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Cryo-electron microscopy as diagnostic tool to reveal the molecular & cellular architecture in healthy & pathogenic cells

Projectomschrijving

Het VUmc-elektronenmicroscopisch centrum diagnosticeert met innovatieve cryoelektronenmicroscopische technieken pathologische afwijkingen in cellen uit patiënten met Alzheimer, Kanker en Hartfalen. Ultrastructureel inzicht in bijv. 3D zal bijdragen aan de zoektocht naar nieuwe medicijnen die pathologische afwijkingen in de intracellulaire architectuur herstellen.

Producten

Titel: C-type lectins and their role in the immune system.
Auteur: K Bloem
Titel: The post-docking role of Synaptotagmin1 and Munc18-1 in mouse chromaffin cells.
Auteur: G.H. Kedar
Titel: Synaptic effects of Munc18-1 alternative splicing and protein modifications
Auteur: M. Meijer
Titel: Water and ion homeostasis in the developing brain: disease models impacting understanding of physiology.
Auteur: M. Dubey
Titel: Kinetic models for synaptic vesicle release
Auteur: S. Schotten
Titel: C2 domain function in healthy and diseased brain
Auteur: A. Giniatullina
Titel: The role of presynaptic proteins in maintaining neuronal viability
Auteur: T. Santos
Titel: Trafficking and secretion of neuropeptides in mouse hippocampal neurons
Auteur: M. Farina
Titel: Dissecting the role of Fbxo41 in neuronal development and signaling.
Auteur: C.R. King
Titel: Mechanisms understanding synapse formation and post-synaptic receptor targeting.
Auteur: F Farzana
Titel: Organizing sites for regulated secretion in adrenal chromaffin cells.
Auteur: J. Kurps
Titel: Comparing and contrasting white matter disorders: a neuropathological approach to pathophysiology
Auteur: M. Bugiani
Titel: Mice with megalencephalic leukoencephalopathy with cysts: A developmental angle
Auteur: Dubey, Mohit, Bugiani, Marianna, Ridder, Margreet C., Postma, Nienke L., Brouwers, Eelke, Polder, Emiel, Jacobs, J. Gerbren, Baayen, Johannes C., Klooster, Jan, Kamermans, Maarten, Aardse, Romy, de Kock, Christiaan P. J., Dekker, Marien P., van Weering, Jan R. T., Heine, Vivi M., Abbink, Truus E. M., Scheper, Gert C., Boor, Ilja, Lodder, Johannes C., Mansvelder, Huibert D., van der Knaap, Marjo S.
Magazine: Annals of Neurology
Titel: VPS35 depletion does not impair presynaptic structure and function
Auteur: Vazquez-Sanchez, Sonia, Bobeldijk, Sander, Dekker, Marien P., van Keimpema, Linda, van Weering, Jan R. T.
Magazine: Scientific Reports
Titel: CNS myelin induces regulatory functions of DC-SIGN expressing antigen-presenting cells via cognate interaction with MOG.
Auteur: J.J. García-Vallejo, J.M. Ilarregui, H. Kalay, S. Chamorro, N. Koning, W.W. Unger, M. Ambrosini, V. Montserrat, R. J. Fernandes, S. C. M. Bruijns, J.R.T. van Weering, N.J. Paauw, T. O’Toole, J. van Horssen, P. van der Valk, K. Nazmi, J.G.M. Bolscher, J. Bajramovic, C.D. Dijkstra, B.A. ’t Hart, and Y. van Kooyk
Magazine: Journal of Experimental Medicine
Titel: The UPR reduces glucose metabolism via IRE1 signaling
Auteur: van der Harg, Judith M., van Heest, Jessica C., Bangel, Fabian N., Patiwael, Sanne, van Weering, Jan R.T., Scheper, Wiep
Magazine: Biochimica et Biophysica Acta (BBA) - Molecular Basis of Disease
Titel: Direct quantitative detection of Doc2b-induced hemifusion in optically trapped membranes
Auteur: Brouwer, Ineke, Giniatullina, Asiya, Laurens, Niels, van Weering, Jan R. T., Bald, Dirk, Wuite, Gijs J. L., Groffen, Alexander J.
Magazine: Nature Communications
Titel: Extension of Helix 12 in Munc18-1 Induces Vesicle Priming
Auteur: Munch, A. S., Kedar, G. H., van Weering, J. R. T., Vazquez-Sanchez, S., He, E., Andre, T., Braun, T., Sollner, T. H., Verhage, M., Sorensen, J. B.
Magazine: Journal of Neuroscience
Titel: MIR137 schizophrenia-associated locus controls synaptic function by regulating synaptogenesis, synapse maturation and synaptic transmission
Auteur: He, Enqi, Lozano, Miguel A Gonzalez, Stringer, Sven, Watanabe, Kyoko, Sakamoto, Kensuke, den Oudsten, Frank, Koopmans, Frank, Giamberardino, Stephanie N, Hammerschlag, Anke, Cornelisse, L Niels, Li, Ka Wan, van Weering, Jan, Posthuma, Danielle, Smit, August B, Sullivan, Patrick F, Verhage, Matthijs
