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At least 577 records · Page 32

Airplane Mesh Development with Grid Density Studies

Automatic Grid Generation Wish List Geometry handling, including CAD clean up and mesh generation, remains a major bottleneck in the application of CFD methods. There is a pressing need for greater automation in several aspects of the geometry preparation in order to reduce set up time and eliminate user intervention as much as possible. Starting from the CAD representation of a configuration, there may be holes or overlapping surfaces which require an intensive effort to establish cleanly abutting surface patches, and collections of many patches may need to be combined for more efficient use of the geometrical representation. Obtaining an accurate and suitable body conforming grid with an adequate distribution of points throughout the flow-field, for the flow conditions of interest, is often the most time consuming task for complex CFD applications. There is a need for a clean unambiguous definition of the CAD geometry. Ideally this would be carried out automatically by smart CAD clean up software. One could also define a standard piece-wise smooth surface representation suitable for use by computational methods and then create software to translate between the various CAD descriptions and the standard representation. Surface meshing remains a time consuming, user intensive procedure. There is a need for automated surface meshing, requiring only minimal user intervention to define the overall density of mesh points. The surface mesher should produce well shaped elements (triangles or quadrilaterals) whose size is determined initially according to the surface curvature with a minimum size for flat pieces, and later refined by the user in other regions if necessary. Present techniques for volume meshing all require some degree of user intervention. There is a need for fully automated and reliable volume mesh generation. In addition, it should be possible to create both surface and volume meshes that meet guaranteed measures of mesh quality (e.g. minimum and maximum angle, stretching ratios, etc.).

Cliff, Susan E.↗

Networked Rectenna Array for Smart Material Actuators

The concept of microwave-driven smart material actuators is envisioned as the best option to alleviate the complexity associated with hard-wired control circuitry. Networked rectenna patch array receives and converts microwave power into a DC power for an array of smart actuators. To use microwave power effectively, the concept of a power allocation and distribution (PAD) circuit is adopted for networking a rectenna/actuator patch array. The PAD circuit is imbedded into a single embodiment of rectenna and actuator array. The thin-film microcircuit embodiment of PAD circuit adds insignificant amount of rigidity to membrane flexibility. Preliminary design and fabrication of PAD circuitry that consists of a few nodal elements were made for laboratory testing. The networked actuators were tested to correlate the network coupling effect, power allocation and distribution, and response time. The features of preliminary design are 16-channel computer control of actuators by a PCI board and the compensator for a power failure or leakage of one or more rectennas.

Choi, Sang H.↗

ROSAT Science Data Center

This report provides a summary of the Smithsonian Astrophysical Observatory (SAO) ROSAT SCIENCE DATA CENTER (RSDC) activities for the recent years of our contract. Details have already been reported in the monthly reports. The SAO was responsible for the High Resolution Imager (HRI) detector on ROSAT. We also provided and supported the HRI standard analysis software used in the pipeline processing (SASS). Working with our colleagues at the Max Planck in Garching Germany (MPE), we fixed bugs and provided enhancements. The last major effort in this area was the port from VMS/VAX to VMS/ALPHA architecture. In 1998, a timing bug was found in the HRI standard processing system which degraded the positional accuracy because events accessed incorrect aspect solutions. The bug was fixed and we developed off-line correction routines and provided them to the community. The Post Reduction Off-line Software (PROS) package was developed by SAO and runs in the IRAF environment. Although in recent years PROS was not a contractual responsibility of the RSDC, we continued to maintain the system and provided new capabilities such as the ability to deal with simulated AXAF data in preparation for the NASA call for proposals for Chandra. Our most recent activities in this area included the debugging necessary for newer versions of IRAF which broke some of our software. At SAO we have an operating version of PROS and hope to release a patch even though almost all functionality that was lost was subsequently recovered via an IRAF patch (i.e. most of our problems were caused by an IRAF bug).

