modulated by a complex network of intracellular factors that co-operate or ant

  8. Expression of Nucleolin Affects Microtubule Dynamics.

    Directory of Open Access Journals (Sweden)

    Xavier Gaume

    Full Text Available Nucleolin is present in diverse cellular compartments and is involved in a variety of cellular processes from nucleolar structure and function to intracellular trafficking, cell adhesion and migration. Recently, nucleolin has been localized at the mature centriole where it is involved in microtubule nucleation and anchoring. Although this new function of nucleolin linked to microtubule regulation has been identified, the global effects of nucleolin on microtubule dynamics have not been addressed yet. In the present study, we analyzed the roles of nucleolin protein levels on global microtubule dynamics by tracking the EB3 microtubule plus end binding protein in live cells. We have found that during microtubule growth phases, nucleolin affects both the speed and life time of polymerization and by analyzing catastrophe events, we showed that nucleolin reduces catastrophe frequency. This new property of nucleolin was then confirmed in a cold induced microtubule depolymerization experiment in which we have found that cold resistant microtubules were totally destabilized in nucleolin depleted cells. Altogether, our data demonstrate a new function of nucleolin on microtubule stabilization, thus bringing novel insights into understanding the multifunctional properties of nucleolin in healthy and cancer cells.

  9. Expression of Nucleolin Affects Microtubule Dynamics.

    Science.gov (United States)

    Gaume, Xavier; Place, Christophe; Delage, Helene; Mongelard, Fabien; Monier, Karine; Bouvet, Philippe

    2016-01-01

    Nucleolin is present in diverse cellular compartments and is involved in a variety of cellular processes from nucleolar structure and function to intracellular trafficking, cell adhesion and migration. Recently, nucleolin has been localized at the mature centriole where it is involved in microtubule nucleation and anchoring. Although this new function of nucleolin linked to microtubule regulation has been identified, the global effects of nucleolin on microtubule dynamics have not been addressed yet. In the present study, we analyzed the roles of nucleolin protein levels on global microtubule dynamics by tracking the EB3 microtubule plus end binding protein in live cells. We have found that during microtubule growth phases, nucleolin affects both the speed and life time of polymerization and by analyzing catastrophe events, we showed that nucleolin reduces catastrophe frequency. This new property of nucleolin was then confirmed in a cold induced microtubule depolymerization experiment in which we have found that cold resistant microtubules were totally destabilized in nucleolin depleted cells. Altogether, our data demonstrate a new function of nucleolin on microtubule stabilization, thus bringing novel insights into understanding the multifunctional properties of nucleolin in healthy and cancer cells. PMID:27309529

  10. Association of Microtubule Dynamics with Chronic Epilepsy.

    Science.gov (United States)

    Xu, Xin; Hu, Yida; Xiong, Yan; Li, Zhonggui; Wang, Wei; Du, Chao; Yang, Yong; Zhang, Yanke; Xiao, Fei; Wang, Xuefeng

    2016-09-01

    Approximately 30 % of epilepsy cases are refractory to current pharmacological treatments through unknown mechanisms. Much work has been done on the role of synaptic components in the pathogenesis of epilepsy, but relatively little attention has been given to the potential role of the microtubules. We investigated the level of microtubule dynamic in 30 human epileptic tissues and two different chronic epilepsy rat models. The administration of microtubule-modulating agent attenuated the progression of chronic epilepsy. By contrast, microtubule-depolymerizing agent aggravated the progression of chronic epilepsy. The electrophysiological index by whole-cell clamp was used to investigate the neuronal excitation and inhibitory synaptic transmission in brain slices after administration of microtubule-modulating agent and microtubule-depolymerizing agent. Interestingly, we found that microtubule-modulating agent significantly increased the frequency of action potential firing in interneurons, and significantly promoted the amplitudes and frequencies of miniature inhibitory postsynaptic currents. Microtubule-depolymerizing agent had an opposite effect. These findings suggest that modulating hyperdynamic microtubules may take an anti-epileptic effect via postsynaptic mechanisms in interneurons. It could represent a potential pharmacologic target in epilepsy treatment. PMID:26377107

  11. Microtubule dynamics: Caps, catastrophes, and coupled hydrolysis

    DEFF Research Database (Denmark)

    Flyvbjerg, H.; Holy, T.E.; Leibler, S.

    1996-01-01

    An effective theory is formulated for the dynamics of the guanosine triphosphate (GTP) cap believed to stabilize growing microtubules. The theory provides a ''coarse-grained'' description of the cap's dynamics. ''Microscopic'' details, such as the microtubule lattice structure and the fate of its...... data. A constant nonzero catastrophe rare, identical for both microtubule ends, is predicted at large growth rates. The delay time for dilution-induced catastrophes is stochastic with a simple distribution that fits the experimental one and, like the experimental one, does not depend on the rate of....... A recent experimental result for the size of the minimal cap that can stabilize a microtubule is shown to agree with the result predicted by the cap model, after its parameters have been extracted from previous experimental results. Thus the effective theory and cap model presented here provide a...

  12. 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.

  13. Integrin-linked kinase regulates interphase and mitotic microtubule dynamics.

    Directory of Open Access Journals (Sweden)

    Simin Lim

    Full Text Available Integrin-linked kinase (ILK localizes to both focal adhesions and centrosomes in distinct multiprotein complexes. Its dual function as a kinase and scaffolding protein has been well characterized at focal adhesions, where it regulates integrin-mediated cell adhesion, spreading, migration and signaling. At the centrosomes, ILK regulates mitotic spindle organization and centrosome clustering. Our previous study showed various spindle defects after ILK knockdown or inhibition that suggested alteration in microtubule dynamics. Since ILK expression is frequently elevated in many cancer types, we investigated the effects of ILK overexpression on microtubule dynamics. We show here that overexpressing ILK in HeLa cells was associated with a shorter duration of mitosis and decreased sensitivity to paclitaxel, a chemotherapeutic agent that suppresses microtubule dynamics. Measurement of interphase microtubule dynamics revealed that ILK overexpression favored microtubule depolymerization, suggesting that microtubule destabilization could be the mechanism behind the decreased sensitivity to paclitaxel, which is known to stabilize microtubules. Conversely, the use of a small molecule inhibitor selective against ILK, QLT-0267, resulted in suppressed microtubule dynamics, demonstrating a new mechanism of action for this compound. We further show that treatment of HeLa cells with QLT-0267 resulted in higher inter-centromere tension in aligned chromosomes during mitosis, slower microtubule regrowth after cold depolymerization and the presence of a more stable population of spindle microtubules. These results demonstrate that ILK regulates microtubule dynamics in both interphase and mitotic cells.

  14. Microtubule-binding agents: a dynamic field of cancer therapeutics

    OpenAIRE

    Dumontet, Charles; Jordan, Mary Ann

    2010-01-01

    International audience Microtubules are dynamic filamentous cytoskeletal proteins composed of tubulin and are an important therapeutic target in tumour cells. Agents that bind to microtubules have been part of the pharmacopoeia of anticancer therapy for decades and until the advent of targeted therapy, microtubules were the only alternative to DNA as a therapeutic target in cancer. The screening of a range of botanical species and marine organisms has yielded promising new antitubulin agen...

  15. Leading at the Front: How EB Proteins Regulate Microtubule Dynamics

    Science.gov (United States)

    Hawkins, Taviare

    2012-02-01

    Microtubules are the most rigid of the cytoskeletal filaments, they provide the cell's scaffolding, form the byways on which motor proteins transport intracellular cargo and reorganize to form the mitotic spindle when the cell needs to divide. These biopolymers are composed of alpha and beta tubulin monomers that create hollow cylindrical nanotubes with an outer diameter of 25 nm and an inner diameter of 17 nm. At steady state concentrations, microtubules undergo a process known as dynamic instability. During dynamic instability the length of individual microtubules is changing as the filament alternates between periods of growth to shrinkage (catastrophe) and shrinkage to growth (rescue). This process can be enhanced or diminished with the addition of microtubule associated proteins (MAPs). MAPs are microtubule binding proteins that stabilize, destabilize, or nucleate microtubules. We will discuss the effects of the stabilizing end-binding proteins (EB1, EB2 and EB3), on microtubule dynamics observed in vitro. The EBs are a unique family of MAPs known to tip track and enhance microtubule growth by stabilizing the ends. This is a different mechanism than those employed by structural MAPs such as tau or MAP4.

  16. 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.

  17. Nonlinear dynamics of C-terminal tails in cellular microtubules

    Science.gov (United States)

    Sekulic, Dalibor L.; Sataric, Bogdan M.; Zdravkovic, Slobodan; Bugay, Aleksandr N.; Sataric, Miljko V.

    2016-07-01

    The mechanical and electrical properties, and information processing capabilities of microtubules are the permanent subject of interest for carrying out experiments in vitro and in silico, as well as for theoretical attempts to elucidate the underlying processes. In this paper, we developed a new model of the mechano-electrical waves elicited in the rows of very flexible C-terminal tails which decorate the outer surface of each microtubule. The fact that C-terminal tails play very diverse roles in many cellular functions, such as recruitment of motor proteins and microtubule-associated proteins, motivated us to consider their collective dynamics as the source of localized waves aimed for communication between microtubule and associated proteins. Our approach is based on the ferroelectric liquid crystal model and it leads to the effective asymmetric double-well potential which brings about the conditions for the appearance of kink-waves conducted by intrinsic electric fields embedded in microtubules. These kinks can serve as the signals for control and regulation of intracellular traffic along microtubules performed by processive motions of motor proteins, primarly from kinesin and dynein families. On the other hand, they can be precursors for initiation of dynamical instability of microtubules by recruiting the proper proteins responsible for the depolymerization process.

  18. Nonlinear dynamics of C-terminal tails in cellular microtubules.

    Science.gov (United States)

    Sekulic, Dalibor L; Sataric, Bogdan M; Zdravkovic, Slobodan; Bugay, Aleksandr N; Sataric, Miljko V

    2016-07-01

    The mechanical and electrical properties, and information processing capabilities of microtubules are the permanent subject of interest for carrying out experiments in vitro and in silico, as well as for theoretical attempts to elucidate the underlying processes. In this paper, we developed a new model of the mechano-electrical waves elicited in the rows of very flexible C-terminal tails which decorate the outer surface of each microtubule. The fact that C-terminal tails play very diverse roles in many cellular functions, such as recruitment of motor proteins and microtubule-associated proteins, motivated us to consider their collective dynamics as the source of localized waves aimed for communication between microtubule and associated proteins. Our approach is based on the ferroelectric liquid crystal model and it leads to the effective asymmetric double-well potential which brings about the conditions for the appearance of kink-waves conducted by intrinsic electric fields embedded in microtubules. These kinks can serve as the signals for control and regulation of intracellular traffic along microtubules performed by processive motions of motor proteins, primarly from kinesin and dynein families. On the other hand, they can be precursors for initiation of dynamical instability of microtubules by recruiting the proper proteins responsible for the depolymerization process. PMID:27475079

  19. 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.

  20. Ibuprofen regulation of microtubule dynamics in cystic fibrosis epithelial cells.

    Science.gov (United States)

    Rymut, Sharon M; Kampman, Claire M; Corey, Deborah A; Endres, Tori; Cotton, Calvin U; Kelley, Thomas J

    2016-08-01

    High-dose ibuprofen, an effective anti-inflammatory therapy for the treatment of cystic fibrosis (CF), has been shown to preserve lung function in a pediatric population. Despite its efficacy, few patients receive ibuprofen treatment due to potential renal and gastrointestinal toxicity. The mechanism of ibuprofen efficacy is also unclear. We have previously demonstrated that CF microtubules are slower to reform after depolymerization compared with respective wild-type controls. Slower microtubule dynamics in CF cells are responsible for impaired intracellular transport and are related to inflammatory signaling. Here, it is identified that high-dose ibuprofen treatment in both CF cell models and primary CF nasal epithelial cells restores microtubule reformation rates to wild-type levels, as well as induce extension of microtubules to the cell periphery. Ibuprofen treatment also restores microtubule-dependent intracellular transport monitored by measuring intracellular cholesterol transport. These effects are specific to ibuprofen as other cyclooxygenase inhibitors have no effect on these measures. Effects of ibuprofen are mimicked by stimulation of AMPK and blocked by the AMPK inhibitor compound C. We conclude that high-dose ibuprofen treatment enhances microtubule formation in CF cells likely through an AMPK-related pathway. These findings define a potential mechanism to explain the efficacy of ibuprofen therapy in CF. PMID:27317686

  1. 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.)

  2. ASTER Mexicali

    Science.gov (United States)

    2000-01-01

    '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 applications include monitoring glacial advances and retreats, potentially active volcanoes, thermal pollution, and coral reef degradation; identifying crop stress; determining cloud morphology and physical properties; evaluating wetlands; mapping surface temperature of soils and geology; and measuring surface heat balance.

  3. 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.

  4. ASTER Andes

    Science.gov (United States)

    2000-01-01

    In this image of the Andes along the Chile-Bolivia border, the visible and infrared data have been computer enhanced to exaggerate the color differences of the different materials. The scene is dominated by the Pampa Luxsar lava complex, occupying the upper right two-thirds of the scene. Lava flows are distributed around remnants of large dissected cones, the largest of which is Cerro Luxsar. On the middle left edge of the image are the Olca and Parumastrato volcanoes, which appear in blue due to a lack of vegetation (colored red in this composite). This image covers an area 60 kilometers (37 miles) wide and 60 kilometers (37 miles) long in three bands of the reflected visible and infrared wavelength region. It was acquired on April 7, 2000.The image is located at 21 degrees south latitude, 68.3 degrees west longitude. 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 applications include monitoring glacial advances and retreats

  5. 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

  6. 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.

  7. 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.

  8. 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.

  9. 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 ...

  10. 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. PMID:27166749

  11. 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.

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

    Science.gov (United States)

    Tian, Xinyong; Tian, Yufeng; Moldobaeva, Nurgul; Sarich, Nicolene; Birukova, Anna A

    2014-01-01

    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. PMID:25198505

  13. 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.

  14. Dynamic microtubules regulate dendritic spine morphology and synaptic plasticity

    NARCIS (Netherlands)

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

    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

  15. 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.

  16. 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.

  17. 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.

  18. Mechanochemical modeling of dynamic microtubule growth involving sheet-to-tube transition

    CERN Document Server

    Ji, Xiang-Ying

    2011-01-01

    Microtubule dynamics is largely influenced by nucleotide hydrolysis and the resultant tubulin configuration changes. The GTP cap model has been proposed to interpret the stabilizing mechanism of microtubule growth from the view of hydrolysis effects. Besides, the microtubule growth involves the closure of a curved sheet at its growing end. The curvature conversion also helps to stabilize the successive growth, and the curved sheet is referred to as the conformational cap. However, there still lacks theoretical investigation on the mechanical-chemical coupling growth process of microtubules. In this paper, we study the growth mechanisms of microtubules by using a coarse-grained molecular method. Firstly, the closure process involving a sheet-to-tube transition is simulated. The results verify the stabilizing effect of the sheet structure, and the minimum conformational cap length that can stabilize the growth is demonstrated to be two dimers. Then, we show that the conformational cap can function independently...

  19. Dictyoceratidan poisons: Defined mark on microtubule-tubulin dynamics.

    Science.gov (United States)

    Gnanambal K, Mary Elizabeth; Lakshmipathy, Shailaja Vommi

    2016-03-01

    Tubulin/microtubule assembly and disassembly is characterized as one of the chief processes during cell growth and division. Hence drugs those perturb these process are considered to be effective in killing fast multiplying cancer cells. There is a collection of natural compounds which disturb microtubule/tubulin dis/assemblage and there have been a lot of efforts concerted in the marine realm too, to surveying such killer molecules. Close to half the natural compounds shooting out from marine invertebrates are generally with no traceable definite mechanisms of action though may be tough anti-cancerous hits at nanogram levels, hence fatefully those discoveries conclude therein without a capacity of translation from laboratory to pharmacy. Astoundingly at least 50% of natural compounds which have definite mechanisms of action causing disorders in tubulin/microtubule kinetics have an isolation history from sponges belonging to the Phylum: Porifera. Poriferans have always been a wonder worker to treat cancers with a choice of, yet precise targets on cancerous tissues. There is a specific order: Dictyoceratida within this Phylum which has contributed to yielding at least 50% of effective compounds possessing this unique mechanism of action mentioned above. However, not much notice is driven to Dictyoceratidans alongside the order: Demospongiae thus dictating the need to know its select microtubule/tubulin irritants since the unearthing of avarol in the year 1974 till date. Hence this review selectively pinpoints all the compounds, noteworthy derivatives and analogs stemming from order: Dictyoceratida focusing on the past, present and future. PMID:26874035

  20. 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.

  1. 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.

  2. 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

    2015-01-01

    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.

  3. 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.

  4. 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.

  5. Titanium dioxide nanoparticles alter cellular morphology via disturbing the microtubule dynamics

    Science.gov (United States)

    Mao, Zhilei; Xu, Bo; Ji, Xiaoli; Zhou, Kun; Zhang, Xuemei; Chen, Minjian; Han, Xiumei; Tang, Qiusha; Wang, Xinru; Xia, Yankai

    2015-04-01

    Titanium dioxide (TiO2) nanoparticles (NPs) have been widely used in our daily lives, for example, in the areas of sunscreens, cosmetics, toothpastes, food products, and nanomedical reagents. Recently, increasing concern has been raised about their neurotoxicity, but the mechanisms underlying such toxic effects are still unknown. In this work, we employed a human neuroblastoma cell line (SH-SY5Y) to study the effects of TiO2 NPs on neurological systems. Our results showed that TiO2 NPs did not affect cell viability but induced noticeable morphological changes until 100 μg ml-1. Immunofluorescence detection showed disorder, disruption, retraction, and decreased intensity of the microtubules after TiO2 NPs treatment. Both α and β tubule expressions did not change in the TiO2 NP-treated group, but the percentage of soluble tubules was increased. A microtubule dynamic study in living cells indicated that TiO2 NPs caused a lower growth rate and a higher shortening rate of microtubules as well as shortened lifetimes of de novo microtubules. TiO2 NPs did not cause changes in the expression and phosphorylation state of tau proteins, but a tau-TiO2 NP interaction was observed. TiO2 NPs could interact with tubule heterodimers, microtubules and tau proteins, which led to the instability of microtubules, thus contributing to the neurotoxicity of TiO2 NPs.Titanium dioxide (TiO2) nanoparticles (NPs) have been widely used in our daily lives, for example, in the areas of sunscreens, cosmetics, toothpastes, food products, and nanomedical reagents. Recently, increasing concern has been raised about their neurotoxicity, but the mechanisms underlying such toxic effects are still unknown. In this work, we employed a human neuroblastoma cell line (SH-SY5Y) to study the effects of TiO2 NPs on neurological systems. Our results showed that TiO2 NPs did not affect cell viability but induced noticeable morphological changes until 100 μg ml-1. Immunofluorescence detection showed disorder

  6. 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.

  7. 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.

  8. 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.

  9. Human chromokinesins promote chromosome congression and spindle microtubule dynamics during mitosis.

    Science.gov (United States)

    Wandke, Cornelia; Barisic, Marin; Sigl, Reinhard; Rauch, Veronika; Wolf, Frank; Amaro, Ana C; Tan, Chia H; Pereira, Antonio J; Kutay, Ulrike; Maiato, Helder; Meraldi, Patrick; Geley, Stephan

    2012-09-01

    Chromokinesins are microtubule plus end-directed motor proteins that bind to chromosome arms. In Xenopus egg cell-free extracts, Xkid and Xklp1 are essential for bipolar spindle formation but the functions of the human homologues, hKID (KIF22) and KIF4A, are poorly understood. By using RNAi-mediated protein knockdown in human cells, we find that only co-depletion delayed progression through mitosis in a Mad2-dependent manner. Depletion of hKID caused abnormal chromosome arm orientation, delayed chromosome congression, and sensitized cells to nocodazole. Knockdown of KIF4A increased the number and length of microtubules, altered kinetochore oscillations, and decreased kinetochore microtubule flux. These changes were associated with failures in establishing a tight metaphase plate and an increase in anaphase lagging chromosomes. Co-depletion of both chromokinesins aggravated chromosome attachment failures, which led to mitotic arrest. Thus, hKID and KIF4A contribute independently to the rapid and correct attachment of chromosomes by controlling the positioning of chromosome arms and the dynamics of microtubules, respectively. PMID:22945934

  10. 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.

  11. 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...

  12. ASTER Flyby of San Francisco

    Science.gov (United States)

    2002-01-01

    The Advanced Spaceborne Thermal Emission and Reflection radiometer, ASTER, is an international project: the instrument was supplied by Japan's Ministry of International Trade and Industry. A joint US/Japan science team developed algorithms for science data products, and is validating instrument performance. With its 14 spectral bands, extremely high spatial resolution, and 15 meter along-track stereo capability, ASTER is the zoom lens of the Terra satellite. The primary mission goals are to characterize the Earth's surface; and to monitor dynamic events and processes that influence habitability at human scales. ASTER's monitoring and mapping capabilities are illustrated by this series of images of the San Francisco area. The visible and near infrared image reveals suspended sediment in the bays, vegetation health, and details of the urban environment. Flying over San Francisco (3.2MB) (high-res (18.3MB)), we see the downtown, and shadows of the large buildings. Past the Golden Gate Bridge and Alcatraz Island, we cross San Pablo Bay and enter Suisun Bay. Turning south, we fly over the Berkeley and Oakland Hills. Large salt evaporation ponds come into view at the south end of San Francisco Bay. We turn northward, and approach San Francisco Airport. Rather than landing and ending our flight, we see this is as only the beginning of a 6 year mission to better understand the habitability of the world on which we live. For more information: ASTER images through Visible Earth ASTER Web Site Image courtesy of MITI, ERSDAC, JAROS, and the U.S./Japan ASTER Science Team

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

    Science.gov (United States)

    Muratov, Alexander; Baulin, Vladimir A

    2015-12-01

    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 microtubule arrays direct the growth and orientation of cellulose microfibrils, forming part of the cell external skeleton and determine the shape of the cell. Reorientation of microtubules is also observed in reaction to light in phototropism and mechanical bending, thus suggesting universality of the proposed mechanism. PMID:26422460

  14. 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.

  15. Regulation of Microtubule Dynamics in Axon Regeneration: Insights from C. elegans [version 1; referees: 3 approved

    Directory of Open Access Journals (Sweden)

    Ngang Heok Tang

    2016-04-01

    Full Text Available The capacity of an axon to regenerate is regulated by its external environment and by cell-intrinsic factors. Studies in a variety of organisms suggest that alterations in axonal microtubule (MT dynamics have potent effects on axon regeneration. We review recent findings on the regulation of MT dynamics during axon regeneration, focusing on the nematode Caenorhabditis elegans. In C. elegans the dual leucine zipper kinase (DLK promotes axon regeneration, whereas the exchange factor for Arf6 (EFA-6 inhibits axon regeneration. Both DLK and EFA-6 respond to injury and control axon regeneration in part via MT dynamics. How the DLK and EFA-6 pathways are related is a topic of active investigation, as is the mechanism by which EFA-6 responds to axonal injury. We evaluate potential candidates, such as the MT affinity-regulating kinase PAR-1/MARK, in regulation of EFA-6 and axonal MT dynamics in regeneration.

  16. 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.

  17. 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.

  18. 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.

  19. The complex dynamic network of microtubule and microfilament cytasters of the leech zygote.

    Science.gov (United States)

    Cantillana, V; Urrutia, M; Ubilla, A; Fernández, J

    2000-12-01

    The organization of the cytoskeleton in the early first interphase zygote and its involvement in organelle redistribution were studied in the glossiphoniid leech Theromyzon trizonare by confocal and electron microscopy, immunofluorescence, and time-lapse video imaging after microinjection of labeled tubulin and/or actin and loading with a mitotracker. The cytoskeleton consists of an inner or endoplasmic and an outer or ectoplasmic domain. The inner domain consists of a monaster whose fibers retract from the zygote periphery by the end of the early first interphase. The outer domain is built upon a network of microtubules and microfilaments cytasters. Short pulses of microinjected labeled actin or tubulin and Taxol treatment demonstrate that cytasters are centers of microtubule and microfilament nucleation. Immunostaining with anti-centrophilin, anti-BX-63, and anti-AH-6 indicates that the network of cytasters includes centrosomal antigens. Cytasters move in an orderly fashion at speeds of 0.5-2 micrometer/min, in an energy-dependent process retarded and finally blocked by the ATP analogue AMP-PNP and high concentrations of Taxol. Colliding cytasters fuse and form larger cytoskeletal nucleation centers. The leech zygote is a highly compartmentalized cell whose cytasters function as articulated components of a very dynamic cytoskeletal system engaged in bulk transportation of organelles during ooplasmic segregation. PMID:11087633

  20. 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.

  1. 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

  2. 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.

  3. Using Photobleaching to Measure Spindle Microtubule Dynamics in Primary Cultures of Dividing Drosophila Meiotic Spermatocytes.

    Science.gov (United States)

    Savoian, Matthew S

    2015-07-01

    In dividing animal cells, a microtubule (MT)-based bipolar spindle governs chromosome movement. Current models propose that the spindle facilitates and/or generates translocating forces by regionally depolymerizing the kinetochore fibers (k-fibers) that bind each chromosome. It is unclear how conserved these sites and the resultant chromosome-moving mechanisms are between different dividing cell types because of the technical challenges of quantitatively studying MTs in many specimens. In particular, our knowledge of MT kinetics during the sperm-producing male meiotic divisions remains in its infancy. In this study, I use an easy-to-implement photobleaching-based assay for measuring spindle MT dynamics in primary cultures of meiotic spermatocytes isolated from the fruit fly Drosophila melanogaster. By use of standard scanning confocal microscopy features, fiducial marks were photobleached on fluorescent protein (FP)-tagged MTs. These were followed by time-lapse imaging during different division stages, and their displacement rates were calculated using public domain software. I find that k-fibers continually shorten at their poles during metaphase and anaphase A through the process of MT flux. Anaphase chromosome movement is complemented by Pac-Man, the shortening of the k-fiber at its chromosomal interface. Thus, Drosophila spermatocytes share the sites of spindle dynamism and mechanisms of chromosome movement with mitotic cells. The data reveal the applicability of the photobleaching assay for measuring MT dynamics in primary cultures. This approach can be readily applied to other systems. PMID:25802491

  4. 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.

  5. Control of microtubule organization and dynamics : two ends in the limelight

    NARCIS (Netherlands)

    Akhmanova, Anna; Steinmetz, Michel O

    2015-01-01

    Microtubules have fundamental roles in many essential biological processes, including cell division and intracellular transport. They assemble and disassemble from their two ends, denoted the plus end and the minus end. Significant advances have been made in our understanding of microtubule plus-end

  6. Evidence for two distinct binding sites for tau on microtubules

    OpenAIRE

    Makrides, Victoria; Massie, Michelle R.; Feinstein, Stuart C.; Lew, John

    2004-01-01

    The microtubule-associated protein tau regulates diverse and essential microtubule functions, from the nucleation and promotion of microtubule polymerization to the regulation of microtubule polarity and dynamics, as well as the spacing and bundling of axonal microtubules. Thermodynamic studies show that tau interacts with microtubules in the low- to mid-nanomolar range, implying moderate binding affinity. At the same time, it is well established that microtubule-bound tau does not undergo ex...

  7. ASTER Expedited Data Services

    Science.gov (United States)

    Duda, K. A.

    2010-12-01

    The Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) on the Terra spacecraft offers near real-time data delivery through an expedited system used to support emergency responders and science field campaigns. Data acquired and processed in this manner can be available for download within six hours of collection, as compared with a lag time of many days before standard products become available at two distribution sites. The orbit revisit frequency is 16 days for nadir views, but this can be reduced substantially through off-nadir pointing, at high latitudes, and by night observations. Scheduling confirmation takes place two to four days prior to imaging. ASTER is a joint mission involving the U.S. and Japan. ASTER expedited data are generated at the Land Processes Distributed Active Archive Center (LP DAAC), where standard products are also archived after receipt from affiliates at the Earth Remote Sensing Data Analysis Center (ERSDAC) in Tokyo. The ASTER instrument operates through scheduled observations, and is very successfully completing a global mapping mission. With three visible and near-infrared (VNIR) bands at 15 m ground resolution, six shortwave infrared (SWIR) bands at 30 m resolution, and five thermal infrared (TIR) bands at 90 m resolution, the 1.7-million scene archive of ASTER data acquired during the last decade provides utility in a wide range of investigations. A back-looking band is also present that enables the generation of digital elevation models (DEM). ASTER expedited data have been requested by national and international emergency response organizations to provide current views of many types of disaster situations, including volcanoes, hurricanes, wildfires, floods, tsunamis, landslides and other events. ASTER data have been provided in response to activations of the International Charter, Space and Major Disasters. Examples of data acquired for a variety of disaster situations will be provided. Recent events supported

  8. 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.

  9. Microtubule dynamics of the centrosome-like polar organizers from the basal land plant Marchantia polymorpha.

    Science.gov (United States)

    Buschmann, Henrik; Holtmannspötter, Michael; Borchers, Agnes; O'Donoghue, Martin-Timothy; Zachgo, Sabine

    2016-02-01

    The liverwort Marchantia employs both modern and ancestral devices during cell division: it forms preprophase bands and in addition it shows centrosome-like polar organizers. We investigated whether polar organizers and preprophase bands cooperate to set up the division plane. To this end, two novel green fluorescent protein-based microtubule markers for dividing cells of Marchantia were developed. Cells of the apical notch formed polar organizers first and subsequently assembled preprophase bands. Polar organizers were formed de novo from multiple mobile microtubule foci localizing to the nuclear envelope. The foci then became concentrated by bipolar aggregation. We determined the comet production rate of polar organizers and show that microtubule plus ends of astral microtubules polymerize faster than those found on cortical microtubules. Importantly, it was observed that conditions increasing polar organizer numbers interfere with preprophase band formation. The data show that polar organizers have much in common with centrosomes, but that they also have specialized features. The results suggest that polar organizers contribute to preprophase band formation and in this way are involved in controlling the division plane. Our analyses of the basal land plant Marchantia shed new light on the evolution of plant cell division. PMID:26467050

  10. N-terminus-modified Hec1 suppresses tumour growth by interfering with kinetochore-microtubule dynamics.

    Science.gov (United States)

    Orticello, M; Fiore, M; Totta, P; Desideri, M; Barisic, M; Passeri, D; Lenzi, J; Rosa, A; Orlandi, A; Maiato, H; Del Bufalo, D; Degrassi, F

    2015-06-01

    Mitotic proteins are attractive targets to develop molecular cancer therapeutics due to the intimate interdependence between cell proliferation and mitosis. In this work, we have explored the therapeutic potential of the kinetochore (KT) protein Hec1 (Highly Expressed in Cancer protein 1) as a molecular target to produce massive chromosome missegregation and cell death in cancer cells. Hec1 is a constituent of the Ndc80 complex, which mediates KT-microtubule (MT) attachments at mitosis and is upregulated in various cancer types. We expressed Hec1 fused with enhanced green fluorescent protein (EGFP) at its N-terminus MT-interaction domain in HeLa cells and showed that expression of this modified Hec1, which localized at KTs, blocked cell proliferation and promoted apoptosis in tumour cells. EGFP-Hec1 was extremely potent in tumour cell killing and more efficient than siRNA-induced Hec1 depletion. In striking contrast, normal cells showed no apparent cell proliferation defects or cell death following EGFP-Hec1 expression. Live-cell imaging demonstrated that cancer cell death was associated with massive chromosome missegregation within multipolar spindles after a prolonged mitotic arrest. Moreover, EGFP-Hec1 expression was found to increase KT-MT attachment stability, providing a molecular explanation for the abnormal spindle architecture and the cytotoxic activity of this modified protein. Consistent with cell culture data, EGFP-Hec1 expression was found to strongly inhibit tumour growth in a mouse xenograft model by disrupting mitosis and inducing multipolar spindles. Taken together, these findings demonstrate that stimulation of massive chromosome segregation defects can be used as an anti-cancer strategy through the activation of mitotic catastrophe after a multipolar mitosis. Importantly, this study represents a clear proof of concept that targeting KT proteins required for proper KT-MT attachment dynamics constitutes a powerful approach in cancer therapy. PMID

  11. Microtubule dynamics and signal transduction at the immunological synapse: new partners and new connections

    OpenAIRE

    Lasserre, Rémi; Alcover, Andrés

    2012-01-01

    Antigen recognition induces T-cell polarization towards antigen presenting cells, generating the immunological synapse at the cell interface. Now, microtubule-mediated polarized vesicle transport is shown to be required for the organization of a signalling-competent synapse and hence T-cell activation.

  12. Distinctions between dynamic characteristics of the single EG5 motor protein along neural vs. cancerous microtubules.

    Science.gov (United States)

    Feizabadi, Mitra Shojania; Jun, Yonggun; Reddy, J N Babu

    2016-09-30

    The kinesin 5 motor contributes critically to mitosis, and is often upregulated in cancer. In vitro motility studies of kinesin 5 moving along bovine brain microtubules indicate that the motors have limited processivity. Cancer cells have abnormal mitotic behavior, so one might wonder whether the functional properties of kinesin 5 change in such a background. Because there could be multiple unknown changes in cancerous vs normal cells, we chose to address this question in a controlled in vitro environment. Specifically, through a series of parallel experiments along bovine brain vs. breast cancer microtubules, we quantified the in vitro motility characteristics of single Eg5 molecular motors along these two types of microtubules, combining the utilization of an optical trapping technique with a study of motion in the unloaded regime. The obtained values indicate that Eg5 processivity is 40% less along MCF7 microtubules, compared to that measured on bovine brain MTs. Interestingly, not all single-molecule properties are altered, as the velocity of the single motor doesn't show any significant changes on either track, though the binding time along MCF7 microtubules is almost 25% shorter. The current results, in conjunction with our previously reported outcomes of the evaluation of the Eg5's characteristics under external load, show that in transition from no-load to high-load regime, the Eg5 binding time has less sensitivity on MCF7 as compared to bovine brain MTs. This finding is intriguing, as it suggests that, potentially, groups of Eg5 motors function more effectively in the cancer background of a large ensemble, possibly contributing to faster mitosis in cancer cells. PMID:27590585

  13. Using Photobleaching to Measure Spindle Microtubule Dynamics in Primary Cultures of Dividing Drosophila Meiotic Spermatocytes

    OpenAIRE

    Savoian, Matthew S.

    2015-01-01

    In dividing animal cells, a microtubule (MT)-based bipolar spindle governs chromosome movement. Current models propose that the spindle facilitates and/or generates translocating forces by regionally depolymerizing the kinetochore fibers (k-fibers) that bind each chromosome. It is unclear how conserved these sites and the resultant chromosome-moving mechanisms are between different dividing cell types because of the technical challenges of quantitatively studying MTs in many specimens. In par...

  14. 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...

  15. 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.

  16. 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.

  17. 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.

  18. High-resolution Time-lapse Imaging and Automated Analysis of Microtubule Dynamics in Living Human Umbilical Vein Endothelial Cells.

    Science.gov (United States)

    Braun, Alexander; Caesar, Nicole M; Dang, Kyvan; Myers, Kenneth A

    2016-01-01

    The physiological process by which new vasculature forms from existing vasculature requires specific signaling events that trigger morphological changes within individual endothelial cells (ECs). These processes are critical for homeostatic maintenance such as wound healing, and are also crucial in promoting tumor growth and metastasis. EC morphology is defined by the organization of the cytoskeleton, a tightly regulated system of actin and microtubule (MT) dynamics that is known to control EC branching, polarity and directional migration, essential components of angiogenesis. To study MT dynamics, we used high-resolution fluorescence microscopy coupled with computational image analysis of fluorescently-labeled MT plus-ends to investigate MT growth dynamics and the regulation of EC branching morphology and directional migration. Time-lapse imaging of living Human Umbilical Vein Endothelial Cells (HUVECs) was performed following transfection with fluorescently-labeled MT End Binding protein 3 (EB3) and Mitotic Centromere Associated Kinesin (MCAK)-specific cDNA constructs to evaluate effects on MT dynamics. PlusTipTracker software was used to track EB3-labeled MT plus ends in order to measure MT growth speeds and MT growth lifetimes in time-lapse images. This methodology allows for the study of MT dynamics and the identification of how localized regulation of MT dynamics within sub-cellular regions contributes to the angiogenic processes of EC branching and migration. PMID:27584860

  19. 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...

