Universite de Bordeaux View Institution's Website 17 articles published in JoVE Biochemistry High-Resolution Neutron Spectroscopy to Study Picosecond-Nanosecond Dynamics of Proteins and Hydration Water Kevin Pounot1,2, Markus Appel2, Christian Beck1,2, Martin Weik3, Giorgio Schirò3, Yann Fichou4, Tilo Seydel2, Frank Schreiber1 1Institut für Angewandte Physik, Universität Tübingen, 2Institut Max von Laue - Paul Langevin (ILL), 3Institut de Biologie Structurale, Université Grenoble Alpes, 4Institut Européen de Chimie et Biologie, Bordeaux INP, Chimie et Biologie des Membranes et des Nanoobjets (CBMN), Université de Bordeaux Neutron backscattering spectroscopy offers a nondestructive and label-free access to the ps-ns dynamics of proteins and their hydration water. The workflow is presented with two studies on amyloid proteins: on the time-resolved dynamics of lysozyme during aggregation and on the hydration water dynamics of tau upon fiber formation. Behavior Handling Techniques to Reduce Stress in Mice Michael Marcotte1, Ashley Bernardo1,2, Nathaniel Linga3, Carmina A. Pérez-Romero4, Jean-Louis Guillou5, Etienne Sibille1,2,3, Thomas D. Prevot1,2 1Campbell Family Mental Health Research Institute of CAMH, 2Department of Psychiatry, University of Toronto, 3Department of Pharmacology and Toxicology, University of Toronto, 4Departamento de Investigación, Universidad Central de Queretaro, 5Centre National de la Recherche Scientifique, UMR 5287, Institut de Neurosciences Cognitives et Intégratives d'Aquitaine, Université de Bordeaux This paper describes a handling technique in mice, the 3D-handling technique, which facilitates routine handling by reducing anxiety-like behaviors and presents details on two existing related techniques (tunnel and tail handling). Chemistry Crystallization of Proteins on Chip by Microdialysis for In Situ X-ray Diffraction Studies Sofia Jaho1, Niels Junius1,3, Franck Borel1, Yoann Sallaz-Damaz1, Jean-Baptiste Salmon2, Monika Budayova-Spano1 1Université Grenoble Alpes, CEA, CNRS, IBS, 2CNRS, Solvay LOF, UMR 5258, Université Bordeaux, 3ELVESYS SAS This paper details the fabrication protocol of microfluidic chips developed for on-chip protein crystallization with the dialysis method and in situ X-ray diffraction experiments. The microfabrication process makes it possible to integrate a semipermeable regenerated cellulose dialysis membrane with any molecular weight cut-off, between two layers of the chip. Cancer Research Co-culture of Glioblastoma Stem-like Cells on Patterned Neurons to Study Migration and Cellular Interactions Joris Guyon1, Pierre-Olivier Strale2,3, Irati Romero-Garmendia4, Andreas Bikfalvi1, Vincent Studer*2, Thomas Daubon*4 1University Bordeaux, INSERM, LAMC, U1029, 33600, Pessac, France, 2Univ. Bordeaux, CNRS, Interdisciplinary Institute for Neuroscience, IINS, UMR 5297, Bordeaux, France, 3Alvéole, Bordeaux, France, 4University Bordeaux, CNRS, IBGC, UMR5095, 33000, Bordeaux, France Here, we present an easy-to-use co-culture assay to analyze glioblastoma (GBM) migration on patterned neurons. We developed a macro in FiJi software for easy quantification of GBM cell migration on neurons, and observed that neurons modify GBM cell invasive capacity. Medicine Assessment of Dependence in Activities of Daily Living Among Older Patients in an Acute Care Unit Vincent Bailly1, Isabelle Bourdel-Marchasson1,2 1Pôle de Gérontologie Clinique, CHU Bordeaux, 2Univ.Bordeaux/CNRS UMR 5536 RMSB During acute medical problems, older people may lose independence in activities of daily living (ADL). Assessment of baseline ADL and ADL on admission can guide personalized treatment plans aimed at preventing nosocomial dependence and ensuring better functional outcomes. Cancer Research A 3D Spheroid Model for Glioblastoma Joris Guyon1,2, Laetitia Andrique1,2, Nadège Pujol1,2, Gro Vatne Røsland3,4, Gaelle Recher1,5,6, Andreas Bikfalvi1,2, Thomas Daubon1,2,4 1University Bordeaux, LAMC, 2INSERM U1029, University Bordeaux, 3Department of Biomedicine, University of Bergen, 4CNRS, IBGC UMR5095, 5LP2N, CNRS UMR 5298, IOA, 6Institut d'Optique Graduate School, IOA Here, we describe