Murray, Stephen↗

Active Wave Propagation and Sensing in Plates

Health monitoring of aerospace structures can be done using an active interrogation approach with diagnostic Lamb waves. Piezoelectric patches are often used to generate the waves, and it is helpful to understand how these waves propagate through a structure. To give a basic understanding of the actual physical process of wave propagation, a model is developed to simulate asymmetric wave propagation in a panel and to produce a movie of the wave motion. The waves can be generated using piezoceramic patches of any size or shape. The propagation, reflection, and interference of the waves are represented in the model. Measuring the wave propagation is the second important aspect of damage detection. Continuous sensors are useful for measuring waves because of the distributed nature of the sensor and the wave. Two sensor designs are modeled, and their effectiveness in measuring acoustic waves is studied. The simulation model developed is useful to understand wave propagation and to optimize the type of sensors that might be used for health monitoring of plate-like structures.

Ghoshal, Anindya↗

Blocked Grid Technologies as Applied to Aerodynamic Configurations

The advantages and disadvantages of the blocked grid methodology are discussed using results from ENSAERO, CNS, and CNS-FV. The first two codes are based on finite differences and the last on cell-centered finite volume formulation. Techniques that enhance the utility of the blocked (or patched) grid methodology are described. These techniques include mesh discontinuous zonal interfaces, sliding zonal interfaces, fast search procedures, and virtual zones. All of these methods are designed with two goals; namely extend the use of patched grids to unsteady aerodynamics, e.g. oscillating control flaps, and provide the user more flexibility in the grid topologies available for gridding complex aerodynamic configurations. For example, the use of virtual zones allows the user the choice of using one grid topology for surface grids, and another for the volume grids. This additional flexibility has a large impact in the amount of calendar time required to block and grid a complex aerodynamic configuration. Several examples are shown demonstrating the new features. Other issues involving grid generation are also discussed. In particular the existing problems of defining grid quality measures which are relevant are also described.

Klopfer, Goetz↗

Coupling Finite Element and Meshless Local Petrov-Galerkin Methods for Two-Dimensional Potential Problems

A coupled finite element (FE) method and meshless local Petrov-Galerkin (MLPG) method for analyzing two-dimensional potential problems is presented in this paper. The analysis domain is subdivided into two regions, a finite element (FE) region and a meshless (MM) region. A single weighted residual form is written for the entire domain. Independent trial and test functions are assumed in the FE and MM regions. A transition region is created between the two regions. The transition region blends the trial and test functions of the FE and MM regions. The trial function blending is achieved using a technique similar to the 'Coons patch' method that is widely used in computer-aided geometric design. The test function blending is achieved by using either FE or MM test functions on the nodes in the transition element. The technique was evaluated by applying the coupled method to two potential problems governed by the Poisson equation. The coupled method passed all the patch test problems and gave accurate solutions for the problems studied.

Chen, T.↗

Light-Toned, Layered Outcrops of Northern Terra Meridiani Mars: Viking, Phobos 2, and Mars Global Surveyor Observations