  20. A Thermodynamic Model of Microtubule Assembly and Disassembly

    OpenAIRE

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

    2009-01-01

    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 mode...

  1. ASTER Images San Francisco Bay Area

    Science.gov (United States)

    2000-01-01

    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. 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.

  2. ASTER Images the Island of Hawaii

    Science.gov (United States)

    2000-01-01

    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. 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.

  3. 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.

  4. Phosphorylation of β-Tubulin by the Down Syndrome Kinase, Minibrain/DYRK1a, Regulates Microtubule Dynamics and Dendrite Morphogenesis.

    Science.gov (United States)

    Ori-McKenney, Kassandra M; McKenney, Richard J; Huang, Hector H; Li, Tun; Meltzer, Shan; Jan, Lily Yeh; Vale, Ronald D; Wiita, Arun P; Jan, Yuh Nung

    2016-05-01

    Dendritic arborization patterns are consistent anatomical correlates of genetic disorders such as Down syndrome (DS) and autism spectrum disorders (ASDs). In a screen for abnormal dendrite development, we identified Minibrain (MNB)/DYRK1a, a kinase implicated in DS and ASDs, as a regulator of the microtubule cytoskeleton. We show that MNB is necessary to establish the length and cytoskeletal composition of terminal dendrites by controlling microtubule growth. Altering MNB levels disrupts dendrite morphology and perturbs neuronal electrophysiological activity, resulting in larval mechanosensation defects. Using in vivo and in vitro approaches, we uncover a molecular pathway whereby direct phosphorylation of β-tubulin by MNB inhibits tubulin polymerization, a function that is conserved for mammalian DYRK1a. Our results demonstrate that phosphoregulation of microtubule dynamics by MNB/DYRK1a is critical for dendritic patterning and neuronal function, revealing a previously unidentified mode of posttranslational microtubule regulation in neurons and uncovering a conserved pathway for a DS- and ASD-associated kinase. PMID:27112495

  5. 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.

  6. Alcohol-Induced Alterations in Hepatic Microtubule Dynamics Can Be Explained by Impaired Histone Deacetylase 6 Function

    OpenAIRE

    Shepard, Blythe D.; Joseph, Rohan A.; Kannarkat, George T.; Rutledge, Tara M.; Dean J. Tuma; Tuma, Pamela L.

    2008-01-01

    We have been using polarized, hepatic WIF-B cells to examine ethanol-induced liver injury. These cells polarize in culture and maintain numerous liver-specific activities including the ability to metabolize alcohol. Previously, we found that microtubules were more highly acetylated and more stable in ethanol-treated WIF-B cells and that increased microtubule acetylation required ethanol metabolism and was likely mediated by acetaldehyde. This study was aimed at identifying the mechanism respo...

  7. 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.

  8. 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.

  9. 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.

  10. Microtubules Have Opposite Orientation in Axons and Dendrites of Drosophila Neurons

    OpenAIRE

    Stone, Michelle C.; Roegiers, Fabrice; Rolls, Melissa M

    2008-01-01

    In vertebrate neurons, axons have a uniform arrangement of microtubules with plus ends distal to the cell body (plus-end-out), and dendrites have equal numbers of plus- and minus-end-out microtubules. To determine whether microtubule orientation is a conserved feature of axons and dendrites, we analyzed microtubule orientation in invertebrate neurons. Using microtubule plus end dynamics, we mapped microtubule orientation in Drosophila sensory neurons, interneurons, and motor neurons. As expec...

  11. Leiodermatolide, a novel marine natural product, has potent cytotoxic and antimitotic activity against cancer cells, appears to affect microtubule dynamics, and exhibits antitumor activity.

    Science.gov (United States)

    Guzmán, Esther A; Xu, Qunli; Pitts, Tara P; Mitsuhashi, Kaoru Ogawa; Baker, Cheryl; Linley, Patricia A; Oestreicher, Judy; Tendyke, Karen; Winder, Priscilla L; Suh, Edward M; Wright, Amy E

    2016-11-01

    Pancreatic cancer, the fourth leading cause of cancer death in the United States, has a negative prognosis because metastasis occurs before symptoms manifest. Leiodermatolide, a polyketide macrolide with antimitotic activity isolated from a deep water sponge of the genus Leiodermatium, exhibits potent and selective cytotoxicity toward the pancreatic cancer cell lines AsPC-1, PANC-1, BxPC-3, and MIA PaCa-2, and potent cytotoxicity against skin, breast and colon cancer cell lines. Induction of apoptosis by leiodermatolide was confirmed in the AsPC-1, BxPC-3 and MIA PaCa-2 cells. Leiodermatolide induces cell cycle arrest but has no effects on in vitro polymerization or depolymerization of tubulin alone, while it enhances polymerization of tubulin containing microtubule associated proteins (MAPs). Observations through confocal microscopy show that leiodermatolide, at low concentrations, causes minimal effects on polymerization or depolymerization of the microtubule network in interphase cells, but disruption of spindle formation in mitotic cells. At higher concentrations, depolymerization of the microtubule network is observed. Visualization of the growing microtubule in HeLa cells expressing GFP-tagged plus end binding protein EB-1 showed that leiodermatolide stopped the polymerization of tubulin. These results suggest that leiodermatolide may affect tubulin dynamics without directly interacting with tubulin and hint at a unique mechanism of action. In a mouse model of metastatic pancreatic cancer, leiodermatolide exhibited significant tumor reduction when compared to gemcitabine and controls. The antitumor activities of leiodermatolide, as well as the proven utility of antimitotic compounds against cancer, make leiodermatolide an interesting compound with potential chemotherapeutic effects that may merit further research. PMID:27376928

  12. Evidence for two distinct binding sites for tau on microtubules

    Science.gov (United States)

    Makrides, Victoria; Massie, Michelle R.; Feinstein, Stuart C.; Lew, John

    2004-01-01

    The microtubule-associated protein tau regulates diverse and essential microtubule functions, from the nucleation and promotion of microtubule polymerization to the regulation of microtubule polarity and dynamics, as well as the spacing and bundling of axonal microtubules. Thermodynamic studies show that tau interacts with microtubules in the low- to mid-nanomolar range, implying moderate binding affinity. At the same time, it is well established that microtubule-bound tau does not undergo exchange with the bulk medium readily, suggesting that the tau-microtubule interaction is essentially irreversible. Given this dilemma, we investigated the mechanism of interaction between tau and microtubules in kinetic detail. Stopped-flow kinetic analysis reveals moderate binding affinity between tau and preassembled microtubules and rapid dissociation/association kinetics. In contrast, when microtubules are generated by copolymerization of tubulin and tau, a distinct population of microtubule-bound tau is observed, the binding of which seems irreversible. We propose that reversible binding occurs between tau and the surface of preassembled microtubules, whereas irreversible binding results when tau is coassembled with tubulin into a tau-microtubule copolymer. Because the latter is expected to be physiologically relevant, its characterization is of central importance. PMID:15096589

  13. Contribution of noncentrosomal microtubules to spindle assembly in Drosophila spermatocytes.

    Directory of Open Access Journals (Sweden)

    Elena Rebollo

    2004-01-01

    Full Text Available Previous data suggested that anastral spindles, morphologically similar to those found in oocytes, can assemble in a centrosome-independent manner in cells that contain centrosomes. It is assumed that the microtubules that build these acentrosomal spindles originate over the chromatin. However, the actual processes of centrosome-independent microtubule nucleation, polymerisation, and sorting have not been documented in centrosome-containing cells. We have identified two experimental conditions in which centrosomes are kept close to the plasma membrane, away from the nuclear region, throughout meiosis I in Drosophila spermatocytes. Time-lapse confocal microscopy of these cells labelled with fluorescent chimeras reveals centrosome-independent microtubule nucleation, growth, and sorting into a bipolar spindle array over the nuclear region, away from the asters. The onset of noncentrosomal microtubule nucleation is significantly delayed with respect to nuclear envelope breakdown and coincides with the end of chromosome condensation. It takes place in foci that are close to the membranes that ensheath the nuclear region, not over the condensed chromosomes. Metaphase plates are formed in these spindles, and, in a fraction of them, some degree of polewards chromosome segregation takes place. In these cells that contain both membrane-bound asters and an anastral spindle, the orientation of the cytokinesis furrow correlates with the position of the asters and is independent of the orientation of the spindle. We conclude that the fenestrated nuclear envelope may significantly contribute to the normal process of spindle assembly in Drosophila spermatocytes. We also conclude that the anastral spindles that we have observed are not likely to provide a robust back-up able to ensure successful cell division. We propose that these anastral microtubule arrays could be a constitutive component of wild-type spindles, normally masked by the abundance of centrosome

  14. 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.

  15. 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....

  16. NO serves as a signaling intermediate downstream of H2O2 to modulate dynamic microtubule cytoskeleton during responses to VD-toxins in Arabidopsis

    OpenAIRE

    Yao, Lin-Lin; Pei, Bao-Lei; Zhou, Qun; Li, Ying-Zhang

    2012-01-01

    Although hydrogen peroxide (H2O2) and nitric oxide (NO) can act as an upstream signaling molecule to modulate the dynamic microtubule cytoskeleton during the defense responses to Verticillium dahliae (VD) toxins in Arabidopsis, it is not known the relationship between these two signaling molecules. Here, we show that VD-toxin-induced NO accumulation was dependent on prior H2O2 production, NO is downstream of H2O2 in the signaling process, and that H2O2 acted synergistically with NO to modulat...

  17. 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.

  18. Insights into Antiparallel Microtubule Crosslinking by PRC1, a Conserved Nonmotor Microtubule Binding Protein

    Energy Technology Data Exchange (ETDEWEB)

    Subramanian, Radhika; Wilson-Kubalek, Elizabeth M.; Arthur, Christopher P.; Bick, Matthew J.; Campbell, Elizabeth A.; Darst, Seth A.; Milligan, Ronald A.; Kapoor, Tarun M. (Scripps); (Rockefeller)

    2010-09-03

    Formation of microtubule architectures, required for cell shape maintenance in yeast, directional cell expansion in plants and cytokinesis in eukaryotes, depends on antiparallel microtubule crosslinking by the conserved MAP65 protein family. Here, we combine structural and single molecule fluorescence methods to examine how PRC1, the human MAP65, crosslinks antiparallel microtubules. We find that PRC1's microtubule binding is mediated by a structured domain with a spectrin-fold and an unstructured Lys/Arg-rich domain. These two domains, at each end of a homodimer, are connected by a linkage that is flexible on single microtubules, but forms well-defined crossbridges between antiparallel filaments. Further, we show that PRC1 crosslinks are compliant and do not substantially resist filament sliding by motor proteins in vitro. Together, our data show how MAP65s, by combining structural flexibility and rigidity, tune microtubule associations to establish crosslinks that selectively mark antiparallel overlap in dynamic cytoskeletal networks.

  19. 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.

  20. Motor-mediated cortical versus astral microtubule organization in lipid-monolayered droplets.

    Science.gov (United States)

    Baumann, Hella; Surrey, Thomas

    2014-08-01

    The correct spatial organization of microtubules is of crucial importance for determining the internal architecture of eukaryotic cells. Microtubules are arranged in space by a multitude of biochemical activities and by spatial constraints imposed by the cell boundary. The principles underlying the establishment of distinct intracellular architectures are only poorly understood. Here, we studied the effect of spatial confinement on the self-organization of purified motors and microtubules that are encapsulated in lipid-monolayered droplets in oil, varying in diameter from 5-100 μm, which covers the size range of typical cell bodies. We found that droplet size alone had a major organizing influence. The presence of a microtubule-crosslinking motor protein decreased the number of accessible types of microtubule organizations. Depending on the degree of spatial confinement, the presence of the motor caused either the formation of a cortical array of bent microtubule bundles or the generation of single microtubule asters in the droplets. These are two of the most prominent forms of microtubule arrangements in plant and metazoan cells. Our results provide insights into the combined organizing influence of spatial constraints and cross-linking motor activities determining distinct microtubule architectures in a minimal biomimetic system. In the future, this simple lipid-monolayered droplet system characterized here can be expanded readily to include further biochemical activities or used as the starting point for the investigation of motor-mediated microtubule organization inside liposomes surrounded by a deformable lipid bilayer. PMID:24966327

  1. 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.

  2. Effect of the microtubule-associated protein tau on dynamics of single-headed motor proteins KIF1A

    Science.gov (United States)

    Sparacino, J.; Farías, M. G.; Lamberti, P. W.

    2014-02-01

    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 nonmotile microtubule-associated proteins, such as tau. Motivated by experiments on kinesin-tau interactions [Dixit et al., Science 319, 1086 (2008), 10.1126/science.1152993] 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 reproduces 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.

  3. 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.

  4. 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.

  5. 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)

  6. Dimer model for Tau proteins bound in microtubule bundles

    Science.gov (United States)

    Hall, Natalie; Kluber, Alexander; Hayre, N. Robert; Singh, Rajiv; Cox, Daniel

    2013-03-01

    The microtubule associated protein tau is important in nucleating and maintaining microtubule spacing and structure in neuronal axons. Modification of tau is implicated as a later stage process in Alzheimer's disease, but little is known about the structure of tau in microtubule bundles. We present preliminary work on a proposed model for tau dimers in microtubule bundles (dimers are the minimal units since there is one microtubule binding domain per tau). First, a model of tau monomer was created and its characteristics explored using implicit solvent molecular dynamics simulation. Multiple simulations yield a partially collapsed form with separate positively/negatively charged clumps, but which are a factor of two smaller than required by observed microtubule spacing. We argue that this will elongate in dimer form to lower electrostatic energy at a cost of entropic ``spring'' energy. We will present preliminary results on steered molecular dynamics runs on tau dimers to estimate the actual force constant. Supported by US NSF Grant DMR 1207624.

  7. 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.

  8. 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

  9. Axis establishment and microtubule-mediated waves prior to first cleavage in Beroe ovata.

    Science.gov (United States)

    Houliston, E; Carré, D; Johnston, J A; Sardet, C

    1993-01-01

    The single axis (oral-aboral) and two planes of symmetry of the ctenophore Beroe ovata become established with respect to the position of zygote nucleus formation and the orientation of first cleavage. Bisection of Beroe eggs at different times revealed that differences in egg organisation are established in relation to the presumptive oral-aboral axis before first cleavage. Lateral fragments produced after but not before the time of first mitosis developed into larvae lacking comb-plates on one side. Time-lapse video demonstrated that waves of cytoplasmic reorganisation spread through the layer of peripheral cytoplasm (ectoplasm) of the egg during the 80 minute period between pronuclear fusion and first cleavage, along the future oral-aboral axis. These waves are manifest as the progressive displacement and dispersal of plaques of accumulated organelles around supernumerary sperm nuclei, and a series of surface movements. Their timing and direction of propagation suggest they may be involved in establishing cytoplasmic differences with respect to the embryonic axis. Inhibitor experiments suggested that the observed cytoplasmic reorganisation involves microtubules. Nocodazole and taxol, which prevent microtubule turnover,blocked plaque dispersal and reduced surface movements. The microfilament-disrupting drug cytochalasin B did not prevent plaque dispersal but induced abnormal surface contractions. We examined changes in microtubule organisation using immunofluorescence on eggs fixed at different times and in live eggs following injection of rhodamine-tubulin. Giant microtubule asters become associated with each male pronucleus after the end of meiosis. Following pronuclear fusion they disappear successively, those nearest the zygote nucleus shrinking first, to establish gradients of aster size within single eggs. Regional differences in microtubule behaviour around the time of mitosis were revealed by brief taxol treatment, which induced the formation of small

  10. 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.

  11. 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.

  12. 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.

  13. 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. PMID:23037062

  14. 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.

  15. 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.

  16. 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.

  17. Molecular and Mechanical Causes of Microtubule Catastrophe and Aging.

    Science.gov (United States)

    Zakharov, Pavel; Gudimchuk, Nikita; Voevodin, Vladimir; Tikhonravov, Alexander; Ataullakhanov, Fazoil I; Grishchuk, Ekaterina L

    2015-12-15

    Tubulin polymers, microtubules, can switch abruptly from the assembly to shortening. These infrequent transitions, termed "catastrophes", affect numerous cellular processes but the underlying mechanisms are elusive. We approached this complex stochastic system using advanced coarse-grained molecular dynamics modeling of tubulin-tubulin interactions. Unlike in previous simplified models of dynamic microtubules, the catastrophes in this model arise owing to fluctuations in the composition and conformation of a growing microtubule tip, most notably in the number of protofilament curls. In our model, dynamic evolution of the stochastic microtubule tip configurations over a long timescale, known as the system's "aging", gives rise to the nonexponential distribution of microtubule lifetimes, consistent with experiment. We show that aging takes place in the absence of visible changes in the microtubule wall or tip, as this complex molecular-mechanical system evolves slowly and asymptotically toward the steady-state level of the catastrophe-promoting configurations. This new, to our knowledge, theoretical basis will assist detailed mechanistic investigations of the mechanisms of action of different microtubule-binding proteins and drugs, thereby enabling accurate control over the microtubule dynamics to treat various pathologies. PMID:26682815

  18. Anti-Microtubule Drugs.

    Science.gov (United States)

    Florian, Stefan; Mitchison, Timothy J

    2016-01-01

    Small molecule drugs that target microtubules (MTs), many of them natural products, have long been important tools in the MT field. Indeed, tubulin (Tb) was discovered, in part, as the protein binding partner of colchicine. Several anti-MT drug classes also have important medical uses, notably colchicine, which is used to treat gout, familial Mediterranean fever (FMF), and pericarditis, and the vinca alkaloids and taxanes, which are used to treat cancer. Anti-MT drugs have in common that they bind specifically to Tb in the dimer, MT or some other form. However, their effects on polymerization dynamics and on the human body differ markedly. Here we briefly review the most-studied molecules, and comment on their uses in basic research and medicine. Our focus is on practical applications of different anti-MT drugs in the laboratory, and key points that users should be aware of when designing experiments. We also touch on interesting unsolved problems, particularly in the area of medical applications. In our opinion, the mechanism by which any MT drug cures or treats any disease is still unsolved, despite decades of research. Solving this problem for particular drug-disease combinations might open new uses for old drugs, or provide insights into novel routes for treatment. PMID:27193863

  19. 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.

  20. 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.

  1. Micromechanical modeling of microtubules

    OpenAIRE

    Arslan, Melis

    2010-01-01

    Microtubules serve as one of the structural components of the cell and take place in some of the important cellular functions such as mitosis and vesicular transport. Microtubules comprise of tubulin subunits tubulin dimers arranged in a cylindrical beta and formed by alpha hollow tube structure with a diameter of 20nm. They are typically comprised of 13 or 14 protofilaments arranged in spiral configurations. The longitudinal bonds between the tubulin dimers are much stiffer and stronger than...

  2. 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.

  3. A New Diterpenoid Glucoside from Aster smithianus

    Institute of Scientific and Technical Information of China (English)

    Shou Jun GUO; Xing Hua ZHAO; Dong Liang CHENG

    2004-01-01

    A new diterpenoid trisaccharide, smithoside A, was isolated from Aster smithianus. Its structure was identified as pimar-15 (16)-β-en-3β, 8β, 1 1α-triol-3-O-β-D-glucopyranosyl (l→3)-[β-glucopyranosyl (1→2)]-β-D-glucopyranoside on the basis of the spectral and chemical methods.

  4. Multiscale modeling and simulation of microtubule-motor-protein assemblies

    Science.gov (United States)

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

    2015-12-01

    Microtubules and motor proteins self-organize into biologically important assemblies including the mitotic spindle and the centrosomal microtubule array. Outside of cells, microtubule-motor mixtures can form novel active liquid-crystalline materials driven out of equilibrium by adenosine triphosphate-consuming motor proteins. Microscopic motor activity causes polarity-dependent interactions between motor proteins and microtubules, but how these interactions yield larger-scale dynamical behavior such as complex flows and defect dynamics is not well understood. We develop a multiscale theory for microtubule-motor systems in which Brownian dynamics simulations of polar microtubules driven by motors are used to study microscopic organization and stresses created by motor-mediated microtubule interactions. We identify polarity-sorting and crosslink tether relaxation as two polar-specific sources of active destabilizing stress. We then develop a continuum Doi-Onsager model that captures polarity sorting and the hydrodynamic flows generated by these polar-specific active stresses. In simulations of active nematic flows on immersed surfaces, the active stresses drive turbulent flow dynamics and continuous generation and annihilation of disclination defects. The dynamics follow from two instabilities, and accounting for the immersed nature of the experiment yields unambiguous characteristic length and time scales. When turning off the hydrodynamics in the Doi-Onsager model, we capture formation of polar lanes as observed in the Brownian dynamics simulation.

  5. 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.

  6. Low-dose laulimalide represents a novel molecular probe for investigating microtubule organization.

    Science.gov (United States)

    Bennett, Melissa J; Chan, Gordon K; Rattner, J B; Schriemer, David C

    2012-08-15

    Laulimalide is a natural product that has strong taxoid-like properties but binds to a distinct site on β-tubulin in the microtubule (MT) lattice. At elevated concentrations, it generates MTs that are resistant to depolymerization, and it induces a conformational state indistinguishable from taxoid-treated MTs. In this study, we describe the effect of low-dose laulimalide on various stages of the cell cycle and compare these effects to docetaxel as a representative of taxoid stabilizers. No evidence of MT bundling in interphase was observed with laulimalide, in spite of the fact that MTs are stabilized at low dose. Cells treated with laulimalide enter mitosis but arrest at prometaphase by generating multiple asters that coalesce into supernumerary poles and interfere with the integrity of the metaphase plate. Cells with a preformed bipolar spindle exist under heightened tension under laulimalide treatment, and chromosomes rapidly shear from the plate, even though the bipolar spindle is well-preserved. Docetaxel generates a similar phenotype for HeLa cells entering mitosis, but when treated at metaphase, cells undergo chromosomal fragmentation and demonstrate reduced centromere dynamics, as expected for a taxoid. Our results suggest that laulimalide represents a new class of molecular probe for investigating MT-mediated events, such as kinetochore-MT interactions, which may reflect the location of the ligand binding site within the interprotofilament groove. PMID:22871740

  7. 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.

  8. Single molecule imaging reveals differences in microtubule track selection between Kinesin motors.

    Directory of Open Access Journals (Sweden)

    Dawen Cai

    2009-10-01

    Full Text Available Cells generate diverse microtubule populations by polymerization of a common alpha/beta-tubulin building block. How microtubule associated proteins translate microtubule heterogeneity into specific cellular functions is not clear. We evaluated the ability of kinesin motors involved in vesicle transport to read microtubule heterogeneity by using single molecule imaging in live cells. We show that individual Kinesin-1 motors move preferentially on a subset of microtubules in COS cells, identified as the stable microtubules marked by post-translational modifications. In contrast, individual Kinesin-2 (KIF17 and Kinesin-3 (KIF1A motors do not select subsets of microtubules. Surprisingly, KIF17 and KIF1A motors that overtake the plus ends of growing microtubules do not fall off but rather track with the growing tip. Selection of microtubule tracks restricts Kinesin-1 transport of VSVG vesicles to stable microtubules in COS cells whereas KIF17 transport of Kv1.5 vesicles is not restricted to specific microtubules in HL-1 myocytes. These results indicate that kinesin families can be distinguished by their ability to recognize microtubule heterogeneity. Furthermore, this property enables kinesin motors to segregate membrane trafficking events between stable and dynamic microtubule populations.

  9. 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. PMID:25897332

  10. Kinks and bell-type solitons in microtubules.

    Science.gov (United States)

    Zdravković, Slobodan; Gligorić, Goran

    2016-06-01

    In the present paper, we study the nonlinear dynamics of microtubules relying on the known u-model. As a mathematical procedure, we use the simplest equation method. We recover some solutions obtained earlier using less general methods. These are kink solitons. In addition, we show that the solution of the crucial differential equation, describing nonlinear dynamics of microtubules, can be a bell-type soliton. The discovery of this new solution is supported by numerical analysis. PMID:27368766

  11. Metallic Glass Wire Based Localization of Kinesin/Microtubule Bio-molecular Motility System

    Science.gov (United States)

    Kim, K.; Sikora, A.; Yaginuma, S.; Nakayama, K. S.; Nakazawa, H.; Umetsu, M.; Hwang, W.; Teizer, W.

    2014-03-01

    We report electrophoretic accumulation of microtubules along metallic glass (Pd42.5Cu30Ni7.5P20) wires free-standing in solution. Microtubules are dynamic cytoskeletal filaments. Kinesin is a cytoskeletal motor protein. Functions of these bio-molecules are central to various dynamic cellular processes. Functional artificial organization of bio-molecules is a prerequisite for transferring their native functions into device applications. Fluorescence microscopy at the individual-microtubule level reveals microtubules aligning along the wire axis during the electrophoretic migration. Casein-treated electrodes are effective for releasing trapped microtubules upon removal of the external field. Furthermore, we demonstrate gliding motion of microtubules on kinesin-treated metallic glass wires. The reversible manner in the local adsorption of microtubules, the flexibility of wire electrodes, and the compatibility between the wire electrode and the bio-molecules are beneficial for spatio-temporal manipulation of the motility machinery in 3 dimensions.

  12. 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.

  13. 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...

  14. Protein 4.1R binds to CLASP2 and regulates dynamics,organization and attachment of microtubules to the cell cortex

    NARCIS (Netherlands)

    A. Ruiz-Saenz (Ana); J. van Haren (Jeffrey); C.L. Sayas (C. Laura); L. Rangel (Laura); J.A.A. Demmers (Jeroen); J. Millán (Jaime); M.A. Alonso (Miguel); N.J. Galjart (Niels); J.M. Correas

    2013-01-01

    textabstractThe microtubule (MT) cytoskeleton is essential for many cellular processes, including cell polarity and migration. Cortical platforms, formed by a subset of MT plus-end-tracking proteins, such as CLASP2, and non-MT binding proteins such as LL5b, attach distal ends of MTs to the cell cort

  15. Regional tsunami vulnerability analysis through ASTER imagery

    Science.gov (United States)

    Dall'Osso, Filippo; Cavalletti, Alessandra; Immordino, Francesco; Gonella, Marco

    2010-05-01

    Analysis of vulnerability to natural hazards is a key issue of prevention measures within ICZM. Knowledge of susceptibility to damage and how this is distributed along the coast allows to optimize possible prevention and mitigation actions. The present study focuses on tsunami vulnerability of a large extension of coastline: the entire westerly Thailand's coast. The work is a follow up of the CRATER project (Coastal Risk Analysis for Tsunamis and Environmental Remediation) carried out on the aftermath of the 26th December 2004 Tsunami event. Vulnerability is analyzed considering an inundation scenario given by a tsunami of seismic origin, causing a maximum run-up of 25m.. An innovative methodology have been here developed and applied, based on the combined use of ASTER (Advanced Spaceborn Thermal Emission and Reflection Radiometer) satellite imagery, SRTM v-3 (Shuttle Radar Topography Mission - version #3) DEMs and GIS. Vulnerability level has been calculated combining information on coastal geomorphology, land use, topography and distance from the shoreline. Land use has been extrapolated from ASTER images through a multi-spectral analysis (a pixel-based and supervised classification process) of ASTER bands 1 to 9, plus one band for the NDVI index (Normalized Difference Vegetation Index). Coastal geomorphology has been obtained through a photo-interpretation process. Results have been organized in a set of vectorial vulnerability maps with horizontal resolution of 90m. The proposed methodology has the great advantage of being repeatable for any case of vulnerability analysis at small-medium scale (i.e. at Regional/National level) with a moderate investment in term of costs and human resources.

  16. 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

  17. 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

  18. 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.

  19. 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.

  20. 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...

  1. 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.

  2. α-Synuclein is a Novel Microtubule Dynamase

    Science.gov (United States)

    Cartelli, Daniele; Aliverti, Alessandro; Barbiroli, Alberto; Santambrogio, Carlo; Ragg, Enzio M.; Casagrande, Francesca V.M.; Cantele, Francesca; Beltramone, Silvia; Marangon, Jacopo; De Gregorio, Carmelita; Pandini, Vittorio; Emanuele, Marco; Chieregatti, Evelina; Pieraccini, Stefano; Holmqvist, Staffan; Bubacco, Luigi; Roybon, Laurent; Pezzoli, Gianni; Grandori, Rita; Arnal, Isabelle; Cappelletti, Graziella

    2016-01-01

    α-Synuclein is a presynaptic protein associated to Parkinson’s disease, which is unstructured when free in the cytoplasm and adopts α helical conformation when bound to vesicles. After decades of intense studies, α-Synuclein physiology is still difficult to clear up due to its interaction with multiple partners and its involvement in a pletora of neuronal functions. Here, we looked at the remarkably neglected interplay between α-Synuclein and microtubules, which potentially impacts on synaptic functionality. In order to identify the mechanisms underlying these actions, we investigated the interaction between purified α-Synuclein and tubulin. We demonstrated that α-Synuclein binds to microtubules and tubulin α2β2 tetramer; the latter interaction inducing the formation of helical segment(s) in the α-Synuclein polypeptide. This structural change seems to enable α-Synuclein to promote microtubule nucleation and to enhance microtubule growth rate and catastrophe frequency, both in vitro and in cell. We also showed that Parkinson’s disease-linked α-Synuclein variants do not undergo tubulin-induced folding and cause tubulin aggregation rather than polymerization. Our data enable us to propose α-Synuclein as a novel, foldable, microtubule-dynamase, which influences microtubule organisation through its binding to tubulin and its regulating effects on microtubule nucleation and dynamics. PMID:27628239

  3. 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.

  4. α-Synuclein is a Novel Microtubule Dynamase.

    Science.gov (United States)

    Cartelli, Daniele; Aliverti, Alessandro; Barbiroli, Alberto; Santambrogio, Carlo; Ragg, Enzio M; Casagrande, Francesca V M; Cantele, Francesca; Beltramone, Silvia; Marangon, Jacopo; De Gregorio, Carmelita; Pandini, Vittorio; Emanuele, Marco; Chieregatti, Evelina; Pieraccini, Stefano; Holmqvist, Staffan; Bubacco, Luigi; Roybon, Laurent; Pezzoli, Gianni; Grandori, Rita; Arnal, Isabelle; Cappelletti, Graziella

    2016-01-01

    α-Synuclein is a presynaptic protein associated to Parkinson's disease, which is unstructured when free in the cytoplasm and adopts α helical conformation when bound to vesicles. After decades of intense studies, α-Synuclein physiology is still difficult to clear up due to its interaction with multiple partners and its involvement in a pletora of neuronal functions. Here, we looked at the remarkably neglected interplay between α-Synuclein and microtubules, which potentially impacts on synaptic functionality. In order to identify the mechanisms underlying these actions, we investigated the interaction between purified α-Synuclein and tubulin. We demonstrated that α-Synuclein binds to microtubules and tubulin α2β2 tetramer; the latter interaction inducing the formation of helical segment(s) in the α-Synuclein polypeptide. This structural change seems to enable α-Synuclein to promote microtubule nucleation and to enhance microtubule growth rate and catastrophe frequency, both in vitro and in cell. We also showed that Parkinson's disease-linked α-Synuclein variants do not undergo tubulin-induced folding and cause tubulin aggregation rather than polymerization. Our data enable us to propose α-Synuclein as a novel, foldable, microtubule-dynamase, which influences microtubule organisation through its binding to tubulin and its regulating effects on microtubule nucleation and dynamics. PMID:27628239

  5. A novel isoform of MAP4 organises the paraxial microtubule array required for muscle cell differentiation.

    Science.gov (United States)

    Mogessie, Binyam; Roth, Daniel; Rahil, Zainab; Straube, Anne

    2015-01-01

    The microtubule cytoskeleton is critical for muscle cell differentiation and undergoes reorganisation into an array of paraxial microtubules, which serves as template for contractile sarcomere formation. In this study, we identify a previously uncharacterised isoform of microtubule-associated protein MAP4, oMAP4, as a microtubule organising factor that is crucial for myogenesis. We show that oMAP4 is expressed upon muscle cell differentiation and is the only MAP4 isoform essential for normal progression of the myogenic differentiation programme. Depletion of oMAP4 impairs cell elongation and cell-cell fusion. Most notably, oMAP4 is required for paraxial microtubule organisation in muscle cells and prevents dynein- and kinesin-driven microtubule-microtubule sliding. Purified oMAP4 aligns dynamic microtubules into antiparallel bundles that withstand motor forces in vitro. We propose a model in which the cooperation of dynein-mediated microtubule transport and oMAP4-mediated zippering of microtubules drives formation of a paraxial microtubule array that provides critical support for the polarisation and elongation of myotubes. PMID:25898002

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

    Science.gov (United States)

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

    2015-08-10

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

  7. 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

  8. ASTER system operating achievement for 15 years on orbit

    Science.gov (United States)

    Inada, Hitomi; Ito, Yoshiyuki; Kikuchi, Masakuni; Sakuma, Fumihiro; Tatsumi, Kenji; Akagi, Shigeki; Ono, Hidehiko

    2015-10-01

    ASTER (Advanced Spaceborne Thermal Emission and Reflection Radiometer) System is operating more than 15 years since launched on board of NASA's Terra spacecraft in December 1999. ASTER System is composed of 3 radiometers (VNIR (Visible and Near Infrared Radiometer), SWIR (Short-Wave Infrared Radiometer), and TIR (Thermal Infrared Radiometer)), CSP (Common Signal Processor) and MSP (Master Power Supply). This paper describes the ASTER System operating history and the achievement of ASTER System long term operation since the initial checkout operation, the normal operation, and the continuous operation. Through the 15 years operation, ASTER system had totally checked the all subsystems (MPS, VNIR, TIR, SWIR, and CSP) health and safety check using telemetry data trend evaluation, and executed the necessary action. The watch items are monitored as the life control items. The pointing mechanics for VNIR, SWIR and TIR, and the cooler for SWIR and TIR are all operating with any problem for over 15 years. In 2003, ASTER was successfully operated for the lunar calibration. As the future plan, ASTER team is proposing the 2nd lunar calibration before the end of mission.

  9. The Microtubule-Associated Protein END BINDING1 Modulates Membrane Trafficking Pathways in Plant Root Cells

    OpenAIRE

    Shahidi, Saeid

    2013-01-01

    EB1 protein preferentially binds to the fast growing ends of microtubules where it regulates microtubule dynamics. In addition to microtubules, EB1 interacts with several additional proteins, and through these interactions modulates various cellular processes. Arabidopsis thaliana eb1 mutants have roots that exhibit aberrant responses to touch/gravity cues. Columella cells in the centre of the root cap are polarized and play key roles in these responses by functioning as sensors.I examined th...

  10. Comparison of ASTER Global Emissivity Database (ASTER-GED) With In-Situ Measurement In Italian Vulcanic Areas

    Science.gov (United States)

    Silvestri, M.; Musacchio, M.; Buongiorno, M. F.; Amici, S.; Piscini, A.

    2015-12-01

    LP DAAC released the Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) Global Emissivity Database (GED) datasets on April 2, 2014. The database was developed by the National Aeronautics and Space Administration's (NASA) Jet Propulsion Laboratory (JPL), California Institute of Technology. The database includes land surface emissivities derived from ASTER data acquired over the contiguous United States, Africa, Arabian Peninsula, Australia, Europe, and China. In this work we compare ground measurements of emissivity acquired by means of Micro-FTIR (Fourier Thermal Infrared spectrometer) instrument with the ASTER emissivity map extract from ASTER-GED and the emissivity obtained by using single ASTER data. Through this analysis we want to investigate differences existing between the ASTER-GED dataset (average from 2000 to 2008 seasoning independent) and fall in-situ emissivity measurement. Moreover the role of different spatial resolution characterizing ASTER and MODIS, 90mt and 1km respectively, by comparing them with in situ measurements. Possible differences can be due also to the different algorithms used for the emissivity estimation, Temperature and Emissivity Separation algorithm for ASTER TIR band( Gillespie et al, 1998) and the classification-based emissivity method (Snyder and al, 1998) for MODIS. In-situ emissivity measurements have been collected during dedicated fields campaign on Mt. Etna vulcano and Solfatara of Pozzuoli. Gillespie, A. R., Matsunaga, T., Rokugawa, S., & Hook, S. J. (1998). Temperature and emissivity separation from Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) images. IEEE Transactions on Geoscience and Remote Sensing, 36, 1113-1125. Snyder, W.C., Wan, Z., Zhang, Y., & Feng, Y.-Z. (1998). Classification-based emissivity for land surface temperature measurement from space. International Journal of Remote Sensing, 19, 2753-2574.