an easy-to-use invasion assay for glioblastoma. This assay is suitable for glioblastoma stem-like cells. A Fiji macro for easy quantification of invasion, migration, and proliferation is also described. Chemistry Obtention of Giant Unilamellar Hybrid Vesicles by Electroformation and Measurement of their Mechanical Properties by Micropipette Aspiration Emmanuel Ibarboure1, Martin Fauquignon1, Jean-François Le Meins1 1Université de Bordeaux, CNRS, Bordeaux INP, LCPO, UMR 5629 The goal of the protocol is to reliably measure membrane mechanical properties of giant vesicles by micropipette aspiration. Neuroscience Assessing Pupil-linked Changes in Locus Coeruleus-mediated Arousal Elicited by Trigeminal Stimulation Maria Paola Tramonti Fantozzi*1, Tommaso Banfi*2,3, Vincenzo De Cicco*1, Massimo Barresi4, Enrico Cataldo5, Davide De Cicco1, Luca Bruschini6, Paola d'Ascanio1, Gastone Ciuti2,3, Ugo Faraguna1,7, Diego Manzoni1 1Department of Translational Research and of New Surgical and Medical Technologies, University of Pisa, 2 To verify whether trigeminal effects on cognitive performance involve locus coeruleus activity, two protocols are presented that aim to evaluate possible correlations between the performance and task-related pupil size changes induced by chewing. These protocols may be applied to conditions in which locus coeruleus contribution is suspected. Immunology and Infection Purification of Extracellular Trypanosomes, Including African, from Blood by Anion-Exchangers (Diethylaminoethyl-cellulose Columns) Pierrette Courtois1,2, Patricia Nabos1,2, Romaric Nzoumbou-Boko1,2, Christine E. Reix3, Frédéric-Antoine Dauchy1,2,4, Sylvie Daulouede1,2, Frédéric Bringaud3, Derrick R. Robinson3, Philippe Vincendeau1,2,4 1Univ. Bordeaux UMR INTERTRYP 177 IRD CIRAD, 2IRD UMR 177 INTERTRYP CIRAD, 3CNRS, UMR 5234, Microbiologie Fondamentale et Pathogénicité, Univ. Bordeaux, 4Centre Hospitalier Université Bordeaux This method of trypanosome separation from blood depends on their surface charge being less negative than mammalian blood cells. Infected blood is placed and treated on an anion-exchanger column. This method, the most fitting diagnostic for African trypanosomiasis, provides purified parasites for immunological, biological, biochemical, pharmaceutical and molecular biology investigations. Neuroscience Functional Magnetic Resonance Spectroscopy at 7 T in the Rat Barrel Cortex During Whisker Activation Jordy Blanc1, Hélène Roumes1, Leslie Mazuel1, Philippe Massot1, Gérard Raffard1, Marc Biran1, Anne-Karine Bouzier-Sore1 1Centre de Résonance Magnétique des Systèmes Biologiques (CRMSB), Unités Mixtes de Recherche (UMR) 5536, Centre National de la Recherche Scientifique (CNRS)/Université Bordeaux After checking by blood-oxygen-level-dependent functional magnetic resonance imaging (BOLD fMRI) that the corresponding somatosensory barrel field cortex area (called S1BF) is correctly activated, the main goal of this study is to quantify lactate content fluctuations in the activated rat brains by localized proton magnetic resonance spectroscopy (1H-MRS) at 7 T. Chemistry Photogeneration of N-Heterocyclic Carbenes: Application in Photoinduced Ring-Opening Metathesis Polymerization Julien Pinaud1, Emeline Placet1, Patrick Lacroix-Desmazes1, Thi Kim Hoang Trinh2,3, Jean Pierre Malval2,3, Abraham Chemtob2,3, Loïc Pichavant4, Valérie Héroguez4 1ICGM, Université de Montpellier, CNRS, ENSCM, 2Institut de Science des Matériaux de Mulhouse (IS2M UMR 7361 CNRS), Université de Haute-Alsace, 3Université de Strasbourg, 4Laboratoire de Chimie des Polymères Organiques (LCPO UMR 5629 ENSCBP), Université de Bordeaux We describe a protocol to photogenerate N-heterocyclic carbenes (NHCs) by UV irradiation of a 2-isopropylthioxanthone/imidazolium tetraphenylborate salt system. Methods to characterize the photoreleased NHC and elucidate the photochemical mechanism are proposed. The protocols for ring-opening metathesis photopolymerization in solution and miniemulsion illustrate the potential of this 2-component NHC photogenerating system. Chemistry In Situ Detection and Single Cell Quantification of Metal Oxide