Locating outcrops of sedimentary rock on Mars is an important step toward deciphering the planet's geologic and climatologic record. Sedimentary rock representing the earliest martian environments, are of particular interest in this context. This is a report about a vast exposure of material proposed to be martian sedimentary rock. The outcrops cover an area (approximately sq 300,000 km) roughly the size of the Colorado Plateau in North America (approximately 260,000 sq km). The materials occur in northern Terra Meridiani, near of one of the four sites being considered for a 2004 NASA Mars Exploration Rover (MER) landing. The landing ellipse, centered at deg S, deg W, lies in a region exhibiting smooth and rough (at meter scale) dark-toned surfaces, with scattered light-toned patches. Stratigraphically, the dark-toned materials at the MER site lie unconformably on top of a previously-eroded, light-toned surface; the light-toned patches in the landing ellipse are geologic windows down to this lower stratigraphic unit. North of the landing ellipse, the light-toned materials are well-exposed because the darker materials have been removed, stranding outlier remnants in a few locations. The light-toned materials are layered, vertically heterogeneous, and exhibit lateral continuity over hundreds of kilometers. Eroded layers produce cliffs; some outcrops are expressed as mesas, buttes, and spires; and impact craters ranging in diameter from a few meters to tens of kilometers are interbedded with the layers. The purpose of this report is to summarize the results of greater than 6 years of photogeologic investigation into the nature of the light-toned outcrops of northern Terra Meridiani. The work is a 'snapshot' of progress made toward eventual geologic mapping and establishment of the stratigraphic sequence for the materials through 30 September 2002, a day prior to the first release of Mars Odyssey Thermal Emission Imaging System (THEMIS) data to the NASA Planetary Data System (PDS). The main body of data examined were Mars Global Surveyor (MGS) Mars Orbiter Camera (MOC) images acquired through 30 September 2002. The data also 2 include Viking orbiter images, a Phobos 2 Termoscan image, MGS Mars Orbiter Laser Altimeter (MOLA) topographic observations, and the products of published Viking Infrared Thermal Mapper (IRTM) and Mars Global Surveyor (MGS) Thermal Emission Spectrometer (TES) analyses. Through September 2002, over 126,000 MOC images had been acquired, and greater than 600 of the MOC narrow angle (1.5-12 m/pixel) images occur within the portions of Terra Meridiani and southwestern Arabia Terra.

Edgett, Kenneth S.↗

A Stochastic Model of Space-Time Variability of Tropical Rainfall: I. Statistics of Spatial Averages

Global maps of rainfall are of great importance in connection with modeling of the earth s climate. Comparison between the maps of rainfall predicted by computer-generated climate models with observation provides a sensitive test for these models. To make such a comparison, one typically needs the total precipitation amount over a large area, which could be hundreds of kilometers in size over extended periods of time of order days or months. This presents a difficult problem since rain varies greatly from place to place as well as in time. Remote sensing methods using ground radar or satellites detect rain over a large area by essentially taking a series of snapshots at infrequent intervals and indirectly deriving the average rain intensity within a collection of pixels , usually several kilometers in size. They measure area average of rain at a particular instant. Rain gauges, on the other hand, record rain accumulation continuously in time but only over a very small area tens of centimeters across, say, the size of a dinner plate. They measure only a time average at a single location. In making use of either method one needs to fill in the gaps in the observation - either the gaps in the area covered or the gaps in time of observation. This involves using statistical models to obtain information about the rain that is missed from what is actually detected. This paper investigates such a statistical model and validates it with rain data collected over the tropical Western Pacific from ship borne radars during TOGA COARE (Tropical Oceans Global Atmosphere Coupled Ocean-Atmosphere Response Experiment). The model incorporates a number of commonly observed features of rain. While rain varies rapidly with location and time, the variability diminishes when averaged over larger areas or longer periods of time. Moreover, rain is patchy in nature - at any instant on the average only a certain fraction of the observed pixels contain rain. The fraction of area covered by rain decreases, as the size of a pixel becomes smaller. This means that within what looks like a patch of rainy area in a coarse resolution view with larger pixel size, one finds clusters of rainy and dry patches when viewed on a finer scale. The model makes definite predictions about how these and other related statistics depend on the pixel size. These predictions were found to agree well with data. In a subsequent second part of the work we plan to test the model with rain gauge data collected during the TRMM (Tropical Rainfall Measuring Mission) ground validation campaign.

Kundu, Prasun K.↗

Morphology and the Strength of Intermolecular Contact in Protein Crystals

The strengths of intermolecular contacts (macrobonds) in four lysozyme crystals were estimated based on the strengths of individual intermolecular interatomic interaction pairs. The periodic bond chain of these macrobonds accounts for the morphology of protein crystals as shown previously. Further in this paper, the surface area of contact, polar coordinate representation of contact site, Coulombic contribution on the macrobond strength, and the surface energy of the crystal have been evaluated. Comparing location of intermolecular contacts in different polymorphic crystal modifications, we show that these contacts can form a wide variety of patches on the molecular surface. The patches are located practically everywhere on this surface except for the concave active site. The contacts frequently include water molecules, with specific intermolecular hydrogen-bonds on the background of non-specific attractive interactions. The strengths of macrobonds are also compared to those of other protein complex systems. Making use of the contact strengths and taking into account bond hydration we also estimated crystal-water interfacial energies for different crystal faces.

