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University of Lausanne

13 ARTICLES PUBLISHED IN JoVE

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Neuroscience

Calcium Imaging of Odor-evoked Responses in the Drosophila Antennal Lobe
Ana F. Silbering 1, Rati Bell 1, C. Giovanni Galizia 2, Richard Benton 1
1Center for Integrative Genomics, University of Lausanne, 2Department of Biology, University of Konstanz

We describe an established technique to measure and analyze odor-evoked calcium responses in the antennal lobe of living Drosophila melanogaster.

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Neuroscience

Imaging Pheromone Sensing in a Mouse Vomeronasal Acute Tissue Slice Preparation
Julien Brechbühl 1, Gaëlle Luyet 1, Fabian Moine 1, Ivan Rodriguez 2, Marie-Christine Broillet 1
1Department of Pharmacology and Toxicology, University of Lausanne, 2Department of Genetics and Evolution, University of Geneva

In mice, the ability to detect pheromones is principally mediated by the vomeronasal organ (VNO). Here, an acute tissue slice preparation of VNO for performing calcium imaging is described. This physiological approach allows observations of subpopulations and/or individual neurons in a living tissue and is convenient for receptor-ligand identification.

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Biology

Temporal Quantification of MAPK Induced Expression in Single Yeast Cells
Serge Pelet 1, Delphine Aymoz 1, Eric Durandau 1
1Department of Fundamental Microbiology, University of Lausanne

Two complementary methods based on flow cytometry and microscopy are presented which enable the quantification, at the single cell level, of the dynamics of gene expression induced by the activation of a MAPK pathway in yeast.

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JoVE Journal

Microscopy of Fission Yeast Sexual Lifecycle
Aleksandar Vjestica *1, Laura Merlini *1, Omaya Dudin *1, Felipe O. Bendezu 1, Sophie G. Martin 1
1Department of Fundamental Microbiology, University of Lausanne

We provide a reproducible basic method for the long-term microscopy of the fission yeast sexual lifecycle. With minor adjustments described, the presented protocol allows research focus on different steps of the reproductive process.

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Immunology and Infection

Using X-ray Crystallography, Biophysics, and Functional Assays to Determine the Mechanisms Governing T-cell Receptor Recognition of Cancer Antigens
Bruce J. MacLachlan *1, Alexander Greenshields-Watson *1, Georgina H Mason *1, Andrea J Schauenburg 1, Valentina Bianchi 1,2,3, Pierre J Rizkallah 1, Andrew K Sewell 1, Anna Fuller 1, David K Cole 1
1Division of Infection and Immunity and Systems Immunity Research Institute, Cardiff University, 2Department of Oncology, University Hospital of Lausanne (CHUV), 3Ludwig Insitutue for Cancer Research, Lausanne Branch, University of Lausanne

Here, we describe methods that we commonly employ in the laboratory to determine how the nature of the interaction between the T-cell receptor and tumor antigens, presented by human leukocyte antigens, governs T-cell functionality; these methods include protein production, X-ray crystallography, biophysics, and functional T-cell experiments.

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JoVE Journal

In Vitro Polymerization of F-actin on Early Endosomes
Olivia Muriel 1,2, Cameron Christopher Scott 1, Jorge Larios 1, Vicent Mercier 1, Jean Gruenberg 1
1Department of Biochemistry, University of Geneva, 2Department of Fundamental Microbiology, University of Lausanne

Early endosome functions depend on F-actin polymerization. Here, we describe a microscopy-based in vitro assay that reconstitutes the nucleation and polymerization of F-actin on early endosomal membranes in test tubes, thus rendering this complex series of reactions amenable to biochemical and genetic manipulations.

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Neuroscience

Quantifying Infra-slow Dynamics of Spectral Power and Heart Rate in Sleeping Mice
Laura M. J. Fernandez 1, Sandro Lecci 1, Romain Cardis 1, Gil Vantomme 1, Elidie Béard 1, Anita Lüthi 1
1Department of Fundamental Neurosciences, University of Lausanne

Here, we present experimental and analytical procedures to describe the temporal dynamics of the neural and cardiac variables of non-REM sleep in mice, which modulate sleep responsiveness to acoustic stimuli.