  11. 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.

  12. 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.

  13. 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.

  14. 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

  15. 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.

  16. 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...

  17. 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...

  18. 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.

  19. Preliminary Attitude Control Studies for the ASTER Mission

    International Nuclear Information System (INIS)

    This work discusses an attitude control study for the ASTER mission, the first Brazilian mission to the deep space. The study is part of a larger scenario that is the development of optimal trajectories to navigate in the 2001 SN263 asteroid system, together with the generation of orbit and attitude controllers for autonomous operation. The spacecraft attitude is defined from the orientation of the body reference system to the Local Vertical Local Horizontal (LVLH) of a circular orbit around the Alpha asteroid. The rotational equations of motion involve the dynamic equations, where the three angular speeds are generated from a set of three reaction wheels and the gravitational torque. The rotational kinematics is represented in the Euler angles format. The controller is developed via the linear quadratic regulator approach with output feedback. It involves the generation of a stability augmentation (SAS) loop and a tracking outer loop, with a compensator of desired structure. It was chosen the feedback of the p, q and r angular speeds in the SAS, one for each reaction wheel. In the outer loop, it was chosen a proportional integral compensator. The parameters are tuned using a numerical minimization that represents a linear quadratic cost, with weightings in the tracking error and controls. Simulations are performed with the nonlinear model. For small angle manoeuvres, the linear results with reaction wheels or thrusters are reasonable, but, for larger manoeuvres, nonlinear control techniques shall be applied, for example, the sliding mode control

  20. 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.

  1. The Membrane-Associated Sec1/Munc18 KEULE is Required for Phragmoplast Microtubule Reorganization During Cytokinesis in Arabidopsis.

    Science.gov (United States)

    Steiner, Alexander; Müller, Lin; Rybak, Katarzyna; Vodermaier, Vera; Facher, Eva; Thellmann, Martha; Ravikumar, Raksha; Wanner, Gerhard; Hauser, Marie-Theres; Assaad, Farhah F

    2016-04-01

    Cytokinesis, the partitioning of the cytoplasm following nuclear division, requires extensive coordination between membrane trafficking and cytoskeletal dynamics. In plants, the onset of cytokinesis is characterized by the assembly of a bipolar microtubule array, the phragmoplast, and of a transient membrane compartment, the cell plate. Little is known about the coordination between membrane deposition at the cell plate and the dynamics of phragmoplast microtubules. In this study, we monitor the localization dynamics of microtubule and membrane markers throughout cytokinesis. Our spatiotemporal resolution is consistent with the general view that microtubule dynamics drive membrane movements. Nonetheless, we provide evidence for active sorting at the cell plate and show that this is, at least in part, mediated by the TRAPPII tethering complex. We also characterize phragmoplast microtubule organization and cell plate formation in a suite of cytokinesis-defective mutants. Of four mutant lines with defects in phragmoplast microtubule organization, only mor1 microtubule-associated mutants exhibited aberrant cell plates. Conversely, the mutants with the strongest impairment in phragmoplast microtubule reorganization are keule alleles, which have a primary defect in membrane fusion. Our findings identify the SEC1/Munc18 protein KEULE as a central regulatory node in the coordination of membrane and microtubule dynamics during plant cytokinesis. PMID:26700031

  2. 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).

  3. 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.

  4. Nuclear and microtubule remodeling and in vitro development of nuclear transferred cat oocytes with skin fibroblasts of the domestic cat (Felis silvestris catus) and leopard cat (Prionailurus bengalensis).

    Science.gov (United States)

    Yin, X J; Lee, Y H; Jin, J Y; Kim, N H; Kong, I K

    2006-10-01

    The leopard cat (Prionailurus bengalensis), a member of the felidae family, is a threatened animal in South Korea. In terms of protecting endangered felids, nuclear transfer (NT) is a potentially valuable technique for assuring the continuation of species with dwindling numbers. In the present experiment, nuclear and microtubule remodeling and the in vitro developmental potential of enucleated domestic cat oocytes reconstructed with nuclei of somatic cells from either domestic cat fibroblast (DCF) or leopard cat fibroblast (LCF) were evaluated. Microtubule aster is allocated to de-condensed chromatin following nuclear transfer (3h after activation) of fibroblast cells from both domestic and leopard cats, suggesting the introduction of a somatic cell centrosome. The transferred fibroblast nuclei formed a large, swollen, pronuclear-like structure in most reconstructed oocytes, in the cat or leopard cat. At 18h following nuclear transfer, mitosis occurred, and according to the photo (F) it appears that spindle microtubules and two asters were observed. The percentages of blastocyst formation from nuclear transfer embryos derived from domestic cat fibroblasts (4/46, 8.6%) were not significantly different than those for nuclear transfer embryos constructed with leopard cat fibroblasts (4/52, 7.6%). These results indicate that nuclear and microtubule remodeling processes and in vitro developmental ability are similar in reconstructed cat oocytes following transfer of nuclei from either domestic or leopard cats. PMID:16310987

  5. 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

    2015-01-01

    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. PMID:26621779

  6. 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.

    2015-01-01

    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. PMID:26621779

  7. An Introduction to ASTER GDEM and Procedure Reading%ASTER GDEM数据介绍与程序读取

    Institute of Scientific and Technical Information of China (English)

    康晓伟; 冯钟葵

    2011-01-01

    2009年6月30日,期待已久的ASTER GDEM数据由日本经济产业省(METI)和美国航天局(NASA)共同发布,其空间分辨率达到了1弧秒×1弧秒(约30m× 30m),相比2003年NASA发布的SRTM数据有了很大的提高,并且其陆地表面覆盖率也大幅提高,达到了陆地面积的99%.本文介绍了ASTER GDEM的相关特性以及使用程序读取该数据的方法.%On 30th,June 2009,METI and NASA distributed the ASTER GDEM(Advanced Space borne Thermal Emission and Reflection Radiometer Global Digital Elevation Model). The space resolution of ASTER GDEM is 1 arc-secX 1 arc-sec(ap-proximately 30m × 30m) and the land surface coverage reached 99%, both have much greater improvement than SRTM which was distributed in 2003 by NASA. This paper introduced characteristics of ASTER GDEM and how to read the data with programming language.

  8. Interactive domains in the molecular chaperone human alphaB crystallin modulate microtubule assembly and disassembly.

    Directory of Open Access Journals (Sweden)

    Joy G Ghosh

    Full Text Available Small heat shock proteins regulate microtubule assembly during cell proliferation and in response to stress through interactions that are poorly understood.Novel functions for five interactive sequences in the small heat shock protein and molecular chaperone, human alphaB crystallin, were investigated in the assembly/disassembly of microtubules and aggregation of tubulin using synthetic peptides and mutants of human alphaB crystallin.The interactive sequence (113FISREFHR(120 exposed on the surface of alphaB crystallin decreased microtubule assembly by approximately 45%. In contrast, the interactive sequences, (131LTITSSLSSDGV(142 and (156ERTIPITRE(164, corresponding to the beta8 strand and the C-terminal extension respectively, which are involved in complex formation, increased microtubule assembly by approximately 34-45%. The alphaB crystallin peptides, (113FISREFHR(120 and (156ERTIPITRE(164, inhibited microtubule disassembly by approximately 26-36%, and the peptides (113FISREFHR(120 and (131LTITSSLSSDGV(142 decreased the thermal aggregation of tubulin by approximately 42-44%. The (131LTITSSLSSDGV(142 and (156ERTIPITRE(164 peptides were more effective than the widely used anti-cancer drug, Paclitaxel, in modulating tubulinmicrotubule dynamics. Mutagenesis of these interactive sequences in wt human alphaB crystallin confirmed the effects of the alphaB crystallin peptides on microtubule assembly/disassembly and tubulin aggregation. The regulation of microtubule assembly by alphaB crystallin varied over a narrow range of concentrations. The assembly of microtubules was maximal at alphaB crystallin to tubulin molar ratios between 1:4 and 2:1, while molar ratios >2:1 inhibited microtubule assembly.Interactive sequences on the surface of human alphaB crystallin collectively modulate microtubule assembly through a dynamic subunit exchange mechanism that depends on the concentration and ratio of alphaB crystallin to tubulin. These are the first

  9. Disruption of cytoplasmic microtubules by ultraviolet radiation

    International Nuclear Information System (INIS)

    Ultraviolet (UV) irradiation of cultured human skin fibroblasts causes the disassembly of their microtubules. Using indirect immunofluorescence microscopy, we have now investigated whether damage to the microtubule precursor pool may contribute to the disruption of microtubules. Exposure to polychromatic UV radiation inhibits the reassembly of microtubules during cellular recovery from cold treatment. In addition, the ability of taxol to promote microtubule polymerization and bundling is inhibited in UV-irradiated cells. However, UV irradiation of taxol-pretreated cells or in situ detergent-extracted microtubules fails to disrupt the microtubule network. These data suggest that damage to dimeric tubulin, or another soluble factor(s) required for polymerization, contributes to the disassembly of microtubules in UV-irradiated human skin fibroblasts

  10. Tau mediates microtubule bundle architectures mimicking fascicles of microtubules found in the axon initial segment

    Science.gov (United States)

    Chung, Peter J.; Song, Chaeyeon; Deek, Joanna; Miller, Herbert P.; Li, Youli; Choi, Myung Chul; Wilson, Leslie; Feinstein, Stuart C.; Safinya, Cyrus R.

    2016-01-01

    Tau, an intrinsically disordered protein confined to neuronal axons, binds to and regulates microtubule dynamics. Although there have been observations of string-like microtubule fascicles in the axon initial segment (AIS) and hexagonal bundles in neurite-like processes in non-neuronal cells overexpressing Tau, cell-free reconstitutions have not replicated either geometry. Here we map out the energy landscape of Tau-mediated, GTP-dependent ‘active' microtubule bundles at 37 °C, as revealed by synchrotron SAXS and TEM. Widely spaced bundles (wall-to-wall distance Dw–w≈25–41 nm) with hexagonal and string-like symmetry are observed, the latter mimicking bundles found in the AIS. A second energy minimum (Dw–w≈16–23 nm) is revealed under osmotic pressure. The wide spacing results from a balance between repulsive forces, due to Tau's projection domain (PD), and a stabilizing sum of transient sub-kBT cationic/anionic charge–charge attractions mediated by weakly penetrating opposing PDs. This landscape would be significantly affected by charge-altering modifications of Tau associated with neurodegeneration. PMID:27452526

  11. 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...

  12. Evaluation of Aster Gdem v3 Using Icesat Laser Altimetry

    Science.gov (United States)

    Carabajal, C. C.; Boy, J.-P.

    2016-06-01

    We have used a set of Ground Control Points (GCPs) derived from altimetry measurements from the Ice, Cloud and land Elevation Satellite (ICESat) to evaluate the quality of the 30 m posting ASTER (Advanced Spaceborne Thermal Emission and Reflection Radiometer) Global Digital Elevation Model (GDEM) V3 elevation products produced by NASA/METI for Greenland and Antarctica. These data represent the highest quality globally distributed altimetry measurements that can be used for geodetic ground control, selected by applying rigorous editing criteria, useful at high latitudes, where other topographic control is scarce. Even if large outliers still remain in all ASTER GDEM V3 data for both, Greenland and Antarctica, they are significantly reduced when editing ASTER by number of scenes (N≥5) included in the elevation processing. For 667,354 GCPs in Greenland, differences show a mean of 13.74 m, a median of -6.37 m, with an RMSE of 109.65 m. For Antarctica, 6,976,703 GCPs show a mean of 0.41 m, with a median of -4.66 m, and a 54.85 m RMSE, displaying smaller means, similar medians, and less scatter than GDEM V2. Mean and median differences between ASTER and ICESat are lower than 10 m, and RMSEs lower than 10 m for Greenland, and 20 m for Antarctica when only 9 to 31 scenes are included.

  13. Tracking the Biogenesis and Inheritance of Subpellicular Microtubule in Trypanosoma brucei with Inducible YFP-α-Tubulin

    Directory of Open Access Journals (Sweden)

    Omar Sheriff

    2014-01-01

    Full Text Available The microtubule cytoskeleton forms the most prominent structural system in Trypanosoma brucei, undergoing extensive modifications during the cell cycle. Visualization of tyrosinated microtubules leads to a semiconservative mode of inheritance, whereas recent studies employing microtubule plus end tracking proteins have hinted at an asymmetric pattern of cytoskeletal inheritance. To further the knowledge of microtubule synthesis and inheritance during T. brucei cell cycle, the dynamics of the microtubule cytoskeleton was visualized by inducible YFP-α-tubulin expression. During new flagellum/flagellum attachment zone (FAZ biogenesis and cell growth, YFP-α-tubulin was incorporated mainly between the old and new flagellum/FAZ complexes. Cytoskeletal modifications at the posterior end of the cells were observed with EB1, a microtubule plus end binding protein, particularly during mitosis. Additionally, the newly formed microtubules segregated asymmetrically, with the daughter cell inheriting the new flagellum/FAZ complex retaining most of the new microtubules. Together, our results suggest an intimate connection between new microtubule formation and new FAZ assembly, consequently leading to asymmetric microtubule inheritance and cell division.

  14. Recent Release of the ASTER Global DEM Product

    Science.gov (United States)

    Behnke, J.; Hall, A.; Meyer, D.; Sohre, T.; Doescher, C.

    2009-12-01

    On June 29th, the ASTER Global Digital Elevation Model (DEM) release was announced to the public and to a very eager audience. ASTER (Advanced Spaceborne Thermal Emission and Reflection Radiometer) is an imaging instrument flying on Terra, a satellite launched in December 1999 as part of NASA's Earth Observing System (EOS). ASTER is a cooperative effort between NASA, Japan's Ministry of Economy, Trade and Industry (METI) and Japan's Earth Remote Sensing Data Analysis Center (ERSDAC). On June 21, NASA Headquarters along with colleagues in Japan (METI) signed a plan for distribution of this product. The global digital elevation model of Earth is available online to users everywhere at no cost from NASA's Land Processes Distributed Active Archive Center (DAAC) located at Sioux Falls, SD. The DAAC is a joint project of NASA and the USGS and is a key component of NASA's EOSDIS. The new ASTER GDEM was created from nearly 1.3 million individual stereo-pair images acquired by the Japanese Advanced Spaceborne Thermal Emission and Reflection Radiometer (Aster) instrument aboard NASA’s Terra satellite. The ASTER elevation model was jointly developed by NASA and METI under contract to Sensor Information Laboratory Corp., Tsukuba, Japan. On June 29, the NASA press release was picked up quickly by numerous news organizations and online sites. Response to the product was incredible! The news of the release of the product was carried on websites across the globe, this fueled a tremendous response from users. Here are a few interesting metrics about the release: - over 41,000 unique visitors to website in first week following release - top countries in order were: US (approx. 20%), Germany, U.K., Brazil, Austria, Canada, Spain, Switzerland, Japan - approximately 29,000 visitors came to the news page in the first week and about 11,000 of these users downloaded the actual press release - by the end of August, over 2 Million ASTER GDEM files had been downloaded from the Land

  15. The plant cytoskeleton: recent advances in the study of the plant microtubule-associated proteins MAP-65, MAP-190 and the Xenopus MAP215-like protein, MOR1.

    Science.gov (United States)

    Hussey, Patrick J; Hawkins, Timothy J; Igarashi, Hisako; Kaloriti, Despina; Smertenko, Andrei

    2002-12-01

    The microtubule cytoskeleton is a dynamic filamentous structure involved in many key processes in plant cell morphogenesis including nuclear and cell division, deposition of cell wall, cell expansion, organelle movement and secretion. The principal microtubule protein is tubulin, which associates to form the wall of the tubule. In addition, various associated proteins bind microtubules either to anchor, cross-link or regulate the microtubule network within cells. Biochemical, molecular biological and genetic approaches are being successfully used to identify these microtubule-associated proteins (MAPs) in plants, and we describe recent progress on three of these proteins. PMID:12516862

  16. 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.

  17. Taking directions: the role of microtubule-bound nucleation in the self-organization of the plant cortical array

    International Nuclear Information System (INIS)

    The highly aligned cortical microtubule array of interphase plant cells is a key regulator of anisotropic cell expansion. Recent computational and analytical work has shown that the non-equilibrium self-organization of this structure can be understood on the basis of experimentally observed collisional interactions between dynamic microtubules attached to the plasma membrane. Most of these approaches assumed that new microtubules are homogeneously and isotropically nucleated on the cortical surface. Experimental evidence, however, shows that nucleation mostly occurs from other microtubules and under specific relative angles. Here, we investigate the impact of directed microtubule-bound nucleations on the alignment process using computer simulations. The results show that microtubule-bound nucleations can increase the degree of alignment achieved, decrease the timescale of the ordering process and widen the regime of dynamic parameters for which the system can self-organize. We establish that the major determinant of this effect is the degree of co-alignment of the nucleations with the parent microtubule. The specific role of sideways branching nucleations appears to allow stronger alignment while maintaining a measure of overall spatial homogeneity. Finally, we investigate the suggestion that observed persistent rotation of microtubule domains can be explained through a handedness bias in microtubule-bound nucleations, showing that this is possible only for an extreme bias and over a limited range of parameters

  18. 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.

  19. 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.

  20. A novel microtubule-modulating agent EM011 inhibits angiogenesis by repressing the HIF-1α axis and disrupting cell polarity and migration

    OpenAIRE

    Karna, Prasanthi; Rida, Padmashree C. G.; Turaga, Ravi Chakra; Gao, Jinmin; Gupta, Meenakshi; Fritz, Andreas; Werner, Erica; Yates, Clayton; Zhou, Jun; Aneja, Ritu

    2012-01-01

    Endothelial tubular morphogenesis relies on an exquisite interplay of microtubule dynamics and actin remodeling to propel directed cell migration. Recently, the dynamicity and integrity of microtubules have been implicated in the trafficking and efficient translation of the mRNA for HIF-1α (hypoxia-inducible factor), the master regulator of tumor angiogenesis. Thus, microtubule-disrupting agents that perturb the HIF-1α axis and neovascularization cascade are attractive anticancer drug candida...

  1. 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.

  2. Spatiotemporal control of microtubule nucleation and assembly using magnetic nanoparticles

    Science.gov (United States)

    Hoffmann, Céline; Mazari, Elsa; Lallet, Sylvie; Le Borgne, Roland; Marchi, Valérie; Gosse, Charlie; Gueroui, Zoher

    2013-03-01

    Decisions on the fate of cells and their functions are dictated by the spatiotemporal dynamics of molecular signalling networks. However, techniques to examine the dynamics of these intracellular processes remain limited. Here, we show that magnetic nanoparticles conjugated with key regulatory proteins can artificially control, in time and space, the Ran/RCC1 signalling pathway that regulates the cell cytoskeleton. In the presence of a magnetic field, RanGTP proteins conjugated to superparamagnetic nanoparticles can induce microtubule fibres to assemble into asymmetric arrays of polarized fibres in Xenopus laevis egg extracts. The orientation of the fibres is dictated by the direction of the magnetic force. When we locally concentrated nanoparticles conjugated with the upstream guanine nucleotide exchange factor RCC1, the assembly of microtubule fibres could be induced over a greater range of distances than RanGTP particles. The method shows how bioactive nanoparticles can be used to engineer signalling networks and spatial self-organization inside a cell environment.

  3. Identificación de metabasaltos alterados mediante datos ASTER

    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.

  4. ASTER temperature and emissivity validation on volcano Teide

    OpenAIRE

    Amici, Stefania; Istituto Nazionale di Geofisica e Vulcanologia, Sezione CNT, Roma, Italia; Piscini, Alessandro; Istituto Nazionale di Geofisica e Vulcanologia, Sezione CNT, Roma, Italia; Buongiorno, Fabrizia; Istituto Nazionale di Geofisica e Vulcanologia, Sezione CNT, Roma, Italia

    2010-01-01

    The Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER ) has operated since 19 December 1999 from NASA’s Terra Earth-orbiting, sun synchronous satellite. Emissivity and temperature standard products are based on the TES algorithms and require periodical validation campaign. In the frame of the EC project PREVIEW (http://www.preview-risk.com/) a field campaign on Volcano Teide was carried on, from the 16th to 24th of September 2007, to validate and to integrate the satellite...

  5. Using ASTER Stereo Images to Quantify Surface Roughness

    Science.gov (United States)

    Mushkin, Amit; Gillespie, Alan

    The unresolved topographic expression of surfaces, surface roughness (SR), is a fundamental surface property that conveys useful information for a wide range of Earth and planetary sciences. Yet, this information is difficult to measure remotely because most spaceborne imagers have resolutions on the order of meters to hundreds of meters and SR can vary significantly below these scales. One way to measure SR is to exploit differential shadowing in stereo images, and in particular, Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER), with its nadir (3N) and backward-looking (3B) near-infrared channels. We have proposed a simple ratio of land-leaving radiance in those two channels as a measure of relative SR at scales photo-interpretation. Moreover, one could calibrate the ratio data to physical parameters, such as RMS height, and translate to SR maps at 15 m/pixel resolution. Two calibration schemes enable this translation: empirical calibration against independent in situ roughness measurements and model-based calibration against forward simulations of ­two-look ratios from very high-resolution (<5 mm) digital elevation models of natural ­surfaces, measured with a ground-based light detection and ranging ­system. Here, we focus on the latter scheme that enables construction of calibration curves for any given viewing and illumination geometries encountered by ASTER. ASTER now ­provides a global archive of images, and the two-look approach with ASTER stereo data enables a unique quantitative mapping capability of SR at 15 m/pixel spatial resolution for almost anywhere on Earth.

  6. Mitosis. Microtubule detyrosination guides chromosomes during mitosis.

    Science.gov (United States)

    Barisic, Marin; Silva e Sousa, Ricardo; Tripathy, Suvranta K; Magiera, Maria M; Zaytsev, Anatoly V; Pereira, Ana L; Janke, Carsten; Grishchuk, Ekaterina L; Maiato, Helder

    2015-05-15

    Before chromosomes segregate into daughter cells, they align at the mitotic spindle equator, a process known as chromosome congression. Centromere-associated protein E (CENP-E)/Kinesin-7 is a microtubule plus-end-directed kinetochore motor required for congression of pole-proximal chromosomes. Because the plus-ends of many astral microtubules in the spindle point to the cell cortex, it remains unknown how CENP-E guides pole-proximal chromosomes specifically toward the equator. We found that congression of pole-proximal chromosomes depended on specific posttranslational detyrosination of spindle microtubules that point to the equator. In vitro reconstitution experiments demonstrated that CENP-E-dependent transport was strongly enhanced on detyrosinated microtubules. Blocking tubulin tyrosination in cells caused ubiquitous detyrosination of spindle microtubules, and CENP-E transported chromosomes away from spindle poles in random directions. Thus, CENP-E-driven chromosome congression is guided by microtubule detyrosination. PMID:25908662

  7. 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.

  8. Short Stop provides an essential link between F-actin and microtubules during axon extension.

    Science.gov (United States)

    Lee, Seungbok; Kolodziej, Peter A

    2002-03-01

    Coordination of F-actin and microtubule dynamics is important for cellular motility and morphogenesis, but little is known about underlying mechanisms. short stop (shot) encodes an evolutionarily conserved, neuronally expressed family of rod-like proteins required for sensory and motor axon extension in Drosophila melanogaster. We identify Shot isoforms that contain N-terminal F-actin and C-terminal microtubule-binding domains, and that crosslink F-actin and microtubules in cultured cells. The F-actin- and microtubule-binding domains of Shot are required in the same molecule for axon extension, though the length of the connecting rod domain can be dramatically reduced without affecting activity. Shot therefore functions as a cytoskeletal crosslinker in axon extension, rather than mediating independent interactions with F-actin and microtubules. A Ca(2+)-binding motif located adjacent to the microtubule-binding domain is also required for axon extension, suggesting that intracellular Ca(2+) release may regulate Shot activity. These results suggest that Shot coordinates regulated interactions between F-actin and microtubules that are crucial for neuronal morphogenesis. PMID:11874915

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

    Directory of Open Access Journals (Sweden)

    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.

  10. 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.

  11. 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.

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

    Directory of Open Access Journals (Sweden)

    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.

  13. Microtubule nucleation and organization in dendrites.

    Science.gov (United States)

    Delandre, Caroline; Amikura, Reiko; Moore, Adrian W

    2016-07-01

    Dendrite branching is an essential process for building complex nervous systems. It determines the number, distribution and integration of inputs into a neuron, and is regulated to create the diverse dendrite arbor branching patterns characteristic of different neuron types. The microtubule cytoskeleton is critical to provide structure and exert force during dendrite branching. It also supports the functional requirements of dendrites, reflected by differential microtubule architectural organization between neuron types, illustrated here for sensory neurons. Both anterograde and retrograde microtubule polymerization occur within growing dendrites, and recent studies indicate that branching is enhanced by anterograde microtubule polymerization events in nascent branches. The polarities of microtubule polymerization events are regulated by the position and orientation of microtubule nucleation events in the dendrite arbor. Golgi outposts are a primary microtubule nucleation center in dendrites and share common nucleation machinery with the centrosome. In addition, pre-existing dendrite microtubules may act as nucleation sites. We discuss how balancing the activities of distinct nucleation machineries within the growing dendrite can alter microtubule polymerization polarity and dendrite branching, and how regulating this balance can generate neuron type-specific morphologies. PMID:27097122

  14. Discrepancy Between ASTER- and MODIS- Derived Land Surface Temperatures: Terrain Effects

    OpenAIRE

    Yasushi Yamaguchi; Yuanbo Liu; Yousuke Noumi

    2009-01-01

    The MODerate resolution Imaging Spectroradiometer (MODIS) and the Advanced Spaceborne Thermal Emission Reflection Radiometer (ASTER) are onboard the same satellite platform NASA TERRA. Both MODIS and ASTER offer routine retrieval of land surface temperatures (LSTs), and the ASTER- and MODIS-retrieved LST products have been used worldwide. Because a large fraction of the earth surface consists of mountainous areas, variations in elevation, terrain slope and aspect angles can cause biases in th...

  15. 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.

  16. Chromosome position at the spindle equator is regulated by chromokinesin and a bipolar microtubule array.

    Science.gov (United States)

    Takagi, Jun; Itabashi, Takeshi; Suzuki, Kazuya; Ishiwata, Shin'ichi

    2013-01-01

    The chromosome alignment is mediated by polar ejection and poleward forces acting on the chromosome arm and kinetochores, respectively. Although components of the motile machinery such as chromokinesin have been characterized, their dynamics within the spindle is poorly understood. Here we show that a quantum dot (Qdot) binding up to four Xenopus chromokinesin (Xkid) molecules behaved like a nanosize chromosome arm in the meiotic spindle, which is self-organized in cytoplasmic egg extracts. Xkid-Qdots travelled long distances along microtubules by changing several tracks, resulting in their accumulation toward and distribution around the metaphase plate. The analysis indicated that the direction of motion and velocity depend on the distribution of microtubule polarity within the spindle. Thus, this mechanism is governed by chromokinesin motors, which is dependent on symmetrical microtubule orientation that may allow chromosomes to maintain their position around the spindle equator until correct microtubule-kinetochore attachment is established. PMID:24077015

  17. Cellulose-Microtubule Uncoupling Proteins Prevent Lateral Displacement of Microtubules during Cellulose Synthesis in Arabidopsis.

    Science.gov (United States)

    Liu, Zengyu; Schneider, Rene; Kesten, Christopher; Zhang, Yi; Somssich, Marc; Zhang, Youjun; Fernie, Alisdair R; Persson, Staffan

    2016-08-01

    Cellulose is the most abundant biopolymer on Earth and is the major contributor to plant morphogenesis. Cellulose is synthesized by plasma membrane-localized cellulose synthase complexes (CSCs). Nascent cellulose microfibrils become entangled in the cell wall, and further catalysis therefore drives the CSC forward through the membrane: a process guided by cortical microtubules via the protein CSI1/POM2. Still, it is unclear how the microtubules can withstand the forces generated by the motile CSCs to effectively direct CSC movement. Here, we identified a family of microtubule-associated proteins, the cellulose synthase-microtubule uncouplings (CMUs), that located as static puncta along cortical microtubules. Functional disruption of the CMUs caused lateral microtubule displacement and compromised microtubule-based guidance of CSC movement. CSCs that traversed the microtubules interacted with the microtubules via CSI1/POM2, which prompted the lateral microtubule displacement. Hence, we have revealed how microtubules can withstand the propulsion of the CSCs during cellulose biosynthesis and thus sustain anisotropic plant cell growth. PMID:27477947

  18. Dilution of individual microtubules observed in real time in vitro: evidence that cap size is small and independent of elongation rate

    OpenAIRE

    1991-01-01

    Although the mechanism of microtubule dynamic instability is thought to involve the hydrolysis of tubulin-bound GTP, the mechanism of GTP hydrolysis and the basis of microtubule stability are controversial. Video microscopy of individual microtubules and dilution protocols were used to examine the size and lifetime of the stabilizing cap. Purified porcine brain tubulin (7-23 microM) was assembled at 37 degrees C onto both ends of isolated sea urchin axoneme fragments in a miniature flow cell ...

  19. 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. PMID:25576435

  20. 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.

  1. 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.

  2. Microtubule networks for plant cell division.

    Science.gov (United States)

    de Keijzer, Jeroen; Mulder, Bela M; Janson, Marcel E

    2014-09-01

    During cytokinesis the cytoplasm of a cell is divided to form two daughter cells. In animal cells, the existing plasma membrane is first constricted and then abscised to generate two individual plasma membranes. Plant cells on the other hand divide by forming an interior dividing wall, the so-called cell plate, which is constructed by localized deposition of membrane and cell wall material. Construction starts in the centre of the cell at the locus of the mitotic spindle and continues radially towards the existing plasma membrane. Finally the membrane of the cell plate and plasma membrane fuse to form two individual plasma membranes. Two microtubule-based cytoskeletal networks, the phragmoplast and the pre-prophase band (PPB), jointly control cytokinesis in plants. The bipolar microtubule array of the phragmoplast regulates cell plate deposition towards a cortical position that is templated by the ring-shaped microtubule array of the PPB. In contrast to most animal cells, plants do not use centrosomes as foci of microtubule growth initiation. Instead, plant microtubule networks are striking examples of self-organizing systems that emerge from physically constrained interactions of dispersed microtubules. Here we will discuss how microtubule-based activities including growth, shrinkage, severing, sliding, nucleation and bundling interrelate to jointly generate the required ordered structures. Evidence mounts that adapter proteins sense the local geometry of microtubules to locally modulate the activity of proteins involved in microtubule growth regulation and severing. Many of the proteins and mechanisms involved have roles in other microtubule assemblies as well, bestowing broader relevance to insights gained from plants. PMID:25136380

  3. 50 ways to build a spindle: the complexity of microtubule generation during mitosis.

    Science.gov (United States)

    Duncan, Tommy; Wakefield, James G

    2011-04-01

    The accurate segregation of duplicated chromosomes, essential for the development and viability of a eukaryotic organism, requires the formation of a robust microtubule (MT)-based spindle apparatus. Entry into mitosis or meiosis precipitates a cascade of signalling events which result in the activation of pathways responsible for a dramatic reorganisation of the MT cytoskeleton: through changes in the properties of MT-associated proteins, local concentrations of free tubulin dimer and through enhanced MT nucleation. The latter is generally thought to be driven by localisation and activation of γ-tubulin-containing complexes (γ-TuSC and γ-TuRC) at specific subcellular locations. For example, upon entering mitosis, animal cells concentrate γ-tubulin at centrosomes to tenfold the normal level during interphase, resulting in an aster-driven search and capture of chromosomes and bipolar mitotic spindle formation. Thus, in these cells, centrosomes have traditionally been perceived as the primary microtubule organising centre during spindle formation. However, studies in meiotic cells, plants and cell-free extracts have revealed the existence of complementary mechanisms of spindle formation, mitotic chromatin, kinetochores and nucleation from existing MTs or the cytoplasm can all contribute to a bipolar spindle apparatus. Here, we outline the individual known mechanisms responsible for spindle formation and formulate ideas regarding the relationship between them in assembling a functional spindle apparatus. PMID:21484448

  4. 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

  5. Talin-KANK1 interaction controls the recruitment of cortical microtubule stabilizing complexes to focal adhesions

    Science.gov (United States)

    Bouchet, Benjamin P; Gough, Rosemarie E; Ammon, York-Christoph; van de Willige, Dieudonnée; Post, Harm; Jacquemet, Guillaume; Altelaar, AF Maarten; Heck, Albert JR; Goult, Benjamin T; Akhmanova, Anna

    2016-01-01

    The cross-talk between dynamic microtubules and integrin-based adhesions to the extracellular matrix plays a crucial role in cell polarity and migration. Microtubules regulate the turnover of adhesion sites, and, in turn, focal adhesions promote the cortical microtubule capture and stabilization in their vicinity, but the underlying mechanism is unknown. Here, we show that cortical microtubule stabilization sites containing CLASPs, KIF21A, LL5β and liprins are recruited to focal adhesions by the adaptor protein KANK1, which directly interacts with the major adhesion component, talin. Structural studies showed that the conserved KN domain in KANK1 binds to the talin rod domain R7. Perturbation of this interaction, including a single point mutation in talin, which disrupts KANK1 binding but not the talin function in adhesion, abrogates the association of microtubule-stabilizing complexes with focal adhesions. We propose that the talin-KANK1 interaction links the two macromolecular assemblies that control cortical attachment of actin fibers and microtubules. DOI: http://dx.doi.org/10.7554/eLife.18124.001 PMID:27410476

  6. 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.

  7. 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.

  8. Prediction of landslides using ASTER imagery and data mining models

    Science.gov (United States)

    Song, Kyo-Young; Oh, Hyun-Joo; Choi, Jaewon; Park, Inhye; Lee, Changwook; Lee, Saro

    2012-03-01

    The aim of this study was to identify landslide-related factors using only remotely sensed data and to present landslide susceptibility maps using a geographic information system, data-mining models, an artificial neural network (ANN), and an adaptive neuro-fuzzy interface system (ANFIS). Landslide-related factors were identified in Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) satellite imagery. The slope, aspect, and curvature of topographic features were calculated from a digital elevation model that was made using the ASTER imagery. Lineaments, land-cover, and normalized difference vegetative index layers were also extracted from the imagery. Landslide-susceptible areas were analyzed and mapped based on occurrence factors using the ANN and ANFIS. The generalized bell-shaped built-in membership function of the ANFIS was applied to landslide susceptibility mapping. Analytical results were validated using landslide test location data. In the validation results, the ANN model showed 80.42% prediction accuracy and the ANFIS model showed 86.55% prediction accuracy. These results suggest that the ANFIS model has a better performance than does the ANN in predicting landslide susceptibility.

  9. 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...

  10. 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

  11. 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...

  12. Stereo Pair with ASTER Image, Iturralde Structure, Bolivia

    Science.gov (United States)

    2002-01-01

    An 8-kilometer (5-mile) wide crater of possible impact origin is shown in this stereoscopic view of an isolated part of the Bolivian Amazon. The view is derived from an Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) satellite image and a Shuttle Radar Topography Mission (SRTM) elevation model. The circular feature covering much of the image, known as the Iturralde Structure, is possibly the Earth's most recent 'big' impact event recording collision with a meteor or comet that might have occurred between 11,000 and 30,000 years ago.Although the structure was identified on satellite photographs in the mid-1980s, its location is so remote that it has only been visited by scientific investigators twice, most recently by a team from NASA's Goddard Space Flight Center in September 2002. Lying in an area of very low relief, the landform is a quasi-circular closed depression only about 20 meters (66 feet) in depth, with sharply defined sub-angular 'rim' materials. It resembles a 'cookie cutter' in that its appearance 'cuts' the heavily vegetated soft-sediments and pampas of this part of Bolivia. The SRTM data have provided investigators with the first topographic map of the site and will allow studies of its three-dimensional structure crucial to determining whether it actually is of impact origin.This stereoscopic image was generated by first draping the ASTER satellite image over the Shuttle Radar Topography Mission digital elevation model. Two differing perspectives were then calculated, one for each eye. They can be seen in 3-D by viewing the left image with the right eye and the right image with the left eye (cross-eyed viewing) or by downloading and printing the image pair and viewing them with a stereoscope. When stereoscopically merged, the result is a vertically exaggerated view of Earth's surface in its full three dimensions.Thick vegetation in part defines the surface that the SRTM radar sees as it maps the terrain. Much of the local

  13. 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)

  14. 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.