Matsuura, Yoshiki↗

Finite Element Modeling of a Cylindrical Contact Using Hertzian Assumptions

The turbine blades in the high-pressure fuel turbopump/alternate turbopump (HPFTP/AT) are subjected to hot gases rapidly flowing around them. This flow excites vibrations in the blades. Naturally, one has to worry about resonance, so a damping device was added to dissipate some energy from the system. The foundation is now laid for a very complex problem. The damper is in contact with the blade, so now there are contact stresses (both normal and tangential) to contend with. Since these stresses can be very high, it is not all that difficult to yield the material. Friction is another non-linearity and the blade is made out of a Nickel-based single-crystal superalloy that is orthotropic. A few approaches exist to solve such a problem and computer models, using contact elements, have been built with friction, plasticity, etc. These models are quite cumbersome and require many hours to solve just one load case and material orientation. A simpler approach is required. Ideally, the model should be simplified so the analysis can be conducted faster. When working with contact problems determining the contact patch and the stresses in the material are the main concerns. Closed-form solutions for non-conforming bodies, developed by Hertz, made out of isotropic materials are readily available. More involved solutions for 3-D cases using different materials are also available. The question is this: can Hertzian1 solutions be applied, or superimposed, to more complicated problems-like those involving anisotropic materials? That is the point of the investigation here. If these results agree with the more complicated computer models, then the analytical solutions can be used in lieu of the numerical solutions that take a very long time to process. As time goes on, the analytical solution will eventually have to include things like friction and plasticity. The models in this report use no contact elements and are essentially an applied load problem using Hertzian assumptions to determine the contact patch dimensions.

Knudsen, Erik↗

Impedance-Based Structural Health Monitoring for Composite Laminates at Cryogenic Environments

One of the important ways of increasing the payload in a reusable launch vehicle (RLV) is to replace heavy metallic materials by lightweight composite laminates. Among various parts and systems of the RLV, this project focuses on tanks containing cryogenic fuel. Historically, aluminum alloys have been used as the materials to construct fuel tanks for launch vehicles. To replace aluminum alloys with composite laminates or honeycomb materials, engineers have to make sure that the composites are free of defects before, during, and after launch. In addition to robust design and manufacturing procedures, the performance of the composite structures needs to be monitored constantly.In recent years, the impedance-based health monitoring technique has shown its promise in many applications. This technique makes use of the special properties of smart piezoelectric materials to identify the change of material properties due to the nucleation and progression of damage. The piezoceramic patch serves as a sensor and an actuator simultaneously. The piezoelectric patch is bonded onto an existing structure or embedded into a new structure and electrically excited at high frequencies. The signature (impedance or admittance) is extracted as a function of the exciting frequency and is compared with the baseline signature of the healthy state. The damage is quantified using root mean square deviation (RMSD) in the impedance signatures with respect to the baseline signature. A major advantage of this technique is that the procedure is nondestructive in nature and does not perturb the properties and performance of the materials and structures. This project aims at applying the impedance-based nondestructive testing technique to the damage identification of composite laminates at cryogenic temperature.

Tseng, Kevin↗

The Variability of Transverse Aeolian Ripples in Troughs on Mars

A precursory glance at MGS images of the surface of Mars show an abundance of aeolian transverse ridges. These ridges are located in a variety of geological terrains. Zimbelman and Wilson have separated the small-scale aeolian features of Syrtis Major into six categories: ripples associated with obstacles, ripple bands, ripple fields, ripple patches, isolated ripple patches and ripples associated with dunes. This paper focuses on one of these categories, that of ripple bands which tend to accumulate within linear troughs. As the origin of these features is still being studied (i.e. ripples versus dunes), we refer to them simply as transverse aeolian ridges.

