Sähkökäyttöjen kaapelivärähtelyt

Taajuudenmuuttajan kytkennän synnyttämä nopea jännitemuutos aiheuttaa pitkään moottorikaapeliin sähköisen värähtelyilmiön. Ilmiö on tullut erityisesti esille uusien nopeasti kytkevien puolijohdetehokytkimien ilmestyttyä markkinoille. Taajuudenmuuttajan lähtöön asennettu jännitteen nousunopeutta rajoittava suodin vähentää kaapelivärähtelyä, mutta riittävän pitkässä kaapelissa värähtely on voimakasta lähtösuotimesta huolimatta. Kaapelivärähtelyilmiön seurauksena moottorikaapelin taajuudenmuuttajan puoleiseen päähän syntyy voimakas virtavärähtely ja moottorin puoleiseen päähän voimakas jännitevärähtely. Sähkökäyttöjen vektorisäätöalgoritmit tekevät ohjauspäätöksiä moottorikaapelin taajuudenmuuttajan päästä tehtyjen virtamittausten perusteella. Säädön päätösväli on niin lyhyt, että kaapelin virtavärähtely ehtii häiritä säädön toimintaa. Tässä työssä on esitetty kaapelivärähtelyä kuvaava taajuudenmuuttajan lähtösuotimen huomioon ottava siirtofunktioperustainen matemaattinen malli. Mallin avulla kaapelivärähtelyilmiötä voi analysoida lineaarisen säätöteorian menetelmillä. Virtavärähtelyn moottorisäätöön tuomiin ongelmiin ratkaisuksi on esitetty virran mittasignaalin käsittelemistä analogisella ja digitaalisella suotimella. Simulointitulosten perusteella ratkaisua voidaan pitää toimivana. Lopuksi esitetään, kuinka avaruusvektoriteorian mukaista induktiomoottorimallia ja kaapelivärähtelymallia voidaan simuloida yhdessä.

Jännitevälipiiritaajuusmuuttajan moottorikaapelin maksimipituuden määrittely

Työssä tutkitaan, kuinka pitkää moottorikaapelia on jännitevälipiiritaajuusmuuttajan kanssa mahdollista käyttää niin, että määritellyt reunaehdot vielä toteutuvat. Tavoitteena on tuottaa tietoa myynnille, markkinoinnille, tuotehallinnalle sekä tuoteylläpidolle siitä, miten taajuus-muuttaja toimii pidemmillä moottorikaapeleilla kuin valmistaja suosittelee. Tutkitut moottori-kaapelipituudet olivat 175…1025 metriä ja tutkitut laitteet nimellislähtövirroiltaan 2,4...25 A. Työssä aihetta käsitellään jänniteheijastusten näkökulmasta. Lisäksi tutkitaan moottorikaapelin pituuden vaikutusta taajuusmuuttajan eri komponenttien lämpenemiseen. Taajuusmuuttajan toiminnallisuutta arvioidaan moottorin suunnanvaihtojen avulla sekä turvallista toimintaa oikosulkutestein. Tutkittujen taajuusmuuttajien kohdalla on mahdollista käyttää taajuusmuuttajavalmistajan suositusta pidempiä moottorikaapeleita. Moottoriliittimien ylijännitteitä aiheuttavat jännite-heijastukset eivät aiheuttaneet raja-arvoja ylittäviä huippujännitteitä tutkituilla laitekokoon-panoilla. Myös lämpötilannousu oli maltillista tai jopa vähäistä taajuusmuuttajasta mitatuilla komponenteilla. Moottorisäätö havaittiin toimintakykyiseksi pidemmilläkin moottorikaape-leilla, tosin moottorin vääntömomentti heikkeni moottorikaapelien pituutta kasvatettaessa. Virranmittaus toimi hyvin myös pitkillä kaapeleilla, tuottaen vikalaukaisun kaikissa tehdyissä oikosulkutilanteissa. Moottorin ja taajuusmuuttajan melutaso nousivat moottorikaapelien pi-tuutta kasvatettaessa, vaikkakin moottorin käynti oli tasaista ja katkotonta Moottorikaapelin pituutta voidaan kasvattaa 325 metriin kaikissa tutkituissa laitteissa ilman, että mikään tutkittu ominaisuus vielä olennaisesti heikkenisi. Vielä 525 metrin moottorikaape-leita on mahdollista käyttää, mutta tällöin vääntömomentin tuotto on jo heikompaa.