Magazine: Human Molecular Genetics
Titel: Development of dendritic tonic GABAergic inhibition regulates excitability and plasticity in CA1 pyramidal neurons
Auteur: M. Groen, O. Paulsen, E. Perez-Garci, T. Nevian, J. Wortel, R. Dekker, H. Mansvelder, A. van Ooyen, and R. Meredith; Journal of Neurophysiology
Magazine: Journal of Neurophysiology
Titel: Differential Maturation of the Two Regulated Secretory Pathways in Human iPSC-Derived Neurons
Auteur: Emperador Melero, Javier, Nadadhur, Aishwarya G., Schut, Desiree, Weering, Jan V., Heine, Vivi M., Toonen, Ruud F., Verhage, Matthijs
Magazine: Stem Cell Reports
Titel: A Post-Docking Role of Synaptotagmin 1-C2B Domain Bottom Residues R398/399 in Mouse Chromaffin Cells
Auteur: Kedar, G. H., Munch, A. S., van Weering, J. R. T., Malsam, J., Scheutzow, A., de Wit, H., Houy, S., Tawfik, B., Sollner, T. H., Sorensen, J. B., Verhage, M.
Magazine: Journal of Neuroscience
Titel: Tyrosine phosphorylation of Munc18-1 inhibits synaptic transmission by preventing SNARE assembly
Auteur: Meijer, Marieke, Dörr, Bernhard, Lammertse, Hanna CA, Blithikioti, Chrysanthi, van Weering, Jan RT, Toonen, Ruud FG, Söllner, Thomas H, Verhage, Matthijs
Magazine: EMBO Journal
Titel: Dense-core vesicle biogenesis and exocytosis in neurons lacking chromogranins A and B
Auteur: Dominguez, Natalia, van Weering, Jan R. T., Borges, Ricardo, Toonen, Ruud F. G., Verhage, Matthijs
Magazine: Journal of Neurochemistry
Titel: Proteomic analysis of cerebrospinal fluid extracellular vesicles: a comprehensive dataset
Auteur: D. Chiasserini, J.R.T. van Weering, S.R. Piersma, T.V. Pham, A . Malekzadeh, C.E. Teunissen, H. de Wit, C.R. Jiménez
Magazine: Journal of Proteomics
Titel: Vitrification of Tokuyasu-style immuno-labeled sections for correlative cryo light microscopy and cryo electron tomography
Auteur: E. Bos, L. Hussaarts, J.R.T. van Weering, M.H. Ellisman, H. de Wit, A.J. Koster
Magazine: Journal of Structural Biology
Titel: Strain Differences in Presynaptic Function
Auteur: Lenselink, A. Mariette, Rotaru, Diana C., Li, Ka Wan, van Nierop, Pim, Rao-Ruiz, Priyanka, Loos, Maarten, van der Schors, Roel, Gouwenberg, Yvonne, Wortel, Joke, Mansvelder, Huibert D., Smit, August B., Spijker, Sabine
Magazine: Journal of Biological Chemistry
Titel: CAPS-1 promotes fusion competence of stationary dense-core vesicles in presynaptic terminals of mammalian neurons
Auteur: Farina, Margherita, van de Bospoort, Rhea, He, Enqi, Persoon, Claudia M, van Weering, Jan RT, Broeke, Jurjen H, Verhage, Matthijs, Toonen, Ruud F
Magazine: eLife
Titel: Tomosyn associates with secretory vesicles in neurons through its N- and C-terminal domains
Auteur: Geerts, Cornelia J., Mancini, Roberta, Chen, Ning, Koopmans, Frank T. W., Li, Ka Wan, Smit, August B., van Weering, Jan R. T., Verhage, Matthijs, Groffen, Alexander J. A.
Magazine: PLoS ONE
Titel: Megalencephalic leukoencephalopathy with cysts: the Glialcam -null mouse model
Auteur: Bugiani, Marianna, Dubey, Mohit, Breur, Marjolein, Postma, Nienke L., Dekker, Marien P., ter Braak, Timo, Boschert, Ursula, Abbink, Truus E. M., Mansvelder, Huibert D., Min, Rogier, van Weering, Jan R. T., van der Knaap, Marjo S.
Magazine: Annals of Clinical and Translational Neurology
Titel: The novel antibiotic rhodomyrtone traps membrane proteins in vesicles with increased fluidity
Auteur: Saeloh, Dennapa, Tipmanee, Varomyalin, Jim, Kin Ki, Dekker, Marien P., Bitter, Wilbert, Voravuthikunchai, Supayang P., Wenzel, Michaela, Hamoen, Leendert W.
Magazine: PLoS Pathogens
Titel: Plasma vesicle miRNAs for therapy response monitoring in Hodgkin lymphoma patients
Auteur: van Eijndhoven, Monique A.J., Zijlstra, Josée M., Groenewegen, Nils J., Drees, Esther E.E., van Niele, Stuart, Baglio, S. Rubina, Koppers-Lalic, Danijela, van der Voorn, Hans, Libregts, Sten F.W.M., Wauben, Marca H.M., de Menezes, Renee X., van Weering, Jan R.T., Nieuwland, Rienk, Visser, Lydia, van den Berg, Anke, de Jong, Daphne, Pegtel, D. Michiel
Magazine: Journal of Clinical Investigation