Bourke, M. C.↗

Crystallization Physics in Biomacromolecular Systems

The crystals are built of molecules of protein, nucleic acid and their complexes, like viruses, approx. 5x10(exp 3)+ 3x10(exp 6) Da in weight and 2 + 20 nm in effective diameter. This size strongly exceeds action range of molecular forces and makes a big difference with inorganic crystals. Intermolecular contacts form patches on the biomacromolecular surface. Each patch may occupy only a small percent of the whole surface and vary from polymorph to polymorph of the same protein. Thus, under different conditions (pH, solution chemistry, temperature, any area on the macromolecular surface may form a contact. The crystal Young moduli, E approx. equals 0.1 + 0.5 GPa are more than 10 times lower than that of inorganics and the biomolecules themselves. Water within biocrystals (30-70%) is unable to flow unless typical deformation time is longer than approx. 10(exp -5)s. This explains the discrepancy between light scattering and static measurements of E. Nucleation and Growth requires typically concentrations exceeding the equilibrium ones up to 100 times - because of the new size scale results in 10 - 10(exp 3) times lower kinetic coefficients than that needed for inorganic solution growth. All phenomena observed in the latter occur with protein crystallization and are even better studied by AFM. Crystals are typically facetted. Among unexpected findings of general significance are - net molecular exchange flux at kinks is much lower than that expected from supersaturation, steps with low (< approx. 10(exp -2)) kink density at steps follow Gibbs-Thomson law only at very low supersaturations, step segment growth rate may be independent of step energy. Crystal perfection is a must of biocrystallization to achieve the major goal to find 3-D atomic structure of biomacromolecules by x-ray diffraction. Poor diffraction resolution (> 3Angstrom) makes crystallization a bottleneck for structural biology. All defects typical of small molecule crystals are found in biocrystals, but the defects responsible for poor resolution are not identified. Conformational changes are one of them. Biocrystallization in microgravity reportedly results in 20% cases of better crystals. The mechanism of how lack of convection can do this is still not clear. Lower supersaturation, self-purification &om preferentially trapped homologous impurities and step bunching are viable hypotheses.

Chernov, A. A.↗

Development of Modal Test Techniques for Validation of a Solar Sail Design

This paper focuses on the development of modal test techniques for validation of a solar sail gossamer space structure design. The major focus is on validating and comparing the capabilities of various excitation techniques for modal testing solar sail components. One triangular shaped quadrant of a solar sail membrane was tested in a 1 Torr vacuum environment using various excitation techniques including, magnetic excitation, and surface-bonded piezoelectric patch actuators. Results from modal tests performed on the sail using piezoelectric patches at different positions are discussed. The excitation methods were evaluated for their applicability to in-vacuum ground testing and to the development of on orbit flight test techniques. The solar sail membrane was tested in the horizontal configuration at various tension levels to assess the variation in frequency with tension in a vacuum environment. A segment of a solar sail mast prototype was also tested in ambient atmospheric conditions using various excitation techniques, and these methods are also assessed for their ground test capabilities and on-orbit flight testing.

Gaspar, James L.↗

Hybrid Active/Passive Control of Sound Radiation from Panels with Constrained Layer Damping and Model Predictive Feedback Control