Power line communication in motor cables of variable-speed electric drives − analysis and

Data transmission between an electric motor and a frequency converter is required in variablespeed electric drives because of sensors installed at the motor. Sensor information can be used for various useful applications to improve the system reliability and its properties. Traditionally, the communication medium is implemented by an additional cabling. However, the costs of the traditional method may be an obstacle to the wider application of data transmission between a motor and a frequency converter. In any case, a power cable is always installed between a motor and a frequency converter for power supply, and hence it may be applied as a communication medium for sensor level data. This thesis considers power line communication (PLC) in inverter-fed motor power cables. The motor cable is studied as a communication channel in the frequency band of 100 kHz−30 MHz. The communication channel and noise characteristics are described. All the individual components included in a variable-speed electric drive are presented in detail. A channel model is developed, and it is verified by measurements. A theoretical channel information capacity analysis is carried out to estimate the opportunities of a communication medium. Suitable communication and forward error correction (FEC) methods are suggested. A general method to implement a broadband and Ethernet-based communication medium between a motor and a frequency converter is proposed. A coupling interface is also developed that allows to install the communication device safely to a three-phase inverter-fed motor power cable. Practical tests are carried out, and the results are analyzed. Possible applications for the proposed method are presented. A speed feedback motor control application is verified in detail by simulations and laboratory tests because of restrictions for the delay in the feedback loop caused by PLC. Other possible applications are discussed at a more general level.