  15. 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

  16. 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.

  17. Three-dimensional tracking of plus-tips by lattice light-sheet microscopy permits the quantification of microtubule growth trajectories within the mitotic apparatus

    Science.gov (United States)

    Yamashita, Norio; Morita, Masahiko; Legant, Wesley R.; Chen, Bi-Chang; Betzig, Eric; Yokota, Hideo; Mimori-Kiyosue, Yuko

    2015-10-01

    Mitotic apparatus, which comprises hundreds of microtubules, plays an essential role in cell division, ensuring the correct segregation of chromosomes into each daughter cell. To gain insight into its regulatory mechanisms, it is essential to detect and analyze the behavior of individual microtubule filaments. However, the discrimination of discrete microtubule filaments within the mitotic apparatus is beyond the capabilities of conventional light microscopic technologies. Recently, we detected three-dimensional (3-D) microtubule growth dynamics within the cellular cytoplasmic space using lattice light-sheet microscopy in conjunction with microtubule growth marker protein end-binding 1, a microtubule plus-end-tracking protein, which was fused to green fluorescent protein (EB1-GFP). This technique enables high-resolution 3-D imaging at subsecond intervals. We adapted mathematical computing and geometric representation techniques to analyze spatial variations in microtubule growth dynamics within the mitotic spindle apparatus. Our analytical approach enabled the different dynamic properties of individual microtubules to be determined, including the direction and speed of their growth, and their growth duration within a 3-D spatial map. Our analysis framework provides an important step toward a more comprehensive understanding of the mechanisms driving cellular machinery at the whole-cell level.

  18. Ship extraction and categorization from ASTER VNIR imagery

    Science.gov (United States)

    Partsinevelos, Panagiotis; Miliaresis, George

    2014-08-01

    We present a methodology for ship extraction and categorization from relatively low resolution multispectral ASTER imagery, corresponding to the sea region south east of Athens in Greece. At a first level, in the radiometrically corrected image, quad tree decomposition and bounding rectangular extraction automatically outline location of objects - possible ships, by statistically evaluating spectral responses throughout the segmented image. Subsequently, the object borders within the rectangular regions are extracted, while connected component labelling combined by size and shape filtering allows ship characterization. The ships' spectral signature is determined in green, red and infrared bands while cluster analysis allows the identification of ship categories on the basis of their size and reflectance. Additional pixel- based measures reveal estimated ship orientation, direction, movement, stability and turning. The results are complemented with additional geographic information and inference tools are formed towards the determination of probable ship type and its destination.

  19. 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.

  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. 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.

  2. Retrieval of a Temporal High-Resolution Leaf Area Index (LAI by Combining MODIS LAI and ASTER Reflectance Data

    Directory of Open Access Journals (Sweden)

    Yonghua Qu

    2014-12-01

    Full Text Available This paper aims to retrieve temporal high-resolution LAI derived by fusing MOD15 products (1 km resolution, field-measured LAI and ASTER reflectance (15-m resolution. Though the inversion of a physically based canopy reflectance model using high-resolution satellite data can produce high-resolution LAI products, the obstacle to producing temporal products is obvious due to the low temporal resolution of high resolution satellite data. A feasible method is to combine different source data, taking advantage of the spatial and temporal resolution of different sensors. In this paper, a high-resolution LAI retrieval method was implemented using a dynamic Bayesian network (DBN inversion framework. MODIS LAI data with higher temporal resolution were used to fit the temporal background information, which is then updated by new, higher resolution data, herein ASTER data. The interactions between the different resolution data were analyzed from a Bayesian perspective. The proposed method was evaluated using a dataset collected in the HiWater (Heihe Watershed Allied Telemetry Experimental Research experiment. The determination coefficient and RMSE between the estimated and measured LAI are 0.80 and 0.43, respectively. The research results suggest that even though the coarse-resolution background information differs from the high-resolution satellite observations, a satisfactory estimation result for the temporal high-resolution LAI can be produced using the accumulated information from both the new observations and background information.

  3. 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.

  4. 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...

  5. 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...

  6. 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)...

  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 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)...

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

    OpenAIRE

    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 ...

  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. 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.

  13. 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.

  14. Observation of microtubule-based motor protein activity.

    Science.gov (United States)

    Sloboda, Roger D

    2015-02-01

    It is possible to detect the presence of motor proteins that have the ability to translocate particles along microtubules. The two procedures described here were developed to detect microtubule-dependent motor protein activity in cell lysates or of purified proteins. In the first procedure, latex beads bound to the putative motor protein are assayed for their ability to translocate along microtubules in an ATP-dependent fashion. If motor protein activity is present, it will bind to the beads and translocate them unidirectionally along the microtubules. In the second procedure, motor proteins induce microtubule gliding over a glass coverslip surface that is coated with active motor protein. Because the mass of a microtubule is negligible compared to that of a coverslip or slide, the microtubule glides over the glass surface when the surface is coated with active motor protein. Also included here are descriptions of assays designed to determine the directionality of movement of microtubule-based motor proteins. PMID:25646501

  15. Hepatic microtubule acetylation and stability induced by chronic alcohol exposure impair nuclear translocation of STAT3 and STAT5B, but not Smad2/3.

    Science.gov (United States)

    Fernandez, David J; Tuma, Dean J; Tuma, Pamela L

    2012-12-15

    Although alcoholic liver disease is clinically well described, the molecular basis for alcohol-induced hepatotoxicity is not well understood. Previously, we found that alcohol exposure led to increased microtubule acetylation and stability in polarized, hepatic WIF-B cells and in livers from ethanol-fed rats. Because microtubules are known to regulate transcription factor nuclear translocation and dynamic microtubules are required for translocation of at least a subset of these factors, we examined whether alcohol-induced microtubule acetylation and stability impair nuclear translocation. We examined nuclear delivery of factors representing the two mechanisms by which microtubules regulate translocation. To represent factors that undergo directed delivery, we examined growth hormone-induced STAT5B translocation and IL-6-induced STAT3 translocation. To represent factors that are sequestered in the cytoplasm by microtubule attachment until ligand activation, we examined transforming growth factor-β-induced Smad2/3 translocation. We found that ethanol exposure selectively impaired translocation of the STATs, but not Smad2/3. STAT5B delivery was decreased to a similar extent by addition of taxol (a microtubule-stabilizing drug) or trichostatin A (a deacetylase inhibitor), agents that promote microtubule acetylation in the absence of alcohol. Thus the alcohol-induced impairment of STAT nuclear translocation can be explained by increased microtubule acetylation and stability. Only ethanol treatment impaired STAT5B activation, indicating that microtubules are not important for its activation by Jak2. Furthermore, nuclear exit was not changed in treated cells, indicating that this process is also independent of microtubule acetylation and stability. Together, these results raise the exciting possibility that deacetylase agonists may be effective therapeutics for the treatment of alcoholic liver disease. PMID:23064763

  16. Microtubule detyrosination guides chromosomes during mitosis

    OpenAIRE

    Barisic, Marin; Silva e Sousa, Ricardo; Tripathy, Suvranta K.; Magiera, Maria M.; Zaytsev, Anatoly V.; Pereira, Ana L.; Janke, Carsten; Grishchuk, Ekaterina L.; Maiato, Helder

    2015-01-01

    Before chromosomes segregate into daughter cells they align at the mitotic spindle equator, a process known as chromosome congression. CENP-E/Kinesin-7 is a microtubule plus-end-directed kinetochore motor required for congression of pole-proximal chromosomes. Because the plus-ends of many astral microtubules in the spindle point to the cell cortex, it remains unknown how CENP-E guides pole-proximal chromosomes specifically towards the equator. Here we found that congression of pole-proximal c...

  17. Remote sensing of volcanic plumes using the Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER)

    Science.gov (United States)

    Henney, Lorna Alison

    The Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) has been used to quantify SO2 emissions from passively degassing volcanoes. This dissertation explores ASTER's capability to detect SO 2 with satellite validation, enhancement techniques and extensive processing of images at a variety of volcanoes. ASTER is compared to the Mini UV Spectrometer (MUSe), a ground based instrument, to determine if reasonable SO2 fluxes can be quantified from a plume emitted from Lascar, Chile. The two sensors were in good agreement with ASTER proving to be a reliable detector of SO2. ASTER illustrated the advantages of imaging a plume in 2D, with better temporal resolution than the MUSe. SO2 plumes in ASTER imagery are not always discernible in the raw TIR data. Principal Component Analysis (PCA) and Decorrelation Stretch (DCS) enhancement techniques were compared to determine how well they highlight a variety of volcanic plumes. DCS produced a consistent output and the composition of the plumes was easy to identify from explosive eruptions. As the plumes became smaller and lower in altitude they became harder to distinguish using DCS. PCA proved to be better at identifying smaller low altitude plumes. ASTER was used to investigate SO2 emissions at Lascar, Chile. Activity at Lascar has been characterized by cyclic behavior and persistent degassing (Matthews et al. 1997). Previous studies at Lascar have primarily focused on changes in thermal infrared anomalies, neglecting gas emissions. Using the SO2 data along with changes in thermal anomalies and visual observations it is evident that Lascar is at the end an eruptive cycle that began in 1993. Declining gas emissions and crater temperatures suggest that the conduit is sealing. ASTER and the Ozone Monitoring Instrument (OMI) were used to determine the annual contribution of SO2 to the troposphere from the Central and South American volcanic arcs between 2000 and 2011. Fluxes of 3.4 Tg/a for Central America and 3

  18. Validation of ASTER Surface Temperature Data with In Situ Measurements to Evaluate Heat Islands in Complex Urban Areas

    OpenAIRE

    Bonggeun Song; Kyunghun Park

    2014-01-01

    This study compared Advanced Spaceborne Thermal Emission Reflection Radiometer (ASTER) surface temperature data with in situ measurements to validate the use of ASTER data for studying heat islands in urban settings with complex spatial characteristics. Eight sites in Changwon, Korea, were selected for analyses. Surface temperature data were extracted from the thermal infrared (TIR) band of ASTER on four dates during the summer and fall of 2012, and corresponding in situ measurements of tempe...

  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. What generates flux of tubulin in kinetochore microtubules?

    Science.gov (United States)

    Forer, Arthur; Pickett-Heaps, Jeremy D; Spurck, Tim

    2008-01-01

    We discuss models for production of tubulin flux in kinetochore microtubules. Current models concentrate solely on microtubules and their associated motors and enzymes. For example, in some models the driving force for flux is enzymes at the poles and the kinetochores; in others the driving force is motor molecules that are associated with a stationary spindle matrix. We present a different viewpoint, that microtubules are propelled poleward by forces arising from the spindle matrix, that the forces on the microtubules "activate" polymerising and depolymerising enzymes at kinetochores and poles, that matrix forces utilise actin, myosin, and microtubule motors, and that the matrix itself may not necessarily be static. PMID:18421550

  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. The role of microtubule movement in bidirectional organelle transport

    CERN Document Server

    Kulić, Igor M; Kim, Hwajin; Kural, Comert; Blehm, Benjamin; Selvin, Paul R; Nelson, Philip C; Gelfand, Vladimir I

    2008-01-01

    We study the role of microtubule movement in bidirectional organelle transport in Drosophila S2 cells and show that EGFP-tagged peroxisomes in cells serve as sensitive probes of motor induced, noisy cytoskeletal motions. Multiple peroxisomes move in unison over large time windows and show correlations with microtubule tip positions, indicating rapid microtubule fluctuations in the longitudinal direction. We report the first high-resolution measurement of longitudinal microtubule fluctuations performed by tracing such pairs of co-moving peroxisomes. The resulting picture shows that motor-dependent longitudinal microtubule oscillations contribute significantly to cargo movement along microtubules. Thus, contrary to the conventional view, organelle transport cannot be described solely in terms of cargo movement along stationary microtubule tracks, but instead includes a strong contribution from the movement of the tracks.

  3. Perturbing microtubule integrity blocks AMP-activated protein kinase-induced meiotic resumption in cultured mouse oocytes.

    Science.gov (United States)

    Ya, Ru; Downs, Stephen M

    2014-02-01

    The oocyte meiotic spindle is comprised of microtubules (MT) that bind chromatin and regulate both metaphase plate formation and karyokinesis during meiotic maturation; however, little information is known about their role in meiosis reinitiation. This study was conducted to determine if microtubule integrity is required for meiotic induction and to ascertain how it affects activation of AMP-activated protein kinase (AMPK), an important participant in the meiotic induction process. Treatment with microtubule-disrupting agents nocodazole and vinblastine suppressed meiotic resumption in a dose-dependent manner in both arrested cumulus cell-enclosed oocytes (CEO) stimulated with follicle-stimulating hormone (FSH) and arrested denuded oocytes (DO) stimulated with the AMPK activator, 5-aminoimidazole-4-carboxamide-1-beta-4-ribofuranoside (AICAR). This effect coincided with suppression of AMPK activation as determined by western blotting and germinal vesicle immunostaining. Treatment with the MT stabilizer paclitaxel also suppressed meiotic induction. Targeting actin filament polymerization had only a marginal effect on meiotic induction. Immunolocalization experiments revealed that active AMPK colocalized with γ-tubulin during metaphase I and II stages, while it localized at the spindle midzone during anaphase. This discrete localization pattern was dependent on MT integrity. Treatment with nocodazole led to disruption of proper spindle pole localization of active AMPK, while paclitaxel induced excessive polymerization of spindle MT and formation of ectopic asters with accentuated AMPK colocalization. Although stimulation of AMPK increased the rate of germinal vesicle breakdown (GVB), spindle formation and polar body (PB) extrusion, the kinase had no effect on peripheral movement of the spindle. These data suggest that the meiosis-inducing action and localization of AMPK are regulated by MT spindle integrity during mouse oocyte maturation. PMID:23199370

  4. 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.

  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. PMID:27238282

  6. Cytoskeletal logic: a model for molecular computation via Boolean operations in microtubules and microtubule-associated proteins.

    Science.gov (United States)

    Lahoz-Beltra, R; Hameroff, S R; Dayhoff, J E

    1993-01-01

    Adaptive behaviors and dynamic activities within living cells are organized by the cytoskeleton: intracellular networks of interconnected protein polymers which include microtubules (MTs), actin, intermediate filaments, microtubule associated proteins (MAPs) and other protein structures. Cooperative interactions among cytoskeletal protein subunit conformational states have been used to model signal transmission and information processing. In the present work we present a theoretical model for molecular computing in which Boolean logic is implemented in parallel networks of individual MTs interconnected by MAPs. Conformational signals propagate on MTs as in data buses and in the model MAPs are considered as Boolean operators, either as bit-lines (like MTs) where a signal can be transported unchanged between MTs ('BUS-MAP'), or as bit-lines where a Boolean operation is performed in one of the two MAP-MT attachments ('LOGIC-MAP'). Three logic MAPs have been defined ('NOT-MAP, 'AND-MAP', 'XOR-MAP') and used to demonstrate addition, subtraction and other arithmetic operations. Although our choice of Boolean logic is arbitrary, the simulations demonstrate symbolic manipulation in a connectionist system and suggest that MT-MAP networks can perform computation in living cells and are candidates for future molecular computing devices. PMID:8318677

  7. Use of ASTER and MODIS thermal infrared data to quantify heat flow and hydrothermal change at Yellowstone National Park

    Science.gov (United States)

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

    2012-01-01

    The overarching aim of this study was to use satellite thermal infrared (TIR) remote sensing to monitor geothermal activity within the Yellowstone geothermal area to meet the missions of both the U.S. Geological Survey and the Yellowstone National Park Geology Program. Specific goals were to: 1) address the challenges of monitoring the surface thermal characteristics of the > 10,000 spatially and temporally dynamic thermal features in the Park (including hot springs, pools, geysers, fumaroles, and mud pots) that are spread out over ~ 5000 km2, by using satellite TIR remote sensing tools (e.g., ASTER and MODIS), 2) to estimate the radiant geothermal heat flux (GHF) for Yellowstone's thermal areas, and 3) to identify normal, background thermal changes so that significant, abnormal changes can be recognized, should they ever occur (e.g., changes related to tectonic, hydrothermal, impending volcanic processes, or human activities, such as nearby geothermal development). ASTER TIR data (90-m pixels) were used to estimate the radiant GHF from all of Yellowstone's thermal features and update maps of thermal areas. MODIS TIR data (1-km pixels) were used to record background thermal radiance variations from March 2000 through December 2010 and establish thermal change detection limits. A lower limit for the radiant GHF estimated from ASTER TIR temperature data was established at ~ 2.0 GW, which is ~ 30–45% of the heat flux estimated through geochemical thermometry. Also, about 5 km2 of thermal areas was added to the geodatabase of mapped thermal areas. A decade-long time-series of MODIS TIR radiance data was dominated by seasonal cycles. A background subtraction technique was used in an attempt to isolate variations due to geothermal changes. Several statistically significant perturbations were noted in the time-series from Norris Geyser Basin, however many of these did not correspond to documented thermal disturbances. This study provides concrete examples of the

  8. 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).

  9. Discrimination of iron ore deposits of granulite terrain of Southern Peninsular India using ASTER data

    Science.gov (United States)

    Rajendran, Sankaran; Thirunavukkarasu, A.; Balamurugan, G.; Shankar, K.

    2011-04-01

    This work describes a new image processing technique for discriminating iron ores (magnetite quartzite deposits) and associated lithology in high-grade granulite region of Salem, Southern Peninsular India using visible, near-infrared and short wave infrared reflectance data of Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER). Image spectra show that the magnetite quartzite and associated lithology of garnetiferrous pyroxene granulite, hornblende biotite gneiss, amphibolite, dunite, and pegmatite have absorption features around spectral bands 1, 3, 5, and 7. ASTER band ratios ((1 + 3)/2, (3 + 5)/4, (5 + 7)/6) in RGB are constructed by summing the bands representing the shoulders of absorption features as a numerator, and the band located nearest the absorption feature as a denominator to map iron ores and band ratios ((2 + 4)/3, (5 + 7)/6, (7 + 9)/8) in RGB for associated lithology. The results show that ASTER band ratios ((1 + 3)/2, (3 + 5)/4, (5 + 7)/6) in a Red-Green-Blue (RGB) color combination identifies the iron ores much better than previously published ASTER band ratios analysis. A Principal Component Analysis (PCA) is applied to reduce redundant information in highly correlated bands. PCA (3, 2, and 1 for iron ores and 5, 4, 2 for granulite rock) in RGB enabled the discrimination between the iron ores and garnetiferrous pyroxene granulite rock. Thus, this image processing technique is very much suitable for discriminating the different types of rocks of granulite region. As outcome of the present work, the geology map of Salem region is provided based on the interpretation of ASTER image results and field verification work. It is recommended that the proposed methods have great potential for mapping of iron ores and associated lithology of granulite region with similar rock units of granulite regions of Southern Peninsular India. This work also demonstrates the ability of ASTER's to provide information on iron ores, which is valuable

  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. 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.

  12. 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.

  13. The Role of Microtubule End Binding (EB) Proteins in Ciliogenesis

    DEFF Research Database (Denmark)

    Schrøder, Jacob Morville

    EB1 is a small microtubule (MT)-binding protein that associates preferentially with MT plus ends. EB1 plays a role in regulating MT dynamics, localizing other MT-associated proteins to the plus end, and in regulating interactions of MTs with the cell cortex, mitotic kinetochores and different...... cellular organelles (Lansbergen and Akhmanova, 2006). EB1 also localizes to centrosomes and is required for centrosomal MT anchoring and organization of the MT network (Askham et al., 2002). Further, EB1 has been shown to localize to the flagellar tip and proximal region of the basal bodies......, are required for assembly of primary cilia in cultured human cells. The EB3 - siRNA ciliary phenotype could be rescued by GFP-EB1 expression, and GFP-EB3 over expression resulted in elongated cilia. Transmission electron microscopy (TEM) revealed that EB3-depleted cells possess stumpy cilia, a disorganized...

  14. 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.

  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. Self-Reduction Rate of a Microtubule

    OpenAIRE

    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 reduce to one of the classical states without measurement if certain conditions are satisfied(self-reductions), and calculate the self-reduction time $\\tau_N$ (the mean interval between two successive self-reductions) of a cluster consisting of more than $N$ neighboring tubulins (basic units composing a mic...

  18. 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。

  19. Observations of microtubules and microtubule-microfilament associations in osmotically treated cells of Micrasterias denticulata Bréb.

    Science.gov (United States)

    Neuhaus-Url, G; Kiermayer, O

    1982-06-01

    As an extension of the observation and interpretation regarding the different microtubule systems of Micrasterias denticulata [12, 19], the existence of intertubular structures, such as microfilaments, which are strongly marked in osmotically treated cells, is especially interesting. The complex of microtubules and microfilaments occurs during post-telophase nuclear migration, probably engaged in the mechanism of movement. The arrangement of microtubules either parallel or perpendicular to the nuclear membrane is characteristic for the stage of nuclear migration. Another microtubule system, the microtubule band in the cortical protoplasm of the isthmus region [12], is described during morphogenesis of the new half cell. Osmotically treated cells in the stage of septum formation demonstrate the presence of cross-linked microtubules near the plasmalemma and microtubule bundles, situated in the protoplasm between the secondary wall and the chloroplast, probably representing the microtubule system in the cortical protoplasm of the old half cell described by Kiermayer [12, 16]. The frequent appearance of microtubules and intertubular structures in differentiating cells of Micrasterias denticulata after osmotic treatment is discussed along with implication for stabilization of microtubules, cross bridges, and microfilaments. PMID:6889505

  20. MICROTUBULE ORGANIZATION 1 Regulates Structure and Function of Microtubule Arrays during Mitosis and Cytokinesis in the Arabidopsis Root1[W

    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°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 mor1-1L174F protein loses function without dissociating from microtubules, providing important insight into the mechanism by which MOR1 may regulate microtubule length. PMID:16377747

  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. GDP-Tubulin Incorporation into Growing Microtubules Modulates Polymer Stability.

    OpenAIRE

    Valiron, Odile; Arnal, Isabelle; Caudron, Nicolas; Job, Didier

    2010-01-01

    Microtubule growth proceeds through the endwise addition of nucleotide-bound tubulin dimers. The microtubule wall is composed of GDP-tubulin subunits, which are thought to come exclusively from the incorporation of GTP-tubulin complexes at microtubule ends followed by GTP hydrolysis within the polymer. The possibility of a direct GDP-tubulin incorporation into growing polymers is regarded as hardly compatible with recent structural data. Here, we have examined GTP-tubulin and GDP-tubulin inco...

  3. 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. 

  4. Producing Conditional Mutants for Studying Plant Microtubule Function

    Energy Technology Data Exchange (ETDEWEB)

    Richard Cyr

    2009-09-29

    The cytoskeleton, and in particular its microtubule component, participates in several processes that directly affect growth and development in higher plants. Normal cytoskeletal function requires the precise and orderly arrangement of microtubules into several cell cycle and developmentally specific arrays. One of these, the cortical array, is notable for its role in directing the deposition of cellulose (the most prominent polymer in the biosphere). An understanding of how these arrays form, and the molecular interactions that contribute to their function, is incomplete. To gain a better understanding of how microtubules work, we have been working to characterize mutants in critical cytoskeletal genes. This characterization is being carried out at the subcellular level using vital microtubule gene constructs. In the last year of funding colleagues have discovered that gamma-tubulin complexes form along the lengths of cortical microtubules where they act to spawn new microtubules at a characteristic 40 deg angle. This finding complements nicely the finding from our lab (which was funded by the DOE) showing that microtubule encounters are angle dependent; high angles encounters results in catastrophic collisions while low angle encounters result in favorable zippering. The finding of a 40 deg spawn of new microtubules from extant microtubule, together with aforementioned rules of encounters, insures favorable co-alignment in the array. I was invited to write a New and Views essay on this topic and a PDF is attached (News and Views policy does not permit funding acknowledgments and so I was not allowed to acknowledge support from the DOE).

  5. The nucleation of microtubules in Aspergillus nidulans germlings

    Directory of Open Access Journals (Sweden)

    Andrade-Monteiro Cristina de

    1999-01-01

    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.

  6. Exploring the limits of identifying sub-pixel thermal features using ASTER TIR data

    Science.gov (United States)

    Vaughan, R.G.; Keszthelyi, L.P.; Davies, A.G.; Schneider, D.J.; Jaworowski, C.; Heasler, H.

    2010-01-01

    Understanding the characteristics of volcanic thermal emissions and how they change with time is important for forecasting and monitoring volcanic activity and potential hazards. Satellite instruments view volcanic thermal features across the globe at various temporal and spatial resolutions. Thermal features that may be a precursor to a major eruption, or indicative of important changes in an on-going eruption can be subtle, making them challenging to reliably identify with satellite instruments. The goal of this study was to explore the limits of the types and magnitudes of thermal anomalies that could be detected using satellite thermal infrared (TIR) data. Specifically, the characterization of sub-pixel thermal features with a wide range of temperatures is considered using ASTER multispectral TIR data. First, theoretical calculations were made to define a "thermal mixing detection threshold" for ASTER, which quantifies the limits of ASTER's ability to resolve sub-pixel thermal mixing over a range of hot target temperatures and % pixel areas. Then, ASTER TIR data were used to model sub-pixel thermal features at the Yellowstone National Park geothermal area (hot spring pools with temperatures from 40 to 90 ??C) and at Mount Erebus Volcano, Antarctica (an active lava lake with temperatures from 200 to 800 ??C). Finally, various sources of uncertainty in sub-pixel thermal calculations were quantified for these empirical measurements, including pixel resampling, atmospheric correction, and background temperature and emissivity assumptions.

  7. 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.

  8. 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...

  9. 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.

  10. ASTER and USGS EROS disaster response: emergency imaging after Hurricane Katrina

    Science.gov (United States)

    Duda, Kenneth A.; Abrams, Michael

    2005-01-01

    The value of remotely sensed imagery during times of crisis is well established, and the increasing spatial and spectral resolution in newer systems provides ever greater utility and ability to discriminate features of interest (International Charter, Space and Major Disasters, 2005). The existing suite of sensors provides an abundance of data, and enables warning alerts to be broadcast for many situations in advance. In addition, imagery acquired soon after an event occurs can be used to assist response and remediation teams in identifying the extent of the affected area and the degree of damage. The data characteristics of the Advanced Spaceborne Thermal Emission and Refl ection Radiometer (ASTER) are well-suited for monitoring natural hazards and providing local and regional views after disaster strikes. For this reason, and because of the system fl exibility in scheduling high-priority observations, ASTER is often tasked to support emergency situations. The Emergency Response coordinators at the United States Geological Survey (USGS) Center for Earth Resources Observation and Science (EROS) work closely with staff at the National Aeronautics and Space Administration (NASA) Land Processes Distributed Active Archive Center (LP DAAC) at EROS and the ASTER Science Team as they fulfi ll their mission to acquire and distribute data during critical situations. This article summarizes the role of the USGS/EROS Emergency Response coordinators, and provides further discussion of ASTER data and the images portrayed on the cover of this issue

  11. Ecosystem engineering effects of Aster tripolium and Salicornia procumbens saltmarsh on macrofaunal community structure

    NARCIS (Netherlands)

    Van der Wal, D.; Herman, P.M.J.

    2012-01-01

    This paper examines how perennial Aster tripolium and annual Salicornia procumbens salt marshes alter the biomass, density, taxon diversity, and community structure of benthic macrofauna, and also examines the role of elevation, sediment grain size, plant cover, and marsh age. Core samples were coll

  12. 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

  13. 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

  14. 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.

  15. Validation of ASTER Surface Temperature Data with In Situ Measurements to Evaluate Heat Islands in Complex Urban Areas

    Directory of Open Access Journals (Sweden)

    Bonggeun Song

    2014-01-01

    Full Text Available This study compared Advanced Spaceborne Thermal Emission Reflection Radiometer (ASTER surface temperature data with in situ measurements to validate the use of ASTER data for studying heat islands in urban settings with complex spatial characteristics. Eight sites in Changwon, Korea, were selected for analyses. Surface temperature data were extracted from the thermal infrared (TIR band of ASTER on four dates during the summer and fall of 2012, and corresponding in situ measurements of temperature were also collected. Comparisons showed that ASTER derived temperatures were generally 4.27°C lower than temperatures collected by in situ measurements during the daytime, except on cloudy days. However, ASTER temperatures were higher by 2.23–2.69°C on two dates during the nighttime. Temperature differences between a city park and a paved area were insignificant. Differences between ASTER derived temperatures and onsite measurements are caused by a variety of factors including the application of emissivity values that do not consider the complex spatial characteristics of urban areas. Therefore, to improve the accuracy of surface temperatures extracted from infrared satellite imagery, we propose a revised model whereby temperature data is obtained from ASTER and emissivity values for various land covers are extracted based on in situ measurements.

  16. NASA and USGS ASTER Expedited Satellite Data Services for Disaster Situations

    Science.gov (United States)

    Duda, K. A.

    2012-12-01

    Significant international disasters related to storms, floods, volcanoes, wildfires and numerous other themes reoccur annually, often inflicting widespread human suffering and fatalities with substantial economic consequences. During and immediately after such events it can be difficult to access the affected areas and become aware of the overall impacts, but insight on the spatial extent and effects can be gleaned from above through satellite images. The Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) on the Terra spacecraft has offered such views for over a decade. On short notice, ASTER continues to deliver analysts multispectral imagery at 15 m spatial resolution in near real-time to assist participating responders, emergency managers, and government officials in planning for such situations and in developing appropriate responses after they occur. The joint U.S./Japan ASTER Science Team has developed policies and procedures to ensure such ongoing support is accessible when needed. Processing and distribution of data products occurs at the NASA Land Processes Distributed Active Archive Center (LP DAAC) located at the USGS Earth Resources Observation and Science Center in South Dakota. In addition to current imagery, the long-term ASTER mission has generated an extensive collection of nearly 2.5 million global 3,600 km2 scenes since the launch of Terra in late 1999. These are archived and distributed by LP DAAC and affiliates at Japan Space Systems in Tokyo. Advanced processing is performed to create higher level products of use to researchers. These include a global digital elevation model. Such pre-event imagery provides a comparative basis for use in detecting changes associated with disasters and to monitor land use trends to portray areas of increased risk. ASTER imagery acquired via the expedited collection and distribution process illustrates the utility and relevancy of such data in crisis situations.

  17. 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.

  18. 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.

  19. Motor Protein Accumulation on Antiparallel Microtubule Overlaps.

    Science.gov (United States)

    Kuan, Hui-Shun; Betterton, Meredith D

    2016-05-10

    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 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. Overlap motor density profiles and motor trajectories resemble experimental measurements. The phase diagram of the model is similar to the single-filament case for low switching rate, while for high switching rate we find a new (to our knowledge) low density-high density-low density-high density phase. The overlap center region, far from the overlap ends, has a constant motor density as one would naïvely expect. However, rather than following a simple binding equilibrium, the center motor density depends on total overlap length, motor speed, and motor switching rate. The size of the crowded boundary layer near the overlap ends is also dependent on the overlap length and switching rate in addition to the motor speed and bulk concentration. The antiparallel microtubule overlap geometry may offer a previously unrecognized mechanism for biological regulation of protein concentration and consequent activity. PMID:27166811

  20. Motor Protein Accumulation on Antiparallel Microtubule Overlaps

    Science.gov (United States)

    Kuan, Hui-Shun; Betterton, Meredith D.

    2016-05-01

    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. Overlap motor density profiles and motor trajectories resemble experimental measurements. The phase diagram of the model is similar to the single-filament case for low switching rate, while for high switching rate we find a new low density-high density-low density-high density phase. The overlap center region, far from the overlap ends, has a constant motor density as one would naively expect. However, rather than following a simple binding equilibrium, the center motor density depends on total overlap length, motor speed, and motor switching rate. The size of the crowded boundary layer near the overlap ends is also dependent on the overlap length and switching rate in addition to the motor speed and bulk concentration. The antiparallel microtubule overlap geometry may offer a previously unrecognized mechanism for biological regulation of protein concentration and consequent activity.

  1. Pattern formation of cortical microtubules and cellulose microfibrils

    NARCIS (Netherlands)

    Lindeboom, J.J.

    2012-01-01

    In this thesis we study the roles of microtubules at the plasma membrane and the cellulose microfibrils in the cell wall and how they are organized. This topic is introduces in chapter 1. In chapter 2 we study the formation of the transverse cortical microtubule array that is characteristic for elon

  2. Ice recovery assay for detection of Golgi-derived microtubules.

    Science.gov (United States)

    Grimaldi, Ashley D; Fomicheva, Maria; Kaverina, Irina

    2013-01-01

    Proper organization of the microtubule cytoskeleton is essential for many cellular processes including maintenance of Golgi organization and cell polarity. Traditionally, the centrosome is considered to be the major microtubule organizing center (MTOC) of the cell; however, microtubule nucleation can also occur through centrosome-independent mechanisms. Recently, the Golgi has been described as an additional, centrosome-independent, MTOC with distinct cellular functions. Golgi-derived microtubules contribute to the formation of an asymmetric microtubule network, control Golgi organization, and support polarized trafficking and directed migration in motile cells. In this chapter, we present an assay using recovery from ice treatment to evaluate the potential of the Golgi, or other MTOCs, to nucleate microtubules. This technique allows for clear separation of distinct MTOCs and observation of newly nucleated microtubules at these locations, which are normally obscured by the dense microtubule network present at steady-state conditions. This type of analysis is important for discovery and characterization of noncentrosomal MTOCs and, ultimately, understanding of their unique cellular functions. PMID:24295320

  3. 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

  4. Modelling the microtubule: towards a better understanding of short-chain fatty acid molecular pharmacology.

    Science.gov (United States)

    Kilner, Josephine; Corfe, Bernard M; Wilkinson, Stephen J

    2011-04-01

    Systems biology combines experimental data with computational modelling to describe complex biological mechanisms and pathways. Short-chain fatty acids (SCFAs-chemopreventive compounds produced in the colon lumen) impair microtubule (MT) function in colon cancer cells by altering the relative expression of β-tubulin isotypes. The β-tubulin isotype composition along MT fibres is believed to contribute to a "tubulin code" defining which microtubule-associated proteins (MAPs) and kinesins are recruited and the arrangement of tubulin post-transcriptional modifications (PTMs) along the fibre, which in turn dictate many critical cellular functions. SCFAs drive acetylation of many proteins by virtue of being histone deacetylase inhibitors (HDACi's). Known acetyl-proteins include transcription factors and cytoplasmic cytoskeletal keratins as well as histones. Disruption of the MT cytoskeleton is a prime target of many cancer therapies including anti-microtubule drugs (AMD). This review focuses on SCFAs as HDACi's and how they might affect tubulin dynamics, modifications and isotypes. It discusses the evolution of mechanistic models that have helped improve understanding of tubulin-MT structure and dynamics and how to develop these models, combined with those describing transcription and the cell cycle, could provide hypotheses for how SCFAs disrupt cytoskeletal function. The review demonstrates how systems biology could offer potentially novel ideas for therapies in the prevention and treatment of cancers through the continued development and elaboration of such models. PMID:21283865

  5. 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.

  6. 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...

  7. 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

  8. 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.

  9. 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. PMID:27611066

  10. Adubação nitrogenada e potássica do Aster ericoides cultivado em ambiente protegido Nitrogen and potassium fertilization of Aster ericoides cultivated in greenhouse

    Directory of Open Access Journals (Sweden)

    Mônica Sartori de Camargo

    2008-06-01

    Full Text Available O Aster ericoides é uma flor de corte recentemente introduzida no Brasil com grande potencial de produção e aceitação pelo mercado consumidor, mas são escassas as informações sobre o manejo da adubação, prática fundamental para desenvolvimento e qualidade da planta. O objetivo deste trabalho foi avaliar adubações nitrogenadas e potássicas na produção e absorção de nutrientes pelo Aster ericoides (White Master cultivado em Latossolo Amarelo eutrófico em condições de estufa. O experimento foi instalado em delineamento de blocos completos em esquema fatorial (4x4 com 4 doses de N (0; 50; 75 e 100 kg ha-1 N, 4 doses de K (0; 52; 78; 104 kg ha-1 K2O e 4 repetições no segundo ciclo da planta. A população foi de 400 mil plantas por hectare. Após 105 dias, foram colhidas dez hastes de cada parcela. A produção de matéria seca de hastes e folhas, número de flores por haste, número de ramificações laterais por haste não foram influenciadas pelos tratamentos. A dose de 46 kg ha-1 de N proporcionou máxima produção de matéria seca de flores (19,40 g planta-1 e a adubação com potássio não influenciou as características avaliadas. Estes resultados mostraram que é preciso ser cauteloso com as altas adubações de N e K no solo estudado para não reduzir a produção de matéria seca de flores. Mais estudos sobre a fertilização nitrogenada e potássica devem ser realizados para definir a recomendação de doses adequadas para o cultivo de Aster ericoides em condições tropicais.The Aster ericoides is a cut flower crop recently introduced into Brazil. It has great production potential and acceptation by consumer market but exist little information about fertilization management, essential to its development and quality. In this work we evaluated the nitrogen and potassium fertilization on dry matter yield and nutrient absorption by Aster ericoides (White Master growing in a Typic Hapludox soil under greenhouse

  11. Mean Lifetime of Microtubules Attached to Nucleating Sites

    Science.gov (United States)

    Rubin, Robert J.

