Bases de données informatisées, inventaire légal et inventaire documentaire

La particularité des musées d’Histoire Naturelle est de conserver dans leurs fonds, de grandes quantités d’ob­jets de nature différente. Ces collections sont gé­né­ra­lement organisées par discipline. Jusqu’au début des années 1980, la gestion des collections reposait essen­tiellement sur les connaissances et la mémoire des per­s­onnels des établissements. C’est à ce moment que l’in­formatique fit son apparition dans les musées. Au muséum d’Histoire Naturelle de Toulouse, cette ar­rivée s’es..

Contact-controlled amoeboid motility induces dynamic cell trapping in 3D-microstructured surfaces.

On flat substrates, several cell types exhibit amoeboid migration, which is characterized by restless stochastic successions of pseudopod protrusions. The orientation and frequency of new membrane protrusions characterize efficient search modes, which can respond to external chemical stimuli as observed during chemotaxis in amoebae. To quantify the influence of mechanical stimuli induced by surface topography on the migration modes of the amoeboid model organism Dictyostelium discoideum, we apply high resolution motion analysis in microfabricated pillar arrays of defined density and geometry. Cell motion is analyzed by a two-state motility-model, distinguishing directed cellular runs from phases of isotropic migration that are characterized by randomly oriented cellular protrusions. Cells lacking myosin II or cells deprived of microtubules show significantly different behavior concerning migration velocities and migrational angle distribution, without pronounced attraction to pillars. We conclude that microtubules enhance cellular ability to react with external 3D structures. Our experiments on wild-type cells show that the switching from randomly formed pseudopods to a stabilized leading pseudopod is triggered by contact with surface structures. These alternating processes guide cells according to the available surface in their 3D environment, which we observed dynamically and in steady-state situations. As a consequence, cells perform "home-runs" in low-density pillar arrays, crawling from pillar to pillar, with a characteristic dwell time of 75 s. At the boundary between a flat surface and a 3D structured substrate, cells preferentially localize in contact with micropillars, due to the additionally available surface in the microstructured arrays. Such responses of cell motility to microstructures might open new possibilities for cell sorting in surface structured arrays

Kinneyia: a flow-induced anisotropic fossil pattern from ancient microbial mats

Kinneyia is the commonly used term to describe a class of trace fossil that is strongly associated with microbial mats. The appearance of Kinneyia (or wrinkle structures) in the fossil record has recently led to a number of possible mechanisms being proposed to explain its formation. Here, we outline, and critically compare, three of these models, involving formation of the characteristic ripple structures (i) in mats over liquefied sediment, (ii) by oscillatory flow of microbial aggregates, and (iii) by a Kelvin–Helmholtz instability of the mat surface. Of these models, our study shows that the hydrodynamic instability compares most favorably with the corresponding structures in the fossil record. Implications for the conditions under which the fossils formed are then further discussed

Persistent Cellular Motion Control and Trapping Using Mechanotactic Signaling

Chemotactic signaling and the associated directed cell migration have been extensively studied owing to their importance in emergent processes of cellular aggregation. In contrast, mechanotactic signaling has been relatively overlooked despite its potential for unique ways to artificially signal cells with the aim to effectively gain control over their motile behavior. The possibility of mimicking cellular mechanotactic signals offers a fascinating novel strategy to achieve targeted cell delivery for in vitro tissue growth if proven to be effective with mammalian cells. Using (i) optimal level of extracellular calcium ([Ca2[superscript +] ][subscript ext] = 3 mM) we found, (ii) controllable fluid shear stress of low magnitude (σ < 0.5 Pa), and (iii) the ability to swiftly reverse flow direction (within one second), we are able to successfully signal Dictyostelium discoideum amoebae and trigger migratory responses with heretofore unreported control and precision. Specifically, we are able to systematically determine the mechanical input signal required to achieve any predetermined sequences of steps including straightforward motion, reversal and trapping. The mechanotactic cellular trapping is achieved for the first time and is associated with a stalling frequency of 0.06 ~ 0.1 Hz for a reversing direction mechanostimulus, above which the cells are effectively trapped while maintaining a high level of directional sensing. The value of this frequency is very close to the stalling frequency recently reported for chemotactic cell trapping [Meier B, et al. (2011) Proc Natl Acad Sci USA 108:11417–11422], suggesting that the limiting factor may be the slowness of the internal chemically-based motility apparatus.SUTD-MIT International Design Centre (Grant IDG31400104

Melatonin Promotes Oligodendroglial Maturation of Injured White Matter in Neonatal Rats