Power semiconductor nonlinearities in active du/dt output filtering

This doctoral thesis introduces an improved control principle for active du/dt output filtering in variable-speed AC drives, together with performance comparisons with previous filtering methods. The effects of power semiconductor nonlinearities on the output filtering performance are investigated. The nonlinearities include the timing deviation and the voltage pulse waveform distortion in the variable-speed AC drive output bridge. Active du/dt output filtering (ADUDT) is a method to mitigate motor overvoltages in variable-speed AC drives with long motor cables. It is a quite recent addition to the du/dt reduction methods available. This thesis improves on the existing control method for the filter, and concentrates on the lowvoltage (below 1 kV AC) two-level voltage-source inverter implementation of the method. The ADUDT uses narrow voltage pulses having a duration in the order of a microsecond from an IGBT (insulated gate bipolar transistor) inverter to control the output voltage of a tuned LC filter circuit. The filter output voltage has thus increased slope transition times at the rising and falling edges, with an opportunity of no overshoot. The effect of the longer slope transition times is a reduction in the du/dt of the voltage fed to the motor cable. Lower du/dt values result in a reduction in the overvoltage effects on the motor terminals. Compared with traditional output filtering methods to accomplish this task, the active du/dt filtering provides lower inductance values and a smaller physical size of the filter itself. The filter circuit weight can also be reduced. However, the power semiconductor nonlinearities skew the filter control pulse pattern, resulting in control deviation. This deviation introduces unwanted overshoot and resonance in the filter. The controlmethod proposed in this thesis is able to directly compensate for the dead time-induced zero-current clamping (ZCC) effect in the pulse pattern. It gives more flexibility to the pattern structure, which could help in the timing deviation compensation design. Previous studies have shown that when a motor load current flows in the filter circuit and the inverter, the phase leg blanking times distort the voltage pulse sequence fed to the filter input. These blanking times are caused by excessively large dead time values between the IGBT control pulses. Moreover, the various switching timing distortions, present in realworld electronics when operating with a microsecond timescale, bring additional skew to the control. Left uncompensated, this results in distortion of the filter input voltage and a filter self-induced overvoltage in the form of an overshoot. This overshoot adds to the voltage appearing at the motor terminals, thus increasing the transient voltage amplitude at the motor. This doctoral thesis investigates the magnitude of such timing deviation effects. If the motor load current is left uncompensated in the control, the filter output voltage can overshoot up to double the input voltage amplitude. IGBT nonlinearities were observed to cause a smaller overshoot, in the order of 30%. This thesis introduces an improved ADUDT control method that is able to compensate for phase leg blanking times, giving flexibility to the pulse pattern structure and dead times. The control method is still sensitive to timing deviations, and their effect is investigated. A simple approach of using a fixed delay compensation value was tried in the test setup measurements. The ADUDT method with the new control algorithm was found to work in an actual motor drive application. Judging by the simulation results, with the delay compensation, the method should ultimately enable an output voltage performance and a du/dt reduction that are free from residual overshoot effects. The proposed control algorithm is not strictly required for successful ADUDT operation: It is possible to precalculate the pulse patterns by iteration and then for instance store them into a look-up table inside the control electronics. Rather, the newly developed control method is a mathematical tool for solving the ADUDT control pulses. It does not contain the timing deviation compensation (from the logic-level command to the phase leg output voltage), and as such is not able to remove the timing deviation effects that cause error and overshoot in the filter. When the timing deviation compensation has to be tuned-in in the control pattern, the precalculated iteration method could prove simpler and equally good (or even better) compared with the mathematical solution with a separate timing compensation module. One of the key findings in this thesis is the conclusion that the correctness of the pulse pattern structure, in the sense of ZCC and predicted pulse timings, cannot be separated from the timing deviations. The usefulness of the correctly calculated pattern is reduced by the voltage edge timing errors. The doctoral thesis provides an introductory background chapter on variable-speed AC drives and the problem of motor overvoltages and takes a look at traditional solutions for overvoltage mitigation. Previous results related to the active du/dt filtering are discussed. The basic operation principle and design of the filter have been studied previously. The effect of load current in the filter and the basic idea of compensation have been presented in the past. However, there was no direct way of including the dead time in the control (except for solving the pulse pattern manually by iteration), and the magnitude of nonlinearity effects had not been investigated. The enhanced control principle with the dead time handling capability and a case study of the test setup timing deviations are the main contributions of this doctoral thesis. The simulation and experimental setup results show that the proposed control method can be used in an actual drive. Loss measurements and a comparison of active du/dt output filtering with traditional output filtering methods are also presented in the work. Two different ADUDT filter designs are included, with ferrite core and air core inductors. Other filters included in the tests were a passive du/dtfilter and a passive sine filter. The loss measurements incorporated a silicon carbide diode-equipped IGBT module, and the results show lower losses with these new device technologies. The new control principle was measured in a 43 A load current motor drive system and was able to bring the filter output peak voltage from 980 V (the previous control principle) down to 680 V in a 540 V average DC link voltage variable-speed drive. A 200 m motor cable was used, and the filter losses for the active du/dt methods were 111W–126 W versus 184 W for the passive du/dt. In terms of inverter and filter losses, the active du/dt filtering method had a 1.82-fold increase in losses compared with an all-passive traditional du/dt output filter. The filter mass with the active du/dt method was 17% (2.4 kg, air-core inductors) compared with 14 kg of the passive du/dt method filter. Silicon carbide freewheeling diodes were found to reduce the inverter losses in the active du/dt filtering by 18% compared with the same IGBT module with silicon diodes. For a 200 m cable length, the average peak voltage at the motor terminals was 1050 V with no filter, 960 V for the all-passive du/dt filter, and 700 V for the active du/dt filtering applying the new control principle.

