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Neuronal polarization shapes the targeting and signaling of G-protein coupled receptors (GPCRs) : type-1 cannabinoid receptors and 5-HT1B serotonin receptors show highly contrasted trafficking and signaling patterns in axons and dendrites / La polarisation neuronale façonne l’adressage et la signalisation des récepteurs couplés aux protéines G (RCPG) : le récepteur canabinoïque de type 1 et le récepteur sérotoninergique 5-HT1B ont un trafic et une signalisation différents dans les axones et les dendritesLadarré, Delphine 03 October 2014 (has links)
L’architecture polarisée des neurones est mise en place est maintenue grâce à un adressage hautement contrôlé de protéines vers l’axone ou vers le compartiment somatodendritique. Parmi ces protéines, les récepteurs aux protéines G (RCPG) neuronaux sont des cibles pharmacologiques clés. Cependant, leur pharmacologie est généralement étudiée dans des lignées cellulaires non polarisées et les résultats obtenus dans ces systèmes ne caractérisent pas correctement les effets physiologiques de l’activation des RCPG présents dans le cerveau. Par conséquent, un des principaux sujets de recherche de notre équipe est de comprendre comment la polarité neuronale influe sur la pharmacologie des RCPG, en étudiant l’un des RCPG les plus abondants dans le cerveau : le récepteur cannabinoïque de type-1 (CB1R). Les études précédentes de notre groupe ont suggéré que CB1R acquiert une polarisation axonale grâce à un adressage transcytotique : après leur synthèse, ces récepteurs apparaissent sur la membrane plasmique somatodendritique d’où ils sont rapidement enlevés par endocytose constitutive puis adressés à la membrane plasmique axonale où ils s’accumulent du fait d’une endocytose réduite. Au début de ma thèse, nous avons directement mesuré cette endocytose différentielle et le transport transcytotique de CB1R en utilisant des neurones de rats mis en culture dans des dispositifs microfluidiques. De plus, nous avons montré que des traitements pharmacologiques prolongés peuvent fortement changer la distribution de RCPG à la surface neuronale. Ces résultats démontrent que l’équilibre endocytotique dépendant du compartiment neuronal, qui est contrôlable pharmacologiquement, est important pour la distribution des RCPG neuronaux. Dans une seconde partie, nous avons étudié si le trafic différentiel de CB1R entre axones et dendrites est corrélé avec une pharmacologie différentielle. CB1R est majoritairement couplé à des protéines de type Gi/o et est connu pour inhiber la production d’AMPc. Nous avons donc développé l’imagerie par Föster Resonance Energy Transfer (FRET) appliqué aux cultures de neurones d’hippocampe de rats afin de mesurer la modulation de la voie de signalisation AMPc/PKA en aval de CB1R endogènes dans l’ensemble des compartiments neuronaux : somata, dendrites, mais aussi dans les axones matures très fins. Nos résultats montrent que CB1R possède une pharmacologie différente entre les dendrites et les axones. Notamment, son activation conduit à une diminution plus forte de l’activité basale de la PKA dans les axones comparé aux dendrites, lié au plus grand nombre de récepteurs présents sur la membrane de ce compartiment. De plus, nous démontrons que, contrairement aux récepteurs axonaux, les CB1R somatodendritiques inhibent constitutivement la voie AMPc/PKA. Cette différence est due à la distribution polarisée de la DAGLipase, l’enzyme synthétisant l’endocannabinoïde principal, le 2-arachidonoyglycerol (2-AG). De plus, l’inhibition pharmacologique de la DAGL modifie l’efficacité de plusieurs agonistes de CB1R dans le compartiment somatodendritique mais pas dans l’axone. Cet effet pourrait être dû à une modulation allostérique. Dans une troisième partie, nous avons étudié si les résultats ci-dessus peuvent être généralisés à d’autres RCPG. Etant donné que l’adressage axonal et la pharmacologie in vitro des récepteurs sérotoninergiques 5-HT1B montrent de fortes similitudes avec ceux de CB1R, nous avons étudié la pharmacologie de ces récepteurs en utilisant la technique de FRET développée précédemment. De façon similaire, nous avons trouvé une pharmacologie différentielle entre l’axone et les dendrites. / Polarized neuronal architecture is achieved and maintained mainly through highly controlled targeting of proteins to axons versus to the somatodendritic compartment. Among these proteins, neuronal G protein coupled receptors (GPCRs) are key therapeutic targets. However, their pharmacology is generally studied in non-polarized cell lines, and results obtained in such systems likely do not fully characterize the physiological effects of brain GPCR activation. Therefore, a main research subject of our group is to understand how neuronal