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Dissertations / Theses on the topic 'Cell organelles'
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Shelby, James Patrick. "The application of microfluidics to the study of biological processes /." Thesis, Connect to this title online; UW restricted, 2004. http://hdl.handle.net/1773/8483.
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Cramer, Louise Pauline. "The biogenesis of secretory organelles in a neuroendocrine cell line." Thesis, Imperial College London, 1991. http://hdl.handle.net/10044/1/46731.
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Deng, Yuping. "Studies of intraorganelle dynamics : the lysosome, the pre-lysosomal compartment, and the golgi apparatus /." Diss., This resource online, 1991. http://scholar.lib.vt.edu/theses/available/etd-07282008-134815/.
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Mortara, R. A. "Microfilament-membrane interactions in isolated P815 filopodia." Thesis, University of Cambridge, 1985. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.372923.
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Hayes, Michael Henry. "Investigating the organization and regulation of aggregated proteins within sub-nuclear organelles." Diss., University of Iowa, 2019. https://ir.uiowa.edu/etd/6759.
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Protein aggregation has long been associated with disease states, such as Alzheimer’s, Huntington’s, and Parkinson’s diseases. This model is challenged by increasing recognition of functional protein aggregates. Our focus has been exploration of intra-aggregate organization, and investigation of the ability of ATP to mediate energy independent aggregate solubilization by acting as a hydrotrope. Furthering our understanding of the composition and regulation of these structures will provide insight into how their dysregulation gives rise to disease, and provide a template for the development of novel therapeutics.
Sub-nuclear organelles are a class of non-membrane bound organelle. These compartments form through liquid-liquid phase separation of their components, resulting in a functional aggregate that demonstrates liquid properties. Many of the factors driving liquid-liquid phase separation also drive the formation of amyloid fibrils. In vitro studies have demonstrated that liquid-liquid phase separation can promote amyloid fibril formation, and that liquid-liquid phase separated droplets can mature through amyloid fibril formation. In vivo, functional amyloids are known to contribute to peptide hormone storage, melanin production, and maintenance of long-term memory. Despite these findings, there has been limited investigation into the relationship between liquid-liquid phase separated droplets and amyloid fibrils in vivo.
Our exploration of intra-aggregate organization took advantage of the numerous protein aggregates found within Xenopus oocytes. Using a dye and antibodies specific to the defining cross-β structure of amyloid, we demonstrated that amyloids form as a normal part of Xenopus oogenesis. Amyloid was detected within nuclear and cytosolic aggregates. In the cytosol, amyloid was observed within yolk platelets and a population of cortical particles whose identity has yet to be determined. In the nucleus, multiple liquid-liquid phase separated sub-nuclear organelles, including those associated with RNA polymerase I, II and III transcription as well as RNA processing, were found to have an amyloid component. Proteomic analysis of aggregate enriched material revealed proteins associated with ribonucleoprotein complex biogenesis, DNA replication, RNA processing and DNA-templated transcription. Our analysis further demonstrated that nuclear amyloids were stable, remaining intact for hours post isolation, but rapidly destabilized following treatment with RNase.
Recently, adenosine triphosphate (ATP) was demonstrated to act as a hydrotrope in vitro. Physiologic levels were shown to maintain protein solubility and solubilize recombinant liquid-liquid phase separated droplets, in the absence of ATP hydrolysis. These observations suggest that this previously unappreciated activity of ATP has a large influence on liquid-liquid phase separated droplet stability. However, these observations were made under conditions that diverge from those found in vivo. The liquid-liquid phase separated droplets tested were devoid of RNA and composed of a single protein. Amyloid containing aggregates required super physiological levels of ATP, suggesting the sub-nuclear organelles we previously described would be resistant to this activity.
By observing nucleoli within isolated Xenopus oocyte nuclei, we demonstrate that ATP has the capacity to act as a hydrotrope in vivo. However, the hydrotropic action alone of ATP is not sufficient to solubilize nucleoli. Instead, solublization requires a sensitization step which is dependent on a soluble factor(s) and requires ATP hydrolysis.
Our studies support an in vivo model of liquid-liquid phase separated aggregate solubilization where a soluble factor(s) sensitizes an aggregate in an energy dependent process. The susceptible aggregate is then solubilized by the hydrotropic action of ATP. We further speculate that liquid-liquid phase separated aggregate formation is a reversal of the above steps. These findings highlight a need to further understand the relationship between liquid-liquid phase separation and amyloid formation, and identify an urgent need to dissect the factors influencing intracellular ATP levels.
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Malchus, Nina Isabelle [Verfasser], and Michael [Akademischer Betreuer] Hausmann. "On the spatial organization of cell organelles and diffusion of proteins in organelle membranes / Nina Isabelle Malchus ; Betreuer: Michael Hausmann." Heidelberg : Universitätsbibliothek Heidelberg, 2011. http://d-nb.info/1179230477/34.
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Distasi, Matthew R. "The 3D characterization of the annulate lamellae : the development of a new methodology incorporating 3D-anaglyph techniques and serial transmission electron microscopy." Virtual Press, 2003. http://liblink.bsu.edu/uhtbin/catkey/1266020.
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So, Ka Chai. "Characterization of mab-22 gene and its role in caenorhabditis elegans sensory ray formation /." View abstract or full-text, 2004. http://library.ust.hk/cgi/db/thesis.pl?BIOL%202004%20SO.
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Thesis (M.Phil.)--Hong Kong University of Science and Technology, 2004.Includes bibliographical references (leaves 116-123). Also available in electronic version. Access restricted to campus users.
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Linka, Marc. "Understanding the origin and function of organellar metabolite transport proteins in photosynthetic eukaryotes Galdieria sulphuraria and Arabidopsis thaliana as model systems /." Diss., Connect to online resource - MSU authorized users, 2008.
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Pance, Alena. "Studies on a variant of the rat PC12 cell line lacking regulated secretory organelles." Thesis, University of Cambridge, 1998. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.624798.
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Wallace, Isha Kimisha. "The kinetochore protein Mif2p is targeted by Cdk1p and development of a selection for regulators of centromere/kinetochore structure/function." Diss., [Riverside, Calif.] : University of California, Riverside, 2010. http://proquest.umi.com/pqdweb?index=0&did=2019869861&SrchMode=2&sid=1&Fmt=2&VInst=PROD&VType=PQD&RQT=309&VName=PQD&TS=1274198666&clientId=48051.
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Thesis (Ph. D.)--University of California, Riverside, 2010.Includes abstract. Available via ProQuest Digital Dissertations. Title from first page of PDF file (viewed May 18, 2010). Includes bibliographical references. Also issued in print.
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Profant, Deborah Ann. "Defining the cis-acting requirements in the HMG-CoA reductase gene for karmellae biogenesis /." Thesis, Connect to this title online; UW restricted, 1999. http://hdl.handle.net/1773/5137.
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Bowman, Amanda. "Lipidomic Analysis of Single Cells and Organelles Using Nanomanipulation Coupled to Mass Spectrometry." Thesis, University of North Texas, 2016. https://digital.library.unt.edu/ark:/67531/metadc849662/.
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The capability to characterize disease states by way of determining novel biomarkers has led to a high demand of single cell and organelle analytical methodologies due to the unexpected heterogeneity present in cells of the same type. Lipids are of particular interest in the search for biomarkers due to their active roles in cellular metabolism and energy storage. Analyzing localized lipid chemistry from individual cells and organelles is challenging however, due to low analyte volume, limited discriminate instrumentation, and common requirements of separation procedures and expenditure of cell sample. Using nanomanipulation in combination with mass spectrometry, individual cells and organelles can be extracted from tissues and cultures in vitro to determine if heterogeneity at the cellular level is present. The discriminate extraction of a single cell or organelle allows the remainder of cell culture or tissue to remain intact, while the high sensitivity and chemical specificity of mass spectrometry provides structural information for limited volumes without the need for chromatographic separation. Mass analysis of lipids extracted from individual cells can be carried out in multiple mass spectrometry platforms through direct-inject mass spectrometry using nanoelectrospray-ionization and through matrix-assisted laser/desorption ionization.
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Gordon, Joe W. "The effect of contractile activity on mitochondrial transcription factor A expression in skeletal muscle." Thesis, National Library of Canada = Bibliothèque nationale du Canada, 2000. http://www.collectionscanada.ca/obj/s4/f2/dsk1/tape2/PQDD_0017/MQ59171.pdf.
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Cooley, Hagans Cristin D. "An Analysis of the Effectiveness of Teacher Versus Student-Generated Science Analogies on Comprehension in Biology and Chemistry." Defiance College / OhioLINK, 2003. http://rave.ohiolink.edu/etdc/view?acc_num=def1281549287.
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Nilsson, Harriet. "The role of nitric oxide in cytoskeleton-mediated organelle transport and cell adhesion /." Linköping : Univ, 2001. http://www.bibl.liu.se/liupubl/disp/disp2001/med660s.pdf.
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Wiking, Mikaela. "Spatial proteome profiling of the compartments of the human cell using an antibody-based approach." Licentiate thesis, KTH, Proteomik och nanobioteknologi, 2017. http://urn.kb.se/resolve?urn=urn:nbn:se:kth:diva-206817.