    1988-01-01

    A simple two-phase (cap, no cap) macroscopic model describing the kinetic behavior at a labile tip of a microtubule has been proposed [Hill, T. L. (1984) Proc. Natl. Acad. Sci. USA 81, 6728-6732]. In the model, a microtubule exists either in a slowly growing phase (first-order rate constant α characterized by the existence of a GTP-tubulin cap at the growing tip; or the same microtubule exists in a rapidly shrinking phase (first-order rate constant, β ),, which is entered if/when the GTP-tubulin cap is lost through a fluctuation, thus exposing GDP-tubulin subunits, which constitute the body of the microtubule. Transition between the two phases-- i.e., loss of a cap (first-order rate constant, k) or formation of a new cap (first-order rate constant, k') occurs very infrequently and in a stochastic manner. In vitro experiments with centrosome-nucleated microtubules by Mitchison and Kirschner and Monte Carlo kinetic simulations, based on a realistic set of microscopic rate constants that apply to the end of a microtubule, suggest this alternation between two ``quasimacroscopic'' phases. In this paper, I outline the calculation of the mean lifetime of a microtubule nucleated on a centrosome by using Hill's model. For a microtubule M units long in the slowly growing phase, the mean lifetime for complete depolymerization is [M(k + k') + α +β ](β k - α k')-1, provided that β k > α k'. If the microtubule is in the rapidly shrinking phase, then the mean lifetime is M(k + k')(β k - α k')-1, provided that β k > α k'. In case β k < α k', the microtubule grows indefinitely, and the mean lifetime is infinite.

  12. 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

  13. Measurements of volcanic SO2 with ASTER. Comparison with automated scanning DOAS measurements

    Science.gov (United States)

    Campion, R. A.; Salerno, G. G.; Bernard, A. M.; Burton, M.; Coheur, P.; Caltabiano, T.

    2009-12-01

    Sulfur dioxide (SO2) emitted by volcanoes has an important impact on the environment and climate and is also a critical parameter for volcano monitoring. A number of satellites operating in the ultra violet and in the Thermal infrared can measure SO2. However a lot of work has still to be done towards a rigorous validation of SO2 measurements from space. ASTER (Advanced Spaceborne Thermal Emission Reflection radiometer) acquires images in the thermal infrared (TIR) with a resolution of 90m/pixel, which enables to quantify the SO2 fluxes emitted in small-scale tropospheric plumes. ASTER images are processed with radiative transfer simulations and a band ratio algorithm to produce maps of SO2 column amounts. The band ratios (B10+B12)/B11 and B14/B11 are used for their insensitivity to variations in ground altitude and atmospheric humidity, two variables that often complicate SO2 retrievals in the TIR. Their sensitivity to surface emissivity is also reduced. So far, the ground validation of satellite SO2 measurements has been complex due to logistics difficulties and the lack of strictly simultaneous measurements. Recently the development of permanent networks of scanning DOAS on several active volcanoes has provide a wealth of ground based SO2 measurements that can be exploited for validating satellite-based measurements. We will present the results of comparisons between SO2 Column Amount (CA) and fluxes measured by ASTER and by the FLAME network of Mt. Etna. The two independent measurements sets are in good agreement in magnitude. Fluxes range from 2000 to 5000 T/days and column amounts from 0 to 4 g/m2. CAs measured by ASTER present a 0.5g/m2 random dispersion and no systematic bias compared to DOAS measurements. However the CAs measured by DOAS are subject to increase at low-scanning angles. These results constitute a rigorous ground validation of ASTER SO2, and provides valuable insights into accuracy and precision on both methodologies. Figure 1: Comparison

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

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    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

  15. 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.

  16. 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.

  17. Stochastic models for plant microtubule self-organization and structure.

    Science.gov (United States)

    Eren, Ezgi C; Dixit, Ram; Gautam, Natarajan

    2015-12-01

    One of the key enablers of shape and growth in plant cells is the cortical microtubule (CMT) system, which is a polymer array that forms an appropriately-structured scaffolding in each cell. Plant biologists have shown that stochastic dynamics and simple rules of interactions between CMTs can lead to a coaligned CMT array structure. However, the mechanisms and conditions that cause CMT arrays to become organized are not well understood. It is prohibitively time-consuming to use actual plants to study the effect of various genetic mutations and environmental conditions on CMT self-organization. In fact, even computer simulations with multiple replications are not fast enough due to the spatio-temporal complexity of the system. To redress this shortcoming, we develop analytical models and methods for expeditiously computing CMT system metrics that are related to self-organization and array structure. In particular, we formulate a mean-field model to derive sufficient conditions for the organization to occur. We show that growth-prone dynamics itself is sufficient to lead to organization in presence of interactions in the system. In addition, for such systems, we develop predictive methods for estimation of system metrics such as expected average length and number of CMTs over time, using a stochastic fluid-flow model, transient analysis, and approximation algorithms tailored to our problem. We illustrate the effectiveness of our approach through numerical test instances and discuss biological insights. PMID:25700800

  18. 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.

  19. Urban Heat Island Analysis Using the Landsat TM Data and ASTER Data: A Case Study in Hong Kong

    OpenAIRE

    Lin Liu; Yuanzhi Zhang

    2011-01-01

    In this paper, the effect of urban heat island is analyzed using the Landsat TM data and ASTER data in 2005 as a case study in Hong Kong. Two algorithms were applied to retrieve the land surface temperature (LST) distribution from the Landsat TM and ASTER data. The spatial pattern of LST in the study area is retrieved to characterize their local effects on urban heat island. In addition, the correlation between LST and the normalized difference vegetation index (NDVI), the normalized differen...

  20. Quinic acids from Aster caucasicus and from transgenic callus expressing a beta-amyrin synthase.

    Science.gov (United States)

    Pecchia, Paola; Cammareri, Maria; Malafronte, Nicola; Consiglio, M Federica; Gualtieri, Maria Josefina; Conicella, Clara

    2011-11-01

    Several different classes of secondary metabolites, including flavonoids, triterpenoid saponins and quinic acid derivatives, are found in Aster spp. (Fam. Asteraceae). Several Aster compounds revealed biological as well as pharmacological activities. In this work, a phytochemical investigation of A. caucasicus evidenced the presence of quinic acid derivatives, as well as the absence of triterpene saponins. To combine in one species the production of different phytochemicals, including triterpenes, an Agrobacterium-mediated transformation of A. caucasicus was set up to introduce A. sedifolius beta-amyrin synthase (AsOXA1)-encoding gene under the control of the constitutive promoter CaMV35S. The quali-quantitative analysis of transgenic calli with ectopic expression of AsOXA1 showed, in one sample, a negligible amount of triterpene saponins combined with higher amount of quinic acid derivatives as compared with the wild type callus. PMID:22224284

  1. 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

  2. Spatiotemporal relationships between growth and microtubule orientation as revealed in living root cells of Arabidopsis thaliana transformed with green-fluorescent-protein gene construct GFP-MBD

    Science.gov (United States)

    Granger, C. L.; Cyr, R. J.

    2001-01-01

    Arabidopsis thaliana plants were transformed with GFP-MBD (J. Marc et al., Plant Cell 10: 1927-1939, 1998) under the control of a constitutive (35S) or copper-inducible promoter. GFP-specific fluorescence distributions, levels, and persistence were determined and found to vary with age, tissue type, transgenic line, and individual plant. With the exception of an increased frequency of abnormal roots of 35S GFP-MBD plants grown on kanamycin-containing media, expression of GFP-MBD does not appear to affect plant phenotype. The number of leaves, branches, bolts, and siliques as well as overall height, leaf size, and seed set are similar between wild-type and transgenic plants as is the rate of root growth. Thus, we conclude that the transgenic plants can serve as a living model system in which the dynamic behavior of microtubules can be visualized. Confocal microscopy was used to simultaneously monitor growth and microtubule behavior within individual cells as they passed through the elongation zone of the Arabidopsis root. Generally, microtubules reoriented from transverse to oblique or longitudinal orientations as growth declined. Microtubule reorientation initiated at the ends of the cell did not necessarily occur simultaneously in adjacent neighboring cells and did not involve complete disintegration and repolymerization of microtubule arrays. Although growth rates correlated with microtubule reorientation, the two processes were not tightly coupled in terms of their temporal relationships, suggesting that other factor(s) may be involved in regulating both events. Additionally, microtubule orientation was more defined in cells whose growth was accelerating and less stringent in cells whose growth was decelerating, indicating that microtubule-orienting factor(s) may be sensitive to growth acceleration, rather than growth per se.

  3. Estimating Evapotranspiration from an Improved Two-Source Energy Balance Model Using ASTER Satellite Imagery

    OpenAIRE

    Qifeng Zhuang; Bingfang Wu

    2015-01-01

    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 al...

  4. 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.

  5. Metabolic potential of microorganisms associated with the halophyte Aster tripolium L. in saline soils

    OpenAIRE

    Szymańska, Sonia; Piernik, Agnieszka; Hrynkiewicz, Katarzyna

    2014-01-01

    Increased soil salinization may be caused by a natural (e.g. climate change) and anthropogenic (e.g. improper fertilization and irrigation of agricultural land) factors. The submitted work assumes that microorganisms associated with plant halophytes have a unique metabolic properties that can stimulate plant growth under salt stress. The aim of the study was to determine the abundance and metabolic biodiversity of endophytic and rhizosphere microorganisms co-existing with Aster tripolium L. a...

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

    OpenAIRE

    Changqing Ke; Yasushi Yamaguchi; Yuanbo Liu

    2007-01-01

    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,...

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

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    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.

  10. 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.

  11. Birefringence of single and bundled microtubules.

    Science.gov (United States)

    Oldenbourg, R; Salmon, E D; Tran, P T

    1998-01-01

    We have measured the birefringence of microtubules (MTs) and of MT-based macromolecular assemblies in vitro and in living cells by using the new Pol-Scope. A single microtubule in aqueous suspension and imaged with a numerical aperture of 1.4 had a peak retardance of 0.07 nm. The peak retardance of a small bundle increased linearly with the number of MTs in the bundle. Axonemes (prepared from sea urchin sperm) had a peak retardance 20 times higher than that of single MTs, in accordance with the nine doublets and two singlets arrangement of parallel MTs in the axoneme. Measured filament retardance decreased when the filament was defocused or the numerical aperture of the imaging system was decreased. However, the retardance "area," which we defined as the image retardance integrated along a line perpendicular to the filament axis, proved to be independent of focus and of numerical aperture. These results are in good agreement with a theory that we developed for measuring retardances with imaging optics. Our theoretical concept is based on Wiener's theory of mixed dielectrics, which is well established for nonimaging applications. We extend its use to imaging systems by considering the coherence region defined by the optical set-up. Light scattered from within that region interferes coherently in the image point. The presence of a filament in the coherence region leads to a polarization dependent scattering cross section and to a finite retardance measured in the image point. Similar to resolution measurements, the linear dimension of the coherence region for retardance measurements is on the order lambda/(2 NA), where lambda is the wavelength of light and NA is the numerical aperture of the illumination and imaging lenses. PMID:9449366

  12. 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.

  13. 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.

  14. 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...

  15. 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

  16. Aged insulin granules display reduced microtubule-dependent mobility and are disposed within actin-positive multigranular bodies

    Science.gov (United States)

    Hoboth, Peter; Müller, Andreas; Ivanova, Anna; Mziaut, Hassan; Dehghany, Jaber; Sönmez, Anke; Lachnit, Martina; Meyer-Hermann, Michael; Kalaidzidis, Yannis; Solimena, Michele

    2015-01-01

    Insulin secretion is key for glucose homeostasis. Insulin secretory granules (SGs) exist in different functional pools, with young SGs being more mobile and preferentially secreted. However, the principles governing the mobility of age-distinct SGs remain undefined. Using the time-reporter insulin-SNAP to track age-distinct SGs we now show that their dynamics can be classified into three components: highly dynamic, restricted, and nearly immobile. Young SGs display all three components, whereas old SGs are either restricted or nearly immobile. Both glucose stimulation and F-actin depolymerization recruit a fraction of nearly immobile young, but not old, SGs for highly dynamic, microtubule-dependent transport. Moreover, F-actin marks multigranular bodies/lysosomes containing aged SGs. These data demonstrate that SGs lose their responsiveness to glucose stimulation and competence for microtubule-mediated transport over time while changing their relationship with F-actin. PMID:25646459

  17. 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

  18. 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.

  19. 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

  20. 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.

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

    Directory of Open Access Journals (Sweden)

    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.

  2. Equilibria of idealized confined astral microtubules and coupled spindle poles.

    Directory of Open Access Journals (Sweden)

    Ivan V Maly

    Full Text Available Positioning of the mitotic spindle through the interaction of astral microtubules with the cell boundary often determines whether the cell division will be symmetric or asymmetric. This process plays a crucial role in development. In this paper, a numerical model is presented that deals with the force exerted on the spindle by astral microtubules that are bent by virtue of their confinement within the cell boundary. It is found that depending on parameters, the symmetric position of the spindle can be stable or unstable. Asymmetric stable equilibria also exist, and two or more stable positions can exist simultaneously. The theory poses new types of questions for experimental research. Regarding the cases of symmetric spindle positioning, it is necessary to ask whether the microtubule parameters are controlled by the cell so that the bending mechanics favors symmetry. If they are not, then it is necessary to ask what forces external to the microtubule cytoskeleton counteract the bending effects sufficiently to actively establish symmetry. Conversely, regarding the cases with asymmetry, it is now necessary to investigate whether the cell controls the microtubule parameters so that the bending favors asymmetry apart from any forces that are external to the microtubule cytoskeleton.

  3. Yeast GSK-3 kinase regulates astral microtubule function through phosphorylation of the microtubule-stabilizing kinesin Kip2.

    Science.gov (United States)

    Drechsler, Hauke; Tan, Ann Na; Liakopoulos, Dimitris

    2015-11-01

    The S. cerevisiae kinesin Kip2 stabilises astral microtubules (MTs) and facilitates spindle positioning through transport of MT-associated proteins, such as the yeast CLIP-170 homologue Bik1, dynein and the adenomatous-polyposis-coli-related protein Kar9 to the plus ends of astral MTs. Here, we show that Kip2 associates with its processivity factor Bim1, the yeast homologue of the plus-end-tracking protein EB1. This interaction requires an EB1-binding motif in the N-terminal extension of Kip2 and is negatively regulated by phosphorylation through Mck1, the yeast glycogen synthase kinase 3. In addition, Mck1-dependent phosphorylation decreases the intrinsic MT affinity of Kip2. Reduction in Kip2 phosphorylation leads to stabilisation of astral MTs, and accumulation of Kip2, dynein and Kar9 at MT plus ends, whereas loss of Mck1 function leads to defects in spindle positioning. Furthermore, we provide evidence that a subpopulation of Mck1 at the bud-cortex phosphorylates Kip2. We propose that yeast GSK-3 spatially controls astral MT dynamics and the loading of dynein and Kar9 on astral MT plus ends by regulating Kip2 interactions with Bim1 and MTs. PMID:26395399

  4. 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.

  5. 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.

  6. 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.

  7. On the significance of microtubule flexural behavior in cytoskeletal mechanics.

    Science.gov (United States)

    Mehrbod, Mehrdad; Mofrad, Mohammad R K

    2011-01-01

    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 exceed the axial energy

  8. 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

  9. 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.

  10. LKB1 destabilizes microtubules in myoblasts and contributes to myoblast differentiation.

    Directory of Open Access Journals (Sweden)

    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.

  11. ASTER spectral analysis and lithologic mapping of the Khanneshin carbonatite volcano, Afghanistan

    Science.gov (United States)

    Mars, J.C.; Rowan, L.C.

    2011-01-01

    Advanced Spaceborne Thermal and Reflection Radiometer (ASTER) data of the early Quaternary Khanneshin carbonatite volcano located in southern Afghanistan were used to identify carbonate rocks within the volcano and to distinguish them from Neogene ferruginous polymict sandstone and argillite. The carbonatitic rocks are characterized by diagnostic CO3 absorption near 11.2 ??m and 2.31-2.33 ??m, whereas the sandstone, argillite, and adjacent alluvial deposits exhibit intense Si-O absorption near 8.7 ??m caused mainly by quartz and Al-OH absorption near 2.20 ??m due to muscovite and illite. Calcitic carbonatite was distinguished from ankeritic carbonatite in the short wave infrared (SWIR) region of the ASTER data due to a slight shift of the CO3 absorption feature toward 2.26 ??m (ASTER band 7) in the ankeritic carbonatite spectra. Spectral assessment using ASTER SWIR data suggests that the area is covered by extensive carbonatite flows that contain calcite, ankerite, and muscovite, though some areas mapped as ankeritic carbonatite on a pre existing geologic map were not identified in the ASTER data. A contact aureole shown on the geologic map was defined using an ASTER false color composite image (R = 6, G = 3, B = 1) and a logical operator byte image. The contact aureole rocks exhibit Fe2+, Al-OH, and Fe, Mg-OH spectral absorption features at 1.65, 2.2, and 2.33 ??m, respectively, which suggest that the contact aureole rocks contain musco vite, epidote, and chlorite. The contact aureole rocks were mapped using an Interactive Data Language (IDL) logical operator. A visible through short wave infrared (VNIR-SWIR) mineral and rock-type map based on matched filter, band ratio, and logical operator analysis illustrates: (1) laterally extensive calcitic carbonatite that covers most of the crater and areas northeast of the crater; (2) ankeritic carbonatite located southeast and north of the crater and some small deposits located within the crater; (3) agglomerate that

  12. Microtubule Protofilament Number Is Modulated in a Stepwise Fashion by the Charge Density of an Enveloping Layer

    OpenAIRE

    Raviv, Uri; Nguyen, Toan; Ghafouri, Rouzbeh; Needleman, Daniel J.; Li, Youli; Miller, Herbert P.; Wilson, Leslie; Bruinsma, Robijn F.; Safinya, Cyrus R.

    2006-01-01

    Microtubules are able to adjust their protofilament (PF) number and, as a consequence, their dynamics and function, to the assembly conditions and presence of cofactors. However, the principle behind such variations is poorly understood. Using synchrotron x-ray scattering and transmission electron microscopy, we studied how charged membranes, which under certain conditions can envelop preassembled MTs, regulate the PF number of those MTs. We show that the mean PF number, 〈N〉, is modulated pri...

  13. Simultaneous Tracking of 3D Actin and Microtubule Strains in Individual MLO-Y4 Osteocytes under Oscillatory Flow

    OpenAIRE

    Baik, Andrew D.; Qiu, Jun; Hillman, Elizabeth M. C.; Dong, Cheng; Guo, X. Edward

    2013-01-01

    Osteocytes in vivo experience complex fluid shear flow patterns to activate mechanotransduction pathways. The actin and microtubule (MT) cytoskeletons have been shown to play an important role in the osteocyte’s biochemical response to fluid shear loading. The dynamic nature of physiologically relevant fluid flow profiles (i.e., 1 Hz oscillatory flow) impedes the ability to image and study both actin and MT cytoskeletons simultaneously in the same cell with high spatiotemporal resolution. To ...

  14. 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.

  15. 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.

  16. 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.

  17. Analysis of ASTER data for mapping bauxite rich pockets within high altitude lateritic bauxite, Jharkhand, India

    Science.gov (United States)

    Guha, Arindam; Singh, Vivek Kr.; Parveen, Reshma; Kumar, K. Vinod; Jeyaseelan, A. T.; Dhanamjaya Rao, E. N.

    2013-04-01

    Bauxite deposits of Jharkhand in India are resulted from the lateritization process and therefore are often associated with the laterites. In the present study, ASTER (Advanced Space borne Thermal Emission and Reflection Radiometer) image is processed to delineate bauxite rich pockets within the laterites. In this regard, spectral signatures of lateritic bauxite samples are analyzed in the laboratory with reference to the spectral features of gibbsite (main mineral constituent of bauxite) and goethite (main mineral constituent of laterite) in VNIR-SWIR (visible-near infrared and short wave infrared) electromagnetic domain. The analysis of spectral signatures of lateritic bauxite samples helps in understanding the differences in the spectral features of bauxites and laterites. Based on these differences; ASTER data based relative band depth and simple ratio images are derived for spatial mapping of the bauxites developed within the lateritic province. In order to integrate the complementary information of different index image, an index based principal component (IPC) image is derived to incorporate the correlative information of these indices to delineate bauxite rich pockets. The occurrences of bauxite rich pockets derived from density sliced IPC image are further delimited by the topographic controls as it has been observed that the major bauxite occurrences of the area are controlled by slope and altitude. In addition to above, IPC image is draped over the digital elevation model (DEM) to illustrate how bauxite rich pockets are distributed with reference to the topographic variability of the terrain. Bauxite rich pockets delineated in the IPC image are also validated based on the known mine occurrences and existing geological map of the bauxite. It is also conceptually validated based on the spectral similarity of the bauxite pixels delineated in the IPC image with the ASTER convolved laboratory spectra of bauxite samples.

  18. Estimation of aboveground biomass in Mediterranean forests by statistical modelling of ASTER fraction images

    Science.gov (United States)

    Fernández-Manso, O.; Fernández-Manso, A.; Quintano, C.

    2014-09-01

    Aboveground biomass (AGB) estimation from optical satellite data is usually based on regression models of original or synthetic bands. To overcome the poor relation between AGB and spectral bands due to mixed-pixels when a medium spatial resolution sensor is considered, we propose to base the AGB estimation on fraction images from Linear Spectral Mixture Analysis (LSMA). Our study area is a managed Mediterranean pine woodland (Pinus pinaster Ait.) in central Spain. A total of 1033 circular field plots were used to estimate AGB from Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) optical data. We applied Pearson correlation statistics and stepwise multiple regression to identify suitable predictors from the set of variables of original bands, fraction imagery, Normalized Difference Vegetation Index and Tasselled Cap components. Four linear models and one nonlinear model were tested. A linear combination of ASTER band 2 (red, 0.630-0.690 μm), band 8 (short wave infrared 5, 2.295-2.365 μm) and green vegetation fraction (from LSMA) was the best AGB predictor (Radj2=0.632, the root-mean-squared error of estimated AGB was 13.3 Mg ha-1 (or 37.7%), resulting from cross-validation), rather than other combinations of the above cited independent variables. Results indicated that using ASTER fraction images in regression models improves the AGB estimation in Mediterranean pine forests. The spatial distribution of the estimated AGB, based on a multiple linear regression model, may be used as baseline information for forest managers in future studies, such as quantifying the regional carbon budget, fuel accumulation or monitoring of management practices.

  19. SRTM DEM与ASTER DEM数据比较研究

    Institute of Scientific and Technical Information of China (English)

    李鹤元; 孙亮

    2014-01-01

    SRTM DEM是利用航天飞机雷达地形测绘获取的全球中低纬度地区地表的高分辨率雷达影像生成的DEM,它具有较高的分辨率和现势性;ASTER DEM是利用高分辨率卫星成像设备ASTER获取的立体影像生成的DEM,全球覆盖范围更广、数据分辨率更高。由于两种DEM获取机理与生产工艺不同,因此存在较大的数据差异。为了利用两种数据进行DEM数据融合与更新,需要对不同地形DEM数据进行比较研究与分析。本文选取具有高山峡谷地貌特征的某山区为实验区,运用总体统计分析、检查点法和剖面法对SRTM DEM与ASTER DEM进行数据比较研究。结果表明,SRTM DEM与ASTER DEM中误差约为45m,误差值在-150m至150m的点数量呈正态分布,在山峰与山谷处高程值差异明显。

  20. 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.

  1. 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?

  2. 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.

  3. 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...

  4. 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

  5. Quantitative Analysis of Tau-Microtubule Interaction Using FRET

    Directory of Open Access Journals (Sweden)

    Isabelle L. Di Maïo

    2014-08-01

    Full Text Available The interaction between the microtubule associated protein, tau and the microtubules is investigated. A fluorescence resonance energy transfer (FRET assay was used to determine the distance separating tau to the microtubule wall, as well as the binding parameters of the interaction. By using microtubules stabilized with Flutax-2 as donor and tau labeled with rhodamine as acceptor, a donor-to-acceptor distance of 54 ± 1 Å was found. A molecular model is proposed in which Flutax-2 is directly accessible to tau-rhodamine molecules for energy transfer. By titration, we calculated the stoichiometric dissociation constant to be equal to 1.0 ± 0.5 µM. The influence of the C-terminal tails of αβ-tubulin on the tau-microtubule interaction is presented once a procedure to form homogeneous solution of cleaved tubulin has been determined. The results indicate that the C-terminal tails of α- and β-tubulin by electrostatic effects and of recruitment seem to be involved in the binding mechanism of tau.

  6. Spatial organization of the Ran pathway by microtubules in mitosis.

    Science.gov (United States)

    Oh, Doogie; Yu, Che-Hang; Needleman, Daniel J

    2016-08-01

    Concentration gradients of soluble proteins are believed to be responsible for control of morphogenesis of subcellular systems, but the mechanisms that generate the spatial organization of these subcellular gradients remain poorly understood. Here, we use a newly developed multipoint fluorescence fluctuation spectroscopy technique to study the ras-related nuclear protein (Ran) pathway, which forms soluble gradients around chromosomes in mitosis and is thought to spatially regulate microtubule behaviors during spindle assembly. We found that the distribution of components of the Ran pathway that influence microtubule behaviors is determined by their interactions with microtubules, resulting in microtubule nucleators being localized by the microtubules whose formation they stimulate. Modeling and perturbation experiments show that this feedback makes the length of the spindle insensitive to the length scale of the Ran gradient, allows the spindle to assemble outside the peak of the Ran gradient, and explains the scaling of the spindle with cell size. Such feedback between soluble signaling pathways and the mechanics of the cytoskeleton may be a general feature of subcellular organization. PMID:27439876

  7. 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.

  8. Highly Transient Molecular Interactions Underlie the Stability of Kinetochore–Microtubule Attachment During Cell Division

    Science.gov (United States)

    Zaytsev, Anatoly V.; Ataullakhanov, Fazly I.; Grishchuk, Ekaterina L.

    2013-01-01

    Chromosome segregation during mitosis is mediated by spindle microtubules that attach to chromosomal kinetochores with strong yet labile links. The exact molecular composition of the kinetochore–microtubule interface is not known but microtubules are thought to bind to kinetochores via the specialized microtubule-binding sites, which contain multiple microtubule-binding proteins. During prometaphase the lifetime of microtubule attachments is short but in metaphase it increases 3-fold, presumably owing to dephosphorylation of the microtubule-binding proteins that increases their affinity. Here, we use mathematical modeling to examine in quantitative and systematic manner the general relationships between the molecular properties of microtubule-binding proteins and the resulting stability of microtubule attachment to the protein-containing kinetochore site. We show that when the protein connections are stochastic, the physiological rate of microtubule turnover is achieved only if these molecular interactions are very transient, each lasting fraction of a second. This “microscopic” time is almost four orders of magnitude shorter than the characteristic time of kinetochore–microtubule attachment. Cooperativity of the microtubule-binding events further increases the disparity of these time scales. Furthermore, for all values of kinetic parameters the microtubule stability is very sensitive to the minor changes in the molecular constants. Such sensitivity of the lifetime of microtubule attachment to the kinetics and cooperativity of molecular interactions at the microtubule-binding site may hinder the accurate regulation of kinetochore–microtubule stability during mitotic progression, and it necessitates detailed experimental examination of the microtubule-binding properties of kinetochore-localized proteins. PMID:24376473

  9. Highly Transient Molecular Interactions Underlie the Stability of Kinetochore-Microtubule Attachment During Cell Division.

    Science.gov (United States)

    Zaytsev, Anatoly V; Ataullakhanov, Fazly I; Grishchuk, Ekaterina L

    2013-12-13

    Chromosome segregation during mitosis is mediated by spindle microtubules that attach to chromosomal kinetochores with strong yet labile links. The exact molecular composition of the kinetochore-microtubule interface is not known but microtubules are thought to bind to kinetochores via the specialized microtubule-binding sites, which contain multiple microtubule-binding proteins. During prometaphase the lifetime of microtubule attachments is short but in metaphase it increases 3-fold, presumably owing to dephosphorylation of the microtubule-binding proteins that increases their affinity. Here, we use mathematical modeling to examine in quantitative and systematic manner the general relationships between the molecular properties of microtubule-binding proteins and the resulting stability of microtubule attachment to the protein-containing kinetochore site. We show that when the protein connections are stochastic, the physiological rate of microtubule turnover is achieved only if these molecular interactions are very transient, each lasting fraction of a second. This "microscopic" time is almost four orders of magnitude shorter than the characteristic time of kinetochore-microtubule attachment. Cooperativity of the microtubule-binding events further increases the disparity of these time scales. Furthermore, for all values of kinetic parameters the microtubule stability is very sensitive to the minor changes in the molecular constants. Such sensitivity of the lifetime of microtubule attachment to the kinetics and cooperativity of molecular interactions at the microtubule-binding site may hinder the accurate regulation of kinetochore-microtubule stability during mitotic progression, and it necessitates detailed experimental examination of the microtubule-binding properties of kinetochore-localized proteins. PMID:24376473

  10. 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.

  11. Relations Between Albedos and Emissivities From MODIS and ASTER Data Over North African Desert

    Science.gov (United States)

    Zhou, L.; Dickinson, R.; Ogawa, K.; Tian, Y.; Jin, M.; Schmugge, T.; Tsvetsinskaya, E.

    2003-12-01

    This paper analyzes relations among MODIS surface albedos, ASTER broadband (3-14 μ m) emissivities, and a soil taxonomy map over the arid areas of Algeria, Libya, and Tunisia in North Africa at 30 second (about 1 km) and 2 minute (about 4 km) spatial resolutions. The MODIS albedo data are from 7 spectral bands and 3 broadbands during dust-free seasons and the emissivity data are derived from a linear combination of the waveband emissivities of the ASTER five thermal infrared channels. Both albedo and emissivity data in the study region show similar considerable spatial variability, larger than assumed by most climate models, and such variability is related to the surface types (sands, rock, and soil orders). Emissivity over bare soils exhibits statistically significant correlations with albedos at both broadbands and most of spectral bands and decreases linearly with albedos. Albedo and emissivity are more strongly correlated with each other than either is to the surface types, apparently because of their higher resolution either spatially or in surface mineralogy. This paper provides guidance for the possible inclusion of such correlation to specify albedo and emissivity in climate models.

  12. 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.

  13. 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.

  14. 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.

  15. Spectral properties and ASTER-based alteration mapping of Masahim volcano facies, SE Iran

    Science.gov (United States)

    Tayebi, Mohammad H.; Tangestani, Majid H.; Vincent, Robert K.; Neal, Devin

    2014-10-01

    This study applies Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) data and the Mixture Tuned Matched Filtering (MTMF) algorithm to map the sub-pixel distribution of alteration minerals associated with the Masahim volcano, SE Iran for understanding the spatial relationship between alteration minerals and volcano facies. Investigations of the alteration mineralogy were conducted using field-spectroscopy, X-ray diffraction (XRD) analysis and ASTER Short Wave Infrared (SWIR) spectral data. In order to spectrally characterize the stratovolcano deposits, lithological units and alteration minerals, the volcano was divided into three facies: the Central, Proximal, and Medial-distal facies. The reflectance spectra of rock samples show absorption features of a number of minerals including white mica, kaolinite, montmorillonite, illite, goethite, hematite, jarosite, opal, and chlorite. The end-members of key alteration minerals including sericite (phyllic zone), kaolinite (argillic zone) and chlorite (propylitic zone) were extracted from imagery using the Pixel Purity Index (PPI) method and were used to map alteration minerals. Accuracy assessment through field observations was used to verify the fraction maps. The results showed that most prominent altered rocks situated at the central facies of volcano. The alteration minerals were discriminated with the coefficient of determination (R2) of 0.74, 0.81, and 0.68 for kaolinite, sericite, and chlorite, respectively. The results of this study have the potential to refine the map of alteration zones in the Masahim volcano.

  16. 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.

  17. Hydrothermal alteration mapping using ASTER data in Baogutu porphyry deposit, China

    International Nuclear Information System (INIS)

    Remote sensing plays an important role in mineral exploration. One of its proven applications is extracting host-rock lithology and alteration zones that are related to porphyry copper deposits. An Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) was used to map the Baogutu porphyry deposit alteration area. A circular alteration mineral zoning pattern was clearly observed in the classification result of potassic, phyllic, argillic, propylitic zones. The potassic is characterized by biotite and anhydrite with an absorption feature centered at 1.94 and 2.1um. The phyllic zone is characterized by illite and sericite that indicates an intense Al-OH absorption feature centered at 2.20um. The narrower argillic zone including kaolinite and alunite displays a secondary Al-OH absorption feature at 2.17 um. The mineral assemblages of the outer propylitic zone are epidote, chlorite and calcite that exhibit absorption features at 2.335um.The performance of Principal Component Analysis(PCA), Minimum Noise Fraction (MNF), band ratio(BR) and Constrained Energy Minimization(CEM) has been evaluated. These techniques identified new prospects of porphyry copper mineralization in the study areas. These results indicate that ASTER is a powerful tool in the initial steps of mineral exploration

  18. 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.

  19. 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...

  20. 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.

  1. 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.

  2. 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

  3. 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

  4. 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.

  5. 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.

  6. Prion protein inhibits microtubule assembly by inducing tubulin oligomerization

    International Nuclear Information System (INIS)

    A growing body of evidence points to an association of prion protein (PrP) with microtubular cytoskeleton. Recently, direct binding of PrP to tubulin has also been found. In this work, using standard light scattering measurements, sedimentation experiments, and electron microscopy, we show for First time the effect of a direct interaction between these proteins on tubulin polymerization. We demonstrate that full-length recombinant PrP induces a rapid increase in the turbidity of tubulin diluted below the critical concentration for microtubule assembly. This effect requires magnesium ions and is weakened by NaCl. Moreover, the PrP-induced light scattering structures of tubulin are cold-stable. In preparations of diluted tubulin incubated with PrP, electron microscopy revealed the presence of ∼50 nm disc-shaped structures not reported so far. These unique tubulin oligomers may form large aggregates. The effect of PrP is more pronounced under the conditions promoting microtubule formation. In these tubulin samples, PrP induces formation of the above oligomers associated with short protofilaments and sheets of protofilaments into aggregates. Noticeably, this is accompanied by a significant reduction of the number and length of microtubules. Hence, we postulate that prion protein may act as an inhibitor of microtubule assembly by inducing formation of stable tubulin oligomers

  7. 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.

  8. 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

  9. Information transport by sine-Gordon solitons in microtubules

    CERN Document Server

    Abdalla, Elcio; Melgar, B C; Sedra, M B; Abdalla, Elcio; Maroufi, Bouchra; Melgar, Bertha Cuadros; Sedra, Moulay Brahim

    2001-01-01

    We study the problem of information propagation in brain microtubules. After considering the propagation of electromagnetic waves in a fluid of permanent electric dipoles, the problem reduces to the sine-Gordon wave equation in one space and one time dimensions. The problem of propagation of information is thus set.

  10. 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.

  11. ASTER and USGS EROS emergency imaging for hurricane disasters: Chapter 4D in Science and the storms-the USGS response to the hurricanes of 2005

    Science.gov (United States)

    Duda, Kenneth A.; Abrams, Michael

    2007-01-01

    Satellite images have been extremely useful in a variety of emergency response activities, including hurricane disasters. This article discusses the collaborative efforts of the U.S. Geological Survey (USGS), the Joint United States-Japan Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) Science Team, and the National Aeronautics and Space Administration (NASA) in responding to crisis situations by tasking the ASTER instrument and rapidly providing information to initial responders. Insight is provided on the characteristics of the ASTER systems, and specific details are presented regarding Hurricane Katrina support.