Nanoparticles Using Nuclear Microprobe Analysis Giovanna Muggiolu*1,2, Marina Simon*1,2, Nathanael Lampe1,2, Guillaume Devès1,2, Philippe Barberet1,2, Claire Michelet1,2, Marie-Hélène Delville3,4, Hervé Seznec1,2 1Centre d'Etudes Nucléaires Bordeaux Gradignan (CENBG), Université de Bordeaux, 2Centre d'Etudes Nucléaires Bordeaux Gradignan (CENBG), CNRS, 3Institut de Chimie de la Matière Condens é e de Bordeaux (ICMCB), CNRS, 4Institut de Chimie de la Matière Condens é e de Bordeaux (ICMCB), Université de Bordeaux We describe a procedure for the detection of chemical elements present in situ in human cells as well as their in vitro quantification. The method is well-suited to any cell type and is particularly useful for quantitative chemical analyses in single cells following in vitro metal oxide nanoparticles exposure. Biochemistry Atomic Scale Structural Studies of Macromolecular Assemblies by Solid-state Nuclear Magnetic Resonance Spectroscopy Antoine Loquet1, James Tolchard1, Melanie Berbon1, Denis Martinez1, Birgit Habenstein1 1Institute of Chemistry, Biology of Membranes, Nanoobjects, UMR5248 CNRS, Université de Bordeaux Structures of supramolecular protein assemblies at atomic resolution are of high relevance because of their crucial roles in a variety of biological phenomena. Herein, we present a protocol to perform high-resolution structural studies on insoluble and non-crystalline macromolecular protein assemblies by magic-angle spinning solid-state nuclear magnetic resonance spectroscopy (MAS SSNMR). Biology Plunge Freezing: A Tool for the Ultrastructural and Immunolocalization Studies of Suspension Cells in Transmission Electron Microscopy Corinne Blancard1, Bénédicte Salin1 1Institut de Biochimie et Génétique Cellulaires, Centre National de la Recherche Scientifique, UMR 5095, Université de Bordeaux This manuscript describes an easy-to-use and low-cost cryofixation method for visualizing suspension cells by transmission electron microscopy. Medicine Procedure for Human Saphenous Veins Ex Vivo Perfusion and External Reinforcement Alban Longchamp*1, Florent Allagnat*2, Xavier Berard3, Florian Alonso2, Jacques-Antoine Haefliger2, Sébastien Deglise*4, Jean-Marc Corpataux*4 1 The mechanisms leading to the development of intimal hyperplasia (IH) and vein graft failure are still poorly understood. This study describes an ex vivo system to perfuse human veins under controlled flow and pressure. Furthermore the efficiency of external mesh reinforcement to limit the development of IH was evaluated. Medicine Transcriptomic Analysis of Human Retinal Surgical Specimens Using jouRNAl Marie-Noëlle Delyfer1,2,3,4, Najate Aït-Ali1,2,3, Hawa Camara1,2,3, Emmanuelle Clérin1,2,3, Jean-François Korobelnik4, José-Alain Sahel1,2,3, Thierry Léveillard1,2,3 1U968, Institut National de la Santé et de la Recherche Médicale, 2UMR S 968, Université Pierre et Marie Curie, 3UMR 7210, Centre National de la Recherche Scientifique, 4Départment d'Ophtalmologie, Centre Hospitalier Universitaire de Bordeaux We used retinal samples from retinectomy for a transcriptomic analysis of retinal detachment. We developed a procedure that allows RNA conservation between the surgical blocks and the laboratory. We standardized a protocol to purify RNA by cesium chloride ultracentrifugation to assure that the purified RNAs are suitable for microarray analysis. Neuroscience Visualization and Genetic Manipulation of Dendrites and Spines in the Mouse Cerebral Cortex and Hippocampus using In utero Electroporation Emilie Pacary1, Matilda A. Haas1, Hendrik Wildner1, Roberta Azzarelli1, Donald M. Bell2, Djoher Nora Abrous3, François Guillemot1 1Division of Molecular Neurobiology, MRC National Institute for Medical Research, 2Confocal and Image Analysis Laboratory, National Institute for Medical Research, 3Physiopathologie de la plasticité neuronale, Neurocentre Magendie, Université de Bordeaux This article describes in detail a protocol to electroporate in utero the cerebral cortex and the hippocampus at E14.5 in mice. We also show that this is a valuable method to study dendrites and spines in these two cerebral regions.