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The human cell is complex, with countless processes ongoing in parallel in specialized compartments, the organelles. Cells can be studied in vitro by using immortalized cell lines that represent cells in vivo to a varying degree. Gene expression varies between cell types and an average cell line expresses around 10,000-12,000 genes, as measured with RNA sequencing. These genes encode the cell’s proteome; the full set of proteins that perform functions in the cell. In paper I we show that RNA sequencing is a necessary tool for studying the proteome of the human cell. By studying the proteome, and proteins’ localization in the cell, information can be assembled on how the cell functions. Image-based methods allow for detailed spatial resolution of protein localization as well as enable the study of temporal events. Visualization of a protein can be accomplished by using either a cell line that is transfected to express the protein with a fluorescent tag, or by targeting the protein with an affinity reagent such as an antibody. In paper II we present subcellular data for a majority of the human proteins, showing that there is a high degree of complexity in regard to where proteins localize in the cell. Cellular energy is generated in the mitochondria, an important organelle that is also active in many other different functions. Today approximately only a third of the estimated mitochondrial proteome has been validated experimentally, indicating that there is much more to understand with regard to the functions of the mitochondria. In paper III we explore the mitochondrial proteome, based on the results of paper II. We also present a method for sublocalizing proteins to subcompartments that can be performed in a high-throughput manner. To conclude, this thesis shows that transcriptomics is a useful tool for proteome-wide subcellular localization, and presents high-resolution spatial distribution data for the human cell with a deeper analysis of the mitochondrial proteome.QC 20170512
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Buks, Ralfs. "Impact of JAK2V617F on Terminal Erythroid Differentiation and Red Blood Cell Electrophysiology in Polycythemia Vera." Thesis, Université de Paris (2019-....), 2020. http://www.theses.fr/2020UNIP7096.
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Les néoplasies myéloprolifératives (NMP) regroupent des pathologies caractérisées par une prolifération et une différenciation anormales des cellules souches hématopoïétiques. La polyglobulie de Vaquez (PV) est un type de NMP caractérisé par un nombre de globules rouges (GR) élevé. Plus de 95% des patients atteints de PV présentent la mutation V617F de la tyrosine kinase JAK2 conduisant à une prolifération érythroïde incontrôlée entraînant un risque élevé de thrombose. Les patients présentant un risque pronostique élevé sont traités par des thérapies cytoréductrices, dont le ruxolitinib, un inhibiteur de JAK1/JAK2. Nous avons analysé le protéome de la membrane des GR PV par spectrométrie de masse et avons montré des niveaux élevés de protéines de liaison au Ca2+ ainsi que la présence anormale de protéines du réticulum endoplasmique.Dans cette thèse, nous avons étudié (1) l'impact de JAK2V617F sur l'expression et le tri des protéines pendant la différenciation érythroïde terminale, l'énucléation et la maturation ; (2) l'impact de JAK2V617F sur l'homéostasie du calcium et l'électrophysiologie des GR dans la PV ; (3) l'import du ruxolitinib dans le compartiment intracellulaire.Nos données montrent que JAK2V617F jouerait un rôle dans la rétention des organelles pendant la phase d'énucléation des érythroblastes, ce qui pourrait perturber la maturation des réticulocytes circulants chez la souris et l'homme. Nos données obtenues par patch-clamp automatisé montrent une homéostasie calcique modifiée dans les GR PV et des lignées cellulaires exprimant JAK2V617F, avec un impact fonctionnel sur l'activité du canal Gárdos, ce qui pourrait contribuer à une déshydratation cellulaire dans un contexte JAK2V617F. Enfin, nous avons étudié le transport du ruxolitinib à l’aide de tests de cytotoxicité et d'apoptose dans des lignées cellulaires humaines, des GR et des progéniteurs érythroïdes humains différenciés in vitro. Nos résultats suggèrent que le ruxolitinib est importé dans le compartiment intracellulaire par la protéine ABCG2, un membre de la superfamille des transporteurs à « ATP binding cassette » (ABC).Étant donné le rôle central du calcium dans la régulation des voies de signalisation, notre étude ouvre de nouvelles perspectives pour explorer la relation entre JAK2V617F, l'homéostasie du calcium et les anomalies cellulaires dans les NMP telles que les interactions cellulaires dans la circulation sanguine en relation avec les événements thrombotiques. Ces interactions pourraient également être déclenchées par les propriétés anormales des GR suite à des défauts de maturation réticulocytaire. En ce qui concerne le traitement des patients, notre étude montre pour la première fois le rôle d'un transporteur spécifique dans l’import intracellulaire du ruxolitinib. Elle ouvre de nouvelles perspectives de recherche sur l'efficacité du ruxolitinib en analysant le polymorphisme d'ABCG2 et son expression sur les cellules cibles des patients traitésMyeloproliferative neoplasms (MPNs) are a group of disorders characterised by abnormal proliferation and differentiation of hematopoietic stem cells in the bone marrow. Polycythemia Vera (PV) is a type of MPN characterised by overproduction of red blood cells (RBCs). Over 95 % of PV patients carry the V617F mutation in the tyrosine kinase Janus kinase 2 (JAK2) resulting in uncontrolled erythroid proliferation and high risk of thrombosis. Patients with high prognostic risk scores are treated with cytoreductive therapies, including ruxolitinib, a JAK1/JAK2 inhibitor. Using mass spectrometry, we analysed the RBC membrane proteome and showed elevated levels of multiple Ca2+ binding proteins as well as endoplasmic reticulum residing proteins in PV RBC membranes compared to RBC membranes from healthy individuals. In this thesis we investigated:(1) the impact of JAK2V617F on protein expression and sorting during terminal erythroid differentiation, enucleation and maturation;(2) the impact of JAK2V617F on calcium homeostasis and RBC electrophysiology in PV;(3) ruxolitinib import in the intracellular compartment.Our data shows that JAK2V617F could play a role in organelle retention during the enucleation step of erythroid differentiation, which may affect maturation of circulating reticulocytes in mouse and human. Our data from automated patch-clamp shows modified calcium homeostasis in PV RBCs and cell lines expressing JAK2V617F, with a functional impact on the activity of the Gárdos channel that could contribute to cellular dehydration. Finally, we investigated ruxolitinib transport using cytotoxicity and apoptosis assays in human cell lines, RBCs and in vitro differentiated human erythroid progenitors. Our findings suggest that ruxolitinib is imported in the intracellular compartment by ABCG2, a member of the ATP-binding cassette (ABC) transmembrane superfamily.Given the central role that calcium plays in the regulation of signalling pathways, our study opens new perspectives to explore the relationship between JAK2V617F, calcium homeostasis and cellular abnormalities in MPNs, including cellular interactions in the bloodstream in relation to thrombotic events. These interactions could also be triggered by the abnormal properties of circulating RBCs secondary to the presence of organelle remnants in their membrane and cytoplasm. Regarding patients’ treatment, our study shows for the first time the role of a specific transporter in ruxolitinib cellular influx. It opens new perspectives in ruxolitinib efficacy research targeting cell types depending on ABCG2 expression and polymorphisms among patients
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Rosfelter, Anne. "Le positionnement du fuseau mitotique chez le zygote d'ascidie et son rôle dans la répartition des organelles." Electronic Thesis or Diss., Sorbonne université, 2023. http://www.theses.fr/2023SORUS063.