OBJECTIVE:To investigate the effects of melatonin treatment in a rat model of white matter damage (WMD) in the developing brain. Additionally, we aim to delineate the cellular mechanisms of melatonin effect on the oligodendroglial cell lineage. METHODS:A unilateral ligation of the uterine artery in pregnant rat at the embryonic day 17 induces fetal hypoxia and subsequent growth restriction (GR) in neonatal pups. GR and control pups received a daily intra-peritoneal injection of melatonin from birth to post-natal day (P) 3. RESULTS:Melatonin administration was associated with a dramatic decrease in microglial activation and astroglial reaction compared to untreated GR pups. At P14, melatonin prevented white matter myelination defects with an increased number of mature oligodendrocytes (APC-immunoreactive) in treated GR pups. Conversely, melatonin was not found to be associated with an increased density of total oligodendrocytes (Olig2-immunoreactive), suggesting that melatonin is able to promote oligodendrocyte maturation but not proliferation. These effects appear to be melatonin-receptor dependent and were reproduced in vitro. INTERPRETATION:These data suggest that melatonin has a strong protective effect on developing damaged white matter through decreased microglial activation and oligodendroglial maturation leading to a normalization of the myelination process. Consequently, melatonin should be a considered as an effective neuroprotective candidate not only in perinatal brain damage but also in inflammatory and demyelinating diseases observed in adults

A Comparison of Mathematical Models for Polarization of Single Eukaryotic Cells in Response to Guided Cues

Polarization, a primary step in the response of an individual eukaryotic cell to a spatial stimulus, has attracted numerous theoretical treatments complementing experimental studies in a variety of cell types. While the phenomenon itself is universal, details differ across cell types, and across classes of models that have been proposed. Most models address how symmetry breaking leads to polarization, some in abstract settings, others based on specific biochemistry. Here, we compare polarization in response to a stimulus (e.g., a chemoattractant) in cells typically used in experiments (yeast, amoebae, leukocytes, keratocytes, fibroblasts, and neurons), and, in parallel, responses of several prototypical models to typical stimulation protocols. We find that the diversity of cell behaviors is reflected by a diversity of models, and that some, but not all models, can account for amplification of stimulus, maintenance of polarity, adaptation, sensitivity to new signals, and robustness

Application de contraintes sur des systèmes complexes artificiels ou vivants : dégonflement de liposomes fonctionnalisés et réorganisation mécanosensible du cytosquelette de cellules Dictyostelium.

Une bourse de l'ARC (Association pour la Recherche sur le Cancer) et deux bourses EMBO (European Molecular Biology Organization) ont été obtenues pour achever ce travail de thèse.During this work, 2 approaches have been explored. First, I quantified the osmotic deswelling of liposomes filled with an agarose gel. The production of such artificial systems aims at mimicking cell behavior under the same constraints. Particularly, I observed that these functionnalized liposomes with a gel concentration between 0.07 and 0.18% w/w adopted crenated morphologies when strongly deswelled. These original shapes look like the ones of echinocytes sometimes seen with red blood cells. The gel is responsible for these shapes, does not affect deswelling kinetics but its elastic pressure stops more rapidly the osmotic deswelling compared to aqueous liposomes. This brings evidence for a water retention effect. In a second approach, I studied the effect of hydrodynamical constraints on Dictyostelium amoebae adhering to a substrate. I quantified the mechanosensitive reorganization of the cytoskeleton of these living cells. To get relocalization kinetics of major cytoskeleton proteins in response to flow forces, I labeled actin and myosin-II with fluorescent proteins (GFP/mRFP) and designed a flow chamber enabling to rapidly change the flow direction. I showed that cells orient against flow forces and after a flow reversal reorient against new forces by inverting their polarity: first, actin depolymerizes, then actin rich protrusions are emitted against new mechanical forces and 15 sec later, the rear edge retracts with a myo-II crescent. Moreover, the actin-myosin contractility is dispensable to sense forces. Similar experiments by inverting the direction of a chemotactic gradient show that this cell reorientation process is not specific of flow experiments. This work proves the existence of a rapid inhibiting signal (leading to actin depolymerization) which is not taken into account in current models of chemotaxis. Finally, the visualization tools that I developped enable to study the role of proteins and cellular structures in mechanotransduction.Durant ce travail, deux approches ont été explorées. Dans la première, j'ai quantifié le dégonflement osmotique de liposomes remplis d'un gel d'agarose. La fabrication de tels systèmes reconstitués vise à permettre de mimer le comportement de cellules soumises aux mêmes contraintes. En particulier, j'ai observé que ces liposomes fonctionnalisés acquièrent des morphologies crénelées lors de leur dégonflement pour une concentration du gel comprise entre 0.07 et 0.18 % en masse. Ces formes originales ressemblent à celles d'échinocytes parfois prises par les globules rouges. Le gel est responsable de l'apparition de ces formes, ne modifie pas les cinétiques de dégonflement mais sa pression élastique arrête précocement le dégonflement comparativement aux liposomes aqueux, mettant en évidence un phénomène de rétention d'eau. Dans la deuxième approche, j'ai étudié l'effet de contraintes hydrodynamiques sur des amibes Dictyostelium adhérentes à un substrat et ai quantifié la réorganisation mécanosensible du cytosquelette de ces cellules vivantes. Pour obtenir les cinétiques de relocalisation de protéines majeures du cytosquelette en réponse aux forces d'un flux, j'ai marqué l'actine et la myosine-II avec des protéines fluorescentes et ai fabriqué une chambre à flux permettant de changer rapidement la direction du flux. J'ai montré que les cellules s'orientent contre les forces du flux et se réorientent contre en inversant leur polarité après une inversion du flux : d'abord l'actine dépolymérise puis des protrusions sont émises contre les nouvelles forces mécaniques, et 15 sec plus tard, l'arrière rétracte en utilisant la myo-II. De plus, la contractilité du système actine-myosine n'est pas nécessaire pour sentir les forces. Des expériences similaires en inversant la direction d'un gradient de chimioattractants montrent que ce processus de réorientation cellulaire n'est pas spécifique d'expériences sous flux. Ce travail met en évidence l'existence d'un signal inhibiteur rapide menant à la dépolymérisation de l'actine, signal qui n'est pas pris en compte dans les modèles actuels expliquant la réponse chimiotactique. Enfin, les outils de visualisation que j'ai développés permettent d'étudier le rôle de protéines et de structures cellulaires dans la mécanotransduction

