Freie Universitat Berlin View Institution's Website 15 articles published in JoVE Neuroscience A Model for Epilepsy of Infectious Etiology using Theiler's Murine Encephalomyelitis Virus Gaelle Batot1, Cameron S. Metcalf1, Laura A. Bell1,2, Alberto Pauletti3, Karen S. Wilcox1,2, Sonja Bröer3 1Department of Pharmacology and Toxicology, University of Utah, 2Interdepartmental Program in Neuroscience, University of Utah, 3Faculty of Veterinary Medicine, Institute of Pharmacology and Toxicology, Freie Universität Berlin Intracerebral infection with the Theiler's murine encephalomyelitis virus (TMEV) in C57BL/6 mice replicates many of the early and chronic clinical symptoms of viral encephalitis and subsequent epilepsy in human patients. This paper describes the virus infection, symptoms, and histopathology of the TMEV model. Biochemistry Visualization and Quantification of TGFβ/BMP/SMAD Signaling under Different Fluid Shear Stress Conditions using Proximity-Ligation-Assay Paul-Lennard Mendez1,2, Leon Obendorf1, Petra Knaus1 1Institute for Chemistry and Biochemistry, Freie Universität Berlin, 2International Max-Planck Research School for Biology and Computation Here, we establish a protocol to simultaneously visualize and analyze multiple SMAD complexes using proximity ligation assay (PLA) in endothelial cells exposed to pathological and physiological fluid shear stress conditions. Biochemistry Workflow and Tools for Crystallographic Fragment Screening at the Helmholtz-Zentrum Berlin Jan Wollenhaupt1, Tatjana Barthel1,2, Gustavo M. A. Lima3, Alexander Metz4, Dirk Wallacher5, Elmir Jagudin3, Franziska U. Huschmann1,4, Thomas Hauß1, Christian G. Feiler1, Martin Gerlach1, Michael Hellmig1, Ronald Förster1, Michael Steffien1, Andreas Heine4, Gerhard Klebe4, Uwe Mueller1, Manfred S. Weiss1 1Macromolecular Crystallography, Helmholtz-Zentrum Berlin, 2Structural Biochemistry Group, Institute for Chemistry and Biochemistry, Freie Universität Berlin, 3BioMAX, MAX IV Laboratory, 4Drug Design Group, Institute of Pharmaceutical Chemistry, Philipps-Universität Marburg, 5Department Sample Environment, Helmholtz-Zentrum Berlin Crystallographic fragment screening at the Helmholtz-Zentrum Berlin is performed using a workflow with dedicated compound libraries, crystal handling tools, fast data collection facilities and largely automated data analysis. The presented protocol intends to maximize the output of such experiments to provide promising starting points for downstream structure-based ligand design. Chemistry Experimental Methods for Efficient Solar Hydrogen Production in Microgravity Environment Katharina Brinkert1,2, Ömer Akay3, Matthias H. Richter1,4, Janine Liedtke2, Katherine T. Fountaine5,6, Hans-Joachim Lewerenz7, Michael Giersig3,8 1Division of Chemistry and Chemical Engineering, California Institute of Technology, 2European Space Agency/ ESTEC, 3Department of Physics, Freie Universitat Berlin, 4Applied Physics and Sensors, Brandenburg University of Technology Cottbus, 5Resnick Sustainability Institute, California Institute of Technology, 6NG Next, Northrop Grumman Corporation, 7Division of Engineering and Applied Science and Joint Center for Artificial Photosynthesis, California Institute of Technology, 8International Academy of Optoelectronics at Zhaoqing, South China Normal University Efficient solar-hydrogen production has recently been realized on functionalized semiconductor-electrocatalyst systems in a photoelectrochemical half-cell in microgravity environment at the Bremen Drop Tower. Here, we report the experimental procedures for manufacturing the semiconductor-electrocatalyst device, details of the experimental set-up in the drop capsule and the experimental sequence during free fall. Behavior Systematic Assessment of Well-Being in Mice for Procedures Using General Anesthesia Katharina Hohlbaum1,2, Bettina Bert2,3, Silke Dietze2, Rupert Palme4, Heidrun Fink2, Christa Thöne-Reineke1 1Institute of Animal Welfare, Animal Behavior and Laboratory Animal Science, Department of Veterinary Medicine, Freie Universität Berlin, 2Institute of Pharmacology and Toxicology, Department of