  12. 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.

  13. 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.

  14. 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.

  15. 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.

    2015-01-01

    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 siRNA to promote wound closure and regeneration. In vitro, depletion of FL2 from mammalian tissue culture cells results in a more than two-fold increase in the rate of cell movement, due in part 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. PMID:25756798

  16. 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. PMID:25756798

  17. 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.

  18. 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

  19. Aster spathulifolius Maxim extract reduces body weight and fat mass in obese humans.

    Science.gov (United States)

    Cho, In-Jin; Choung, Se Young; Hwang, You-Cheol; Ahn, Kyu Jeung; Chung, Ho Yeon; Jeong, In-Kyung

    2016-07-01

    Aster spathulifolius Maxim (AS), a perennial herb of the genus Aster within the family Asteraceae, induced weight loss in a rat model of diet-induced obesity. We hypothesized that AS could also reduce body weight in obese humans. Therefore, we performed a randomized, double-blind, placebo-controlled clinical trial in Korea to evaluate the effect of AS extract (ASE) on body weight and fat mass and its safety in obese humans. Forty-four obese participants (body mass index [BMI], 25-30 kg/m(2)) aged ≥20 years were randomly assigned to the placebo or ASE group (700 mg/d of ASE) and were instructed to take a once-daily pill for 12 weeks. Weight, BMI, waist circumference, fat mass (measured using bioimpedance, dual-energy X-ray absorptiometry, and computed tomography), and laboratory tests were assessed at baseline and at 12 weeks. Body weight significantly decreased after 12 weeks of treatment in the ASE group (placebo vs ASE: -0.08 ± 2.11 kg vs -3.30 ± 3.15 kg, P bioimpedance method: -0.51 ± 1.89 kg vs -2.38 ± 2.30 kg, P < .05; dual-energy X-ray absorptiometry: 0.38 ± 1.59 kg vs -2.26 ± 2.37 kg, P < .05). Changes in lipid profiles, fasting plasma glucose, and hemoglobin A1c did not differ between the 2 groups. No drug-related adverse events were observed during the study. In conclusion, ASE significantly decreases body weight and fat mass in obese humans, suggesting that ASE may be a potential therapeutic candidate for reducing obesity. PMID:27333958

  20. 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.

  1. 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.

  2. 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.

  3. Visual interpretation of ASTER satellite data, Part 1: Geologic mapping in the Cradle of Humankind World Heritage Site

    Directory of Open Access Journals (Sweden)

    Elna van Niekerk

    2007-09-01

    Full Text Available Since the first earth observing satellite was launched in 1972, remote sensing has become a powerful tool in the arsenal of geoscientists. This satellite became known as Landsat 1 and carried the Multispectral Scanner (MSS delivering imagery at a spatial resolution of 80, and spectral resolution from blue to near infrared. Ongoing satellite and sensor development to the end of the century produced the Landsat Thematic Mapper (TM with improved spatial and spectral resolution, as well as the SPOT series of satellites delivering the highest spatial but limited spectral resolution. These developments culminated in the SPOT 4 (1998 and Landsat Enhanced Thematic Mapper (1999 sensors. While Landsat ETM in particular provided much improved spatial and spectral resolutions, on the basis of which a large amount of geoscientific remote sensing was conducted world wide, the data did not provide adequate spectral and spatial sensitivity to be optimally effective for geological mapping at the local scale. On 18 December 1999 the Terra platform was launched, carrying five remote sensing instruments, including ASTER (Advanced Space borne Thermal Emission and Reflection Radiometer. ASTER consists of three separate instrument subsystems, each operating in a different spectral region, and using separate optical systems. These are the Visible and Very Near Infrared (VNIR subsystem with a 15m-spatial resolution, the Short Wave Infrared (SWIR subsystem with a 30m-spatial resolution and the Thermal Infrared (TIR subsystem with a 90m-spatial resolution. ASTER effectively offers an improvement on Landsat MSS, Landsat TM, Landsat ETM+ and SPOT spectral and spatial resolutions. Given the paucity of published research on geological remote sensing at the local scale in South Africa, and particularly on the use of ASTER for geological mapping in South Africa, it is imperative that the value of ASTER be investigated. This article reports on the improved detail and scale

  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. The resolution-dependence of satellite-based cloud retrievals: First results from ASTER and MODIS observations

    Science.gov (United States)

    Werner, F.; Wind, G.; Zhang, Z.; Platnick, S. E.; Di Girolamo, L.

    2015-12-01

    The spatial resolution dependence of retrieved optical and microphysical cloud properties of marine shallow convective water clouds is presented using data from the Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER), as well as the Moderate-resolution Imaging Spectroradiometer (MODIS) onboard the scientific research satellite Terra. Both instruments are characterized by vastly different spatial resolutions of 15m (ASTER) and 1000m (MODIS), respectively. Cloud optical thickness (τ) and effective droplet radius (reff) are derived by means of the Cross-platform HIgh resolution Multi-instrument AtmosphEric Retrieval Algorithms (CHIMAERA) system which yields MODIS-like cloud property retrievals via a shared-core architecture. The retrieval algorithm employs a standard bi-spectral retrieval scheme with two reflectances (ρ) in the visible to near-infrared spectral wavelength range (VNIR, 0.86μm) and shortwave infrared spectral wavelength range (SWIR, 2.1μm), respectively. For an exemplary granule the high-resolution ρ sampled by the ASTER instrument are aggregated from 15m to an increasingly coarse spatial resolution between (30-1000m). Subsequently, retrieved τ and reff from aggregated ρ are compared to the mean of the high-resolution cloud properties within the aggregated pixels. The differences in retrieved τ and reff are related to the sub-pixel covariance of ρ in the VNIR and SWIR band, as well as the inhomogeneity index (i.e., the ratio of standard deviation to mean value of ρ in the VNIR). This analysis highlights the impact of sub-pixel inhomogeneity and plane-parallel assumptions in the cloud property retrieval. CHIMAERA also allows for a comparison of ASTER and MODIS retrievals without introducing biases due to individual instrument algorithms. Retrieved τ and reff from the 1000m aggregated ρ sampled by ASTER are compared to the retrieved cloud properties provided by MODIS. The presented results highlight the different

  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. 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.

  8. Variation and significance of microtubules in rat volume overload cardiac hypertrophy

    Institute of Scientific and Technical Information of China (English)

    刘华胜; 马爱群; 王春梅; 刘勇; 田红燕; 白玲

    2003-01-01

    Objective To investigate the function of microtubules in volume overload cardiac hypertrophy of rat. Methods The structure of microtubules was observed using an immunofluorescent microscope, while the pixel intensity and distribution of microtubule imaging was estimated from laser scanning confocal images of left ventricular cardiocytes immuno-labeled with an antibody to β-tubulin. Results The pixels of the microtubule image taken just after volume overload were not evenly distributed. At 6 hours after overload, the pixel intensity of the microtubule image was decreased to less than 150 (arbitrary units), which was the same as the pixel intensity and distribution of the colchicine depolymerized microtubule image. The changes were partially recovered to 200 (arbitrary units) after 4 more days. The pixel intensity of the control microtubule image was 250 (arbitrary units) and had an even distribution. The structuring of the microtubules was more disordered as volume overload hypertrophy developed. Conclusions There are disorders in the signal transduction pathways governing the hypertrophic response of cardiomyocytes in the hypertrophic myocardium and microtubule is one of the members of the signal transduction pathways governing the hypertrophic response of cardiomyocytes in the hypertrophic myocardium. The disordered microtubule array may be targeted during heart failure treatment.

  9. 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.

  10. 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.

  11. 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.

  12. Disruption of microtubules uncouples budding and nuclear division in Toxoplasma gondii.

    Science.gov (United States)

    Morrissette, Naomi S; Sibley, L David

    2002-03-01

    The tachyzoite stage of the protozoan parasite Toxoplasma gondii has two populations of microtubules: spindle microtubules and subpellicular microtubules. To determine how these two microtubule populations are regulated, we investigated microtubule behavior during the cell cycle following treatment with microtubule-disrupting drugs. Previous work had established that the microtubule populations are individually nucleated by two distinct microtubule-organizing centers (MTOCs): the apical polar ring for the subpellicular microtubules and spindle pole plaques/centrioles for the spindle microtubules. When replicating tachyzoites were treated with 0.5 microM oryzalin or 1.0 mM colchicine they retained the capacity to form a spindle and undergo nuclear division. Although these parasites could complete budding, they lost the bulk of their subpellicular microtubules and the ability to reinvade host cells. Both nascent spindle and subpellicular microtubules were disrupted in 2.5 microM oryzalin or 5.0 mM colchicine. Under these conditions, parasites grew in size and replicated their genome but were incapable of nuclear division. After removal from 0.5 microM oryzalin, Toxoplasma tachyzoites were able to restore normal subpellicular microtubules and a fully invasive phenotype. When oryzalin was removed from Toxoplasma tachyzoites treated with 2.5 microM drug, the parasites attempted to bud as crescent-shaped tachyzoites. Because the polyploid nuclear mass could not be correctly segregated, many daughter parasites lacked nuclei altogether although budding and scission from the maternal mass was able to be completed. Multiple MTOCs permit Toxoplasma tachyzoites to control nuclear division independently from cell polarity and cytokinesis. This unusual situation grants greater cell cycle flexibility to these parasites but abolishes the checks for coregulation of nuclear division and cytokinesis found in other eukaryotes. PMID:11870220

  13. Topographic profile of a target with use of laser pulses. A survey directed to the Brazilian deep space mission ASTER

    International Nuclear Information System (INIS)

    This work is directly related to the development of the laser altimeter for the ASTER mission, named ALR. The Brazilian deep space mission ASTER plans to send a small spacecraft to encounter and investigate the triple asteroid 2001-SN263. The launch is scheduled to occur in 2017 and the ALR is now under development in partnership with UNICAMP, UFABC and aerospace companies. In this work, the environment and the operation of the instrument were modeled and simulations were carried out in order to better understand and define the instrument parameters. The creation of the simulation software to control the operation of the instrument was the main purpose of this work, and the software so far created is the main result of it. The software was successfully tested with respect to some common expected situations

  14. Mapping Hydrothermal Alterations in the Muteh Gold Mining Area in Iran by using ASTER satellite Imagery data

    Science.gov (United States)

    Asadi Haroni, Hooshang; Hassan Tabatabaei, Seyed

    2016-04-01

    Muteh gold mining area is located in 160 km NW of Isfahan town. Gold mineralization is meso-thermal type and associated with silisic, seresitic and carbonate alterations as well as with hematite and goethite. Image processing and interpretation were applied on the ASTER satellite imagery data of about 400 km2 at the Muteh gold mining area to identify hydrothermal alterations and iron oxides associated with gold mineralization. After applying preprocessing methods such as radiometric and geometric corrections, image processing methods of Principal Components Analysis (PCA), Least Square Fit (Ls-Fit) and Spectral Angle Mapper (SAM) were applied on the ASTER data to identify hydrothermal alterations and iron oxides. In this research reference spectra of minerals such as chlorite, hematite, clay minerals and phengite identified from laboratory spectral analysis of collected samples were used to map the hydrothermal alterations. Finally, identified hydrothermal alteration and iron oxides were validated by visiting and sampling some of the mapped hydrothermal alterations.

  15. 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 ...

  16. 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

  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. Surface Heat Balance Analysis of Tainan City on March 6, 2001 Using ASTER and Formosat-2 Data

    Science.gov (United States)

    Kato, Soushi; Yamaguchi, Yasushi; Liu, Cheng-Chien; Sun, Chen-Yi

    2008-01-01

    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.

  19. Untersuchungen zum Entwicklungsrhythmus und zur Kultursteuerung belaubt überwinternder Stauden am Beispiel von Aster alpinus 'Happy End'

    OpenAIRE

    Jende, Andreas

    2000-01-01

    Am Beispiel von Aster alpinus 'Happy End' wird der Entwicklungsverlauf vom Zeitpunkt der Vermehrung bis zur Ausbildung von Infloreszenzen beschrieben, um daraus Empfehlungen für die Kultur als blühende Topfstaude abzuleiten. Dazu wurden über einen Zeitraum von drei Jahren Versuche mit unterschiedlich vermehrten Pflanzen durchgeführt. Verwendet wurden hierbei Pflanzen aus generativer und vegetativer Vermehrung (Stecklinge, in vitro). Untersucht wurde der Verlauf der vegetativen und generativen...

  20. Using ASTER Imagery in Land Use/cover Classification of Eastern Mediterranean Landscapes According to CORINE Land Cover Project

    OpenAIRE

    Recep Gundogan; Abdullah E. Akay; Alaaddin Yüksel

    2008-01-01

    The satellite imagery has been effectively utilized for classifying land cover types and detecting land cover conditions. The Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) sensor imagery has been widely used in classification process of land cover. However, atmospheric corrections have to be made by preprocessing satellite sensor imagery since the electromagnetic radiation signals received by the satellite sensors can be scattered and absorbed by the atmospheric gases...

  1. An approach of surface coal fire detection from ASTER and Landsat-8 thermal data: Jharia coal field, India

    Science.gov (United States)

    Roy, Priyom; Guha, Arindam; Kumar, K. Vinod

    2015-07-01

    Radiant temperature images from thermal remote sensing sensors are used to delineate surface coal fires, by deriving a cut-off temperature to separate coal-fire from non-fire pixels. Temperature contrast of coal fire and background elements (rocks and vegetation etc.) controls this cut-off temperature. This contrast varies across the coal field, as it is influenced by variability of associated rock types, proportion of vegetation cover and intensity of coal fires etc. We have delineated coal fires from background, based on separation in data clusters in maximum v/s mean radiant temperature (13th band of ASTER and 10th band of Landsat-8) scatter-plot, derived using randomly distributed homogeneous pixel-blocks (9 × 9 pixels for ASTER and 27 × 27 pixels for Landsat-8), covering the entire coal bearing geological formation. It is seen that, for both the datasets, overall temperature variability of background and fires can be addressed using this regional cut-off. However, the summer time ASTER data could not delineate fire pixels for one specific mine (Bhulanbararee) as opposed to the winter time Landsat-8 data. The contrast of radiant temperature of fire and background terrain elements, specific to this mine, is different from the regional contrast of fire and background, during summer. This is due to the higher solar heating of background rocky outcrops, thus, reducing their temperature contrast with fire. The specific cut-off temperature determined for this mine, to extract this fire, differs from the regional cut-off. This is derived by reducing the pixel-block size of the temperature data. It is seen that, summer-time ASTER image is useful for fire detection but required additional processing to determine a local threshold, along with the regional threshold to capture all the fires. However, the winter Landsat-8 data was better for fire detection with a regional threshold.

  2. Google Earth's derived digital elevation model: A comparative assessment with Aster and SRTM data

    International Nuclear Information System (INIS)

    This paper presents a statistical analysis showing additional evidence that Digital Elevation Model (DEM) derived from Google Earth is commendable and has a good correlation with ASTER (Advanced Space-borne Thermal Emission and Reflection Radiometer) and SRTM (Shuttle Radar Topography Mission) elevation data. The accuracy of DEM elevation points from Google Earth was compared against that of DEMs from ASTER and SRTM for flat, hilly and mountainous sections of a pre-selected rural watershed. For each section, a total of 5,000 DEM elevation points were extracted as samples from each type of DEM data. The DEM data from Google Earth and SRTM for flat and hilly sections are strongly correlated with the R2 of 0.791 and 0.891 respectively. Even stronger correlation is shown for the mountainous section where the R2 values between Google Earth's DEM and ASTER's and between Google Earth's DEM and SRTM's DEMs are respectively 0.917 and 0.865. Further accuracy testing was carried out by utilising the DEM dataset to delineate Muar River's watershed boundary using ArcSWAT2009, a hydrological modelling software. The result shows that the percentage differences of the watershed size delineated from Google Earth's DEM compared to those derived from Department of Irrigation and Drainage's data (using 20m-contour topographic map), ASTER and SRTM data are 9.6%, 10.6%, and 7.6% respectively. It is therefore justified to conclude that the DEM derived from Google Earth is relatively as acceptable as DEMs from other sources

  3. 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.

  4. 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.

  5. 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.

  6. 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.

  7. 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.

  8. 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.

  9. Generating daily high spatial land surface temperatures by combining ASTER and MODIS land surface temperature products for environmental process monitoring.

    Science.gov (United States)

    Wu, Mingquan; Li, Hua; Huang, Wenjiang; Niu, Zheng; Wang, Changyao

    2015-08-01

    There is a shortage of daily high spatial land surface temperature (LST) data for use in high spatial and temporal resolution environmental process monitoring. To address this shortage, this work used the Spatial and Temporal Adaptive Reflectance Fusion Model (STARFM), Enhanced Spatial and Temporal Adaptive Reflectance Fusion Model (ESTARFM), and the Spatial and Temporal Data Fusion Approach (STDFA) to estimate high spatial and temporal resolution LST by combining Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) LST and Moderate Resolution Imaging Spectroradiometer (MODIS) LST products. The actual ASTER LST products were used to evaluate the precision of the combined LST images using the correlation analysis method. This method was tested and validated in study areas located in Gansu Province, China. The results show that all the models can generate daily synthetic LST image with a high correlation coefficient (r) of 0.92 between the synthetic image and the actual ASTER LST observations. The ESTARFM has the best performance, followed by the STDFA and the STARFM. Those models had better performance in desert areas than in cropland. The STDFA had better noise immunity than the other two models. PMID:26165141

  10. 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.

  11. 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.

  12. 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.

  13. 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.

  14. The distribution of microtubules in differentiating cells of Micrasterias denticulata bréb.

    Science.gov (United States)

    Kiermayer, O

    1968-09-01

    As an extension of earlier cytophysiological and morphological studies on differentiating cells of Micrasterias denticulata, a fine structural investigation of glutaraldehyde-osmium tetroxide fixed material has been made. Special emphasis has been placed on the distribution of cytoplasmic microtubules and on their possible role in the processes of growth and differentiation. Four distinct systems of microtubules were found: (a) a band in the cortical protoplasm of the isthmus region which surrounds the nucleus; (b) several bands in the cortical protoplasm of the old half cells, with rod-like cross bridges between individual microtubules and between the microtubules and the plasmalemma; (c) clusters of microtubules near the posttelophase nucleus, some separated by "intertubular structures" possibly fibrils; and (d) microtubules in the internal and cortical protoplasm of differentiating half cells. PMID:24519210

  15. Advanced Spaceborne Thermal Emission and Reflection Radometer (ASTER) Enhanced Vegetation Index (EVI) Products from Global Earth Observation (GEO) Grid: An Assessment Using Moderate Resolution Imaging Spectroradiometer (MODIS) for Synergistic Applications

    OpenAIRE

    Satoshi Tsuchida; Tomoaki Miura; Hirokazu Yamamoto

    2012-01-01

    We assessed the compatibility of three Advanced Spaceborne Thermal Emission and Reflection Radometer (ASTER) based Enhanced Vegetation Index (EVI) products generated in the GEO Grid system to Moderate Resolution Imaging Spectroradiometer (MODIS) EVI. The three products were two forms of the two-band EVI with ASTER red and NIR bands but without a blue band and the original, three-band EVI computed with ASTER red and NIR, and MODIS blue reflectances. Our assessment results showed good compatibi...

  16. 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

    2016-01-01

    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. PMID:27461361

  17. 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.

  18. Modulating microtubule stability enhances the cytotoxic response of cancer cells to paclitaxel

    OpenAIRE

    Ahmed, Ahmed Ashour; Wang, Xiaoyan; Lu, Zhen; Goldsmith, Juliet,; Le, Xiao-Feng; Grandjean, Geoffrey; Bartholomeusz, Geoffrey; Broom, Bradley; Bast, Robert C.

    2011-01-01

    The extracellular matrix protein TGFBI enhances the cytotoxic response of cancer cells to paclitaxel by affecting integrin signals that stabilize microtubules. Extending the implications of this knowledge, we tested the more general hypothesis that cancer cell signals which increase microtubule stability before exposure to paclitaxel may increase its ability to stablize microtubules and thereby enhance its cytotoxicity. Toward this end, we performed an siRNA screen to evaluate how genetic dep...

  19. Assessment Of ASTER GDEM And SRTM Performance By Comparing With Survey Control Points And Icesat/GLAS Data

    Science.gov (United States)

    Jarihani, A. A.; Callow, J. N.

    2011-12-01

    Topographic data plays a critical role in water resources modeling with determining watershed hydrologic characteristics from raster-based digital elevation models (DEM). Raster-based DEMs have been widely used to derive topographic attributes used in hydraulic and hydrologic modeling such as slope, stream network, basins boundary and area. Accurate models of floodplain topography are essential for having accurate output of hydrologic models. The Shuttle Radar Topography Mission (SRTM) provides near-global topographic coverage of the Earth's surface with unprecedented consistency and accuracy with the resolution of 1-3 arc sec. A new Global Digital Elevation Model (GDEM) from optical stereo data acquired by the Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) was recently released with the resolution of 1 arc sec. This study the performance of the ASTER GDEM and SRTM DEMs is assessed by comparing with ground-based survey control points and point data from ICESat/GLAS (Ice, Cloud, and land Elevation Satellite/Geoscience Laser Altimeter System) for a 7x7degree area in Queensland Australia. This area contains large and remote river basins, where these datasets provide an invaluable resource from which river floodplain inundation can be measured and modelled. Our study focuses on both data and datum issues, which are required to provide a realistic assessment of the achievable relative and absolute ground topographic accuracies. We assessed ground-based survey control points' with ICESat satellite altimetry points and in turn assessed accuracy of the ASTER GDEM and SRTM DEMs over the study area. ICESat provides globally-distributed elevation data of high accuracy (2.47.3 m horizontal error and 0.040.13 m (per degree of incidence angle) vertical error)We applied the GLA14 elevation products (Land/Canopy elevations) for the Laser 3a observation period and Release 31 to compare with 5000 ground survey control points in study area.Our ICESat

  20. 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.

  1. Feature level fusion for enhanced geological mapping of ophiolile complex using ASTER and Landsat TM data

    International Nuclear Information System (INIS)

    Chromite ore deposit occurrence is related to ophiolite complexes as a part of the oceanic crust and provides a good opportunity for lithological mapping using remote sensing data. The main contribution of this paper is a novel approaches to discriminate different rock units associated with ophiolite complex using the Feature Level Fusion technique on ASTER and Landsat TM satellite data at regional scale. In addition this study has applied spectral transform approaches, consisting of Spectral Angle Mapper (SAM) to distinguish the concentration of high-potential areas of chromite and also for determining the boundary between different rock units. Results indicated both approaches show superior outputs compared to other methods and can produce a geological map for ophiolite complex rock units in the arid and the semi-arid region. The novel technique including feature level fusion and Spectral Angle Mapper (SAM) discriminated ophiolitic rock units and produced detailed geological maps of the study area. As a case study, Sikhoran ophiolite complex located in SE, Iran has been selected for image processing techniques. In conclusion, a suitable approach for lithological mapping of ophiolite complexes is demonstrated, this technique contributes meaningfully towards economic geology in terms of identifying new prospects

  2. 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.

  3. The complete chloroplast genome sequence of Aster spathulifolius (Asteraceae); genomic features and relationship with Asteraceae.

    Science.gov (United States)

    Choi, Kyoung Su; Park, SeonJoo

    2015-11-10

    Aster spathulifolius, a member of the Asteraceae family, is distributed along the coast of Japan and Korea. This plant is used for medicinal and ornamental purposes. The complete chloroplast (cp) genome of A. sphathulifolius consists of 149,473 bp that include a pair of inverted repeats of 24,751 bp separated by a large single copy region of 81,998 bp and a small single copy region of 17,973 bp. The chloroplast genome contains 78 coding genes, four rRNA genes and 29 tRNA genes. When compared to other cpDNA sequences of Asteraceae, A. spathulifolius showed the closest relationship with Jacobaea vulgaris, and its atpB gene was found to be a pseudogene, unlike J. vulgaris. Furthermore, evaluation of the gene compositions of J. vulgaris, Helianthus annuus, Guizotia abyssinica and A. spathulifolius revealed that 13.6-kb showed inversion from ndhF to rps15, unlike Lactuca of Asteraceae. Comparison of the synonymous (Ks) and nonsynonymous (Ka) substitution rates with J. vulgaris revealed that synonymous genes related to a small subunit of the ribosome showed the highest value (0.1558), while nonsynonymous rates of genes related to ATP synthase genes were highest (0.0118). These findings revealed that substitution has occurred at similar rates in most genes, and the substitution rates suggested that most genes is a purified selection. PMID:26164759

  4. 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.

  5. 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.

  6. 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.

  7. Astral microtubule pivoting promotes their search for cortical anchor sites during mitosis in budding yeast.

    Directory of Open Access Journals (Sweden)

    Stephan Baumgärtner

    Full Text Available Positioning of the mitotic spindle is crucial for proper cell division. In the budding yeast Saccharomyces cerevisiae, two mechanisms contribute to spindle positioning. In the Kar9 pathway, astral microtubules emanating from the daughter-bound spindle pole body interact via the linker protein Kar9 with the myosin Myo2, which moves the microtubule along the actin cables towards the neck. In the dynein pathway, astral microtubules off-load dynein onto the cortical anchor protein Num1, which is followed by dynein pulling on the spindle. Yet, the mechanism by which microtubules target cortical anchor sites is unknown. Here we quantify the pivoting motion of astral microtubules around the spindle pole bodies, which occurs during spindle translocation towards the neck and through the neck. We show that this pivoting is largely driven by the Kar9 pathway. The microtubules emanating from the daughter-bound spindle pole body pivot faster than those at the mother-bound spindle pole body. The Kar9 pathway reduces the time needed for an astral microtubule inside the daughter cell to start pulling on the spindle. Thus, we propose a new role for microtubule pivoting: By pivoting around the spindle pole body, microtubules explore the space laterally, which helps them search for cortical anchor sites in the context of spindle positioning in budding yeast.

  8. 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.

  9. 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.

  10. 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.

  11. 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.

  12. 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. PMID:1681420

  13. 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.

  14. Interaction of microtubules with active principles of Xanthium strumarium.

    Science.gov (United States)

    Menon, G S; Kuchroo, K; Dasgupta, D

    2001-01-01

    Indigenous variety of Xanthium strumarium (X. strumarium) was screened for its antimitotic activity using the microtubule-tubulin system isolated from mammalian tissue. A preliminary phytochemical screening of the whole extracts of the plant was carried out followed by partial purification of the whole extract of X.strumarium. The separated fractions obtained were identified and used for in vitro polymerization studies. The whole as well as partially separated chemical constituents of X. strumarium showed effective inhibition of tubulin polymerization. The results thus suggest that X. strumarium may possess antimitotic components. PMID:12002689

  15. Fluid mechanics in dentinal microtubules provides mechanistic insights into the difference between hot and cold dental pain.

    Directory of Open Access Journals (Sweden)

    Min Lin

    Full Text Available 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.

  16. 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

    2011-01-01

    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. PMID:21448459

  17. An Improved Method to Retrieve Surface Emissivity in a Canary Pine Forest Using Aster Data

    Science.gov (United States)

    Barreto-Velasco, Africa; Hernandez-Leal, Pedro A.; Arbelo, Manuel; Podesta, Guillermo P.

    2012-07-01

    The widely-used Temperature and Emissivity Separation (TES) algorithm has been shown to provide reliable temperature and emissivity estimates from land-leaving thermal data. Nevertheless, TES has some important limitations mainly related to its inability to correct important inaccuracies for gray bodies, such as vegetation. In this study, this problem is addressed through an improvement to the current NEM (Normalized Emissivity Method) module included in the TES algorithm. The proposed method, called Modified NEM (ModNEM), has been specifically designed to retrieve an accurate surface emissivity for bodies with a spectral behavior typical of gray-bodies, i. e., flat and near spectrally invariant. ModNEM selects two different approximations of surface temperature instead of the usual maximum temperature, with the aim of accounting for the peculiar spectral behavior of brightness temperature and emissivity for vegetation. The NEM and TES, as well as the ModNEM, have been used to recover surface emissivity in a pine forest located in Tenerife Island (Canary Islands, Spain) using data from the Advanced Spaceborne Thermal Emission Reflection Radiometer (ASTER). Results have been compared and validated using reference emissivity values obtained from the pines by means of the box method. This validation study showed that high uncertainties are associated with the standard techniques (up to 0.034), whereas ModNEM results in lower uncertainties in emissivity estimates (NEM module are not suitable for surfaces with a spectral behavior similar to a grey body. This work was supported by MICINN under Grant CGL2010-22189-C02.

  18. ALR - Laser altimeter for the ASTER deep space mission. Simulated operation above a surface with crater

    Science.gov (United States)

    de Brum, A. G. V.; da Cruz, F. C.; Hetem, A., Jr.

    2015-10-01

    To assist in the investigation of the triple asteroid system 2001-SN263, the deep space mission ASTER will carry onboard a laser altimeter. The instrument was named ALR and its development is now in progress. In order to help in the instrument design, with a view to the creation of software to control the instrument, a package of computer programs was produced to simulate the operation of a pulsed laser altimeter with operating principle based on the measurement of the time of flight of the travelling pulse. This software Simulator was called ALR_Sim, and the results obtained with its use represent what should be expected as return signal when laser pulses are fired toward a target, reflect on it and return to be detected by the instrument. The program was successfully tested with regard to some of the most common situations expected. It constitutes now the main workbench dedicated to the creation and testing of control software to embark in the ALR. In addition, the Simulator constitutes also an important tool to assist the creation of software to be used on Earth, in the processing and analysis of the data received from the instrument. This work presents the results obtained in the special case which involves the modeling of a surface with crater, along with the simulation of the instrument operation above this type of terrain. This study points out that the comparison of the wave form obtained as return signal after reflection of the laser pulse on the surface of the crater with the expected return signal in the case of a flat and homogeneous surface is a useful method that can be applied for terrain details extraction.

  19. IMÁGENES ASTER EN LA DISCRIMINACIÓN DE ÁREAS DE USO AGRÍCOLA EN COLOMBIA ASTER IMAGES FOR DISCRIMINATION OF AGRICULTURAL USE AREAS IN COLOMBIA

    OpenAIRE

    Nidia Esperanza Ortiz Lozano; Uriel Pérez Gómez

    2009-01-01

    El avance de las tecnologías de la información geográfica ha llevado a la puesta de nuevos sensores para observación de la tierra. ASTER (Advanced Spaceborne Termal Emission and Reflection Radiometer), considerado como sensor de última generación, posee características especiales que lo colocan como una alternativa para estudios de la cobertura vegetal de la tierra. El presente trabajo se basó en sus características espaciales, espectrales y radiométricas con el fin de discriminar las áreas d...

  20. Clasificación de yeso mediante imágenes ASTER en la Cordillera Principal Mendocina Gypsum classification based on ASTER images in the Principal Cordillera of Mendoza

    OpenAIRE

    José F Mescua

    2010-01-01

    Se presenta una metodología para obtener el mapa de distribución de yeso con imágenes satelitales ASTER. Se utilizaron imágenes con nivel de procesamiento ASTERL3A correspondientes a dos áreas de la Cordillera Principal mendocina. La estratigrafía de las áreas de estudio se encuentra compuesta por una espesa sucesión sedimentaria mesozoica, volcanitas cenozoicas y depósitos cuaternarios. Dentro de los depósitos mesozoicos se encuentra la Formación Auquilco, integrada por yeso y anhidrita. El ...

  1. 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 aplicação do modelo SEBAL em estimar os fluxos de energia em superfície e a evapotranspiração diária, numa extensa área de cultivo de arroz irrigado, no município de Paraíso do Sul, RS, tendo como parâmetros dados do sensor ASTER. As variáveis estudadas constituem importantes parâmetros do tempo e do clima em estudos agrometeorológicos e de racionalização no uso da água. As metodologias convencionais de estimativa desses parâmetros são pontuais e geralmente apresentam incertezas, que aumentam quando o interesse é o comportamento espacial desses parâmetros. Aplicou-se o algoritmo "Surface Energy Balance Algorithm for Land" (SEBAL, em uma imagem do sensor "Advanced Spaceborne Thermal Emission and Reflection Radiometer" (ASTER. As estimativas obtidas foram comparadas com medições em campo, realizadas por uma estação micrometeorológica localizada no interior da área de estudo. As estimativas mais precisas foram as de fluxo de calor sensível e de evapotranspiração diária, e a estimativa que apresentou maior erro foi a do fluxo de calor no solo. A metodologia empregada foi capaz de reproduzir os fluxos de energia em superfície de maneira satisfatória para estudos agrometeorológicos e de rendimento de culturas.The objective of this study was to evaluate the efficiency of SEBAL model in estimating soil surface energy fluxes and daily evapotranspiration for a large area of irrigated rice farms, near the municipality of Paraíso do Sul, RS, Brazil, using data from ASTER sensor. The evaluated variables are important weather and climatic parameters for agrometeorological studies and rationalization of water use. The conventional methodologies for estimating these parameters generally present uncertainties, which increase when concern is in the spatial behavior of such parameters. The Surface Energy Balance Algorithm for Land (SEBAL was applied in an Advanced Spaceborne

  2. Atividade antibacteriana e antifúngica de extratos etanólicos de Aster lanceolatus Willd., Asteraceae Antibacterial and antifungal activity of ethanolics extracts from Aster lanceolatus Willd., Asteraceae

    OpenAIRE

    Josiane F. Gaspari Dias; Suzane Virtuoso; Aline Davet; Miriam M. Cunico; Marilis D. Miguel; Obdúlio G. Miguel; Celso G. Auer; Albino Grigoletti-Júnior; Andressa B. Oliveira; Marlene L. Ferronato

    2006-01-01

    Conhecida popularmente como áster-arbustiva, margarida-de-são-miguel e monte-cassino, Aster lanceolatus é uma planta ornamental de corte. Sabendo-se que não existem estudos que atestem a atividade biológica desta espécie, procurou-se neste trabalho atividades antibacteriana e antifúngica. Para tanto se utilizaram extratos brutos etanólicos de duas porções distintas, das flores e dos caules com folhas. Para a atividade antibacteriana, oito bactérias patogênicas foram submetidas a ensaio de dif...

  3. CAMSAP3 orients the apical-to-basal polarity of microtubule arrays in epithelial cells.

    Science.gov (United States)

    Toya, Mika; Kobayashi, Saeko; Kawasaki, Miwa; Shioi, Go; Kaneko, Mari; Ishiuchi, Takashi; Misaki, Kazuyo; Meng, Wenxiang; Takeichi, Masatoshi

    2016-01-12

    Polarized epithelial cells exhibit a characteristic array of microtubules that are oriented along the apicobasal axis of the cells. The minus-ends of these microtubules face apically, and the plus-ends face toward the basal side. The mechanisms underlying this epithelial-specific microtubule assembly remain unresolved, however. Here, using mouse intestinal cells and human Caco-2 cells, we show that the microtubule minus-end binding protein CAMSAP3 (calmodulin-regulated-spectrin-associated protein 3) plays a pivotal role in orienting the apical-to-basal polarity of microtubules in epithelial cells. In these cells, CAMSAP3 accumulated at the apical cortices, and tethered the longitudinal microtubules to these sites. Camsap3 mutation or depletion resulted in a random orientation of these microtubules; concomitantly, the stereotypic positioning of the nucleus and Golgi apparatus was perturbed. In contrast, the integrity of the plasma membrane was hardly affected, although its structural stability was decreased. Further analysis revealed that the CC1 domain of CAMSAP3 is crucial for its apical localization, and that forced mislocalization of CAMSAP3 disturbs the epithelial architecture. These findings demonstrate that apically localized CAMSAP3 determines the proper orientation of microtubules, and in turn that of organelles, in mature mammalian epithelial cells. PMID:26715742

  4. Motility and microtubule depolymerization mechanisms of the Kinesin-8 motor, KIF19A

    Science.gov (United States)

    Wang, Doudou; Nitta, Ryo; Morikawa, Manatsu; Yajima, Hiroaki; Inoue, Shigeyuki; Shigematsu, Hideki; Kikkawa, Masahide; Hirokawa, Nobutaka

    2016-01-01

    The kinesin-8 motor, KIF19A, accumulates at cilia tips and controls cilium length. Defective KIF19A leads to hydrocephalus and female infertility because of abnormally elongated cilia. Uniquely among kinesins, KIF19A possesses the dual functions of motility along ciliary microtubules and depolymerization of microtubules. To elucidate the molecular mechanisms of these functions we solved the crystal structure of its motor domain and determined its cryo-electron microscopy structure complexed with a microtubule. The features of KIF19A that enable its dual function are clustered on its microtubule-binding side. Unexpectedly, a destabilized switch II coordinates with a destabilized L8 to enable KIF19A to adjust to both straight and curved microtubule protofilaments. The basic clusters of L2 and L12 tether the microtubule. The long L2 with a characteristic acidic-hydrophobic-basic sequence effectively stabilizes the curved conformation of microtubule ends. Hence, KIF19A utilizes multiple strategies to accomplish the dual functions of motility and microtubule depolymerization by ATP hydrolysis. DOI: http://dx.doi.org/10.7554/eLife.18101.001 PMID:27690357

  5. Feeding cells induced by phytoparasitic nematodes require γ-tubulin ring complex for microtubule reorganization.

    Directory of Open Access Journals (Sweden)

    Mohamed Youssef Banora

    2011-12-01

    Full Text Available Reorganization of the microtubule network is important for the fast isodiametric expansion of giant-feeding cells induced by root-knot nematodes. The efficiency of microtubule reorganization depends on the nucleation of new microtubules, their elongation rate and activity of microtubule severing factors. New microtubules in plants are nucleated by cytoplasmic or microtubule-bound γ-tubulin ring complexes. Here we investigate the requirement of γ-tubulin complexes for giant feeding cells development using the interaction between Arabidopsis and Meloidogyne spp. as a model system. Immunocytochemical analyses demonstrate that γ-tubulin localizes to both cortical cytoplasm and mitotic microtubule arrays of the giant cells where it can associate with microtubules. The transcripts of two Arabidopsis γ-tubulin (TUBG1 and TUBG2 and two γ-tubulin complex proteins genes (GCP3 and GCP4 are upregulated in galls. Electron microscopy demonstrates association of GCP3 and γ-tubulin as part of a complex in the cytoplasm of giant cells. Knockout of either or both γ-tubulin genes results in the gene dose-dependent alteration of the morphology of feeding site and failure of nematode life cycle completion. We conclude that the γ-tubulin complex is essential for the control of microtubular network remodelling in the course of initiation and development of giant-feeding cells, and for the successful reproduction of nematodes in their plant hosts.