Sähköverkkotiedonsiirtomenetelmän kehittäminen taajuusmuuttajaohjattuihin sähkökäyttöihin

Sähkömoottorin jatkuva-aikainen kunnonvalvonta vaatii tiedonsiirtoa moottorilta ylemmälle tietojärjestelmätasolle, kuten taajuusmuuttajalle. Uusien kaapeleiden asennus on työlästä ja kallista. Moottorin ja taajuusmuuttajan välillä on kuitenkin aina kaapeli, jota käytetään moottorin tehonsyöttöön. Tätä kaapelia on mahdollista käyttää myös tiedonsiirtokanavana. Tässä diplomityössä käsitellään ja tutkitaan sähköverkkotiedonsiirtomenetelmää, jonka avulla voidaan muodostaa Ethernet-yhteys moottorilta taajuusmuuttajalle tai toisinpäin. Työssä kehitetään simulointimalli tiedonsiirtokanavan taajuusriippuvan vaimennuksen simuloimiseen. Lisäksi kehitetään kytkentärajapinta, joka mahdollistaa tiedonsiirron kolmivaiheisessa taajuusmuuttajan syöttämässä moottorikaapelissa. Työssä suunnitellaan ja toteutetaan tiedonsiirtoa varten pilot-laitteisto, jolla tehdään laboratoriotestit. Laboratoriotesteillä varmistetaan menetelmän soveltuvuus. Laboratoriotestien avulla etsitään myös reunaehdot tiedonsiirron toiminnalle ja tiedonsiirtonopeuteen vaikuttavia tekijöitä taajuusmuuttajakäytössä. Lisäksi tarkastellaan menetelmän aiheuttamaa viivettä, joka on tärkeäsäätösovellusten kannalta. Lopuksi arvioidaan menetelmän soveltuvuus käytännön sovelluksiin.

Jännitteen reunan hallinta taajuusmuuttajissa

Diplomityössä perehdyttiin taajuusmuuttajien toimintaan ja ohjaukseen. Lisäksi työssä tarkasteltiin vaihtosuuntaajan nopeiden transienttitilojen aiheuttamaa moottorin ylijännitettä. Moottorikaapelin heijastuksia käsiteltiin vertaamalla moottorikaapelia siirtolinjaan ja todennettiin ylijännitteen syyt. Ylijännitteen vähentämiseksi on kehitetty useita suodatusmenetelmiä. Työssä vertailtiin näitä menetelmiä ja kartoitettiin kaupallisia vaihtoehtoja. Taajuusmuuttajan ohjaus on tähän päivään asti tehty yleensä käyttäen mikroprosessoria sekä logiikkapiiriä. Tulevaisuudessa ohjaukseen käytetään todennäköisesti uudelleenohjelmoitavia FPGA-piirejä (Field Programmable Gate Array). FPGA-piirin etuihin kuuluu uudelleenohjelmoitavuus sekä ohjauksen keskittäminen yhdelle piirille.

Hybrid LC Filter for Power Electronic Drives: Theory and Implementation

Power electronic converter drives use, for the sake of high efficiency, pulse-width modulation that results in sequences of high-voltage high-frequency steep-edged pulses. Such a signal contains a set of high harmonics not required for control purposes. Harmonics cause reflections in the cable between the motor and the inverter leading to faster winding insulation ageing. Bearing failures and problems with electromagnetic compatibility may also result. Electrical du/dt filters provide an effective solution to problems caused by pulse-width modulation, thereby increasing the performance and service life of the electrical machines. It is shown that RLC filters effectively decrease the reflection phenomena in the cable. Improved (simple, but effective) solutions are found for both differential- and common-mode signals; these solutions use a galvanic connection between the RLC filter star point and the converter DC link. Foil chokes and film capacitors are among the most widely used components in high-power applications. In actual applications they can be placed in different parts of the cabinet. This fact complicates the arrangement of the cabinet and decreases the reliability of the system. In addition, the inductances of connection wires may prevent filtration at high frequencies. This thesis introduces a new hybrid LC filter that uses a natural capacitance between the turns of the foil choke based on integration of an auxiliary layer into it. The main idea of the hybrid LC filter results from the fact that both the foil choke and the film capacitors have the same roll structure. Moreover, the capacitance between the turns (“intra capacitance”) of the foil inductors is the reason for the deterioration of their properties at high frequencies. It is shown that the proposed filter has a natural cancellation of the intra capacitance. A hybrid LC filter may contain two or more foil layers isolated from each other and coiled on a core. The core material can be iron or even air as in the filter considered in this work. One of the foils, called the main foil, can be placed between the inverter and the motor cable. Other ones, called auxiliary foils, may be connected in star to create differential-mode noise paths, and then coupled to the DC link midpoint to guarantee a traveling path, especially for the common-mode currents. This way, there is a remarkable capacitance between the main foil and the auxiliary foil. Investigations showed that such a system can be described by a simple equivalent LC filter in a wide range of frequencies. Because of its simple hybrid construction, the proposed LC filter can be a cost-effective and competitive solution for modern power drives. In the thesis, the application field of the proposed filter is considered and determined. The basics of hybrid LC filter design are developed further. High-frequency behaviour of the proposed filter is analysed by simulations. Finally, the thesis presents experimental data proving that the hybrid LC filter can be used for du/dt of PWM pulses and reduction of common-mode currents.