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Après la fécondation d’un ovocyte, un aster de microtubules se forme autour de l’ADN mâle. Cet aster spermatique permet d’amener le pro-noyau femelle jusqu’au pro-noyau mâle pour qu’ils puissent fusionner. Il permet aussi de déplacer l’ADN fusionné jusqu’au centre de la cellule pour assurer une division cellulaire équitable. Les mécanismes de centration d’un aster ou d’un fuseau ont donné lieu à de nombreuses recherches, que ce soit par modélisation, expérimentalement chez des espèces telles C. elegans, P. lividus, M.musculus ou in vitro sur des extraits de Xenopus laevis. Trois mécanismes principaux se dégagent : le pushing, le cortical pulling et le cytoplasmic pulling (ou bulk pulling). En étudiant le déplacement de l’aster et du fuseau mitotique chez le zygote de l’ascidie P. mammillata j’ai découvert un système qui combine ces trois mécanismes en s’appuyant sur l’alternance des étapes du cycle cellulaire. En méiose, l’aster utilise la polymérisation des microtubules qui le composent pour pousser contre le cortex d’actine et s’en décoller (pushing). Arrivé en interphase, l’aster retourne contre le cortex grâce à une traction qu’exerce la membrane sur les microtubules (cortical pulling). Enfin à l’entrée en mitose, la traction membranaire cesse et libère les asters du fuseau mitotique, qui cèdent donc aux forces exercées par le transport d’organelles vers le centre de l’aster (cytoplasmic pulling) qui semblent constantes durant le cycle cellulaire. Cela permet de centrer le fuseau. En même temps que l’aster se forme et se déplace, une réorganisation des compartiments intracellulaires se met en place. Pour comprendre de quelle manière l’organisation intracellulaire peut être perturbée par la formation de l’aster, j’ai étudié le cas du vitellus. En effet, le vitellus, qui est présent sous forme de vésicules, est initialement abondant et homogène dans l’ovocyte non fécondé. Cependant, dès que l’aster apparaît, sa répartition change et les vésicules de vitellus sont exclues de la zone contenant l’aster. Cette exclusion générée à la formation de l’aster chez le zygote, est maintenue au cours du développement. Dans mes travaux, j’ai pu observer qu’elle est majoritairement due à l’accumulation à l’aster d’autres organelles comme le réticulum endoplasmique. La fonction de transport des microtubules de l’aster suffit donc à réorganiser complètement la cellule en excluant certaines organelles et en en accumulant d’autres. Les déplacements de l’aster et du fuseau mitotique, leur régulation par le cycle cellulaire, et la réorganisation intracellulaire, identifiés ici chez le zygote d’ascidie, s’appuient sur le fonctionnement d’éléments fondamentaux d’une cellule, à savoir : les microtubules, le cortex d’actine, le réticulum endoplasmique, les protéines du cycle cellulaire, etc. Les découvertes présentées revêtent ainsi une portée universelle, adaptable aux spécificités de différents types cellulairesAfter oocyte fertilization, a microtubule aster forms around the male DNA. The sperm aster brings the female pro-nucleus to the male pro-nucleus so they can fuse, but it also moves the fused nuclei to the cell center to ensure an equitable cell division. Numerous studies performed in vitro, by modeling or experimentally in species such as C. elegans, P. lividus, and M. musculus, addressed the aster and spindle centration mechanisms. Three main mechanisms emerged; pushing, cortical pulling, and cytoplasmic pulling. By studying aster centration in the zygote of the ascidian P. mammillata, I discovered a system that combines these three mechanisms based on the cell cycle stages. In meiosis, the aster uses the polymerization of its microtubules to push against the actin cortex and move away from it (pushing). Once in interphase, the aster returns to the cortex by a pull exerted by the membrane on the microtubules (cortical pulling). At mitosis entry, cortical pulling stops, and releases the mitotic spindle's asters. In consequence, the asters give in to the forces exerted by the transport of organelles to the aster center (cytoplasmic pulling), that appeared constant during the cell cycle. Cytoplasmic pulling hence participate in centering the spindle While the aster forms and moves, the intracellular compartments reorganize. To understand how intracellular organization can be disrupted by aster formation, I studied the case of yolk. The yolk, in the form of vesicles (called granules or platelets), is initially abundant and homogeneous in the unfertilized oocyte. However, as soon as the aster appears, its distribution changes and the yolk platelets are excluded from the region containing the aster. This exclusion generated by the aster formation in the zygote is maintained during development. I observed that yolk exclusion is mainly due to the accumulation at the aster of other organelles such as the endoplasmic reticulum. The transport function of the aster microtubules is therefore sufficient to completely reorganize the cell by excluding some organelles and accumulating others. The movements of the aster and the spindle, their regulation by cell cycle, and the intracellular reorganization, identified here in the ascidian zygote, rely on basic elements of a cell, namely: the microtubules, the actin cortex, the endoplasmic reticulum, the proteins of the cell cycle, etc. Thus, the discoveries presented here cover a broad scope, and seem adaptable to the specificities of different cell types
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Besnault, Pierre. "Impact pathologique du stress chronique dans la maladie de Parkinson : rôle des récepteurs aux glucocorticoïdes dans la régulation des organelles de signalisation immunitaire innée." Electronic Thesis or Diss., Sorbonne université, 2023. http://www.theses.fr/2023SORUS534.
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Ces travaux s’intéressent aux conséquences pathologiques du stress chronique (SC) et à l'altération de la signalisation médiée par les glucocorticoïdes (GCs) et leurs récepteurs (GRs) dans la maladie de Parkinson (MP). Le stress, via l’activation de l’axe hypothalamo-hypophysaire-surrénalien (HHS) et la sécrétions de GCs, déclenche un grand nombre de réponses physiologiques qui sont bénéfiques à court terme. En revanche le SC perturbe l’axe HHS, altère ces réponses physiologiques et conduit au développement de maladies neuropsychiatriques comme la dépression, qui constitue par ailleurs un facteur de risque important de la MP. En outre, un faisceau d’arguments suggèrent que l’axe HHS est perturbé chez les patients parkinsoniens, qui présentent également une altération de la signalisation GC/GR au niveau central. Dans quelle mesure ces altérations peuvent impacter les mécanismes physiopathologiques de la maladie et contribuer à sa progression ? La MP est une affection neurologique pour laquelle il n’existe toujours pas de traitement curatif. Ceci vient en partie du fait que l’origine de la maladie reste généralement mal comprise. En effet, les formes sporadiques représentent une très large majorité des cas de MP, et résultent vraisemblablement d’interactions complexes entre des facteurs de risque génétiques et des facteurs de risque environnementaux. Le diagnostic clinique de la MP s’appuie sur l’identification de symptômes moteurs qui résultent d’une perte importante des neurones dopaminergiques de la substance noire. Cette dégénérescence neuronale est par ailleurs associée à l’agrégation et au dépôt de la protéine alpha-Synucléine (aSyn) au sein d’inclusions intracytoplasmiques neuronales, les corps et neurites de Lewy. La mort des neurones dopaminergiques est également accompagnée d’une réponse immunitaire orchestrant des processus neuroinflammatoires qui semble jouer un rôle important dans la progression de la maladie. L’une des découvertes les plus remarquables ces dernières années concernant le mécanisme d’activation des cellules immunitaires dans la MP a été celle révélant les propriétés structurelles des assemblages d’aSyn capables d’agir comme de véritables motifs moléculaires associées aux microbes, et de stimuler l’activation des centres organisateurs supramoléculaires (SMOCs), qui sont considérés comme de véritables organelles de la signalisation immunitaire innée. Etant donné que l’une des propriétés du GR consiste à réguler les réponses inflammatoires, nous formulons l’hypothèse que l’altération de l’axe HHS et de la signalisation GC/GR par le stress chronique provoquerait une réponse inflammatoire exacerbée et délétère via la suractivation de la signalisation SMOC-dépendante. Nos travaux montrent que le SC altère d’une part la signalisation GC/GR microgliale et d’autre part aggrave les paramètres neuropathologiques (mort neuronale et neuroinflammation) dans un modèle murin de MP. Comprendre précisément le lien entre l’altération de la signalisation GC/GR et l’augmentation des réponses inflammatoires en s’intéressant à la signalisation SMOC-dépendante permettra de mieux appréhender les différents mécanismes moléculaires physiopathologiques pouvant être impliqués dans la maladie, mais aussi d’identifier de nouvelles cibles thérapeutiquesThis project focuses on the pathological consequences of chronic stress (CS) and the alteration of signaling mediated by glucocorticoids (GCs) and their receptors (GRs) in Parkinson's disease (PD). Stress stimulates the hypothalamic-pituitary-adrenal (HPA) axis and the secretion of GCs that triggers a large number of physiological responses which are beneficial in the short term. On the other hand, CS disrupts the HPA axis, alters these physiological responses and leads to the development of neuropsychiatric disorders such as depression, a known risk factor for PD. In line with this, evidence suggests that the HPA axis is altered in PD patients together with a central alteration of GC/GR signaling. How could these alterations impact the pathomechanisms and contribute to disease progression? PD is an uncurable neurological disorder of still unknown origin. Sporadic form of the disease represents a very large majority of PD cases, and probably results from complex interactions between genetic and environmental risk factors. Clinically, PD is diagnosed upon identification of motor symptoms resulting from a massive loss of dopaminergic neurons in the substantia nigra. Neurodegeneration is associated with the aggregation of alpha-synuclein (aSyn) that deposits into intraneuronal inclusions known as Lewy bodies and neurites. Associated with neurodegeneration, immune responses orchestrating inflammatory processes are mounted and likely involved in disease progression. In the past years, one of the greatest finding regarding the mechanism of immune cell activation in PD has been the discovery of the structural properties of aSyn assemblies being able to act as true pathogen-associated molecular pattern, and to mediate the activation of supramolecular organizing centers (SMOCs), which are considered as important innate immune signaling platforms. Given that one of the major properties of GR is to regulate inflammatory response in immune cells, we hypothesize that chronic stress-mediated HPA axis and GC/GR signaling alterations could exacerbate inflammatory responses through an overactivation of SMOC-dependent signaling. Our work shows that CS not only alters microglial GC/GR signaling, but also increases neuronal death and microglia-associated inflammatory response in a mouse model of PD. Understanding precisely the link between the alteration of GC/GR signaling and the increase in the inflammatory response by focusing on SMOC-dependent signaling will allow us to better understand the different pathophysiological molecular mechanisms, but also identify new therapeutic targets
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Morgan, Gloria Yvonne. "The expression of immunophilins in cells and organelles." Thesis, University of Sussex, 1994. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.282145.
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Mayer, Jürgen. "Investigation of the biophysical basis for cell organelle morphology." Master's thesis, Saechsische Landesbibliothek- Staats- und Universitaetsbibliothek Dresden, 2010. http://nbn-resolving.de/urn:nbn:de:bsz:14-qucosa-26600.
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It is known that fission yeast Schizosaccharomyces pombe maintains its nuclear envelope during mitosis and it undergoes an interesting shape change during cell division - from a spherical via an ellipsoidal and a peanut-like to a dumb-bell shape. However, the biomechanical system behind this amazing transformation is still not understood. What we know is, that the shape must change due to forces acting on the membrane surrounding the nucleus and the microtubule based mitotic spindle is thought to play a key role. To estimate the locations and directions of the forces, the shape of the nucleus was recorded by confocal light microscopy. But such data is often inhomogeneously labeled with gaps in the boundary, making classical segmentation impractical. In order to accurately determine the shape we developed a global parametric shape description method, based on a Fourier coordinate expansion. The method implicitly assumes a closed and smooth surface. We will calculate the geometrical properties of the 2-dimensional shape and extend it to 3-dimensional properties, assuming rotational symmetry.