  6. Configuration of the microtubule cytoskeleton in elongating fibers of flax (Linum usitatissimum L.)

    NARCIS (Netherlands)

    Lammeren, van A.A.M.; Ageeva, M.; Kieft, H.; Lhuissier, F.G.P.; Vos, J.; Gorshkova, T.; Emons, A.M.C.

    2003-01-01

    There are three basic types of plant cell growth: isodiametric, unidirectional diffuse, and tip growth. During plant cell growth, microtubules are present in the cell cortex, appressed against the plasma membrane. It is well documented that these cortical microtubules determine the orientation of ce

  7. Specific polar subpopulations of astral microtubules control spindle orientation and symmetric neural stem cell division.

    Science.gov (United States)

    Mora-Bermúdez, Felipe; Matsuzaki, Fumio; Huttner, Wieland B

    2014-01-01

    Mitotic spindle orientation is crucial for symmetric vs asymmetric cell division and depends on astral microtubules. Here, we show that distinct subpopulations of astral microtubules exist, which have differential functions in regulating spindle orientation and division symmetry. Specifically, in polarized stem cells of developing mouse neocortex, astral microtubules reaching the apical and basal cell cortex, but not those reaching the central cell cortex, are more abundant in symmetrically than asymmetrically dividing cells and reduce spindle orientation variability. This promotes symmetric divisions by maintaining an apico-basal cleavage plane. The greater abundance of apical/basal astrals depends on a higher concentration, at the basal cell cortex, of LGN, a known spindle-cell cortex linker. Furthermore, newly developed specific microtubule perturbations that selectively decrease apical/basal astrals recapitulate the symmetric-to-asymmetric division switch and suffice to increase neurogenesis in vivo. Thus, our study identifies a novel link between cell polarity, astral microtubules, and spindle orientation in morphogenesis. PMID:24996848

  8. ASTER Observations of 2000-2007 Thermal Features at Pavlof Volcano and Mt. Hague (Emmons Lake Volcanic Center), Alaska

    Science.gov (United States)

    Wessels, R. L.; Schneider, D.; Ramsey, M.; Mangan, M. T.

    2007-12-01

    Emmons Lake Volcanic Center (ELVC) is a 15 km by 30 km area of nested calderas, stratovolcanoes, lava domes, hyaloclastite rings, and cinder cones aligned along the arc axis. Pavlof Volcano is the most active volcano along the ELVC, with more than 40 historic eruptions since 1790. The most recent eruption of Pavlof Volcano began in August 2007 after almost 11 years of quiescence. Mount Hague is a prominent intracaldera vent with no known historical eruptions that lies approximately 7 kilometers to the southwest of Pavlof. The southern crater of Mount Hague commonly fluctuates between a crater-filling lake to a dry crater floor with vigorously steaming fumaroles. Mount Hague has another fumarole field on the southeast flank at nearly the same elevation as the crater floor. To better document the behavior of persistent thermal features at these remote volcanoes, we have compiled temperature and dimension data using a seven-year long time series of satellite data. Over 25 daytime and 40 nighttime clear thermal infrared (TIR) images (90 m resolution) from the Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) have recorded variations in the thermal activity at both volcanic vents since July 2000. All cloud-free ASTER TIR observations document persistent low- temperature features at both Pavlof Volcano and Mount Hague during this period. The size and temperature of each thermal feature varies throughout the study period. The data show that the 2518 m summit of Pavlof Volcano is occasionally snow-free in early summer whereas neighboring peaks at lower elevations are still snow-clad. FLIR data acquired near the summit of Pavlof in 2004 show that the majority of warm ground was at 20°C to 40°C. These warm areas commonly persist snow-free into the winter. Temperature variations observed at Mt Hague crater usually correlate to the size of the ephemeral crater lake. As the lake grows, the pixel-integrated ASTER TIR temperature increases. Measurements

  9. Clasificación de yeso mediante imágenes ASTER en la Cordillera Principal Mendocina

    OpenAIRE

    José F Mescua

    2010-01-01

    Se presenta una metodología para obtener el mapa de distribución de yeso con imágenes satelitales ASTER. Se utilizaron imágenes con nivel de procesamiento ASTERL3A correspondientes a dos áreas de la Cordillera Principal mendocina. La estratigrafía de las áreas de estudio se encuentra compuesta por una espesa sucesión sedimentaria mesozoica, volcanitas cenozoicas y depósitos cuaternarios. Dentro de los depósitos mesozoicos se encuentra la Formación Auquilco, integrada por yeso y anhidrita. El ...

  10. Microtubules restrict plastid sedimentation in protonemata of the moss Ceratodon

    Science.gov (United States)

    Schwuchow, J.; Sack, F. D.

    1994-01-01

    Apical cells of protonemata of the moss Ceratodon purpureus are unusual among plant cells with sedimentation in that only some amyloplasts sediment and these do not fall completely to the bottom of vertical cells. To determine whether the cytoskeleton restricts plastid sedimentation, the effects of amiprophos-methyl (APM) and cytochalasin D (CD) on plastid position were quantified. APM treatments of 30-60 min increased the plastid sedimentation that is normally seen along the length of untreated or control cells. Longer APM treatments often resulted in more dramatic plastid sedimentation, and in some cases almost all plastids sedimented to the lowermost point in the cell. In contrast, the microfilament inhibitor CD did not affect longitudinal plastid sedimentation compared to untreated cells, although it did disturb or eliminate plastid zonation in the tip. These data suggest that microtubules restrict the sedimentation of plastids along the length of the cell and that microtubules are load-bearing for all the plastids in the apical cell. This demonstrates the importance of the cytoskeleton in maintaining organelle position and cell organization against the force of gravity.

  11. Cytomagnetometric study of interactions between microfilaments and microtubules by measuring the energy imparted to magnetic particles within the cells

    Energy Technology Data Exchange (ETDEWEB)

    Nemoto, Iku [Tokyo Denki University School of Information Environment 2-1200, Muzai-Gakuendai, Inzai, Chiba 270-1382 (Japan)]. E-mail: nemoto@sie.dendai.ac.jp; Kawamura, Kazuhisa [Tokyo Denki University School of Science and Engineering Hatoyama, Saitama 350-0394 (Japan)

    2005-05-15

    Cytomagnetometric measurements of the energy imparted to intracellular organelles were made to study the relationship between microtubules and microfilaments. Depolymerization of microtubules by colchicine resulted in an increase in the energy suggesting that microtubules in control condition suppress the activity of microfilaments.

  12. Phospholipase d activation correlates with microtubule reorganization in living plant cells.

    Science.gov (United States)

    Dhonukshe, Pankaj; Laxalt, Ana M; Goedhart, Joachim; Gadella, Theodorus W J; Munnik, Teun

    2003-11-01

    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 of PLD, cortical microtubules were released from the plasma membrane and partially depolymerized, as visualized with four-dimensional confocal imaging. The isomers sec- and tert-butanol, which did not activate PLD, did not affect microtubule organization. The effect of treatment on PLD activation was monitored by the in vivo formation of phosphatidylbutanol, a specific reporter of PLD activity. Tobacco cells also were treated with mastoparan, xylanase, NaCl, and hypoosmotic stress as reported activators of PLD. We confirmed the reports and found that all treatments induced microtubule reorganization and PLD activation within the same time frame. PLD still was activated in microtubule-stabilized (taxol) and microtubule-depolymerized (oryzalin) situations, suggesting that PLD activation triggers microtubular reorganization and not vice versa. Exogenously applied water-soluble synthetic phosphatidic acid did not affect the microtubular cytoskeleton. Cell cycle studies revealed that n-butanol influenced not just interphase cortical microtubules but also those in the preprophase band and phragmoplast, but not those in the spindle structure. Cell growth and division were inhibited in the presence of n-butanol, whereas sec- and tert-butanol had no such effects. Using these novel insights, we propose a model for the mechanism by which PLD activation triggers microtubule reorganization in plant cells.

  13. The Penetration Depth Derived from the Synthesis of ALOS/PALSAR InSAR Data and ASTER GDEM for the Mapping of Forest Biomass

    Directory of Open Access Journals (Sweden)

    Wenjian Ni

    2014-08-01

    Full Text Available The Global Digital Elevation Model produced from stereo images of Advanced Spaceborne Thermal Emission and Reflection Radiometer data (ASTER GDEM covers land surfaces between latitudes of 83°N and 83°S. The Phased Array type L-band Synthetic Aperture Radar (PALSAR onboard Advanced Land Observing Satellite (ALOS collected many SAR images since it was launched on 24 January 2006. The combination of ALOS/PALSAR interferometric data and ASTER GDEM should provide the penetration depth of SAR data assuming ASTER GDEM was the elevation of vegetation canopy top. It would be correlated with forest biomass because penetration depth could be affected by forest density and forest canopy height. Their combination held great promises for the forest biomass mapping over large area. The feasibility of forest biomass mapping through the data synthesis of ALOS/PALSAR InSAR data and ASTER GDEM was investigated in this study. A procedure for the extraction of penetration depth was firstly proposed. Then three models were built for biomass estimation: (I model only using backscattering coefficients of ALOS/PALSAR data; (II model only using penetration depth; (III model using both of them. The biomass estimated from Lidar data was taken as reference data to evaluate the three different models. The results showed that the combination of backscattering coefficients and penetration depth gave the best accuracy. The forest disturbance has to be considered in forest biomass estimation because of the long time span of ASTER data for generating ASTER GDEM. The spatial homogeneity could be used to improve estimation accuracy.

  14. Tools for the rational design of bivalent microtubule-targeting drugs.

    Science.gov (United States)

    Marangon, Jacopo; Christodoulou, Michael S; Casagrande, Fancesca V M; Tiana, Guido; Dalla Via, Lisa; Aliverti, Alessandro; Passarella, Daniele; Cappelletti, Graziella; Ricagno, Stefano

    2016-10-01

    Microtubule (MT) dynamic behaviour is an attractive drug target for chemotherapy, whose regulation by MT-stabilizing and destabilizing agents has been fruitfully applied in treating several types of cancers. MT-stabilizing agents are also emerging as potential remedies for neurodegenerative conditions, such as Alzheimer's and Parkinson's disease, although single-target drugs are not expected to fully cure these complex pathologies. Drug combination often displays enhanced efficacy with respect to mono-therapies. In particular, MT-targeting bivalent compounds (MTBCs) represent a promising class of molecules; however, surprisingly, the majority of MTBCs reported so far exhibit equal if not less efficacy than their building monomers. In order to shed light on MTBCs poor performance, we characterised through a set of complementary approaches thiocolchine (TH) and two bivalent TH-homodimers as prototype molecules. First, the binding affinities of these three molecules were assessed, then we obtained the crystallographic structure of a tubulin-TH complex. The binding affinities were interpreted in light of structural data and of molecular dynamics simulations. Finally, their effects on MT cytoskeleton and cell survival were validated on HeLa cells. The ensemble of these data provides chemical and structural considerations on how a successful rational design of MTBCs should be conceived.

  15. 天睿子公司Aster Data的专利技术打开大数据商机大门

    Institute of Scientific and Technical Information of China (English)

    2011-01-01

    中国,北京—全球最大、专注于数据仓库,大数据分析和整合营销管理解决方案的厂商Teradata天睿公司(Teradata Corporation,美国纽约证券交易所交易代码:TDC)的子公司Aster Data日前宣布,其SQL—Map Reduce技术获得了美国第7966340号专利。Teradata天睿公司于今年四月收购了Aster Data公司。

  16. Clasificación de yeso mediante imágenes ASTER en la Cordillera Principal Mendocina Gypsum classification based on ASTER images in the Principal Cordillera of Mendoza

    Directory of Open Access Journals (Sweden)

    José F Mescua

    2010-06-01

    Full Text Available Se presenta una metodología para obtener el mapa de distribución de yeso con imágenes satelitales ASTER. Se utilizaron imágenes con nivel de procesamiento ASTERL3A correspondientes a dos áreas de la Cordillera Principal mendocina. La estratigrafía de las áreas de estudio se encuentra compuesta por una espesa sucesión sedimentaria mesozoica, volcanitas cenozoicas y depósitos cuaternarios. Dentro de los depósitos mesozoicos se encuentra la Formación Auquilco, integrada por yeso y anhidrita. El reconocimiento y distribución areal de esta unidad es el objetivo de este trabajo. Se aplicaron dos métodos de procesamiento a las imágenes. Por un lado se utilizó el índice de cuarzo (Qi, uno de los índices de determinación de minerales definidos para el subsistema TIR de ASTER. Este índice puede usarse también para reconocer sulfatos, y en particular yeso, ya que estos minerales proporcionan una respuesta muy baja en el Qi. El segundo método consistió en la aplicación combinada de los cocientes de bandas 4/5 y 7/5. Este método permite identificar al yeso por su elevada respuesta en el cociente 4/5 y baja respuesta en el cociente 7/5. A fin de simplificar la metodología, se generó un nuevo cociente a partir de los dos mencionados: (4/5/(7/5, de manera de destacar el yeso con alta respuesta. A partir del relevamiento de campo se observó que en ambos métodos los errores por exceso fueron más importantes que los errores por defecto, y que los píxeles determinados como yeso erróneamente por cada método son diferentes. Este hecho permite la aplicación de la técnica de "Decision Tree" del programa ENVI 4.2 con el objetivo de eliminar estos errores. En cada una de las imágenes se determinaron los valores de umbral para los píxeles reconocidos como yeso. Luego se generó un "árbol de decisiones" combinando los umbrales de cada imagen, seleccionando en una nueva clase los píxeles determinados como yeso por ambos métodos y

  17. Digital Mapping of Soil Drainage Classes Using Multitemporal RADARSAT-1 and ASTER Images and Soil Survey Data

    Directory of Open Access Journals (Sweden)

    Mohamed Abou Niang

    2012-01-01

    Full Text Available Discriminant analysis classification (DAC and decision tree classifiers (DTC were used for digital mapping of soil drainage in the Bras-d’Henri watershed (QC, Canada using earth observation data (RADARSAT-1 and ASTER and soil survey dataset. Firstly, a forward stepwise selection was applied to each land use type identified by ASTER image in order to derive an optimal subset of soil drainage class predictors. The classification models were then applied to these subsets for each land use and merged to obtain a digital soil drainage map for the whole watershed. The DTC method provided better classification accuracies (29 to 92% than the DAC method (33 to 79% according to the land use type. A similarity measure (S was used to compare the best digital soil drainage map (DTC to the conventional soil drainage map. Medium to high similarities (0.6≤S<0.9 were observed for 83% (187 km2 of the study area while 3% of the study area showed very good agreement (S≥0.9. Few soil polygons showed very weak similarities (S<0.3. This study demonstrates the efficiency of combining radar and optical remote sensing data with a representative soil dataset for producing digital maps of soil drainage.

  18. Microtubule +TIP protein EB1 binds to GTP and undergoes dissociation from dimer to monomer on binding GTP.

    Science.gov (United States)

    Gireesh, K K; Sreeja, Jamuna S; Chakraborti, Soumyananda; Singh, Puja; Thomas, Geethu Emily; Gupta, Hindol; Manna, Tapas

    2014-09-01

    The +TIP protein EB1 autonomously tracks the growing plus end of microtubules and regulates plus-end dynamics. Previous studies have indicated that EB1 can recognize GTP-bound tubulin structures at the plus end, and it localizes on the microtubule surface at a site close to the exchangeable GTP-binding site of tubulin. Although the GTP-dependent structural change in tubulin has been demonstrated to be a critical determinant for recognition of plus ends by EB1, the effect of GTP on the structure of EB1 has remained unclear. Here, we have used spectroscopic, calorimetric, and biochemical methods to analyze the effect of GTP on EB1 in vitro. Isothermal titration calorimetry and tryptophan fluorescence quenching experiments demonstrated that EB1 binds to GTP with a dissociation constant ~30 μM. Circular dichroism measurements showed that EB1 undergoes changes in its secondary structure on binding GTP. Size-exclusion chromatography and urea-induced unfolding analyses revealed that GTP binding induces dissociation of the EB1 dimer to monomers. Size-exclusion chromatography followed by biochemical analysis further determined that EB1-GTP binding involves association of approximately one molecule of GTP per EB1 monomer. The results reveal a hitherto unknown GTP-dependent mechanism of dimer-to-monomer transition in EB1 and further implicate its possible role in regulating the stability of the EB1 dimer vs monomer as well as plus-end regulation in cells. PMID:25111064

  19. Synthesis and SAR requirements of adamantane-colchicine conjugates with both microtubule depolymerizing and tubulin clustering activities.

    Science.gov (United States)

    Zefirova, Olga N; Nurieva, Evgeniya V; Shishov, Dmitrii V; Baskin, Igor I; Fuchs, Fabian; Lemcke, Heiko; Schröder, Fabian; Weiss, Dieter G; Zefirov, Nikolay S; Kuznetsov, Sergei A

    2011-09-15

    A series of analogues of conjugate 1, combining an adamantane-based paclitaxel (taxol) mimetic with colchicine was synthesized and tested for cytotoxicity in a cell-based assay with the human lung carcinoma cell line A549. The most active compounds (10 EC(50) 2 ± 1.0 nM, 23 EC(50) 6 ± 1.4 nM, 26 EC(50) 5 ± 1.8 nM, 28 EC(50) 11 ± 1.7 nM, 30 EC(50) 4.8 ± 0.5 nM) were found to interfere with the microtubule dynamics in an interesting manner. Treatment of the cells with these compounds promoted disassembly of microtubules followed by the formation of stable tubulin clusters. Structure-activity relationships for the analogues of 23 revealed the sensitivity of both cytotoxicity and tubulin clustering ability to the linker length. The presence of adamantane (or another bulky hydrophobic and non-aromatic moiety) in 23 was found to play an important role in the formation of tubulin clusters. Structural requirements for optimal activity have been partially explained by molecular modeling. PMID:21873068

  20. Septin 9 induces lipid droplets growth by a phosphatidylinositol-5-phosphate and microtubule-dependent mechanism hijacked by HCV

    Science.gov (United States)

    Akil, Abdellah; Peng, Juan; Omrane, Mohyeddine; Gondeau, Claire; Desterke, Christophe; Marin, Mickaël; Tronchère, Hélène; Taveneau, Cyntia; Sar, Sokhavuth; Briolotti, Philippe; Benjelloun, Soumaya; Benjouad, Abdelaziz; Maurel, Patrick; Thiers, Valérie; Bressanelli, Stéphane; Samuel, Didier; Bréchot, Christian; Gassama-Diagne, Ama

    2016-01-01

    The accumulation of lipid droplets (LD) is frequently observed in hepatitis C virus (HCV) infection and represents an important risk factor for the development of liver steatosis and cirrhosis. The mechanisms of LD biogenesis and growth remain open questions. Here, transcriptome analysis reveals a significant upregulation of septin 9 in HCV-induced cirrhosis compared with the normal liver. HCV infection increases septin 9 expression and induces its assembly into filaments. Septin 9 regulates LD growth and perinuclear accumulation in a manner dependent on dynamic microtubules. The effects of septin 9 on LDs are also dependent on binding to PtdIns5P, which, in turn, controls the formation of septin 9 filaments and its interaction with microtubules. This previously undescribed cooperation between PtdIns5P and septin 9 regulates oleate-induced accumulation of LDs. Overall, our data offer a novel route for LD growth through the involvement of a septin 9/PtdIns5P signalling pathway. PMID:27417143

  1. Characterization of tub4P287L, a b-tubulin mutant, revealed new aspects of microtubule regulation in shade

    Institute of Scientific and Technical Information of China (English)

    Jie Yu; Hong Qiu; Xin Liu; Meiling Wang; Yongli Gao; Joanne Chory; Yi Tao

    2015-01-01

    When sun plants, such as Arabidopsis thaliana, are under canopy shade, elongation of stems/petioles will be induced as one of the most prominent responses. Plant hormones mediate the elongation growth. However, how environmental and hormonal signals are translated into cell expansion activity that leads to the elongation growth remains elusive. Through forward genetic study, we identi-fied shade avoidance2 (sav2) mutant, which contains a P287L mutation in b-TUBULIN 4. Cortical microtubules (cMTs) play a key role in anisotropic cell growth. Hypocotyls of sav2 are wild type-like in white light, but are short and highly swollen in shade and dark. We showed that shade not only induces cMT rearrangement, but also affects cMT stability and dynamics of plus ends. Even though auxin and brassinosteroids are required for shade-induced hypocotyl elongation, they had little effect on shade-induced rearrangement of cMTs. Blocking auxin transport suppressed dark phenotypes of sav2, while overexpressing EB1b-GFP, a microtubule plus-end binding protein, rescued sav2 in both shade and dark, suggesting that tub4P287L represents a unique type of tubulin mutation that does not affect cMT function in supporting cell elongation, but may affect the ability of cMTs to respond properly to growth promoting stimuli.

  2. Vertical Accuracy Assessment of 30-M Resolution Alos, Aster, and Srtm Global Dems Over Northeastern Mindanao, Philippines

    Science.gov (United States)

    Santillan, J. R.; Makinano-Santillan, M.

    2016-06-01

    The ALOS World 3D - 30 m (AW3D30), ASTER Global DEM Version 2 (GDEM2), and SRTM-30 m are Digital Elevation Models (DEMs) that have been made available to the general public free of charge. An important feature of these DEMs is their unprecedented horizontal resolution of 30-m and almost global coverage. The very recent release of these DEMs, particularly AW3D30 and SRTM- 30 m, calls for opportunities for the conduct of localized assessment of the DEM's quality and accuracy to verify their suitability for a wide range of applications in hydrology, geomorphology, archaelogy, and many others. In this study, we conducted a vertical accuracy assessment of these DEMs by comparing the elevation of 274 control points scattered over various sites in northeastern Mindanao, Philippines. The elevations of these control points (referred to the Mean Sea Level, MSL) were obtained through 3rd order differential levelling using a high precision digital level, and their horizontal positions measured using a global positioning system (GPS) receiver. These control points are representative of five (5) land-cover classes namely brushland (45 points), built-up (32), cultivated areas (97), dense vegetation (74), and grassland (26). Results showed that AW3D30 has the lowest Root Mean Square Error (RMSE) of 5.68 m, followed by SRTM-30 m (RMSE = 8.28 m), and ASTER GDEM2 (RMSE = 11.98 m). While all the three DEMs overestimated the true ground elevations, the mean and standard deviations of the differences in elevations were found to be lower in AW3D30 compared to SRTM-30 m and ASTER GDEM2. The superiority of AW3D30 over the other two DEMS was also found to be consistent even under different landcover types, with AW3D30's RMSEs ranging from 4.29 m (built-up) to 6.75 m (dense vegetation). For SRTM-30 m, the RMSE ranges from 5.91 m (built-up) to 10.42 m (brushland); for ASTER GDEM2, the RMSE ranges from 9.27 m (brushland) to 14.88 m (dense vegetation). The

  3. MTB-3, a microtubule plus-end tracking protein (+TIP of Neurospora crassa.

    Directory of Open Access Journals (Sweden)

    Rosa R Mouriño-Pérez

    Full Text Available The microtubule (MT "plus end" constitutes the platform for the accumulation of a structurally and functionally diverse group of proteins, collectively called "MT plus-end tracking proteins" (+TIPs. +TIPs control MT dynamics and link MTs to diverse sub-cellular structures. Neurospora crassaMicroTubule Binding protein-3 (MTB-3 is the homolog of yeast EB1, a highly conserved +TIP. To address the function of MTB-3, we examined strains with mtb-3 deletions, and we tagged MTB-3 with GFP to assess its dynamic behavior. MTB-3-GFP was present as comet-like structures distributed more or less homogeneously within the hyphal cytoplasm, and moving mainly towards the apex at speeds up to 4× faster than the normal hyphal elongation rates. MTB-3-GFP comets were present in all developmental stages, but were most abundant in mature hyphae. MTB-3-GFP comets were observed moving in anterograde and retrograde direction along the hypha. Retrograde movement was also observed as originating from the apical dome. The integrity of the microtubular cytoskeleton affects the presence and dynamics of MTB-3-GFP comets, while actin does not seem to play a role. The size of MTB-3-GFP comets is affected by the absence of dynactin and conventional kinesin. We detected no obvious morphological phenotypes in Δmtb-3 mutants but there were fewer MTs in Δmtb-3, MTs were less bundled and less organized. Compared to WT, both MT polymerization and depolymerization rates were significantly decreased in Δmtb-3. In summary, the lack of MTB-3 affects overall growth and morphological phenotypes of N. crassa only slightly, but deletion of mtb-3 has strong effect on MT dynamics.

  4. Probing a self-assembled fd virus membrane with a microtubule

    Science.gov (United States)

    Xie, Sheng; Pelcovits, Robert A.; Hagan, Michael F.

    2016-06-01

    The self-assembly of highly anisotropic colloidal particles leads to a rich variety of morphologies whose properties are just beginning to be understood. This article uses computer simulations to probe a particle-scale perturbation of a commonly studied colloidal assembly, a monolayer membrane composed of rodlike fd viruses in the presence of a polymer depletant. Motivated by experiments currently in progress, we simulate the interaction between a microtubule and a monolayer membrane as the microtubule "pokes" and penetrates the membrane face-on. Both the viruses and the microtubule are modeled as hard spherocylinders of the same diameter, while the depletant is modeled using ghost spheres. We find that the force exerted on the microtubule by the membrane is zero either when the microtubule is completely outside the membrane or when it has fully penetrated the membrane. The microtubule is initially repelled by the membrane as it begins to penetrate but experiences an attractive force as it penetrates further. We assess the roles played by translational and rotational fluctuations of the viruses and the osmotic pressure of the polymer depletant. We find that rotational fluctuations play a more important role than the translational ones. The dependence on the osmotic pressure of the depletant of the width and height of the repulsive barrier and the depth of the attractive potential well is consistent with the assumed depletion-induced attractive interaction between the microtubule and viruses. We discuss the relevance of these studies to the experimental investigations.

  5. Deposition features of Ni on self-assembled microtubule template from biolipid by electroless method

    Institute of Scientific and Technical Information of China (English)

    FU; Yubin; ZHANG; Lide; ZHENG; Jiyong; FU; Shangang; ZHU; M

    2004-01-01

    Diacetylenic glycero-phosphatidylcholine is a chiral molecule with amphiphilic property, and it can self-assembly into a lipid microtubular structure. The lipid microtubule is a stable structure formed by tightly wound helical ribbons, and the ribbon-wrapping patterns have a significant effect on their chemical deposition on the microtubules. The deposition of colloidal Pd catalyst occurs mainly on the helical edge of the wound helical ribbons to form helical deposition lines of colloidal Pd particles in the interior and exterior of the lipid microtubules, resulting in an uneven chemical deposition of Ni on the microtubules. Catalyzed by as-deposited colloidal Pd, metallized Ni microtubules are characterized by a helical form, which may be in relation to inner stress due to the thickness difference or the different deposition processes. The observation of microtom shows that metallized tubules have a hollow structure. Some metallized tubules have a kind of coaxial double layer structure observed in the direct experiment evidence, indicating that metallization can occur in the inner and outer surface of the lipid tubules. Both lipid microtubules and metallized microtubules can be used as vehicles for encapsulating biological active molecules to control their release and to develop micro-components in biological and mechanical systems.

  6. Anillin interacts with microtubules and is part of the astral pathway that defines cortical domains.

    Science.gov (United States)

    van Oostende Triplet, Chloe; Jaramillo Garcia, Melina; Haji Bik, Husni; Beaudet, Daniel; Piekny, Alisa

    2014-09-01

    Cytokinesis occurs by the ingression of an actomyosin ring that separates the cell into two daughter cells. The mitotic spindle, comprising astral and central spindle microtubules, couples contractile ring ingression with DNA segregation. Cues from the central spindle activate RhoA, the upstream regulator of the contractile ring. However, additional cues from the astral microtubules also reinforce the localization of active RhoA. Using human cells, we show that astral and central spindle microtubules independently control the localization of contractile proteins during cytokinesis. Astral microtubules restrict the accumulation and localization of contractile proteins during mitosis, whereas the central spindle forms a discrete ring by directing RhoA activation in the equatorial plane. Anillin stabilizes the contractile ring during cytokinesis. We show that human anillin interacts with astral microtubules and that this interaction is competed by the cortical recruitment of anillin by active RhoA. Anillin restricts the localization of myosin to the equatorial cortex and that of NuMA (part of the microtubule-tethering complex that regulates spindle position) to the polar cortex. The sequestration of anillin by astral microtubules might alter the organization of cortical proteins to polarize cells for cytokinesis. PMID:24994938

  7. Arl2- and Msps-dependent microtubule growth governs asymmetric division.

    Science.gov (United States)

    Chen, Keng; Koe, Chwee Tat; Xing, Zhanyuan Benny; Tian, Xiaolin; Rossi, Fabrizio; Wang, Cheng; Tang, Quan; Zong, Wenhui; Hong, Wan Jin; Taneja, Reshma; Yu, Fengwei; Gonzalez, Cayetano; Wu, Chunlai; Endow, Sharyn; Wang, Hongyan

    2016-03-14

    Asymmetric division of neural stem cells is a fundamental strategy to balance their self-renewal and differentiation. It is long thought that microtubules are not essential for cell polarity in asymmetrically dividing Drosophila melanogaster neuroblasts (NBs; neural stem cells). Here, we show that Drosophila ADP ribosylation factor like-2 (Arl2) and Msps, a known microtubule-binding protein, control cell polarity and spindle orientation of NBs. Upon arl2 RNA intereference, Arl2-GDP expression, or arl2 deletions, microtubule abnormalities and asymmetric division defects were observed. Conversely, overactivation of Arl2 leads to microtubule overgrowth and depletion of NBs. Arl2 regulates microtubule growth and asymmetric division through localizing Msps to the centrosomes in NBs. Moreover, Arl2 regulates dynein function and in turn centrosomal localization of D-TACC and Msps. Arl2 physically associates with tubulin cofactors C, D, and E. Arl2 functions together with tubulin-binding cofactor D to control microtubule growth, Msps localization, and NB self-renewal. Therefore, Arl2- and Msps-dependent microtubule growth is a new paradigm regulating asymmetric division of neural stem cells.

  8. Altered nucleotide-microtubule coupling and increased mechanical output by a kinesin mutant.

    Directory of Open Access Journals (Sweden)

    Hong-Lei Liu

    Full Text Available Kinesin motors hydrolyze ATP to produce force and do work in the cell--how the motors do this is not fully understood, but is thought to depend on the coupling of ATP hydrolysis to microtubule binding by the motor. Transmittal of conformational changes from the microtubule- to the nucleotide-binding site has been proposed to involve the central β-sheet, which could undergo large structural changes important for force production. We show here that mutation of an invariant residue in loop L7 of the central β-sheet of the Drosophila kinesin-14 Ncd motor alters both nucleotide and microtubule binding, although the mutated residue is not present in either site. Mutants show weak-ADP/tight-microtubule binding, instead of tight-ADP/weak-microtubule binding like wild type--they hydrolyze ATP faster than wild type, move faster in motility assays, and assemble long spindles with greatly elongated poles, which are also produced by simulations of assembly with tighter microtubule binding and faster sliding. The mutated residue acts like a mechanochemical coupling element--it transmits changes between the microtubule-binding and active sites, and can switch the state of the motor, increasing mechanical output by the motor. One possibility, based on our findings, is that movements by the residue and the loop that contains it could bend or distort the central β-sheet, mediating free energy changes that lead to force production.

  9. Arabidopsis FH1 Formin Affects Cotyledon Pavement Cell Shape by Modulating Cytoskeleton Dynamics.

    Science.gov (United States)

    Rosero, Amparo; Oulehlová, Denisa; Stillerová, Lenka; Schiebertová, Petra; Grunt, Michal; Žárský, Viktor; Cvrčková, Fatima

    2016-03-01

    Plant cell morphogenesis involves concerted rearrangements of microtubules and actin microfilaments. We previously reported that FH1, the main Arabidopsis thaliana housekeeping Class I membrane-anchored formin, contributes to actin dynamics and microtubule stability in rhizodermis cells. Here we examine the effects of mutations affecting FH1 (At3g25500) on cell morphogenesis and above-ground organ development in seedlings, as well as on cytoskeletal organization and dynamics, using a combination of confocal and variable angle epifluorescence microscopy with a pharmacological approach. Homozygous fh1 mutants exhibited cotyledon epinasty and had larger cotyledon pavement cells with more pronounced lobes than the wild type. The pavement cell shape alterations were enhanced by expression of the fluorescent microtubule marker GFP-microtubule-associated protein 4 (MAP4). Mutant cotyledon pavement cells exhibited reduced density and increased stability of microfilament bundles, as well as enhanced dynamics of microtubules. Analogous results were also obtained upon treatments with the formin inhibitor SMIFH2 (small molecule inhibitor of formin homology 2 domains). Pavement cell shape in wild-type (wt) and fh1 plants in some situations exhibited a differential response towards anti-cytoskeletal drugs, especially the microtubule disruptor oryzalin. Our observations indicate that FH1 participates in the control of microtubule dynamics, possibly via its effects on actin, subsequently influencing cell morphogenesis and macroscopic organ development. PMID:26738547

  10. HSPB1 facilitates the formation of non-centrosomal microtubules.

    Directory of Open Access Journals (Sweden)

    Leonardo Almeida-Souza

    Full Text Available The remodeling capacity of microtubules (MT is essential for their proper function. In mammals, MTs are predominantly formed at the centrosome, but can also originate from non-centrosomal sites, a process that is still poorly understood. We here show that the small heat shock protein HSPB1 plays a role in the control of non-centrosomal MT formation. The HSPB1 expression level regulates the balance between centrosomal and non-centrosomal MTs. The HSPB1 protein can be detected specifically at sites of de novo forming non-centrosomal MTs, while it is absent from the centrosomes. In addition, we show that HSPB1 binds preferentially to the lattice of newly formed MTs in vitro, suggesting that its function occurs by stabilizing MT seeds. Our findings open new avenues for the understanding of the role of HSPB1 in the development, maintenance and protection of cells with specialized non-centrosomal MT arrays.