Active du/dt Filtering for Variable-Speed AC Drives

An oscillating overvoltage has become a common phenomenon at the motor terminal in inverter-fed variable-speed drives. The problem has emerged since modern insulated gate bipolar transistors have become the standard choice as the power switch component in lowvoltage frequency converter drives. Theovervoltage phenomenon is a consequence of the pulse shape of inverter output voltage and impedance mismatches between the inverter, motor cable, and motor. The overvoltages are harmful to the electric motor, and may cause, for instance, insulation failure in the motor. Several methods have been developed to mitigate the problem. However, most of them are based on filtering with lossy passive components, the drawbacks of which are typically their cost and size. In this doctoral dissertation, application of a new active du/dt filtering method based on a low-loss LC circuit and active control to eliminate the motor overvoltages is discussed. The main benefits of the method are the controllability of the output voltage du/dt within certain limits, considerably smaller inductances in the filter circuit resulting in a smaller physical component size, and excellent filtering performance when compared with typical traditional du/dt filtering solutions. Moreover, no additional components are required, since the active control of the filter circuit takes place in the process of the upper-level PWM modulation using the same power switches as the inverter output stage. Further, the active du/dt method will benefit from the development of semiconductor power switch modules, as new technologies and materials emerge, because the method requires additional switching in the output stage of the inverter and generation of narrow voltage pulses. Since additional switching is required in the output stage, additional losses are generated in the inverter as a result of the application of the method. Considerations on the application of the active du/dt filtering method in electric drives are presented together with experimental data in order to verify the potential of the method.

Monimoottorikäyttöjen suojaukset, mitoitus ja ohjeistus

Työssä tutkitaan pienitehoisen monimoottoritaajuusmuuttajakäytön yhteismuotovirtojen aiheuttamia ongelmia. Työn tavoitteena on muodostaa monimoottorikäyttöjen toteuttajille opastus, jonka avulla sähkökäytön suunnittelun alkuvaiheessa voidaan ottaa huomioon monimoottorikäytön erikoistarpeita. Työn teoriaosuudessa esitellään taajuusmuuttajan vaihtosuuntaustekniikan aiheuttamia ongelmia, moottorikaapelin mallintamista ja vertaillaan ongelmien ratkaisukeinoja pienitehoisten taajuusmuuttajien näkökulmasta. Yhteismuotoista virtaa tutkitaan mittaamalla rakennetulla monimoottorikäytöllä sekä simuloimalla vaihdellen kaapelin pituutta ja moottorien lukumäärää. Mittaamalla vertaillaan myös virran suodatusratkaisuja. Muodostettu simulointimalli on hyvin yksinkertainen ja luotettavammat virta-arvot saadaankin mittauksella. Kuitenkin työn perusteella voidaan antaa ohjeita monimoottorikäytön suunnittelulle. Esimerkiksi jokaiselle moottorille täytyy suunnitella erillinen suojaus ja taajuusmuuttaja on syytä mitoittaa todellisen kuormavirran mukaan. Yhteismuotoisen virran aiheuttamat ongelmat ovat sitä pienempiä, mitä vähemmän moottoreita on rinnakkain ja mitä lyhyempi moottorikaapelien yhteispituus on.