There has been considerable interest over the past several years in applying feedback control methods to problems of structural acoustics. One problem of particular interest is the control of sound radiation from aircraft panels excited on one side by a turbulent boundary layer (TBL). TBL excitation appears as many uncorrelated sources acting on the panel, which makes it difficult to find a single reference signal that is coherent with the excitation. Feedback methods have no need for a reference signal, and are thus suited to this problem. Some important considerations for the structural acoustics problem include the fact that the required controller bandwidth can easily extend to several hundred Hertz, so a digital controller would have to operate at a few kilohertz. In addition, aircraft panel structures have a reasonably high modal density over this frequency range. A model based controller must therefore handle the modally dense system, or have some way to reduce the bandwidth of the problem. Further complicating the problem is the fact that the stiffness and dynamic properties of an aircraft panel can vary considerably during flight due to altitude changes resulting in significant resonant frequency shifts. These considerations make the tradeoff between robustness to changes in the system being controlled and controller performance especially important. Recent papers concerning the design and implementation of robust controllers for structural acoustic problems highlight the need to consider both performance and robustness when designing the controller. While robust control methods such as H1 can be used to balance performance and robustness, their implementation is not easy and requires assumptions about the types of uncertainties in the plant being controlled. Achieving a useful controller design may require many tradeoff studies of different types of parametric uncertainties in the system. Another approach to achieving robustness to plant changes is to make the controller adaptive. For example, a mathematical model of the plant could be periodically updated as the plant changes, and the feedback gains recomputed from the updated model. To be practical, this approach requires a simple plant model that can be updated quickly with reasonable computational requirements. A recent paper by the authors discussed one way to simplify a feedback controller, by reducing the number of actuators and sensors needed for good performance. The work was done on a tensioned aircraft-style panel excited on one side by TBL flow in a low speed wind tunnel. Actuation was provided by a piezoelectric (PZT) actuator mounted on the center of the panel. For sensing, the responses of four accelerometers, positioned to approximate the response of the first radiation mode of the panel, were summed and fed back through the controller. This single input-single output topology was found to have nearly the same noise reduction performance as a controller with fifteen accelerometers and three PZT patches. This paper extends the previous results by looking at how constrained layer damping (CLD) on a panel can be used to enhance the performance of the feedback controller thus providing a more robust and efficient hybrid active/passive system. The eventual goal is to use the CLD to reduce sound radiation at high frequencies, then implement a very simple, reduced order, low sample rate adaptive controller to attenuate sound radiation at low frequencies. Additionally this added damping smoothes phase transitions over the bandwidth which promotes robustness to natural frequency shifts. Experiments were conducted in a transmission loss facility on a clamped-clamped aluminum panel driven on one side by a loudspeaker. A generalized predictive control (GPC) algorithm, which is suited to online adaptation of its parameters, was used in single input-single output and multiple input-single output configurations. Because this was a preliminary look at the potential constrained layer damping for adaptive control, static feedback control with no online adaptation was used. Two configurations of CLD in addition to a bare panel configuration were studied. For each CLD configuration, two sensor arrangements for the feedback controller were compared. The first arrangement used fifteen accelerometers on the panel to estimate the responses of the first six radiation modes of the panel. The second sensor arrangement was simpler, using the summed responses of only four accelerometers to approximate the response of the first radiation mode of the panel. In all cases a PZT patch was mounted at the center of the panel for control input. The performance of the controller was quantified using the responses of the fifteen accelerometers on the panel to estimate radiated sound power. The paper begins with a brief discussion of the GPC algorithm and the experimental setup. The experimental results are discussed next, comparing the CLD and sensor configurations, followed by discussion and conclusions.

Cabell, Randolph H.↗

Bone morphogenic protein 4 produced in endothelial cells by oscillatory shear stress stimulates an inflammatory response

Atherosclerosis is now viewed as an inflammatory disease occurring preferentially in arterial regions exposed to disturbed flow conditions, including oscillatory shear stress (OS), in branched arteries. In contrast, the arterial regions exposed to laminar shear (LS) are relatively lesion-free. The mechanisms underlying the opposite effects of OS and LS on the inflammatory and atherogenic processes are not clearly understood. Here, through DNA microarrays, protein expression, and functional studies, we identify bone morphogenic protein 4 (BMP4) as a mechanosensitive and pro-inflammatory gene product. Exposing endothelial cells to OS increased BMP4 protein expression, whereas LS decreased it. In addition, we found BMP4 expression only in the selective patches of endothelial cells overlying foam cell lesions in human coronary arteries. The same endothelial patches also expressed higher levels of intercellular cell adhesion molecule-1 (ICAM-1) protein compared with those of non-diseased areas. Functionally, we show that OS and BMP4 induced ICAM-1 expression and monocyte adhesion by a NFkappaB-dependent mechanism. We suggest that BMP4 is a mechanosensitive, inflammatory factor playing a critical role in early steps of atherogenesis in the lesion-prone areas.