Using a mechanical model for the lipid bilayer and the so called Helfrich-Canham free energy we want to calculate the minimum energy shape while respecting system-specific constraints to the surface and the enclosed volume. Comparing it with the observed shape leads to the forces. This provides the needed research tools to study forces based on images.
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Mayer, Jürgen. "Investigation of the biophysical basis for cell organelle morphology." Master's thesis, Max-Planck-Institut für Molekulare Zellbiologie und Genetik, 2008. https://tud.qucosa.de/id/qucosa%3A25225.
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It is known that fission yeast Schizosaccharomyces pombe maintains its nuclear envelope during mitosis and it undergoes an interesting shape change during cell division - from a spherical via an ellipsoidal and a peanut-like to a dumb-bell shape. However, the biomechanical system behind this amazing transformation is still not understood. What we know is, that the shape must change due to forces acting on the membrane surrounding the nucleus and the microtubule based mitotic spindle is thought to play a key role. To estimate the locations and directions of the forces, the shape of the nucleus was recorded by confocal light microscopy. But such data is often inhomogeneously labeled with gaps in the boundary, making classical segmentation impractical. In order to accurately determine the shape we developed a global parametric shape description method, based on a Fourier coordinate expansion. The method implicitly assumes a closed and smooth surface. We will calculate the geometrical properties of the 2-dimensional shape and extend it to 3-dimensional properties, assuming rotational symmetry.
Using a mechanical model for the lipid bilayer and the so called Helfrich-Canham free energy we want to calculate the minimum energy shape while respecting system-specific constraints to the surface and the enclosed volume. Comparing it with the observed shape leads to the forces. This provides the needed research tools to study forces based on images.
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Dolman, Nicholas James. "Polarised signalling and organelle distribution in the pancreatic acinar cell." Thesis, University of Liverpool, 2004. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.406669.
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Robertson, Elizabeth Jean. "Relationships between the cytoskeleton and cytoplasmic organelles in bryophyte cells." Thesis, Queen Mary, University of London, 1991. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.294284.
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Bai, Seoung-Jai. "Nanoscale probes for electrochemical measurements on single cells and organelles /." May be available electronically:, 2007. http://proquest.umi.com/login?COPT=REJTPTU1MTUmSU5UPTAmVkVSPTI=&clientId=12498.
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Fujisawa, Alma. "Development of chemical labeling methods for organelle molecule analysis." Kyoto University, 2019. http://hdl.handle.net/2433/243315.
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Lu, Hang 1977. "Microfluidic biomechanical and electrical devices for rapid analysis of cells and organelles." Thesis, Massachusetts Institute of Technology, 2003. http://hdl.handle.net/1721.1/7996.
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Thesis (Ph. D.)--Massachusetts Institute of Technology, Dept. of Chemical Engineering, 2003.Includes bibliographical references (p. 139-145).
This thesis focuses on micro devices aimed at rapid analysis of cells and subcellular organelles. These devices take advantage of microfabrication techniques to create environment suitable for biomechanical and biochemical stimulation of cells, to break cell membranes to extract the intracellular materials, and to separate or concentrate organelles and proteins of interest. These procedures greatly reduce the amounts of samples and reagents necessary and the process time required from their macro counterparts. Moreover, they demonstrate operational advantages, such as lower voltages, less heating, and no significant gas formation in electrolysis, over their macroscopic counterparts. First in line of the process stream are a series of microfluidic devices developed for the purpose of studying cell adhesion on biomaterials. Numerical models are developed to aid the quantitative analysis of fluid shear stresses on cells in these devices. The experimental results demonstrate that these devices are capable of capturing ligand-density-dependent, shear-dependent, and growth-factor-dependent adhesion behavior of cell cultures. Next, two electrical microfluidic devices are developed for the purpose of cell lysis and organelle separation. Both devices are fabricated using electroplating techniques to create three-dimensional electrodes, and lithography to accommodate flexible designs in the fluid channels. Simple electrical models for cells and organelles are used to guide the design and operation of the miniaturized electroporation device that can successfully break open cells and release their content.
(cont.) To study the isoelectric focusing field flow fractionation phenomena, another model combining flow calculation and reactive transport of amphoteric particles is used. The experimental results of organelle separation are in good agreement with predicted focusing behavior in the model. This technique is able to separate or concentrate organelles such as mitochondria and peroxisomes in a few minutes, which is greatly reduced from conventional techniques. Experiments demonstrate the separation of nuclei and whole cells from mitochondria. Additionally, the devices are used to distinguish mitochondria with intact membrane potential from those that have lost transmembrane potential. This ability to distinguish the mitochondria populations could potentially be used to assay whether cells have committed apoptosis via the mitochondrial pathway. The modules developed in this thesis demonstrate advantages of scaling down bioanalytical processes. Further developing and combining the technologies demonstrated in this thesis will enable a platform for parallel, fast, and automated cell dynamics and proteomics studies for systems biology.
by Hang Lu.
Ph.D.
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Westmoreland, D. A. "Lysosome-related organelles : an investigation into clinical disorders of endothelial cells and platelets." Thesis, University College London (University of London), 2017. http://discovery.ucl.ac.uk/1561240/.
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Lysosome-related organelles (LROs) are a heterogeneous group of organelles that have important functions in a number of specialised cell types. LROs, despite their distinct features and morphology, have been grouped together due to the observation that they are simultaneously functionally perturbed by single mutations in a number of genetic disorders, yet as a group they are still poorly understood. Firstly, it was investigated whether the genes that are important for the formation/maturation of other LROs can also affect Weibel-Palade bodies (WPBs) an endothelial LRO that is critical to haemostasis and inflammation. In the genetic disorder Hermansky Pudlak syndrome (HPS) a number of LROs are affected, but the effect of these mutations on WPBs is not yet established. It was investigated whether these genes are indeed important for the biogenesis and function of WPBs, potentially revealing a new aspect of the disease phenotype. siRNA ablation in human endothelial cells of genes identified as involved in LRO biogenesis proved to give inconclusive results as to their importance in WPB formation and function. Secondly, the understanding of LRO-related genetic disorders would be aided by an improvement in diagnostics. The diagnosis of platelet storage disorders (PSDs) is currently limited to the observation of symptoms (e.g. a bleeding disorder or albinism) that are often shared with other, more common diseases. Most HPS patients are initially misdiagnosed and many see 4 to 6 specialists before being correctly identified. I investigated whether Super Resolution Microscopy, allowing images to be taken with a higher resolution than the diffraction limit (< 200 nm), has the potential for improving the imaging of platelet granules and thereby the diagnosis and characterisation of LRO-related disorders. The use of structured illumination microscopy, coupled with automated image analysis bioinformatics allowed for a highly efficient differentiation between control and patient platelets.
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Rivalin, Romain. "Intégration de la régulation post-transcriptionnelle et des interactions avec le cytosquelette clans les voies de contrôle du métabolisme mitochondrial." Angers, 2013. https://tel.archives-ouvertes.fr/tel-01022923/document.
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La mitochondrie fournit l'énergie nécessaire au fonctionnement cellulaire, grâce au mécanisme de phosphorylation oxydative. Cette fonction nécessite une expression coordonnée des génomes nucléaires et mitochondriaux assurée par la famille de coactivateurs transcriptionnels PGC-I (Peroxisome proliferatoractivated receptor V Coactivator-l), sensibles aux signaux endogènes et/ou environnementaux. Une régulatioh plus fine de la phosphorylation oxydative par des miRNAs est maintenant soupçonnée. Afin de préciser ces différents modes de régulation dans des modèles cellulaires de carcinomes thyroïdiens, nous avons exploré la voie PRC-dépendante (PGC-related coactivator) et les miRNAs spécifiquement exprimés dans ces modèles présentant une richesse en mitochondries et des niveaux de PRC et de PGC-Ia différents. Ce travail a permis de mettre en évidence miR-218 comme marqueur clé de régulation de la fonction mitochondriale. Au-delà de la régulation de l'expression génique, une fourniture énergétique adéquate nécessite également une répartition optimale des mitochondries au sein de la cellule, grâce à d'étroites connexions entre le cytosquelette et la mitochondrie. Des peptides issus de la sous-unité légère des neurofilaments, dont le NFL-TBS. 40-63, sont capables d'entrer spécifiquement dans les cellules de glioblastomes humains et d'y déstabiliser le réseau microtubulaire, conduisant à la mort cellulaire par apoptose. Pour étudier I'impact de ce peptide sur le réseau de mitochondries et leurs fonctions, nous avons traité le modèle cellulaire de glioblastomes humains T98G, par différentes concentrations de NFL-TBS. 40-63. Ce travail révèle une perturbation du réseau de mitochondries et une diminution de la respiration mitochondriale dans les cellules exposées. L'ensemble de ces travaux doit permettre le développement de traitements ciblés de la fonction mitochondrialeMitochondrion provides energy for cell metabolism through the mechanism of oxidative phosphorylation. This function requires a coordinated expression of nuclear and mitochondrial genomes provided by the family of transcriptional coactivators PGC -1 (peroxisome proliferator-activated receptor V Coactivator -1), responding to endogenous and/or environmental signals. A fine regulation of the oxidative phosphorylation by miRNAs is now suspected. To specify these regulatory pathways in cellular models of human follicular thyroid carcinomas, we have explored the PRC-related (PGC- related coactivator) pathway and specific microRNAs, in models presenting various mitochondrial abundance and differences in PRC and PGC- la expression levels. We have highlighted the role of miR-218 as a key regulatory factor of mitochondrial functions. An adequate energy supply also requires close connections between cytoskeleton and mitochondria to ensure an optimal distribution of mitochondria within the cell. Peptides derived from the light neurofilament subunit, as NFL - TBS. 40 -63, are able to specifically enter into human glioblastoma cells and destabilize the microtubule network, leading to cell death by apoptosis. To study the impact of this peptide on mitochondrial network and oxidative phosphorylation, we have treated the T98G human glioblastoma cells by different concentrations of NFL- TBS. 40 -63. Our work showed disturbance in mitochondrial network and reduction in mitochondrial respiration rate in the treated cells. All these results should allow the development of therapy targeting the mitochondrial function
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Fisher, Joshua. "Mitochondrial adaptions in the placenta during pregnancy." Thesis, Griffith University, 2020. http://hdl.handle.net/10072/394314.