Active inverter output filtering methods

Frequency converters are widely used in the industry to enable better controllability and efficiency of variable speed AC motor drives. Despite these advantages, certain challenges concerning the inverter and motor interfacing have been present for decades. As insulated gate bipolar transistors entered the market, the inverter output voltage transition rate significantly increased compared with their predecessors. Inverters operate based on pulse width modulation of the output voltage, and the steep voltage edge fed by the inverter produces a motor terminal overvoltage. The overvoltage causes extra stress to the motor insulation, which may lead to a prematuremotor failure. The overvoltage is not generated by the inverter alone, but also by the sum effect of the motor cable length and the impedance mismatch between the cable and the motor. Many solutions have been shown to limit the overvoltage, and the mainstream products focus on passive filters. This doctoral thesis studies an alternative methodology for motor overvoltage reduction. The focus is on minimization of the passive filter dimensions, physical and electrical, or better yet, on operation without any filter. This is achieved by additional inverter control and modulation. The studied methods are implemented on different inverter topologies, varying in nominal voltage and current.For two-level inverters, the studied method is termed active du/dt. It consists of a small output LC filter, which is controlled by an independent modulator. The overvoltage is limited by a reduced voltage transition rate. For multilevel inverters, an overvoltage mitigation method operating without a passive filter, called edge modulation, is implemented. The method uses the capability of the inverter to produce two switching operations in the same direction to cancel the oscillating voltages of opposite phases. For parallel inverters, two methods are studied. They are both intended for two-level inverters, but the first uses individual motor cables from each inverter while the other topology applies output inductors. The overvoltage is reduced by interleaving the switching operations to produce a similar oscillation accumulation as with the edge modulation. The implementation of these methods is discussed in detail, and the necessary modifications to the control system of the inverter are presented. Each method is experimentally verified by operating industrial frequency converters with the modified control. All the methods are found feasible, and they provide sufficient overvoltage protection. The limitations and challenges brought about by the methods are discussed.

Detection of sub-8-nm movements of kinesin by high-resolution optical-trap microscopy.

Kinesin is a molecular motor that transports organelles along microtubules. This enzyme has two identical 7-nm-long motor domains, which it uses to move between consecutive tubulin binding sites spaced 8 nm apart along a microtubular protofilament. The molecular mechanism of this movement, which remains to be elucidated, may be common to all families of motor proteins. In this study, a high-resolution optical-trap microscope was used to measure directly the magnitude of abrupt displacements produced by a single kinesin molecule transporting a microscopic bead. The distribution of magnitudes reveals that kinesin not only undergoes discrete 8-nm movements, in agreement with previous work [Svoboda, K., Schmidt, C. F., Schnapp, B. J. & Block, S.M. (1993) Nature (London) 365, 721-727], but also frequently exhibits smaller movements of about 5 nm. A possible explanation for these unexpected smaller movements is that kinesin's movement from one dimer to the next along a protofilament involves at least two distinct events in the mechanical cycle.

Long-term Spinal Ventral Root Reimplantation, But Not Bone Marrow Mononuclear Cell Treatment, Positively Influences Ultrastructural Synapse Recovery And Motor Axonal Regrowth.