polarity influences GPCR pharmacology, by studying one of the most abundant GPCR in the brain: the type-1 cannabinoid receptor (CB1R). Previous studies of the group suggested that CB1Rs achieve axonal polarization through transcytotic targeting: after their synthesis, these receptors appear on the somatodendritic plasma membrane from where they are removed rapidly by constitutive endocytosis and then targeted to the axonal plasma membrane where they accumulate due to relatively reduced endocytosis rate. At the beginning of my PhD project we directly demonstrated this differential endocytosis and transcytotic transport of CB1Rs by using cultured neurons in microfluidic devices. Moreover, we showed that chronic pharmacological treatments may strongly change neuronal GPCR distribution on the neuronal surface. These results demonstrate that subdomain-dependent steady-state endocytosis, which is pharmacologically controllable, is important for GPCR distribution in neurons. In a second part, we asked if differential traffic of CB1Rs between axons and dendrites is correlated with differential pharmacology. CB1R is predominantly coupled to Gi/o proteins and is known to inhibit cAMP production. Thus, we developed live Föster Resonance Energy Transfer (FRET) imaging in cultured hippocampal neurons in order to measure basal cAMP/PKA pathway modulation downstream of endogenous CB1Rs in all neuronal compartments: in somata, in dendrites but also in the very thin mature axons. Our results show that CB1R displays differential pharmacology between axon and dendrites. Notably, its activation leads to a stronger decrease of PKA activity in axons compared to dendrites, due to increased number of membrane receptors in this compartment. Moreover, we demonstrate that somatodendritic CB1Rs constitutively inhibit cAMP/PKA pathway, while axonal receptors do not. This difference is due to polarized distribution of DAGLipase, the enzyme that synthesizes the major endocannabinoid 2-arachidonoylglycerol (2-AG). Moreover, blocking DAGL by pharmacological treatment modifies somatodendritic, but not axonal effects of several CB1R agonists, possibly through allosteric action. In a third part, we asked if the above results may be generalized to other GPCRs. Because the axonal targeting and in vitro pharmacology of 5-HT1B serotonin receptors demonstrate strong similarities with CB1Rs, we studied their neuronal pharmacology by using the previously developed FRET technique. We found similar differential responses to pharmacological treatments between axon and dendrites. In a fourth part, we investigated the role of the threonine 210 (T210) residue in the constitutive activity of neuronal CB1R. We showed that the hypoactive mutant T210A-CB1R do not constitutively recruit signaling pathways even in somatodendritic compartment, where 2-AG is present. This result demonstrates that T210 is necessary for constitutive CB1R activation by 2-AG.Finally, previous results of our group demonstrated the involvement of CB1R in neuronal development. Notably, CB1R activation was shown to have an overall inhibitory effect on the development of polarized neuronal morphology. We established a bibliographic review on this subject. The published literature data suggest that not only neuronal polarization influences both CB1R traffic and pharmacology but CB1Rs also contribute to the achievement of neuronal polarization. (...)
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Micropatterning of hippocampal neurons : characterization and implications for studying synaptogenesisBelkaid, Wiam, 1983- January 2008 (has links)
During development of the nervous system, formation of specific connections between nerve cells depends on the stability of growing axons to reach appropriate target cells and form synapses. In culture, hippocampal neurons form numerous synapses by developing axonal and dendritic extensions. To elucidate principles of neuronal signaling and network establishment, creation of neuronal networks in which connectivity and pathways can be experimentally controlled is of great interest. In the present study we used a microcontact printing technique to control and study neurite outgrowth of hippocampal neurons in vitro. My preliminary results show that hippocampal neurons follow the microcontact printed pattern of poly-D-lysine (PDL). In doing so, neurons retain their morphology with normal subcellular distribution of various cell adhesion and synaptic molecules. However, the distribution of various axonal or dendrite components is altered. Hence we have developed a system in which isolated axons and dendrites align with inputs from very few neurons. With this technique we intend to study axon-dendrite communications on a spatially restricted and defined substrate.