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The placenta is a unique organ critical for the growth and development of the fetus. The placenta provides the link between maternal and fetal circulations, supplying nutrients and oxygen, removing waste and regulating metabolic and hormonal responses in both mother and fetus. The placenta develops from a single cell mass with a common cell progenitor diverging down multiple pathways responsible for the development of many cell lineages. Among these cell lineages is the villous trophoblast which form the junction for the aforementioned maternal and fetal interface. This junction consists of underlying cytotrophoblast cells which fuse and differentiate into a multinucleated syncytium. During this transformation from cytotrophoblast to syncytiotrophoblast, the organelles which accompany these cell types also undergo morphological and functional changes. This PhD will investigate these changes in organelle morphology and function.
Cell studies on transformed placental trophoblast cells, Swan-71’s, in Chapter 2 examined the effect of endoplasmic reticular (ER) stress on mitochondrial function and dynamics under acute and chronic exposure. The aim of this work was to address the interactions between mitochondria and endoplasmic reticular stress which converges on pathways often associated with mitochondria dysfunction, that have been proposed to play a role in gestational disorders. It has been established that alterations in ER stress effects mitochondrial functionality in different ways dependent on exposure time and the pathway of ER stress initiated. Mitochondrial alterations in bioenergetics, reactive oxygen species production, mitochondrial dynamics and key metabolic proteins associated with differentiation were assessed under normal conditions and after induction of ER stress.
While finding that ER stress does initiate mitochondrial dysfunction, the translatability of such a finding was difficult to ascertain due to examining a trophoblast precursor cell (most similar to the cytotrophoblast) containing a mostly uniform mitochondrial population in contrast to the vastly different populations observed in the placenta. Investigations in Chapter 3 aimed to optimise isolation methodology enabling the characterisation of mitochondria from the cytotrophoblast and syncytiotrophoblast cell lineages. These studies provided an experimental model to assess the bioenergetic and metabolic alterations between these different mitochondria, laying a foundation for future studies to be translated and the continued examination of mitochondrial interactions in pathologies. Chapter 4 utilised the optimized protocol to generate a proteomic profile of the mitochondrial populations from the placenta post differentiation, identifying key proteins involved in the mechanisms which may drive this previously speculative process. Through this characterisation we established a more comprehensive understanding of mitochondrial transformations associated with trophoblast differentiation. We further investigated altered protein expression patterns in carbohydrate, fatty acid and amino acid metabolism, in combination with key structural proteins within the electron transport chain that were significantly altered. These findings expanded our knowledge and proposed mechanisms by which the previously observed alterations in mitochondrial functionality may occur.
This was followed by an investigation into gestational disorders in Chapter 5 through examination of gestational diabetes mellitus (GDM) in placental samples. The aim of this investigation was to examine bioenergetic and metabolic pathways to determine if mitochondrial dysfunction occurred as a result of alterations in the previously identified mechanisms and proteins; with the specific intent to assess mitochondria from both cell lineages individually. The purpose of this was to ascertain if contradictory findings on mitochondrial function in GDM were as a result of whole tissue analysis, therefore not accounting for the differing mitochondrial populations present in the placenta.
Overall, the findings presented in this thesis establishes that mitochondria and the ER are intrinsically linked through functionality, the implication of which require further investigation in the placenta. However, with the comprehensive characterisation of the two mitochondrial populations present in the cytotrophoblast and syncytiotrophoblast, this interaction can now be more thoroughly studied. This body of work identifies key mitochondrial pathways which are altered between the two populations, identifying mechanisms which may drive mitochondrial transformation within the placenta. Observations which have previously been speculative based on morphological appearance alone. This thesis provides a well characterised physiological basis for future work into placental mitochondria. Further, this body of work forms a critical foundation when assessing the importance of mitochondria in pathologies. While often linked to pathologies through mitochondrial dysfunction, this thesis highlights the importance of a sessing isolated mitochondria from pathological placentae in order to minimise the often contradicting results in the literature obtained due to examination of whole tissue, thereby, not accounting for the different physiology of the mitochondrion in the placenta. This body of work identified that mitochondrial appear dysfunctional in GDM and proposed mechanisms which may be involved in the pathogenesis of the disorder although mechanisms underpinning this require further examination.Thesis (PhD Doctorate)
Doctor of Philosophy (PhD)
School of Medical Science
Griffith Health
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Kroeger, Benjamin Robert. "The genetic regulation and subcellular dynamics of secretory and endolysosomal organelles of Drosophila secondary cells." Thesis, University of Oxford, 2017. https://ora.ox.ac.uk/objects/uuid:dce9ae14-b03d-4fca-8429-de839cc40d6a.
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Secretory processes underpin the emergence of cellular diversity in complex multicellular organisms. However, our understanding of the basic mechanisms controlling the different secretory and endosomal compartments involved remains surprisingly incomplete. During my DPhil I have studied a specialised epithelial cell type in the male Drosophila accessory glands, the secondary cell, which contains unusually large intracellular compartments that are accessible to detailed morphological study. I characterise the organisation, ultrastructure and molecular composition of this cell's secretory and endosomal compartments, and I employ specific Rab GTPases, conserved coordinators of membrane trafficking and identity, to define multiple compartmental subtypes. By developing super-resolution and time-lapse microscopy approaches in these cells, I show that numerous intraluminal vesicles (ILVs) are formed within Rab11-labelled secretory compartments and released into the accessory gland lumen as exosomes, the first clear demonstration in eukaryotic cells of exosome biogenesis within a non-late endosomal compartment. Biogenesis of these ILVs is dependent on evolutionarily conserved Endosomal Sorting Complexes Required for Transport (ESCRT) 0-III genes and involves loading of compartment-specific cargoes. Work by others, some in collaboration with me, has shown that these novel mechanisms are conserved in human cells. I show that dense-core granules, the structures employed to package proteins and other molecules destined for regulated secretion, form within large non-cored Rab6- positive compartments, in a process that seems to involve inputs from both the Golgi and recycling endosomal pathways. Further analysis has revealed roles for specific Rabs, for ILVs, and for the conserved fibrillar protein Mfas/TGFBI in different aspects of DCG formation. I also show that DCGs are not only secreted, but can also be degraded by fusion to acidic endosomal compartments. Remarkably, there is evidence that mammalian cells may employ all of these mechanisms and defects in these processes may be linked to diseases like cancer, diabetes and neurodegenerative disorders. Hence my work has established a new system to study complex secretory mechanisms, which can now be developed to model specific disease processes in the future. In summary, I have discovered several novel cell biological mechanisms controlling exosome biology, dense-core granule biogenesis, regulated secretion, and endolysosomal trafficking. Some of these already appear relevant to human health and disease, suggesting that the secondary cell system has considerable further potential for unravelling the fundamental processes underlying eukaryotic secretion in the future.
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Ahmadi, Shiva [Verfasser]. "Application of BioID to in vitro organelle and in vivo cell-type-specific proteomics / Shiva Ahmadi." Bonn : Universitäts- und Landesbibliothek Bonn, 2020. http://nbn-resolving.de/urn:nbn:de:hbz:5-59196.
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Sunderij, Abid. "The effect of Epstein-Barr virus lytic cycle on the secretory pathway organelles in B cells." Thesis, London School of Hygiene and Tropical Medicine (University of London), 2011. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.549747.
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Qattan, A. T. M. "Large scale quantitative organelle proteomics of protein distribution in breast cancer MCF-7 cells." Thesis, University College London (University of London), 2013. http://discovery.ucl.ac.uk/1415963/.