Application de contraintes sur des systèmes complexes artificiels ou vivants (dégonflement de liposomes fonctionnalisés et réorganisation mécanosensible du cytosquelette de cellules dictyostelium)

Dans la première approche de ce travail, j'ai quantifié le dégonflement osmotique de liposomes remplis d'un gel d'agarose. La fabrication de tels systèmes reconstitués vise à permettre de mimer le comportement de cellules soumises aux mêmes contraintes. En particulier, j'ai observé que ces liposomes fonctionnalisés acquièrent des morphologies crénelées lors de leur dégonflement pour une concentration du gel comprise entre 0.07 et 0.18 % en masse. Ces formes originales ressemblent à celles d'échinocytes parfois prises par les globules rouges. Le gel est responsable de l'apparition de ces formes, ne modifie pas les cinétiques de dégonflement mais sa pression élastique arrête précocement le dégonflement comparativement aux liposomes aqueux, mettant en évidence un phénomène de rétention d'eau. Dans la deuxième approche, j'ai étudié l'effet de contraintes hydrodynamiques sur des amibes Dictyostelium adhérentes à un substrat et ai quantifié la réorganisation mécanosensible du cytosquelette de ces cellules vivantes. Pour obtenir les cinétiques de relocalisation de protéines majeures du cytosquelette en réponse aux forces d'un flux, j'ai marqué l'actine et la myosine-II avec des protéines fluorescentes et ai fabriqué une chambre à flux permettant de changer rapidement la direction du flux. Les cellules étudiées s'orientent contre les forces du flux et se réorientent contre en inversant leur polarité après une inversion du flux: d'abord l'actine dépolymérise puis des protrusions sont émises contre les nouvelles forces mécaniques, et 15 sec plus tard, l'arrière rétracte en utilisant la myo-II. La contractilité du système actine-myosine n'est pas nécessaire pour sentir les forces.GRENOBLE1-BU Sciences (384212103) / SudocSudocFranceF

Giant Lipid Vesicles Filled with a Gel: Shape Instability Induced by Osmotic Shrinkage

We report the properties of giant lipid vesicles enclosing an agarose gel. In this system, the lipid bilayer retains some basic properties of biological membranes and the internal fluid exhibits viscoelastic properties, thus permitting us to address the question of the deformation of a cell membrane in relation to the mechanical properties of its cytoskeleton. The agarose gel (concentration c(0gel) = 0.07%, 0.18%, 0.36%, and 1% w/w), likely not anchored to the membrane, confers to the internal volume elastic moduli in the range of 10–10(4) Pa. Shapes and kinetics of de-swelling of gel-filled and aqueous solution-filled vesicles are compared upon either a progressive or a fast osmotic shrinkage. Both systems exhibit similar kinetics. Shapes of solution-filled vesicles are well described using the area difference elasticity model, whereas gel-filled vesicles present original patterns: facets, bumps, spikes (c(0gel) < 0.36%), or wrinkles (c(0gel) ≥ 0.36%). These shapes partially vanish upon re-swelling, and some of them are reminiscent of echinocytic shapes of erythrocytes. Their characteristic size (microns) decreases upon increasing c(0gel). A possible origin of these patterns, relying on the formation of a dense impermeable gel layer at the vesicle surface and associated with a transition toward a collapsed gel phase, is advanced