NASA Program Fundamental Space Biology↗

A geostatistical analysis of small-scale spatial variability in bacterial abundance and community structure in salt marsh creek bank sediments

Small-scale variations in bacterial abundance and community structure were examined in salt marsh sediments from Virginia's eastern shore. Samples were collected at 5 cm intervals (horizontally) along a 50 cm elevation gradient, over a 215 cm horizontal transect. For each sample, bacterial abundance was determined using acridine orange direct counts and community structure was analyzed using randomly amplified polymorphic DNA fingerprinting of whole-community DNA extracts. A geostatistical analysis was used to determine the degree of spatial autocorrelation among the samples, for each variable and each direction (horizontal and vertical). The proportion of variance in bacterial abundance that could be accounted for by the spatial model was quite high (vertical: 60%, horizontal: 73%); significant autocorrelation was found among samples separated by 25 cm in the vertical direction and up to 115 cm horizontally. In contrast, most of the variability in community structure was not accounted for by simply considering the spatial separation of samples (vertical: 11%, horizontal: 22%), and must reflect variability from other parameters (e.g., variation at other spatial scales, experimental error, or environmental heterogeneity). Microbial community patch size based upon overall similarity in community structure varied between 17 cm (vertical) and 35 cm (horizontal). Overall, variability due to horizontal position (distance from the creek bank) was much smaller than that due to vertical position (elevation) for both community properties assayed. This suggests that processes more correlated with elevation (e.g., drainage and redox potential) vary at a smaller scale (therefore producing smaller patch sizes) than processes controlled by distance from the creek bank. c2002 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.

Non-NASA Center↗

Furrow microtubules and localized exocytosis in cleaving Xenopus laevis embryos

In dividing Xenopus eggs, furrowing is accompanied by expansion of a new domain of plasma membrane in the cleavage plane. The source of the new membrane is known to include a store of oogenetically produced exocytotic vesicles, but the site where their exocytosis occurs has not been described. Previous work revealed a V-shaped array of microtubule bundles at the base of advancing furrows. Cold shock or exposure to nocodazole halted expansion of the new membrane domain, which suggests that these microtubules are involved in the localized exocytosis. In the present report, scanning electron microscopy revealed collections of pits or craters, up to approximately 1.5 micro m in diameter. These pits are evidently fusion pores at sites of recent exocytosis, clustered in the immediate vicinity of the deepening furrow base and therefore near the furrow microtubules. Confocal microscopy near the furrow base of live embryos labeled with the membrane dye FM1-43 captured time-lapse sequences of individual exocytotic events in which irregular patches of approximately 20 micro m(2) of unlabeled membrane abruptly displaced pre-existing FM1-43-labeled surface. In some cases, stable fusion pores, approximately 2 micro m in diameter, were seen at the surface for up to several minutes before suddenly delivering patches of unlabeled membrane. To test whether the presence of furrow microtubule bundles near the surface plays a role in directing or concentrating this localized exocytosis, membrane expansion was examined in embryos exposed to D(2)O to induce formation of microtubule monasters randomly under the surface. D(2)O treatment resulted in a rapid, uniform expansion of the egg surface via random, ectopic exocytosis of vesicles. This D(2)O-induced membrane expansion was completely blocked with nocodazole, indicating that the ectopic exocytosis was microtubule-dependent. Results indicate that exocytotic vesicles are present throughout the egg subcortex, and that the presence of microtubules near the surface is sufficient to mobilize them for exocytosis at the end of the cell cycle.

Non-NASA Center↗