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This thesis analyzes the dynamic complexity involved in the subcellular distribution of the proteins for the malignant breast epithelial MCF-7 cell line, using mass spectrometry based quantitative proteomics for the indirect measurement of the subcellular dispersion of the constituent proteins of core cellular functions. The thesis demonstrates that there are many proteins which can be present in more than one subcellular organelle. Quantitative proteomics using LFQP (Label-Free Quantitative Proteomics) methodology based on mass spectrometry was shown to be suitable for the indirect measurement of the distribution of the proteins in the malignant breast epithelial cell line. The study used partial purification by means of dynamic sucrose gradient centrifugation to avoid the need of multiple purification procedures for different organelles and loss of proteins during purification. This was followed by proteomics identification and analysis of the protein content from the sucrose gradient fractions corresponding to the major organellar compartment. These included the nucleus, cytosol, mitochondria, plasma membrane and endoplasmi c reti cul um. The first part of the thesis indicates that 50.00% - 75.00% of the proteins detected showed multiple-locations. Out of the total quantified proteome using LFQP methodology, 2184 proteins were securely identified. 481 proteins (22.00%) were found in unique sucrose gradient fractions which suggest that they may have unique locations, while 454 proteins (20.80%) were found to be ubiquitously distributed and the remaining 1249 proteins (57.20%) were consistent with intermediate distribution over multiple locations. 94 proteins implicated in breast cancer and 478 other proteins which share the same major cellular biological processes with most of the breast cancer proteins were observed in 334 and 1223 subcellular locations respectively. The second part of the thesis concentrates on the spatial distribution of proteins between two organelles of particular interest for cancer: the nucleus and mitochondria. Two important characteristics of cancer cellular function include high degrees of genetic instability and major changes in cellular energy metabolism. The genetic instability, which is associated with the cell nucleus, allows cells to escape from a variety of normal restrictions on proliferation, whereas the changes in energy metabolism are associated with mitochondria and the need for new cellular components to be produced in the proliferating cells. The large scale proteomics analysis of the partitioning of proteins between mitochondria and the nucleus reveals that 40.00% of all the proteins were shared between the mitochondria and the nucleus. The observed partitioning of these proteins between these two organelles showed a functional distribution which is consistent with the first part of the thesis. The analysis of the distribution between the nucleus and mitochondria of specific subgroups of the proteins involved in oxidative phosphorylation, the tricarboxylic acid cycle, RNA processing/translation, glycolysis and Ras-related signalling suggests that the spatial distribution of numerous proteins over multiple sites is critical to cellular function and that there are unrecognized aspects of functional coordination between the mitochondria and the nuclei which need further investigation. In addition, the large number of proteins identified to be in multiple locations indicates that subcellular spatial integration of function may be a vital aspect of cancer. The findings in this study also reveal that most of the observed proteins with multiple subcellular locations had current annotations of location which are still sparse in public databases. In general terms, the extensive subcellular dispersion of the constitute proteins of core cellular functions may be a fundamental feature of the cell which may be constituent with the requirements for robustness in a complex system.
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Hatchel, Jennifer M. "Structure and Function of the Electron-dense Core in Mycoplasma pneumoniae and its Relatives." Miami University / OhioLINK, 2009. http://rave.ohiolink.edu/etdc/view?acc_num=miami1248183957.
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Matsuzaki, Satoshi. "Hole Burning Imaging Studies of Cancerous and Analogous Normal Ovarian Tissues Utilizing Organelle Specific Dyes." Ames, Iowa : Oak Ridge, Tenn. : Ames Laboratory ; distributed by the Office of Scientific and Technical Information, U.S. Dept. of Energy, 2004. http://www.osti.gov/servlets/purl/837275-3aN4nd/webviewable/.
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Thesis (Ph.D.); Submitted to Iowa State Univ., Ames, IA (US); 19 Dec 2004.Published through the Information Bridge: DOE Scientific and Technical Information. "IS-T 2692" Satoshi Matsuzaki. US Department of Energy 12/19/2004. Report is also available in paper and microfiche from NTIS.
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Asiri, Sumayyah. "Role of Cu metabolism in the cisplatin-sensitive and resistant ovarian cancer cells." Thesis, The University of Sydney, 2022. https://hdl.handle.net/2123/29652.
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Ovarian cancer is the fourth most common cancer among women worldwide, leading to high mortality rates. Platinum anti-cancer drugs have been widely used in the treatment of ovarian cancers, but the development of resistance to these drugs is a rapidly growing impediment for their clinical use. Studies of the cell morphology using IncuCyte Zoom and HoloMonitor M4 have shown that A2780.CisR cells contained heterogeneous cell populations that partially underwent epithelial-to-mesenchymal transition (EMT). Mesenchymal A2780.CisR cells were likely to be more resistant to a cytotoxic natural killer (NKL) cell line, compared with the epithelial A2780 cells. Mesenchymal phenotype was associated with lower total Cu content and higher total Fe content in A2780.CisR compared with A2780 cells under physiologically relevant conditions (2.0 M Cu(II) or 10 M Fe(III) for 24 h). Conversely, live cell confocal microscopy studies with a novel ratiometric Cu(I)-sensitive fluorescent dye, InCCu1, revealed higher cellular content of labile Cu in A2780.CisR cells compared with A2780 cells. Moreover, the InCCu1 dye showed promise for differential staining of multiple cellular organelles, including mitochondria, endosomes/lysosomes, fat droplets and budding extracellular vesicles. Fat droplets were more abundant in A2780.CisR compared with A2780 cells and concentrated at the advancing edges of cellular protrusions (shown by InCCu1 and Nile Red staining). Comparison of the biochemical content of cell membranes using ATR-FTIR (attenuated total reflection Fourier transform infrared) spectroscopy technique showed higher relative content of fatty acids and cholesterol in the membranes and surrounding environments of A2780.CisR compared with A2780 cells.
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Latge, Bruno. "Rôle des moteurs moléculaires dans l'établissement de la polarité cellulaire." Electronic Thesis or Diss., Paris Sciences et Lettres (ComUE), 2017. http://www.theses.fr/2017PSLET023.
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La polarité cellulaire est une caractéristique fondamentale du fonctionnement des cellules et de l’homéostasie tissulaire. Elle se définit par une distribution asymétrique des constituants cellulaires le long d’un axe de polarité et est le résultat d’une interaction complexe entre des mécanismes intrinsèques et extrinsèques. Les voies de signalisation œuvrant pour l’établissement de la polarité cellulaire et l’organisation anisotrope du cortex cellulaire ont été intensément étudiés depuis des décennies. Cependant, l’implication du positionnement des organites dans la mise en place de la polarité cellulaire et les mécanismes sous-jacents restent encore méconnus. Les moteurs moléculaires sont responsables de l’attachement des organites au cytosquelette et de leurs mouvements. Ils jouent aussi des rôles prépondérants dans l’établissement et le maintien de la polarité cellulaire. Cette thèse a pour but d’étudier les mécanismes moléculaires par lesquels les protéines motrices de la famille des kinésines régulent le positionnement des organites et leur rôle dans la mise en place de la polarité cellulaire. Nous avons combiné l’utilisation de micro-patrons contrôlant l’adhésion des cellules et l’analyse quantitative de l’organisation intracellulaire pour identifier les kinésines impliquées dans le positionnement des organites. Les résultats de notre crible de petits ARN interférents ciblant 43 kinésines humaines suggérèrent un rôle important des kinésines dans la distribution des organites intracellulaires le long de l’axe de polarité antéro-postérieur. Le phénotype le plus marquant a été observé après inhibition de l’expression de la kinésine-1 Kif5B. En permutant les organites autour du centrosome, l’organisation des cellules micro-patronnées en forme d’arbalète était inversée. De plus, lors de la migration cellulaire, l’absence de Kif5B ralentissait le réalignement des organites le long de l’axe de polarité antéro-postérieur, survenant suite à un changement de direction. Ces cellules se sont aussi révélées être moins persistantes. Nous avons alors émis l’hypothèse que les mouvements d’organites induits par Kif5B s’intégraient dans une réponse cellulaire globale, liée à l’alignement des organites le long de l’axe de polarité antéro-postérieur. Disséquant le mécanisme, nous avons montré que les défauts de positionnement des organites observés en l’absence de Kif5B étaient indépendants de l’organisation anisotrope du cortex cellulaire. Il était en revanche corrélé à une augmentation significative de la distance entre le noyau et le centrosome. Etonnamment, l’inhibition de l’expression de RanBP2, un partenaire de Kif5B localisé à l’enveloppe nucléaire, entrainait aussi la permutation des organites. Nous proposons un modèle selon lequel Kif5B, en lien avec RanBP2, coordonnerait le positionnement du noyau, en organisant une cage de microtubules autour. De plus, nous avons mis en évidence le rôle indirect de KifC3 dans le positionnement des organites intracellulaires. Révélé dans notre crible des kinésines, KifC3 contrôle le mouvement centripète des lysosomes, mais localise au centrosome. Le recrutement de KifC3 au centrosome a été caractérisé. Nous avons observé une accumulation préférentielle de KifC3 au centriole père qui était dépendant de Cep170. Nous avons alors proposé que KifC3 soit localisé aux appendices subdistaux du centrosome, grâce à Cep170, et régule l’ancrage des microtubules. KifC3 agirait alors sur l’organisation globale du cytosquelette de microtubules. Ensemble les résultats de cette thèse ont permis d’attirer l’attention sur les fonctions cellulaires générales de deux kinésines, Kif5B et KifC3, dans le control de l’organisation intracellulaire sur laquelle s’appuie la polarité cellulaireCellular polarity is instrumental for normal cell function and tissue homeostasis. It is defined by an asymmetrical distribution of cellular constituents along a polarity axis and results from a complex interplay of intrinsic and extrinsic mechanisms. Cell polarity signaling and cortex anisotropy have been extensively studied in the last decades. However, the role of organelle positioning in cell polarity establishment and the mechanisms by which polarized positioning is achieved are still not well understood. Molecular motor proteins link organelles to cellular cytoskeleton and play an essential role in cell polarity establishment and maintenance. This PhD thesis aimed at the study of the molecular mechanisms by which motors of the kinesin family regulate polarized organelle positioning and their role in cell polarity establishment. We took advantage of our approach that combines cell micropatterning and quantitative analysis of the intracellular organization to identify kinesins that contribute to organelle positioning. Results from our siRNA-based screening targeting 43 members of the kinesin family in human suggested a central role of kinesins in front-rear polarized alignment of intracellular organelles. The strongest phenotype was observed upon kinesin-1 Kif5B depletion that inverted the organization of crossbow-shaped micropatterned cells by flipping organelles around the centrosome. Additionally, Kif5B-depleted cells were constrained in their abilities to re-align organelles along the front-rear polarity axis after directional changes during migration and were less persistent. We therefore hypothesized that Kif5B-dependent organelle movement integrates into a global cellular response, relying on the polarized organelle alignment along the front-rear axis. Dissecting the mechanism, we showed that organelle positioning defects upon Kif5B depletion were independent of the cell cortex anisotropy and correlated with substantial increase in the distance between the nucleus and the centrosome. Unexpectedly, depletion of the Kif5B-interacting partner RanBP2, which is localized at the nuclear envelope, copied the organelle inversion phenotype. We propose a model in which Kif5B, together with RanBP2, controls nucleus positioning by organizing a microtubule scaffold around it. We have additionally evidenced the indirect role of KifC3 in organelle positioning. Identified as a hit in our screen analysis, KifC3 controled the centripetal movement of lysosomes, but localizes to the centrosome. Characterizing the recruitment of KifC3 to the centrosome, we showed its preferential accumulation at the mother centriole that was dependent on Cep170. We propose that KifC3 is localized at the subdistal appendages of centrosomes through Cep170 and regulates microtubule anchorage, and thus,the global microtubule organization. Together the results of this PhD shed light on the global cellular functions of two kinesins, Kif5B and KifC3, in controlling the intracellular organization that supports cell polarity
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Ozanne, Angelica. "Investigations into the formation of pdu microcompartments in mammalian and bacterial cells as novel 'organelles' for protein folding and post-translational modifications." Thesis, University of Kent, 2013. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.594270.