We recently proposed a new surgical approach to treat ventral root avulsion, resulting in motoneuron protection. The present work combined such a surgical approach with bone marrow mononuclear cells (MC) therapy. Therefore, MC were added to the site of reimplantation. Female Lewis rats (seven weeks old) were subjected to unilateral ventral root avulsion (VRA) at L4, L5 and L6 levels and divided into the following groups (n = 5 for each group): Avulsion, sealant reimplanted roots and sealant reimplanted roots plus MC. After four weeks and 12 weeks post-surgery, the lumbar intumescences were processed by transmission electron microscopy, to analyze synaptic inputs to the repaired α motoneurons. Also, the ipsi and contralateral sciatic nerves were processed for axon counting and morphometry. The ultrastructural results indicated a significant preservation of inhibitory pre-synaptic boutons in the groups repaired with sealant alone and associated with MC therapy. Moreover, the average number of axons was higher in treated groups when compared to avulsion only. Complementary to the fiber counting, the morphometric analysis of axonal diameter and g ratio demonstrated that root reimplantation improved the motor component recovery. In conclusion, the data herein demonstrate that root reimplantation at the lesion site may be considered a therapeutic approach, following proximal lesions in the interface of central nervous system (CNS) and peripheral nervous system (PNS), and that MC therapy does not further improve the regenerative recovery, up to 12 weeks post lesion.

ASSOCIATION OF A MYOSIN MOTOR WITH MEMBRANE-BOUNDED PIGMENT GRANULES IN FRESHWATER SHRIMP CHROMATOPHORES: EVIDENCE FROM THE NITELLA ACTIN-CABLE ASSAY

We have adapted an actin-mosin motility assay to examine the interactions in vitro between actin cables isolated from the giant internodal cells of the freshwater alga, Nitella, and pigment granules extracted from red ovarian chromatophores of the freshwater palaemonid shrimp, Macrobrachium olfersi. The chromatophore pigment mass consists of large (0.5-1.0-mu m diameter) membrane-bounded granules, and small (140-nm diameter), a membranous granules, both structurally continuous with the abundant smooth endoplasmic reticulum. Our previous immunocytochemical studies show a myosin motor to be stably associated with the pigment mass; however, to which granule type or membrane the myosin motor is attached is unclear. Here, we show that sodium vanadate, a myosin ATPase inhibitor, markedly increases the affinity of isolated, large, membrane-bounded granules for Nitella actin cables to which they become permanently attached. This interaction does not occur in granule preparations containing ATP with uninhibited, active myosin without vanadate. We propose that a stable state of elevated affinity is established between the granule-located myosin motor and the Nitella actin cables, resulting from a vanadate-inhibited acto-myosin-ADP complex. This finding provides further evidence for a myosin motor positioned on the surface of the membrane-bounded pigment granules in shrimp ovarian chromatophores.