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Samenvatting van de aanvraag

With the deciphering of the human genome many proteins are isolated but from most of these proteins their function remains unknown. In order to validate the function of these proteins high-resolution in situ studies are required. Electron microscopy (EM) offers a resolution which is about two orders of magnitude higher than that of light microscopy. Using EM it is possible to analyze the subcellular immunolocalization (IEM) of proteins and the morphology/distribution of cell organelles in health or diseased conditions. Therefore, EM studies are important for a fundamental understanding of protein (dys)function and unraveling ultrastructural consequences linked to most prevalent and debilitating diseases like brain disorders, as well as cancer and cardiovascular failures. Although, EM gives the resolution to investigate the exact macromolecular atlas of the cell under normal and pathological conditions, however, conventional EM can cause artefacts caused by chemical fixation and dehydration procedures and have important repercussions for EM studies in life sciences and diagnostics. Nowadays many EM studies start from cells that are structurally stabilized within a few milliseconds, by fast freezing facilitated by the development of high-pressure freezing (HPF) equipment. Combined with correlative light- and electron microscopy (CLEM) fast freezing allows the rapid capture of dynamic cellular processes (i.e. vesicle transport) as observed by real-time imaging in a living cell for ultrastructural analyses in a near-native state. In addition, cryo-electron tomography (ET) provides a view in 3 dimensions (3D) of the molecular architecture in cells at nanometer resolution and as such is complementary to lower resolution medical tomographic techniques (e.g. CAT, PET and MRI). The Netherlands has a strong tradition in EM but to compete with other cryo-EM research groups in the world and lower the costs of buying expensive cryo-EM equipment it is important to collaborate and share cryo-EM equipment/experience to the highest degree. This is for example illustrated by the recent initiative of the ‘Netherlands Center for Electron Nanoscopy’ (NeCEN) consortium in which major cryo-EM centers have joined forces to perform a 3D study of cellular macromolecular complexes. As such, complimentary to the work in NeCEN now we aim to invest in a multidisciplinary cryo-EM-unit especially suited for researchers and physicians working with biomedical and clinical research questions and the intention to work from fundamental approach (from basic) to therapeutic (bedside) implication. Recently, a new EM support facility (coordinated by De Wit) has been started at the VU campus in the new institute Neuroscience Campus Amsterdam, however within the EM support facility of NCA cryo-EM infrastructure is lacking. Therefore new investments in cryo-EM equipment (e.g. HPF, freeze-substitution, cryo-ET, vitreous cryo-sectioning) are absolutely urgent. With these investments we propose to join forces with other EM researchers on the VU campus, such that an excellent level of EM infrastructure and expertise can be realized for the coming decade. This is exactly the focus of the present application in which 3 important institutes of the VUmc will heavily participate: NCA (affiliated to Alzheimer- and MS Center and Nederlandse Hersenbank), Cancer Center Amsterdam/VU Institute of Cancer and Immunology (CCA-VICI) and Institute for Cardiovascular Research VU (ICAR-VU). All participants have the intention to work from fundamental approach (from basic) to therapeutic (bedside) implication. To succeed participants want to develop an excellent cryo-EM infrastructure that allow visualization of the molecular architecture in their near-native state in healthy and pathogenic cells to improve our efforts for understanding and finding good treatments against brain disorders (Alzheimer’s disease, fragile X, Multiple sclerosis), cancer and heart failure. In order to boost collaborative cryo-EM research in life sciences and diagnostics on the VU campus a new core EM facility will be build in the coming years at the ground floor of the new research tower of VUmc (annex 1). The requested cryo-EM equipment will be housed in this central EM facility of VU/VUmc to create an excellent cryo-EM-infrastructure and ensure the use of this innovative cryo-EM equipment at the cutting edge of science and diagnostics covering brain diseases, cancer and cardiovascular failures by several institutes within VU/VUmc as well as researcher from other regions in Amsterdam (NKI-AVL/AMC) and the NeCEN consortium.

Onderwerpen

Kenmerken

Projectnummer:
91111009
Looptijd: 100%
Looptijd: 100 %
2011
2020
Onderdeel van programma:
Gerelateerde subsidieronde:
Projectleider en penvoerder:
Drs. J.R.T. van Weering
Verantwoordelijke organisatie:
Amsterdam UMC - locatie VUmc