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In some bacteria proteinaceous polyhedral structures known as microcompartments (MCPs) are formed that encapsulate particular metabolic processes. One such bacterial Mep is the 1,2-propanediol utilisation (Pdu) MCP expressed in Citrobacter freundii and it is possible to synthesise functional pdu MCPs in Esherichia coli (Eeoli) by transferring the pdu operon from Citrobaeter freundii into Ecolf. Subsequent investigations have shown that if only the genes encoding for the pdu MCP shell proteins are transferred into E.coli empty pdu MCP structures are formed. The minimum number of genes thought to be necessary to form a recombinant MCP structure is five. In addition, if only one of these essential genes (PduA) is expressed in E-coli then large structures are observed which are either hexagonallattice like in appearance or long and thin spanning much of the E.coli cell. Importantly for this project, recent work has also demonstrated that proteins can be selectively targeted to within the pdu MCPs using specific N-terminal sequence tags. This IJ project set out to investigate the formation of MCPs in mammalian and bacterial cells as novel 'organelles' for protein folding and post-translational modifications. Initially, it was investigated whether the essential pdu shell proteins required to form the MCP can be expressed in CHOKI cells and whether upon expression these can subsequently form proteinaceous structures within such cells. Using western blotting and fluorescent immunostaining it was apparent that individual pdu shell proteins with a V5 tag could be transiently expressed in CHOKI cells but stably expressing cell lines could not be established. However, stable cell lines expressing OFP tagged pdu proteins could be generated although the level of expression was much less than OFP alone cell lines. Electron microscopy analysis revealed that the transient expression of OFP-pduA or all five pdu shell protein with a OFP tag generated in some cells structures that resembled those seen in E.coli when the ratio of subunits were manipulated. This data suggests that pdu protein based structures can be formed in mammalian cells, however a threshold of expression must be obtained before organised structure are obseIVed which only occurred upon transient expression. Stable cell lines did not presumably exceed this threshold and hence organised structures were not observed. Following on from these studies, MCPs were successfully generated in E.coli and specific proteins targeted to within these using N-terminal tags. To determine whether the targeting of proteins to the MCPs could enhance protein folding, the protein vtPA, which requires the correct formation of a number of disulphide bridges to be functionally active, was targeted to the MCP with or without PDL When vtPA alone was targeted to the MCP an increase in vtPA activity was observed over the control and although the additional targeting ofPDI also led to an increase in activity over the control, the activity was less than when vtPA alone was targeted. This is likely to be the result of dilution of expression as the bacteria was making two proteins (vtPA and PDI) as opposed to one alone and hence development of bacterial strains into which the pdu and PDI genes were integrated into the genome would in all likelihood be more beneficial for enhanced expression and folding of a single protein. In conclusion, the work presented here has demonstrated that pdu proteins and organised structures can be formed in mammalian cells and that the targeting of specific proteins to MCPs in bacterial systems offers the potential to develop a novel environment for improved protein folding.
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Ludewig-Klingner, Ann-Kathrin Verfasser], and Jörn [Akademischer Betreuer] [Petersen. "From Malaria to the Sparkling of the Sea - Organelle and Host Cell Evolution in Alveolates (Apicomplexa, Dinoflagellates, Ciliates) / Ann-Kathrin Ludewig-Klingner ; Betreuer: Jörn Petersen." Braunschweig : Technische Universität Braunschweig, 2017. http://d-nb.info/1175816825/34.
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Danylchuk, Dmytro. "Environment-sensitive targeted fluorescent probes for live-cell imaging." Thesis, Strasbourg, 2021. http://www.theses.fr/2021STRAF012.
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Le ciblage, l'imagerie et le sondage spécifiques des membranes plasmiques et des organites intracellulaires peuvent être faits par des sondes fluorescentes à façon sensibles à la polarité. Ici, un nouveau fragment ciblant la membrane plasmique à été développé et testé dans cinq colorants cyanines, montrant d'excellentes performances en microscopie cellulaire et in vivo. Le fragment à été greffé à un fluorophore solvatochrome Prodan, donnant une sonde de membrane plasmique avec une sensibilité élevée à l'ordre lipidique. Le rouge de Nil, greffé aux fragments avec les chaînes alkyles C12 et C4, à donné deux sondes solvatochromes à membrane plasmique : NR12A pour la microscopie conventionnelle, et NR4A pour la microscopie à super-résolution PAINT. Le rouge de Nil avec des groupes ciblant les organites à donné un éventail de sondes sensibles à la polarité et à l'ordre lipidique dans les membranes des organites. Les sondes synthétisées trouveront des applications en bioimagerie, biologie cellulaire, biophysique ou mécanobiologieSpecific targeting, imaging and probing of cell plasma membranes and intracellular organelles can be addressed by rationally designed polarity-sensitive fluorescent probes. Here, a new efficient plasma membrane-targeting moiety was developed and tested in five cyanine dyes, showing excellent performance in cellular and in vivo microscopy. Next, the targeting moiety was grafted to a solvatochromic dye Prodan, yielding a plasma membrane probe with high lipid order sensitivity. Modifying a Nile Red using the moieties with varied alkyl chain lengths resulted in two solvatochromic plasma membrane probes: NR12A with high affinity to membranes for conventional microscopy, and NR4A, a low-affinity probe for PAINT super-resolution microscopy. Tethering Nile Red with organelle-targeted groups yielded an array of probes, able to sense polarity and lipid order in organelle membranes. The synthesized probes will find applications in bioimaging, cell biology, biophysics or mechanobiology
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Murray, Iain Colquhoun. "The immunohistochemical localization of basement membrane components to secretory organelles is observed in the youngest endothelial cells of the rat incisor, suggesting that the synthesis of basement membrane components occurs mainly in young cells." Thesis, McGill University, 1986. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=65989.
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Marguet, Maïté. "Vésicules polymères biomimétiques : du virus à la cellule." Thesis, Bordeaux 1, 2012. http://www.theses.fr/2012BOR14717/document.