Role of myosin V in actin cable organization in fission yeast

Functional specialization is tightly linked to the ability of eukaryotic cells to acquire a particular shape. Cell morphogenesis, in turn, relies on the capacity to establish and maintain cell "polarity", which is achieved by orienting the trafficking of signaling molecules and organelles towards specific cellular locations and/or membrane domains. The "oriented" transport is based upon cytoskeletal polymers, microtubules and actin filaments, which serve as tracks for molecular motors. These latter generate motion that is translated either into pulling forces or directed transport. Fission yeast, a rod-like unicellular eukaryote, shapes itself by restricting growth at cell tips through the concerted activity of microtubules and actin cables. Microtubules, which assemble into 2-6 bundles and run parallel to the long axis of the cell, serve to orient growth to the tips. Growth is supported by the actin cytoskeleton, which provides tracks, the cables, for motor-based transport of secretory vesicles. The molecular motors, which bind cargos and deliver them to the tips along cables, are also known as type V myosins (hereafter indicated as myosin V). How the bundles of parallel actin filaments, i.e. the cables, extend from the tips through the cell and whether they serve any other purpose, besides providing tracks, is poorly understood. It is also unclear how the crosstalk between the two cytoskeletal systems is achieved. These are the basic questions I addressed during my PhD. The first part of the thesis work (Chapter two) suggests that the sole function of actin cables in polarized growth is to serve as tracks for motors. The data indicate that cells may have evolved two cytoskeletal systems to provide robustness to the polarization process but in principle a unique cytoskeleton might have been able to direct and support polarized growth. How actin cables are organized within the cell to optimize cargo transport is addressed later on (Chapter three). The major finding, based on the actin cable defect of cells lacking myosin Vs, is that actin filaments self-organize through the activity of the transport motors. In fact, by delivering cargos to cell tips and exerting physical pulling forces on actin filaments, Myosin Vs contribute not only to polarize cargo transport but also actin tracks. Among the cargos transported by Myosin V, which may be relevant to its function in organizing cables, there is likely the endoplasmic reticulum (ER). Actin cables, which run parallel to cortical ER, may serve as tracks for Myosin V. Myosin V-driven displacement, in turn, may account for the dynamic expansion and organization of ER during polarized growth as suggested in Chapter four. The last part of the work (Chapter five) highlights the existence of a crosstalk between actin and microtubules. In absence of myosin V, indeed, microtubules contribute to actin cable organization, likely playing a scaffolding/tethering function. Whether or not the kinesin 1, Klp3, plays any role in such process has to be demonstrated. In conclusion the work proposes a novel role for myosin Vs in actin organization, besides its transport function, and provides molecular tools to further dissect the role of this type of myosin in fission yeast. - La spécialisation fonctionnelle est étroitement connectée à la capacité des cellules eucaryotes d'acquérir une forme particulière. La morphogenèse cellulaire à son tour, est basée sur la capacité d'établir et de maintenir la polarité cellulaire, polarité réalisée en orientant le trafic des molécules signales et des organelles vers des zones cellulaires spécifiques. Ce transport directionnel dépend des polymères du cytosquelette, microtubules et microfilaments, qui servent comme des voies pour les moteurs moléculaires. Ces derniers engendrent du mouvement, traduit soit en force de traction soit en transport directionnel. La levure fissipare, un eucaryote unicellulaire en forme de bâtonnet, acquière sa forme en limitant sa croissance aux extrémités par l'action concertée des microtubules et de l'actine. Les microtubules, qui s'assemblent de façon antiparallèle et parcourent la cellule parallèlement à l'axe longitudinal, servent à orienter la croissance aux extrémités. Cette croissance est permise par le cytosquelette d'actine, fournissant des voies, les câbles, pour le transport actif des vésicules de sécrétion. Les moteurs moléculaires, responsables de ce transport actif sont aussi appelés myosines de type V (par la suite appelés myosines V). La manière dont ces câbles s'étendent depuis l'extrémité jusqu'à l'intérieur de la cellule est peu connue. De plus, on ignore également si ces câbles présentent une fonction autre que le transport. L'interaction entre les deux cytosquelettes est également obscure. Ce sont ces questions de base auxquelles j'ai tenté de répondre lors de ma thèse. La première partie de cette thèse (chapitre II) suggère que les câbles d'actine, pendant la croissance polarisée, fonctionnent uniquement comme des voies pour les moteurs moléculaires. Les données indiqueraient que les cellules ont fait évoluer deux systèmes de cytosquelette pour assurer plus de robustesse au processus de polarisation, bien que, comme nous le verrons, un système unique est suffisant. Au chapitre III, nous verrons comment les câbles d'actine sont organisés à l'intérieur de la cellule afin d'optimiser le transport des cargo. La découverte majeure, réalisée en observant des cellules dont la myosine V fait défaut, est que ces filaments d'actine s'auto organisent grâce au passage des moteurs moléculaires le long de ces voies. En réalité, en délivrant les cargos aux extrémités de la cellule et en exerçant des forces de traction sur les câbles, les myosines V contribuent non seulement à polariser le transport mais également à polariser les voies elles mêmes. Nous verrons également au chapitre IV, que parmi les cargos importants pour l'organisation des câbles, il y aurait le réticulum endoplasmique (RE). En effet, les câbles d'actine, qui s'étalent parallèlement au RE cortical, pourraient servir comme voie pour la myosine V. Cette dernière en retour pourrait être responsable de l'expansion dynamique et de l'organisation du RE pendant la croissance polarisée.

Long-Term Spinal Ventral Root Reimplantation, but not Bone Marrow Mononuclear Cell Treatment, Positively Influences Ultrastructural Synapse Recovery and Motor Axonal Regrowth

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)