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Microstructural features and mechanical behaviour of lead free solders for microelectronic packagingGong, Jicheng January 2007 (has links)
The demands for high density, fine pitch interconnections in electronics systems has seen solder-based approaches for such interconnections miniaturized to the scale of tens of micro meters. At such a small scale, such 'micro joints' may contain only one or a few grains and the resultant mechanical behaviour may not be that for a polycrystalline aggregate, but rather for a single crystal. Since the ~-Sn matrix of SnAgCu solder has a contracted body-centred tetragonal (BCT) structure, such a solder grain is expected to demonstrate a considerably anisotropic behaviour. In such cases the reliability of a Phfree solder is strongly dependent on the local microstructural features, such as the size and orientation of the grains. This thesis presents the investigation of the evolution of microstructure within a joint or at the interface and, the influence of such microstructural features on the meso-scale mechanical behaviour of the Ph-free solder. It includes Evolution of the interface between a molten solder and the Cu substrate To form a joint, the solder alloy is heated and molten, wetting a solid under-bump metallization. After solidification, layers of brittle intermetallic compounds (IMCs) are formed at the interface. In this project, facilities were set up to obtain interfacial reactants at an arbitrary moment of the liquid/solid reaction. Formation and evolution ~ during reflow of SnCu IMCs at the interface between the molten SnAgCu alloy and the Cu UBM was captured and presented for the first time. Formation of phases and IMCs with the body of a liquid SnAgCu solder during solidification The formation behaviour of basic components for a SnAgCu grain (including Sn dendrites, AIDSn and Cu6Sns IMCs) during solidification was investigated. Relationships between the growth behaviour of these components and their internal lattice orientation were studied. The characteristic growth and coupling of AIDSn IMCs and the Sn matrix to form eutectics has been elaborated and presented in this study for - 1- the first time. Based on the results, the forming process of a eutectic SnAgCu grain under the non-equilibrioum solidification condition was illustrated; and major factors that determine the lattice-orientation, size and substructure of the grain were discussed. Meso- and Micro- scale mechanical behaviour of a SnAgCu solder joint To study the size effect on the microstructure, and subsequently, the meso-scale mechanical behaviour, solder joints were manufactured with varying geometries. Shearing tests were performed on these meso-scale joints. The results first demonstrated that the anisotropic characteristics of a SnAgCu grain play an important role in the mechanical behaviour of both a meso-scale solder joint and the adjacent interfacial IMCs. To further investigate the micro-scale deformation and damage mechanisms, micro-mechanical tests were preformed within a SnAgCu grain. Constitutive equations for a SnAgCu grain Based on the experimental results, a crystal model was established to describe the local microstructure-dependent mechanical behaviour. The constitutive equation was implemented by means of the finite element approach, and applied in solder joints of a Flip Chip (FC) package by a multi-scale method. To describe the crystal behaviour at the higher temperature, the model was improved to account for deformations due to vacancy diffusion and thermal expansion. This model was integrated by an implicit approach, and implemented in a full three dimension (3D) finite element (FE) model.
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Dissection of protein-protein interactions that regulate dendritic growth and synaptic transmission /Pradhan, Anuradha January 2005 (has links) (PDF)
Thesis (Ph. D.)--University of Oklahoma. / Bibliography: leaves 117-135.