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Les polymersomes, obtenus par auto-assemblage en solution aqueuse de copolymères à blocs amphiphiles en structure vésiculaire, sont présentés comme d’excellent mimes synthétiques des virus, dont les propriétés membranaires – principalement élasticité, perméabilité, fonctionnalité- peuvent être très proches. Il y a ainsi un fort engouement quant à leur utilisation en biotechnologie et surtout en vectorisation d’actifs pharmaceutiques ou cosmétiques. Afin d’aller encore plus loin dans le biomimétisme ou la bio-inspiration, une étape devait être franchie : encapsuler ces polymersomes les uns dans les autres. Ce cloisonnement ou multi-compartimentalisation permet de mimer cette fois la structure d’une cellule dite eukaryote, elle-même constituée de compartiments internes (organelles) et d’un cytoplasme (lui conférant entre autres une certaine stabilité mécanique) contenues dans le compartiment externe représenté par la membrane cellulaire. Toutefois, l’obtention d’un simple mime structural d’une structure si complexe représente déjà un challenge en soi, nécessitant maîtrise de la physico-chimie des systèmes, de la stabilisation des interfaces et des outils de formulation. Une méthode d’émulsion-centrifugation a été développée et a permis d’obtenir de telles structures compartimentalisées (mimes d’organelles) à cavité gélifiée (mime de cytoplasme). Finalement, différentes voies d’exploitation de ces systèmes sont présentées, allant de l’encapsulation multiple, la libération contrôlée jusqu’au développement de réactions enzymatiques en cascade confinées, mimant ainsi le métabolisme cellulaireAmphiphilic block copolymers self-assemble in water into vesicles, coined “polymersomes”; these vesicles are described as excellent synthetic mimics of viral capsids due to the resemblance of their respective membrane properties (in terms of elasticity, permeability, and functionality). As a result, they were massively investigated over the last years regarding applications in biotechnology and more particularly for the targeted delivery of pharmaceutical or cosmetic actives.In order to go further towards bio-inspiration and cell biomimicry, the next step required the encapsulation of polymersomes in other polymersomes. This multicompartmentalization indeed enables to mimic the structure of an eukaryotic cell; an outer cellular membrane compartment encloses internal compartments (organelles) and a cytoplasm responsible amongst others for a certain mechanic stability. However, alone the controlled formation of a system mimicking such a complex structure represents a technological challenge in terms of control over the physical chemistry of these systems, the stabilization of their interfaces and their formulation. A formation method based upon an emulsion-centrifugation has been developed and enabled the formation of such multicompartmentalized structures (organelle mimics) with a gelified lumen (cytoplasm mimic). Finally, various potential applications of these systems are presented: from multiple encapsulation, controlled drug release, to the development of enzymatic and confined cascade reactions that mimick the cellular metabolism
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Arias, Hidalgo Mariela Eugenia [Verfasser], GEROLF [Akademischer Betreuer] GROS, and Anaclet [Akademischer Betreuer] Ngezahayo. "Adaptation of the CO2 permeability of various cells and organelles to their specific metabolic needs / Mariela Eugenia Arias Hidalgo ; Akademische Betreuer: Gerolf Gros, Anaclet Ngezahayo ; Abteilung Molekular- und Zellphysiologie, AG Vegetative Physiologie 4220." Hannover : Bibliothek der Medizinischen Hochschule Hannover, 2018. http://d-nb.info/1151400629/34.
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Arias, Hidalgo Mariela Eugenia [Verfasser], Gerolf [Akademischer Betreuer] Gros, and Anaclet [Akademischer Betreuer] Ngezahayo. "Adaptation of the CO2 permeability of various cells and organelles to their specific metabolic needs / Mariela Eugenia Arias Hidalgo ; Akademische Betreuer: Gerolf Gros, Anaclet Ngezahayo ; Abteilung Molekular- und Zellphysiologie, AG Vegetative Physiologie 4220." Hannover : Bibliothek der Medizinischen Hochschule Hannover, 2018. http://nbn-resolving.de/urn:nbn:de:gbv:354-2017111592.
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Stadler, Charlotte. "Towards subcellular localization of the human proteome using bioimaging." Doctoral thesis, KTH, Proteomik, 2012. http://urn.kb.se/resolve?urn=urn:nbn:se:kth:diva-103616.
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Since the publication of the complete sequence of the human genome in 2003 there has been great interest in exploring the functions of the proteins encoded by the genes. To reveal the function of each and every protein, investigation of protein localization at the subcellular level has become a central focus in this research area, since the localization and function of a protein is closely related. The objective of the studies presented in this doctoral thesis was to systematically explore the human proteome at the subcellular level using bioimaging and to develop techniques for validation of the results obtained. A common imaging technique for protein detection is immunofluorescence (IF), where antibodies are used to target proteins in fixated cells. A fixation protocol suitable for large-scale IF studies was developed and optimized to work for a broad set of proteins. As the technique relies on antibodies, validation of their specificity to the target protein is crucial. A platform based on siRNA gene silencing in combination with IF was set-up to evaluate antibody specificity by quantitative image analysis before and after suppression of its target protein. As a proof of concept, the platform was then used for validation of 75 antibodies, proving it to be applicable for validation of antibodies in a systematic manner. Because of the fixation, there is a common concern about how well IF data reflects the in vivo subcellular distribution of proteins. To address this, 500 proteins were tagged with green fluorescent protein (GFP) and used to compare protein localization results between IF to those achieved using GFP tagged proteins in live cells. It was concluded that protein localization data from fixated cells satisfactory represented the situation in vivo and together exhibit a powerful approach for confirming localizations of yet uncharacterized proteins. Finally, a global analysis based on IF data of approximately 20 % of the human proteome was performed, providing a first overview of the subcellular landscape in three different cell lines. It was found that the intracellular distribution of proteins is complex, with many proteins occurring in several organelles. The results also confirmed the close relationship between protein function and localization, which in a way further strengthens the accuracy of the IF approach for detection of proteins at the subcellular level.QC 20121017
The Human Protein Atlas
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Vieira, Moema Queiroz. "Análise ultraestrutural de células-tronco mesenquimais humanas derivadas de tecido adiposo (hADSC) durante a diferenciação adipogênica : interações entre as gotas lipídicas citoplasmáticas e outras organelas." reponame:Biblioteca Digital de Teses e Dissertações da UFRGS, 2013. http://hdl.handle.net/10183/107632.
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As células-tronco mesenquimais humanas derivadas de tecido adiposo, do inglês human adipose-derived stem cells (hADSC), são células progenitoras que residem entre adipócitos e armazenam lipídios neutros, principalmente triglicerídeos e ésteres de colesterol (TG e EC), em gotas lipídicas citoplasmáticas (GLC), contribuindo para o turnover do tecido adiposo. As GLC são organelas que desempenham um papel crucial na homeostasia energética e no metabolismo dessas células. Caveolas são invaginações de 50-100 nm que foram inicialmente caracterizadas por microscopia eletrônica. A forma e a organização estrutural das caveolas deve-se a proteínas especificas da família das caveolinas (caveolina-1, -2 e -3) que se associam em oligômeros para formar cavidades/invaginações na membrana plasmática. A função das caveolinas na formação das GLC está relacionada com a captação de ácidos graxos e seu metabolismo, e a relação existente entre estes dois componentes celulares parece ser crucial para manutenção da homeostasia celular. Muitas organelas que são funcionalmente conectadas ao metabolismo de lipídeos são encontradas justapostas as GLC. Distinções morfológicas observadas reforçam diferenças que podem existir na maneira pelas quais as GLC interagem com outras organelas em adipócitos. Sítios de contatos entre membranas, do inglês membrane contact sites (MCS), são descritos para muitas organelas e parecem funcionalmente importantes nos processos de interações entre as GLC e outras organelas celulares. O presente trabalho avaliou as diferenças ultraestruturais entre hADSC diferenciadas ou não para pré-adipócitos, comparando com células 3T3-L1. Também foram avaliadas, através de microscopia eletrônica de transmissão, as interações das GLC com outras organelas celulares durante a diferenciação adipogênica, pois apesar de sua importância no metabolismo energético e em várias doenças, as GLC são pouco compreendidas como organelas celulares. De fato, o tamanho, a composição e a regulação das GLC variam consideravelmente entre organismos e tipos celulares. A complexidade das interações das GLC com outras organelas também varia consideravelmente em adipócitos e não adipócitos. Este trabalho mostrou a importância de estudos que visam esclarecer como as GLC são formadas, modificadas e reguladas. Através destes estudos poderemos ter uma melhor compreensão acerca da relação existente entre o acúmulo excessivo de lipídios no organismo e a chamada síndrome metabólica (obesidade, diabetes e aterosclerose).Human adipose-derived stem cells (hADSC) are progenitor cells that reside between adipocytes, store neutral lipids, especially triglycerides and cholesterol esters (TG and CE) into cytoplasmic lipid droplets (CLD), contributing to the turnover of the adipose tissue. The CLD are organelles that play a crucial role in energy homeostasis and cell metabolism. Caveolae are invaginations of 50-100 nm that were initially characterized by transmission electron microscopy. The shape and structural organization of caveolae are held by specific proteins of the family of caveolinas (caveolin-1, -2 and -3) that associate to form oligomers in cavities/invaginations on the plasma membrane. The caveolin functions on the CLD development are related to the fatty acid uptake and its metabolism. The relationship between these cell components seems to be pivotal for the cellular homeostasis maintenance. Many organelles that are functionally connected to lipid metabolism are found juxtaposed to the CLD. Morphologic distinctions reveal the differences that may exist in the way through which the CLD interact with other organelles within adipocytes. The membrane contact sites (MCS) have been described for many organelles and seems to be functionally important in the interaction processes with CLD. This work evaluated the ultrastructural differences between hASDC differentiated or not to pre-adipocyte compared to the 3T3-L1 cells. It was also evaluated through transmission electron microscopy, the CLD interactions to the others cellular organelles in hASDC during adipogenesis induction because, despite its importance in energy metabolism and in various diseases, the CLD are poorly understood as cell organelles. Indeed, the size, composition and regulation of GLC vary considerably between organisms and cell types. The complexity of CLD interactions with other organelles also ranged considerably between adipocytes and undifferentiated. This work showed the importance of studies that aim the clarify how the CLD are formed, modified and regulated. Through these studies it is possible to get a better understanding of the relationship between the excessive accumulation of body lipids and the metabolic syndrome (obesity, diabetes and atherosclerosis).
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Tourmente, Sylvette. "Evolution des mitochondries pendant l'ovogenese de drosophile : morphologie, distribution, replication et expression du genome." Clermont-Ferrand 2, 1987. http://www.theses.fr/1987CLF21073.
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Chemudupati, Mahesh. "Investigating the effects of nuclear envelope proteins on nuclear structure and organization in Aspergillus nidulans." The Ohio State University, 2016. http://rave.ohiolink.edu/etdc/view?acc_num=osu148009978216118.
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