  11. Studies on the role of microtubules in myofibrillogenesis

    Institute of Scientific and Technical Information of China (English)

    LINZHONGXIANG; HOWARDHOLTZER

    1990-01-01

    Co-localization of microtubule (MT) and muscle myosin (MHC) myofibril immunofluoresoonoe in developing myotubes of chicken skeletal muscle cultures was observed by using double staining of tubulin and MHC indirect immunofluorescence.120-tetradecanoyl-phorbol-12-acetate (TPA) selectively and reversibly blocks myofibrillogenesis and alters the morphology of myotubes in to myosacs where MTs are present in radiating pattern.When the arrested myogenic cells recover and start myofibrillogenesis after released from TPA,prior to the emergence of myofibrils,the pre-ecisting MTs become bipolarly aligned coincidently with the tubular restoration of cell shape.Single nascent myofibrils overlapping with MTs extend into the base of growth tips where MTs go farther to the end of the tips.That MT might act as scaffold in guiding the bipolar elongation of the growing myofibrils was suggested.Taxol and colcemid disturbed MT polymerization and disposition,and interfered with the normal spatial assembly of myofibrils in developing myotubes.

  12. Conformational mechanism for the stability of microtubule-kinetochore attachments

    CERN Document Server

    Bertalan, Zsolt; Maiato, Helder; Zapperi, Stefano

    2014-01-01

    Regulating the stability of microtubule(MT)-kinetochore attachments is fundamental to avoiding mitotic errors and ensure proper chromosome segregation during cell division. While biochemical factors involved in this process have been identified, its mechanics still needs to be better understood. Here we introduce and simulate a mechanical model of MT-kinetochore interactions in which the stability of the attachment is ruled by the geometrical conformations of curling MT-protofilaments entangled in kinetochore fibrils. The model allows us to reproduce with good accuracy in vitro experimental measurements of the detachment times of yeast kinetochores from MTs under external pulling forces. Numerical simulations suggest that geometrical features of MT-protofilaments may play an important role in the switch between stable and unstable attachments.

  13. A Mechanism for Cytoplasmic Streaming: Kinesin-Driven Alignment of Microtubules and Fast Fluid Flows.

    Science.gov (United States)

    Monteith, Corey E; Brunner, Matthew E; Djagaeva, Inna; Bielecki, Anthony M; Deutsch, Joshua M; Saxton, William M

    2016-05-10

    The transport of cytoplasmic components can be profoundly affected by hydrodynamics. Cytoplasmic streaming in Drosophila oocytes offers a striking example. Forces on fluid from kinesin-1 are initially directed by a disordered meshwork of microtubules, generating minor slow cytoplasmic flows. Subsequently, to mix incoming nurse cell cytoplasm with ooplasm, a subcortical layer of microtubules forms parallel arrays that support long-range, fast flows. To analyze the streaming mechanism, we combined observations of microtubule and organelle motions with detailed mathematical modeling. In the fast state, microtubules tethered to the cortex form a thin subcortical layer and undergo correlated sinusoidal bending. Organelles moving in flows along the arrays show velocities that are slow near the cortex and fast on the inward side of the subcortical microtubule layer. Starting with fundamental physical principles suggested by qualitative hypotheses, and with published values for microtubule stiffness, kinesin velocity, and cytoplasmic viscosity, we developed a quantitative coupled hydrodynamic model for streaming. The fully detailed mathematical model and its simulations identify key variables that can shift the system between disordered (slow) and ordered (fast) states. Measurements of array curvature, wave period, and the effects of diminished kinesin velocity on flow rates, as well as prior observations on f-actin perturbation, support the model. This establishes a concrete mechanistic framework for the ooplasmic streaming process. The self-organizing fast phase is a result of viscous drag on kinesin-driven cargoes that mediates equal and opposite forces on cytoplasmic fluid and on microtubules whose minus ends are tethered to the cortex. Fluid moves toward plus ends and microtubules are forced backward toward their minus ends, resulting in buckling. Under certain conditions, the buckling microtubules self-organize into parallel bending arrays, guiding varying directions

  14. Cytoskeletal dynamics in and traumatic injury of cerebellar and hippocampal neurons

    NARCIS (Netherlands)

    J.E. Slemmer (Jennifer)

    2005-01-01

    markdownabstract__Abstract__ This thesis addresses two separate, yet overlapping, physiological processes, namely traumatic brain injury (TBI) and microtubule (MT) dynamics, primarily through the use of cultured cells derived from embryonic mice. The difficulties which arise through the experimenta

  15. Polypyrrole microtubules and their use in the construction of a third generation biosensor

    Energy Technology Data Exchange (ETDEWEB)

    Koopal, C.G.J.; Feiters, M.C.; Nolte, R.J.M. (Nijmegen SON Research Center, Univ. of Nijmegen (Netherlands)); Ruiter, B. de (TNO Industrial Research, Plastics and Rubber Research Inst., Delft (Netherlands)); Schasfoort, R.B.M. (TNO Food Research, Inst. of Biotechnology and Chemistry, Zeist (Netherlands)); Czajka, R.; Kempen, H. van (Univ. of Nijmegen, Research Inst. for Materials (Netherlands))

    1992-09-01

    Conducting polypyrrole microtubules have been prepared by template synthesis inside track-etch membranes. The interiors of these microtubules can adsorb the redox enzyme, glucose oxidase. The enzyme-coated tubules have been employed in the construction of a third generation amperometric biosensor for the determination of glucose. With this biosensor, glucose concentrations in the range 0.1 - 250 mM can be measured easily. The polypyrrole microtubules have been characterized by different microscopic techniques, including scanning tunneling microscopy. Based on the microscopy data, a model is presented for the interaction between the conducting polymer and the glucose oxidase molecules. (orig.).

  16. The Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER): Data Products for the High Spatial Resolution Imager on NASA's EOS-AMI Platform

    Science.gov (United States)

    Abrams, M.

    1999-01-01

    The Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) is a high spatial resolution, multispectral imager with along-track stereo capabilities scheduled for launch on the first NASA spacecraft of the Earth Observing System (EOS AM-1) in mid-1999.

  17. Crescimento e absorção de nutrientes pelo Aster ericoides cultivado em solo sob estufa Growth and nutrients absorption by Aster ericoides cultivated in soil at greenhouse conditions

    Directory of Open Access Journals (Sweden)

    Mônica S. de Camargo

    2005-06-01

    Full Text Available O Aster ericoides é flor de corte recente no Brasil com grande potencial de produção e aceitação pelo mercado consumidor, mas são escassas as informações sobre sua nutrição e adubação. O objetivo foi avaliar o crescimento, produção de matéria seca e a absorção de nutrientes pelo Aster ericodes (cv. White Master cultivado em estufa comercial. Plântulas dessa espécie foram transplantadas para canteiros após 30 dias da semeadura, em outubro de 1999. As avaliações do material vegetal foram feitas aos 15; 30; 45; 60; 75; 90 e 96 dias após o transplantio. O crescimento inicial foi lento, mas aos 60 dias as plantas já haviam atingido 69% do crescimento em altura e, respectivamente, 27 e 50% do peso final da matéria seca da parte aérea e das raízes. O aparecimento dos botões florais ocorreu no período de 75 a 90 dias. No final do ciclo, as plantas atingiram 137 cm e 24,21 g de matéria seca. A absorção de macronutrientes (kg ha-1 pela parte aérea aos 96 dias foi de: 316,32 de K > 230,52 de N > 35,30 de P > 18,54 de S > 16,14 de Mg > 8,58 de Ca e de micronutrientes (g ha-1 3464,89 de Mn > 1603,23 de Fe > 1104,12 de Zn > 308,40 de B > 61,02 de Cu.Aster ericoides is a cut flower crop with great market and production potential that was only recently introduced into Brazil. Thus, little information is available regarding fertilization and nutrition under tropical conditions. The growth, dry matter production and nutrient absorption of Aster ericoides (cv. White Master cultivated in an UDOX soil under greenhouse conditions was evaluated. Seedlings were transplanted 30 days after sowing date, in October 99. Plant samples were taken at 15; 30; 45; 60; 75; 90 and 96 days after transplanting. Plant growth and dry matter production were slow in the beginning but at 60 days (before flowering 69% of the height, 27% of the top and 50% of the roots dry matter was reached. Plants reached 137 cm in height and 24.21 g of dry weight at the

  18. Diacylglycerol Guides the Hopping of Clathrin-Coated Pits along Microtubules for Exo-Endocytosis Coupling.

    Science.gov (United States)

    Yuan, Tianyi; Liu, Lin; Zhang, Yongdeng; Wei, Lisi; Zhao, Shiqun; Zheng, Xiaolu; Huang, Xiaoshuai; Boulanger, Jerome; Gueudry, Charles; Lu, Jingze; Xie, Lihan; Du, Wen; Zong, Weijian; Yang, Lu; Salamero, Jean; Liu, Yanmei; Chen, Liangyi

    2015-10-12

    Many receptor-mediated endocytic processes are mediated by constitutive budding of clathrin-coated pits (CCPs) at spatially randomized sites before slowly pinching off from the plasma membrane (60-100 s). In contrast, clathrin-mediated endocytosis (CME) coupled with regulated exocytosis in excitable cells occurs at peri-exocytic sites shortly after vesicle fusion (∼10 s). The molecular mechanism underlying this spatiotemporal coupling remains elusive. We show that coupled endocytosis makes use of pre-formed CCPs, which hop to nascent fusion sites nearby following vesicle exocytosis. A dynamic cortical microtubular network, anchored at the cell surface by the cytoplasmic linker-associated protein on microtubules and the LL5β/ELKS complex on the plasma membrane, provides the track for CCP hopping. Local diacylglycerol gradients generated upon exocytosis guide the direction of hopping. Overall, the CCP-cytoskeleton-lipid interaction demonstrated here mediates exocytosis-coupled fast recycling of both plasma membrane and vesicular proteins, and it is required for the sustained exocytosis during repetitive stimulations. PMID:26439397

  19. A posteriori error analysis for hydro-mechanical couplings and implementation in Code-Aster; Analyse d'erreur a posteriori pour les couplages hydro-mecaniques et mise en oeuvre dans Code-Aster

    Energy Technology Data Exchange (ETDEWEB)

    Meunier, S

    2007-11-15

    We analyse approximations by finite elements in space and finite differences in time of coupled Hydro-Mechanical (HM) problems related to the quasi-static linear poro-elasticity theory. The physical bases of this theory are briefly restated and an abstract setting is proposed to perform the mathematical study of the stationary and un-stationary versions of the HM problem. For the stationary version, the well-posedness of the continuous and discrete problems are established and the a priori error analysis is performed. Then, we propose the a posteriori error analysis by using two different techniques suited to estimate the displacement error and the pressure error, respectively, both in the H{sub x}{sup 1}-norm. The classical properties of reliability and optimality are proved for the associated error estimators. Some numerical experiments using Code-Aster illustrate the theoretical results. For the un-stationary version, we first establish a stability result for the continuous problem. Then, we present an optimal a priori error analysis using elliptic projection techniques. Finally, the a posteriori error analysis is performed by using two different approaches: a direct approach and an elliptic reconstruction approach. The first is suited to estimate the pressure error in the L{sub t}{sup 2}(H{sub x}{sup 1})-norm and the second is suited to estimate the displacement error in the L{sub t}{sup {infinity}}(H{sub x}{sup 1})-norm and the pressure error in the L{sub t}{sup {infinity}}(H{sub x}{sup 1})-norm. Numerical experiments using Code-Aster complete the theoretical results. (author)

  20. Shot and Patronin polarise microtubules to direct membrane traffic and biogenesis of microvilli in epithelia.

    Science.gov (United States)

    Khanal, Ichha; Elbediwy, Ahmed; Diaz de la Loza, Maria Del Carmen; Fletcher, Georgina C; Thompson, Barry J

    2016-07-01

    In epithelial tissues, polarisation of microtubules and actin microvilli occurs along the apical-basal axis of each cell, yet how these cytoskeletal polarisation events are coordinated remains unclear. Here, we examine the hierarchy of events during cytoskeletal polarisation in Drosophila melanogaster epithelia. Core apical-basal polarity determinants polarise the spectrin cytoskeleton to recruit the microtubule-binding proteins Patronin (CAMSAP1, CAMSAP2 and CAMPSAP3 in humans) and Shortstop [Shot; MACF1 and BPAG1 (also known as DST) in humans] to the apical membrane domain. Patronin and Shot then act to polarise microtubules along the apical-basal axis to enable apical transport of Rab11 endosomes by the Nuf-Dynein microtubule motor complex. Finally, Rab11 endosomes are transferred to the MyoV (also known as Didum in Drosophila) actin motor to deliver the key microvillar determinant Cadherin 99C to the apical membrane to organise the biogenesis of actin microvilli. PMID:27231092

  1. Computational Study of Pseudo-phosphorylation of the Microtubule associated Protein Tau

    Science.gov (United States)

    Prokopovich, Dmitriy; Larini, Luca

    This computational study focuses on the effect of pseudo-phosphorylation on the aggregation of the microtubule associated protein tau. In the axon of the neuron, tau regulates the assembly of microtubules in the cytoskeleton. This is important for both stabilization of and transport across the microtubules. One of the hallmarks of the Alzheimer's disease is that tau is hyper-phosphorylated and aggregates into neurofibrillary tangles that lay waste to the neurons. It is not known if hyper-phosphorylation directly causes the aggregation of tau into tangles. Experimentally, pseudo-phosphorylation mimics the effects of phosphorylation by mutating certain residues of the protein chain into charged residues. In this study, we will consider the fragment called PHF43 that belongs to the microtubule binding region and has been shown to readily aggregate.

  2. Clostridium difficile toxin CDT hijacks microtubule organization and reroutes vesicle traffic to increase pathogen adherence.

    Science.gov (United States)

    Schwan, Carsten; Kruppke, Anna S; Nölke, Thilo; Schumacher, Lucas; Koch-Nolte, Friedrich; Kudryashev, Mikhail; Stahlberg, Henning; Aktories, Klaus

    2014-02-11

    Clostridium difficile causes antibiotic-associated diarrhea and pseudomembranous colitis by the actions of Rho-glucosylating toxins A and B. Recently identified hypervirulent strains, which are associated with increased morbidity and mortality, additionally produce the actin-ADP-ribosylating toxin C. difficile transferase (CDT). CDT depolymerizes actin, causes formation of microtubule-based protrusions, and increases pathogen adherence. Here we show that CDT-induced protrusions allow vesicle traffic and contain endoplasmic reticulum tubules, connected to microtubules via the calcium sensor Stim1. The toxin reroutes Rab11-positive vesicles containing fibronectin, which is involved in bacterial adherence, from basolateral to the apical membrane sides in a microtubule- and Stim1-dependent manner. The data yield a model of C. difficile adherence regulated by actin depolymerization, microtubule restructuring, subsequent Stim1-dependent Ca(2+) signaling, vesicle rerouting, and secretion of ECM proteins to increase bacterial adherence.

  3. Microtubule Disruption in Keratinocytes Induces Cell-Cell Adhesion through Activation of Endogenous E-Cadherin

    OpenAIRE

    Kee, Sun-Ho; Steinert, Peter M.

    2001-01-01

    The association of the cytoskeleton with the cadherin–catenin complex is essential for strong cell-cell adhesion in epithelial cells. In this study, we have investigated the effect of microtubule organization on cell-cell adhesion in differentiating keratinocytes. When microtubules of normal human epidermal keratinocytes (NHEKs) grown in low calcium media (0.05 mM) were disrupted with nocodazole or colcemid, cell-cell adhesion was induced through relocalization of the ...

  4. Structural Basis for Induction of Peripheral Neuropathy by Microtubule-Targeting Cancer Drugs.

    Science.gov (United States)

    Smith, Jennifer A; Slusher, Barbara S; Wozniak, Krystyna M; Farah, Mohamed H; Smiyun, Gregoriy; Wilson, Leslie; Feinstein, Stuart; Jordan, Mary Ann

    2016-09-01

    Peripheral neuropathy is a serious, dose-limiting side effect of cancer treatment with microtubule-targeting drugs. Symptoms present in a "stocking-glove" distribution, with longest nerves affected most acutely, suggesting a length-dependent component to the toxicity. Axonal transport of ATP-producing mitochondria along neuronal microtubules from cell body to synapse is crucial to neuronal function. We compared the effects of the drugs paclitaxel and ixabepilone that bind along the lengths of microtubules and the drugs eribulin and vincristine that bind at microtubule ends, on mitochondrial trafficking in cultured human neuronal SK-N-SH cells and on axonal transport in mouse sciatic nerves. Antiproliferative concentrations of paclitaxel and ixabepilone significantly inhibited the anterograde transport velocity of mitochondria in neuronal cells, whereas eribulin and vincristine inhibited transport only at significantly higher concentrations. Confirming these observations, anterogradely transported amyloid precursor protein accumulated in ligated sciatic nerves of control and eribulin-treated mice, but not in paclitaxel-treated mice, indicating that paclitaxel inhibited anterograde axonal transport, whereas eribulin did not. Electron microscopy of sciatic nerves of paclitaxel-treated mice showed reduced organelle accumulation proximal to the ligation consistent with inhibition of anterograde (kinesin based) transport by paclitaxel. In contrast, none of the drugs significantly affected retrograde (dynein based) transport in neuronal cells or mouse nerves. Collectively, these results suggest that paclitaxel and ixabepilone, which bind along the lengths and stabilize microtubules, inhibit kinesin-based axonal transport, but not dynein-based transport, whereas the microtubule-destabilizing drugs, eribulin and vincristine, which bind preferentially to microtubule ends, have significantly less effect on all microtubule-based axonal transport. Cancer Res; 76(17); 5115-23.

  5. ICP0 dismantles microtubule networks in herpes simplex virus-infected cells.

    Directory of Open Access Journals (Sweden)

    Mingyu Liu

    Full Text Available Infected-cell protein 0 (ICP0 is a RING finger E3 ligase that regulates herpes simplex virus (HSV mRNA synthesis, and strongly influences the balance between latency and replication of HSV. For 25 years, the nuclear functions of ICP0 have been the subject of intense scrutiny. To obtain new clues about ICP0's mechanism of action, we constructed HSV-1 viruses that expressed GFP-tagged ICP0. To our surprise, both GFP-tagged and wild-type ICP0 were predominantly observed in the cytoplasm of HSV-infected cells. Although ICP0 is exclusively nuclear during the immediate-early phase of HSV infection, further analysis revealed that ICP0 translocated to the cytoplasm during the early phase where it triggered a previously unrecognized process; ICP0 dismantled the microtubule network of the host cell. A RING finger mutant of ICP0 efficiently bundled microtubules, but failed to disperse microtubule bundles. Synthesis of ICP0 proved to be necessary and sufficient to disrupt microtubule networks in HSV-infected and transfected cells. Plant and animal viruses encode many proteins that reorganize microtubules. However, this is the first report of a viral E3 ligase that regulates microtubule stability. Intriguingly, several cellular E3 ligases orchestrate microtubule disassembly and reassembly during mitosis. Our results suggest that ICP0 serves a dual role in the HSV life cycle, acting first as a nuclear regulator of viral mRNA synthesis and acting later, in the cytoplasm, to dismantle the host cell's microtubule network in preparation for virion synthesis and/or egress.

  6. Genetic evidence that cellulose synthase activity influences microtubule cortical array organization

    OpenAIRE

    Paredez, A.; S. Persson; Ehrhardt, D; Somerville, C

    2008-01-01

    To identify factors that influence cytoskeletal organization we screened for Arabidopsis ( Arabidopsis thaliana) mutants that show hypersensitivity to the microtubule destabilizing drug oryzalin. We cloned the genes corresponding to two of the 131 mutant lines obtained. The genes encoded mutant alleles of PROCUSTE1 and KORRIGAN, which both encode proteins that have previously been implicated in cellulose synthesis. Analysis of microtubules in the mutants revealed that both mutants have altere...

  7. Lis1 mediates planar polarity of auditory hair cells through regulation of microtubule organization

    OpenAIRE

    Sipe, Conor W.; Liu, Lixia; Lee, Jianyi; Grimsley-Myers, Cynthia; Lu, Xiaowei

    2013-01-01

    The V-shaped hair bundles atop auditory hair cells and their uniform orientation are manifestations of epithelial planar cell polarity (PCP) required for proper perception of sound. PCP is regulated at the tissue level by a conserved core Wnt/PCP pathway. However, the hair cell-intrinsic polarity machinery is poorly understood. Recent findings implicate hair cell microtubules in planar polarization of hair cells. To elucidate the microtubule-mediated polarity pathway, we analyzed Lis1 functio...

  8. Interactions of the HSV-1 UL25 Capsid Protein with Cellular Microtubule-associated Protein

    Institute of Scientific and Technical Information of China (English)

    Lei GUO; Ying ZHANG; Yan-chun CHE; Wen-juan WU; Wei-zhong LI; Li-chun WANG; Yun LIAO; Long-ding LIU; Qi-han LI

    2008-01-01

    An interaction between the HSV-1 UL25 capsid protein and cellular microtubule-associated protein was found using a yeast two-hybrid screen and β-D-galactosidase activity assays. Immunofluorescence microscopy of the UL25 protein demonstrated its co-localization with cellular microtubule-associated protein in the plasma membrane. Further investigations with deletion mutants suggest that UL25 is likely to have a function in the nucleus.

  9. Katanin maintains meiotic metaphase chromosome alignment and spindle structure in vivo and has multiple effects on microtubules in vitro.

    Science.gov (United States)

    McNally, Karen; Berg, Evan; Cortes, Daniel B; Hernandez, Veronica; Mains, Paul E; McNally, Francis J

    2014-04-01

    Assembly of Caenorhabditis elegans female meiotic spindles requires both MEI-1 and MEI-2 subunits of the microtubule-severing ATPase katanin. Strong loss-of-function mutants assemble apolar intersecting microtubule arrays, whereas weaker mutants assemble bipolar meiotic spindles that are longer than wild type. To determine whether katanin is also required for spindle maintenance, we monitored metaphase I spindles after a fast-acting mei-1(ts) mutant was shifted to a nonpermissive temperature. Within 4 min of temperature shift, bivalents moved off the metaphase plate, and microtubule bundles within the spindle lengthened and developed a high degree of curvature. Spindles eventually lost bipolar structure. Immunofluorescence of embryos fixed at increasing temperature indicated that MEI-1 was lost from spindle microtubules before loss of ASPM-1, indicating that MEI-1 and ASPM-1 act independently at spindle poles. We quantified the microtubule-severing activity of purified MEI-1/MEI-2 complexes corresponding to six different point mutations and found a linear relationship between microtubule disassembly rate and meiotic spindle length. Previous work showed that katanin is required for severing at points where two microtubules intersect in vivo. We show that purified MEI-1/MEI-2 complexes preferentially sever at intersections between two microtubules and directly bundle microtubules in vitro. These activities could promote parallel/antiparallel microtubule organization in meiotic spindles. PMID:24501424

  10. A deterministic oscillatory model of microtubule growth and shrinkage for differential actions of short chain fatty acids.

    Science.gov (United States)

    Kilner, Josephine; Corfe, Bernard M; McAuley, Mark T; Wilkinson, Stephen J

    2016-01-01

    Short chain fatty acids (SCFA), principally acetate, propionate, butyrate and valerate, are produced in pharmacologically relevant concentrations by the gut microbiome. Investigations indicate that they exert beneficial effects on colon epithelia. There is increasing interest in whether different SCFAs have distinct functions which may be exploited for prevention or treatment of colonic diseases including colorectal cancer (CRC), inflammatory bowel disease and obesity. Based on experimental evidence, we hypothesised that odd-chain SCFAs may possess anti-mitotic capabilities in colon cancer cells by disrupting microtubule (MT) structural integrity via dysregulation of β-tubulin isotypes. MT dynamic instability is an essential characteristic of MT cellular activity. We report a minimal deterministic model that takes a novel approach to explore the hypothesised pathway by triggering spontaneous oscillations to represent MT dynamic behaviour. The dynamicity parameters in silico were compared to those reported in vitro. Simulations of untreated and butyrate (even-chain length) treated cells reflected MT behaviour in interphase or untreated control cells. The propionate and valerate (odd-chain length) simulations displayed increased catastrophe frequencies and longer periods of MT-fibre shrinkage. Their enhanced dynamicity was dissimilar to that observed in mitotic cells, but parallel to that induced by MT-destabilisation treatments. Antimicrotubule drugs act through upward or downward modulation of MT dynamic instability. Our computational modelling suggests that metabolic engineering of the microbiome may facilitate managing CRC risk by predicting outcomes of SCFA treatments in combination with AMDs. PMID:26562762

  11. Spectro-radiometers ASTER and MODIS - character of data, their accessibility and exploitability in area of environment

    International Nuclear Information System (INIS)

    In this presentation some basic information about spectro-radiometers ASTER and MODIS are presented. Relative wide opportunities of exploitation of these products in area of environment, their high spectral and in case of MODIS time resolution are discussed. These parameters create starting-point for building-up of regional monitoring systems of different biophysical characteristics of terrestrial ecosystems and monitoring of time and spatial variability. Next effort in this area should be aimed on development and optimisation of regional models based on monitoring of time and spatial changes of vegetable and foliar indexes (NDVI, EVI, LAI), photosynthetically active part of radiation absorbed by vegetation (FPAR) and likewise, as well as detail analyses of these data in context of global climatic changes. Perspectives of remote sensing earth in the Slovak republic are discussed

  12. Microtubule-Destabilizing Agents: Structural and Mechanistic Insights from the Interaction of Colchicine and Vinblastine with Tubulin

    Science.gov (United States)

    Gigant, B.; Cormier, A.; Dorléans, A.; Ravelli, R. B. G.; Knossow, M.

    Microtubules (MTs) are dynamic structures of the eukaryotic cytoskeleton that, during cell division, form the mitotic spindle. Perturbing them leads to mitotic arrest and ultimately to cell death. Consistently, MTs and their building block, αβ tubulin, are one of the best characterized targets in anti-cancer chemotherapy. Drugs that interfere with MTs either stabilize or destabilize them. The latter class is the subject of this review. These ligands bind to the colchicine site or to the vinca domain, two distinct sites located at a distance from each other on tubulin. Nevertheless the effects of both classes of ligands share a common theme, they prevent the formation of MT specific contacts, therefore triggering their disassembly.

  13. Emerging roles for microtubules in angiosperm pollen tube growth highlight new research cues

    Directory of Open Access Journals (Sweden)

    Alessandra eMoscatelli

    2015-02-01

    Full Text Available In plants, actin filaments have an important role in organelle movement and cytoplasmic streaming. Otherwise microtubules have a role in restricting organelles to specific areas of the cell and in maintaining organelle morphology. In somatic plant cells, microtubules also participate in cell division and morphogenesis, allowing cells to take their definitive shape in order to perform specific functions. In the latter case, microtubules influence assembly of the cell wall, controlling the delivery of enzymes involved in cellulose synthesis and of wall modulation material to the proper sites.In angiosperm pollen tubes, organelle movement is generally attributed to the acto-myosin system, the main role of which is in distributing organelles in the cytoplasm and in carrying secretory vesicles to the apex for polarized growth. Recent data on membrane trafficking suggests a role of microtubules in fine delivery and repositioning of vesicles to sustain pollen tube growth. This review examines the role of microtubules in secretion and endocytosis, highlighting new research cues regarding cell wall construction and pollen tube-pistil crosstalk, that help unravel the role of microtubules in polarized growth.

  14. A mitotic SKAP isoform regulates spindle positioning at astral microtubule plus ends.

    Science.gov (United States)

    Kern, David M; Nicholls, Peter K; Page, David C; Cheeseman, Iain M

    2016-05-01

    The Astrin/SKAP complex plays important roles in mitotic chromosome alignment and centrosome integrity, but previous work found conflicting results for SKAP function. Here, we demonstrate that SKAP is expressed as two distinct isoforms in mammals: a longer, testis-specific isoform that was used for the previous studies in mitotic cells and a novel, shorter mitotic isoform. Unlike the long isoform, short SKAP rescues SKAP depletion in mitosis and displays robust microtubule plus-end tracking, including localization to astral microtubules. Eliminating SKAP microtubule binding results in severe chromosome segregation defects. In contrast, SKAP mutants specifically defective for plus-end tracking facilitate proper chromosome segregation but display spindle positioning defects. Cells lacking SKAP plus-end tracking have reduced Clasp1 localization at microtubule plus ends and display increased lateral microtubule contacts with the cell cortex, which we propose results in unbalanced dynein-dependent cortical pulling forces. Our work reveals an unappreciated role for the Astrin/SKAP complex as an astral microtubule mediator of mitotic spindle positioning. PMID:27138257

  15. Quantitative Changes in Microtubule Distribution Correlate with Guard Cell Function in Arabidopsis

    Institute of Scientific and Technical Information of China (English)

    William R. Eisinger; Viktor Kirik; Charlotte Lewis; David W. Ehrhardt; Winslow R. Briggs

    2012-01-01

    Radially arranged cortical microtubules are a prominent feature of guard cells.We observed guard cells expressing GFP-tubulin (GFP-TUA6) with confocal microscopy and found recognizable changes in the appearance of microtubules when stomata open or close (Eisinger et al.,2012).In the present study,analysis of fluorescence distribution showed a dramatic increase in peak intensities of microtubule bundles within guard cells as stomata open.This increase was correlated with an increase in the total fluorescence that could be attributed to polymerized tubulin.Adjacent pavement cells did not show similar changes in peak intensities or integrated fluorescence when stomatal apertures changed.Imaging of RFP-tagged end binding protein 1 (EB1) and YFP-tagged α-tubulin expressed in the same cell revealed that the number of microtubules with growing ends remained constant,although the total amount of polymerized tubulin was higher in open than in closed guard cells.Taken together,these results indicate that the changes in microtubule array organization that are correlated with and required for normal guard cell function are characterized by changes in microtubule clustering or bundling.

  16. ATPase Cycle of the Nonmotile Kinesin NOD Allows Microtubule End Tracking and Drives Chromosome Movement

    Energy Technology Data Exchange (ETDEWEB)

    Cochran, J.; Sindelar, C; Mulko, N; Collins, K; Kong, S; Hawley, R; Kull, F

    2009-01-01

    Segregation of nonexchange chromosomes during Drosophila melanogaster meiosis requires the proper function of NOD, a nonmotile kinesin-10. We have determined the X-ray crystal structure of the NOD catalytic domain in the ADP- and AMPPNP-bound states. These structures reveal an alternate conformation of the microtubule binding region as well as a nucleotide-sensitive relay of hydrogen bonds at the active site. Additionally, a cryo-electron microscopy reconstruction of the nucleotide-free microtubule-NOD complex shows an atypical binding orientation. Thermodynamic studies show that NOD binds tightly to microtubules in the nucleotide-free state, yet other nucleotide states, including AMPPNP, are weakened. Our pre-steady-state kinetic analysis demonstrates that NOD interaction with microtubules occurs slowly with weak activation of ADP product release. Upon rapid substrate binding, NOD detaches from the microtubule prior to the rate-limiting step of ATP hydrolysis, which is also atypical for a kinesin. We propose a model for NOD's microtubule plus-end tracking that drives chromosome movement.

  17. Hypothesis: NDL Proteins Function in Stress Responses by Regulating Microtubule Organization

    Directory of Open Access Journals (Sweden)

    Nisha eKhatri

    2015-10-01

    Full Text Available N-MYC DOWNREGULATED-LIKE proteins (NDL, members of the alpha/beta hydrolase superfamily were recently rediscovered as interactors of G-protein signaling in Arabidopsis thaliana. Although the precise molecular function of NDL proteins is still elusive, in animals these proteins play protective role in hypoxia and expression is induced by hypoxia and nickel, indicating role in stress. Homology of NDL1 with animal counterpart NDRG suggests similar functions in animals and plants. It is well established that stress responses leads to the microtubule depolymerization and reorganization which is crucial for stress tolerance. NDRG is a microtubule-associated protein (MAP which mediates the microtubule organization in animals by causing acetylation and increases the stability of α-tubulin. As NDL1 is highly homologous to NDRG, involvement of NDL1 in the microtubule organization during plant stress can also be expected. Discovery of interaction of NDL with protein kinesin light chain- related 1, enodomembrane family protein 70, syntaxin-23, tubulin alpha-2 chain, as a part of G protein interactome initiative encourages us to postulate microtubule stabilizing functions for NDL family in plants. Our search for NDL interactors in G protein interactome also predicts the role of NDL proteins in abiotic stress tolerance management. Based on published report in animals and predicted interacting partners for NDL in G protein interactome lead us to hypothesize involvement of NDL in the microtubule organization during abiotic stress management in plants.

  18. Single Vesicle Analysis of Endocytic Fission on Microtubules In Vitro

    Science.gov (United States)

    Wolkoff, Allan W.

    2016-01-01

    Following endocytosis, internalized molecules are found within intracellular vesicles and tubules that move along the cytoskeleton and undergo fission, as demonstrated here using primary cultured rat hepatocytes. Although the use of depolymerizing drugs has shown that the cytoskeleton is not required to segregate endocytic protein, many studies suggest that the cytoskeleton is involved in the segregation of protein in normal cells. To investigate whether cytoskeletal-based movement results in the segregation of protein, we tracked the contents of vesicles during in vitro microscopy assays. These studies showed that the addition of ATP causes fission of endocytic contents along microtubules, resulting in the segregation of proteins that are targeted for different cellular compartments. The plasma membrane proteins, sodium (Na+) taurocholate cotransporting polypeptide (ntcp) and transferrin receptor, segregated from asialoorosomucoid (ASOR), an endocytic ligand that is targeted for degradation. Epidermal growth factor receptor, which is degraded, and the asialoglycoprotein receptor, which remains partially bound to ASOR, segregated less efficiently from ASOR. Vesicles containing ntcp and transferrin receptor had reduced fission in the absence of ASOR, suggesting that fission is regulated to allow proteins to segregate. A single round of fission resulted in 6.5-fold purification of ntcp from ASOR, and 25% of the resulting vesicles were completely depleted of the endocytic ligand. PMID:18284582

  19. Toxicity and interaction of titanium dioxide nanoparticles with microtubule protein

    Institute of Scientific and Technical Information of China (English)

    Zahra Naghdi Gheshlaghi; Gholam Hossein Riazi; Shahin Ahmadian; Mahmoud Ghafari; Roya Mahinpour

    2008-01-01

    Titanium dioxide (TiO2) nanoparticles (NPs) are widely used in several manufactured products. The small size of NPs facilitates their uptake into cells as well as transcytosis across epithelial cells into blood and lymph circulation to reach different sites, such as the central nervous system. Different studies have shown the risks that TiO2 NPs in the neuronal system and other organs present. As membranebound layer aggregates or single particles, TiO2 NPs can enter not only cells, but also mitochondria and nuclei.Therefore these particles can interact with cytoplasmic proteins such as microtubules (MTs). MTs are cytoskeletal proteins that are essential in eukaryotic cells for a variety of functions, such as cellular transport, cell motility and mitosis. MTs in neurons are used to transport substances such as neurotransmitters. Single TiO2 NPs in cytoplasm can interact with these proteins and affect their crucial functions in different tissues. In this study, we showed the effects of TiO2 NPs on MT polymerization and structure using ultraviolet spectrophotometer and fluorometry. The fluorescent spectroscopy showed a significant tubulin conformational change in the presence of TiO2 NPs and the ultraviolet spectroscopy results showed that TiO2 NPs affect tubulin polymerization and decrease it. The aim of this study was to find the potential risks that TiO2 NPs pose to human organs and cells.

  20. The rise and fall of the phragmoplast microtubule array.

    Science.gov (United States)

    Lee, Yuh-Ru Julie; Liu, Bo

    2013-12-01

    The cytokinetic apparatus, the phragmoplast, contains a bipolar microtubule (MT) framework that has the MT plus ends concentrated at or near the division site. This anti-parallel MT array provides tracks for the transport of Golgi-derived vesicles toward the plus ends so that materials enclosed are subsequently deposited at the division site. Here we will discuss a proposed model of the centrifugal expansion of the phragmoplast that takes place concomitantly with the assembly of the cell plate, the ultimate product of vesicle fusion. The expansion is a result of continuous MT assembly at the phragmoplast periphery while the MTs toward the center of the phragmoplast are disassembled. These events are the result of MT-dependent MT polymerization, bundling of anti-parallel MTs coming from opposite sides of the division plane that occurs selectively at the phragmoplast periphery, positioning of the plus ends of cross-linked MTs at or near the division site by establishing a minimal MT-overlapping zone, and debundling of anti-parallel MTs that is triggered by phosphorylation of MT-associated proteins. The debundled MTs are disassembled at last by factors including the MT severing enzyme katanin. PMID:24172707