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Análise microestrutural de revestimentos usados no reparo de turbinas hidráulicas danificadas pela cavitaçãoMusardo, Gustavo Borges [UNESP] 27 October 2006 (has links) (PDF)
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musardo_gb_me_ilha.pdf: 3996445 bytes, checksum: ce4f6124da4b6648c41885a10eb3e368 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / No Brasil, a maior parte da energia elétrica disponível é gerada por usinas hidrelétricas as quais contam com grande número de turbinas hidráulicas. Durante sua operação estas turbinas sofrem sérios danos provenientes tanto de natureza mecânica como também hidráulica. Fenômenos de perda de massa e também trincas e rachaduras são alguns dos principais problemas que ocorrem devido a um fenômeno chamado de erosão cavitacional, que comumente é recuperado por soldagem. Sendo assim, grandes níveis de resistência à cavitação vêm sendo obtidos com a deposição de aços inoxidáveis com a presença de cobalto pelo processo de soldagem a arco com proteção gasosa. Neste trabalho, seis amostras feitas de aço-carbono ASTM A36, as quais foram extraídas de retalhos de turbinas, foram usadas como metal de base para as amostras. Usando um processo a arco elétrico com proteção gasosa (GMAW) em posição plana, duas camadas de aço AWS E70-S6 (1,2 mm de diâmetro) foi depositada em todas as amostras, e, somente em três das amostras foram depositadas duas camadas de amanteigamento com o arame de aço inoxidável AWS E309-T1 (1,6 mm de diâmetro) usando o mesmo processo, onde foi utilizado como proteção gasosa uma mistura de 75% de dióxido de carbono e 25% de argônio. A energia de soldagem nominal nestes casos foi de 0,5 kJ/mm. Por final, mais duas camadas de aço inoxidável com cobalto, liga resistente à cavitação foram depositadas por arco pulsado, com energia de soldagem nominal de 0,5 a 0,8kJ/mm, com uma mistura de gases de 98% de argônio e 2% de oxigênio como proteção. Secções transversais das amostras foram preparadas, devidamente lixadas e depois polidas com alumina 1,0μm, seguido de ataque químico moderado com Villela para observação de microestruturas. Foi feito o estudo de microdureza Vickers com carga padrão de 0,4 N e espaçamento regular (0,4 mm)... / Nowadays most of the power supply used in Brazil is provided for a large number of hydraulic turbines. During its operation in hydroelectric power plants, these turbines usually have been damaged either mechanical or hydraulic reasons. So, catastrophic cracking and loss-of-mass due to cavitation erosion are main problems which are commonly repaired by welding. Higher levels of resistance to cavitation erosion have been attained with surface deposition of a cobalt-alloyed stainless steel coating by gas-protected arc welding techniques. In the present work six plates, which were machined from blades of hydraulic turbines made with ASTM A36 carbon steel grade, were used as base metal. Using manual gas-metal arc welding (GMAW) in flat weld position two layers of AWS E70-S6 carbon steel (1.2 mm diameter) were deposited on all samples. Only 3 samples two buttering layers were deposited with AWS E309-T1 flux-cored wire (1.6 mm diameter) using same processing, where were applied a 75% carbon dioxide - 25% argon mixture as protection gas. The nominal heat input used in all layers was approximately 0.5 kJ/mm. At the last welding deposition another two cobalt-alloyed, cavitation resistant, stainless steel cladding layers were deposited under pulsed arc with a nominal heat input of 0.5 to 0.8kJ/mm, being used for them a protective gas mixture of 98% argon - 2% oxygen. Transverse sections of weld deposit were prepared according standard grinding method (up to 1200-grit SiC paper) and final mechanical polishing using 1.0μm alumina, followed by moderate etching in Villela reagent for microstructural observation. Vickers microhardness measurements were carried out at standard load (0.4 N) and regular spacings (0,4mm) from surface to base metal. Light microscopy (LM) was used to determine average size of inclusions, which was measured from digitalized images using a freeware image analysis ...(Complete abstract, click electronic access below)
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The effect of filler metal on the corrosion resistance of stainless steel weldments in a hot organic acid environmentOrsmond, Charles Petrus Marais 30 August 2010 (has links)
Selective corrosion of type 316L austenitic stainless steel welds during the production of organic acids resulted in losses in production due to unscheduled downtimes to perform repairs. Estimated corrosion rates of type 316L filler material welds were an order of magnitude higher than that of the base material. Alternative higher alloyed commercial filler materials were evaluated under actual production conditions. The evaluated filler materials were types 316L, 317L, 309L, 309MoL, 2205, 2507, 625, 825 and 904L. The effect of nitrogen on the corrosion properties of type 309L filler material was evaluated by manipulating the nitrogen concentration of the shielding gas during MIG welding. These changes in nitrogen concentration did not influence the corrosion resistance of the type 309L filler material. No correlation could be established between the corrosion rates, analysed chemical composition of the product and operating temperature during production. In almost all the cases where the chemical composition of the filler material was comparable with that of the base material the corrosion rates of the filler materials were higher than that base material. It might be expected that the ferrite phase with higher molybdenum and chromium should be more corrosion resistant while the austenite should be less resistant. This was, however, not the case with the corrosion of type 309L filler material. It would thus appear that in this case nickel enrichment of the austenite phase had a larger influence on the corrosion resistance