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Behavior

Supramaximal Intensity Hypoxic Exercise and Vascular Function Assessment in Mice
Jessica Lavier 1,2, Manon Beaumann 3, Steeve Ménetrey 3, Lucia Mazzolai 1, Anne-Christine Peyter 3, Maxime Pellegrin 1, Grégoire P. Millet 2
1Division of Angiology, Heart and Vessel Department, Lausanne University Hospital (CHUV), 2Institute of Sport Sciences, Faculty of Biology and Medicine, University of Lausanne, 3Neonatal Research Laboratory, Clinic of Neonatology, Department Woman-Mother-Child, Lausanne University Hospital

High-intensity training in hypoxia is a protocol that has been proven to induce vascular adaptations potentially beneficial in some patients and to improve athletes' repeated sprint ability. Here, we test the feasibility of training mice using that protocol and identify those vascular adaptations using ex vivo vascular function assessment.

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Environment

Combined Size and Density Fractionation of Soils for Investigations of Organo-Mineral Interactions
Fanny Viret 1, Stephanie Grand 1
1Faculty of Geosciences and Environment, University of Lausanne

Combined size and density fractionation (CSDF) is a method to physically separate soil into fractions differing in texture (particle size) and mineralogy (density). The purpose is to isolate fractions with different reactivities towards soil organic matter (SOM), in order to better understand organo-mineral interactions and SOM dynamics.

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Biology

Measurement of Mitochondrial Mass and Membrane Potential in Hematopoietic Stem Cells and T-cells by Flow Cytometry
Mukul Girotra *1,2, Anne-Christine Thierry 3, Alexandre Harari 1,3, George Coukos 1, Olaia Naveiras 2,4, Nicola Vannini *1
1Department of Oncology UNIL CHUV, Ludwig Institute for Cancer Research Lausanne, University of Lausanne, 2Swiss Institute for Experimental Cancer Research (ISREC), School of Life Sciences, Swiss Federal Institute of Technology Lausanne (EPFL), 3Center of Experimental Therapeutics, Department of Oncology, Centre Hospitalier Universitaire Vaudois, 4Hematology Service, Department of Oncology, Centre Hospitalier Universitaire Vaudois (CHUV)

Here we describe a reliable method to measure mitochondrial mass and membrane potential in ex vivo cultured hematopoietic stem cells and T cells.

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Genetics

Detection of Human Immunodeficiency Virus Type 1 (HIV-1) Antisense Protein (ASP) RNA Transcripts in Patients by Strand-Specific RT-PCR
Antonio Mancarella 1, Francesco A. Procopio 1, Tilmann Achsel 2, Elisa De Crignis 3, Brian T. Foley 4, Giampietro Corradin 5, Claudia Bagni 2, Giuseppe Pantaleo 1, Cecilia Graziosi 1
1Division of Immunology and Allergy, Lausanne University Hospital, 2Department of Fundamental Neuroscience, University of Lausanne, 3Department of Biochemistry, Erasmus Medical Center, 4Theoretical Biology and Biophysics Group, Los Alamos National Laboratories, 5Department of Biochemistry, University of Lausanne

RNA hairpins and loops can function as primers for reverse transcription (RT) in absence of sequence-specific primers, interfering with the study of overlapping antisense transcripts. We have developed a technique able to identify strand-specific RNA, and we have used it to study HIV-1 antisense protein ASP.

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Neuroscience

Morphological and Functional Evaluation of Axons and their Synapses during Axon Death in Drosophila melanogaster
Maria Paglione *1, Arnau Llobet Rosell *1, Jean-Yves Chatton 1, Lukas J. Neukomm 1
1Department of Fundamental Neurosciences, University of Lausanne

Here, we provide protocols to perform three simple injury-induced axon degeneration (axon death) assays in Drosophila melanogaster to evaluate the morphological and functional preservation of severed axons and their synapses.

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Biology

Two-Step Tag-Free Isolation of Mitochondria for Improved Protein Discovery and Quantification
Joan Blanco-Fernandez 1, Alexis A. Jourdain 1
1Department of Immunobiology, University of Lausanne

We present a two-step protocol for high-quality mitochondria isolation that is compatible with protein discovery and quantification at a proteome scale. Our protocol does not require genetic engineering and is thus suitable for studying mitochondria from any primary cells and tissues.

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