of the austenite phase than the chromium and molybdenum had on the corrosion resistance of the ferrite phase. It appears that nickel and molybdenum had the largest contribution to the corrosion resistance of stainless steels welds under these operating conditions. It is, however, believed that a certain minimum concentration of chromium is also required to provide corrosion resistance to these alloys in hot organic acid environments. In contrast with the fact that a substantial alloying content is required to improve corrosion resistance of the filler material, the small difference in composition between ferrite and austenite phases, due to micro segregation, appeared to affect the corrosion resistance on micro scale. This is illustrated by the micrographs, which show corrosion to etch out the dendrite structure. Since the morphology of the austenite and ferrite phases is so similar, it could not always be conclusively established which one of the two phases corroded selectively. Analyses performed on the austenite and ferrite phases did not indicate a concentration difference within the phases itself. However, there were significant differences in the concentration of elements between the phases, with the austenite stabilising elements reporting to the austenite phase and the ferrite stabilizing elements reporting to the ferrite phase, in line with thermodynamic predictions. In the case of the filler materials following the austenite mode of solidification, no significant concentration differences were detected within the matrix. Although all highly alloyed high nickel alloyed filler materials (types 904L, 825 and 625) corroded at a lower rate than the type 316L base material, type 625 filler material was the filler material of choice due to the lack of any pitting of the weld. Pitting was detected in both the 825 and 904L filler materials. Galvanic corrosion was not noted at any of the weld/HAZ interfaces and in no case did the type 316L parent metal adjacent to the weld corrode preferentially to the material further away from the weld. Copyright / Dissertation (MEng)--University of Pretoria, 2010. / Materials Science and Metallurgical Engineering / unrestricted
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Studium jevů limitujících životnost sekundárních článků Ni-Zn / A Study of Phenomena that Limit the Life-Span of Ni-Zn Secondary CellsChladil, Ladislav January 2014 (has links)
This dissertation is focused on the study of selected additives and their effect on positive and negative electrodes of Ni-Zn secondary cells. The first group of additives consists of the inorganic compounds that reduce zinc oxide solubility and thus prevents shape changes of the zinc electrode. The second group contains organic additives that have the beneficial effect of dendrite growth prevention. All additives were examined in relationship to their compatibility with the positive electrode and to their effect on the zinc electrode dendrites inhibition, hydrogen evolution and corrosion inhibition. Moreover, the study of decomposition kinetics of supersaturated zincate solution with the first group of additives was performed. Finally, a short study of pulse regime charging mode with regard to dendrites inhibition was performed and evaluated.
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Experimental and Modeling Studies of Dendrite Initiation during Lithium ElectrodepositionMaraschky, Adam M. 07 September 2020 (has links)
No description available.
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Understanding the molecular, cellular, and circuit defects characterizing the early stages of Alzheimer’s diseaseVirga, Daniel Michael January 2023 (has links)
One of the most foundational and personal philosophical questions one can ask is what makes you, you? In large part, you are made up of your relationships, experiences, and memories. The hippocampus, a brain region which is critical for the formation of memories, has been the focus of neuroscience research for decades due partially to this function, which is foundational to our individuality. In Alzheimer’s disease (AD), one of the most common and well-researched neurodegenerative diseases in the world, the hippocampus is one of the earliest targets. Despite extensive work on AD, we still lack a coherent understanding of what is causing the disease, the mechanisms by which it is causing neuronal dysfunction and death within the hippocampus and other brain regions, and how it ultimately causes deficits in cognition and behavior, leading to an erosion of our selves.
In this thesis, I explore three independent but related questions: 1) what molecular mechanisms are causing early synaptic loss in AD, specifically within the hippocampus, 2) what molecular effectors are responsible for establishing and maintaining intracellular architecture in hippocampal neurons, which are exploited in early AD, and 3) how and when does the hippocampal circuit dysfunction in AD progression?
Using a variety of experimental techniques, ranging from in utero and ex utero electroporation, primary murine and human neuronal cell culture, longitudinal confocal microscopy, immunohistochemistry, biochemistry, cell and molecular biology, in vivo two-photon calcium imaging, and behavioral assays, I have found that, within CA1 of the hippocampus, synapse loss requires degradation of the dendritic mitochondrial network, activity and input specificity are driving mitochondrial compartmentalization within CA1 neurons through the same pathway that is aberrantly overactivated in AD, and the hippocampal circuit is overly rigid in encoding the environment as the disease progresses.
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Suppressing Dendritic Growth during Zinc Electrodeposition using Polyethylenimine as an Electrolyte Additive for Rechargeable Zinc BatteriesBanik, Stephen John, II 31 May 2016 (has links)
No description available.
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