Veterinary Medicine, Freie Universität Berlin, 3German Federal Institute for Risk Assessment (BfR), 4Unit of Physiology, Pathophysiology and Experimental Endocrinology, Department of Biomedical Sciences, University of Veterinary Medicine We developed a protocol to assess well-being in mice during procedures using general anesthesia. A series of behavioral parameters indicating levels of well-being as well as glucocorticoid metabolites were analyzed. The protocol can serve as a general aid to estimate the degree of severity in a scientific, animal-centered manner. Developmental Biology Processing of Human Cardiac Tissue Toward Extracellular Matrix Self-assembling Hydrogel for In Vitro and In Vivo Applications Matthias Becker1,2, Janita A. Maring1,2, Barbara Oberwallner2, Benjamin Kappler2, Oliver Klein1,2, Volkmar Falk2,3,4,5, Christof Stamm2,3,4 1Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, 2Berlin-Brandenburg Center for Regenerative Therapies (BCRT), 3German Center for Cardiovascular Research (DZHK), 4Deutsches Herzzentrum Berlin (DHZB), 5Department of Cardiovascular Surgery, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health This protocol describes the complete decellularization of human myocardium while preserving its extracellular matrix components. Further processing of the extracellular matrix results in the production of microparticles and a cytoprotective self-assembling hydrogel. Behavior Introducing Clicker Training as a Cognitive Enrichment for Laboratory Mice Charlotte Leidinger1,2, Felix Herrmann1, Christa Thöne-Reineke*2, Nadine Baumgart*1, Jan Baumgart*1 1Translational Animal Research Center, University Medical Centre, Johannes Gutenberg-Universität Mainz, 2Department of Veterinary Medicine, Institute of Animal Welfare, Animal Behavior and Laboratory Animal Science, Freie Universität Berlin The development of new refinement strategies for laboratory mice is a challenging task that contributes towards fulfilling the 3R principle. This protocol introduces clicker training as a cognitive enrichment program for laboratory mice. Engineering Proton Transfer and Protein Conformation Dynamics in Photosensitive Proteins by Time-resolved Step-scan Fourier-transform Infrared Spectroscopy Víctor A. Lórenz-Fonfría1, Joachim Heberle1 1Experimental Molecular Biophysics, Freie Universität Berlin Key steps of protein function, in particular backbone conformational changes and proton transfer reactions, often take place in the microsecond to millisecond time scale. These dynamical processes can be studied by time-resolved step-scan Fourier-transform infrared spectroscopy, in particular for proteins whose function is triggered by light. Biology Who is Who? Non-invasive Methods to Individually Sex and Mark Altricial Chicks Iris Adam1, Constance Scharff1, Mariam Honarmand1 1Department of Animal Behavior, Freie Universität Berlin This protocol provides a convenient set of methods, which enables extremely fast, easy, non-invasive, reliable and low-cost, molecular sex determination of birds and their non-invasive, quick, safe and easily recognizable marking shortly after hatching. Only limited handling of chicks is required. This convenient toolbox of methods complies entirely with the RRR-guidelines. Behavior Assessment of Age-related Changes in Cognitive Functions Using EmoCogMeter, a Novel Tablet-computer Based Approach Philipp Fuge*1,2, Simone Grimm*1,2,3,4, Anne Weigand1,2,3, Yan Fan1,3, Matti Gärtner1,3, Melanie Feeser1,3, Malek Bajbouj1,2,3 1Cluster of Excellence "Languages of Emotion", Freie Universität Berlin, 2Department of Psychiatry, Campus Benjamin Franklin, Charité Berlin, 3Dahlem Institute for Neuroimaging of Emotion, Freie Universität Berlin, 4Clinic for Affective Disorders and General Psychiatry, Psychiatric University Hospital Zurich We tested the usability of a tablet-computer-based application (EmoCogMeter) in investigating the effects of age on cognition. Results show an age-related cognitive decline, thereby proving the usability of our application. Findings underline the great clinical and practical potential of a tablet-based application for detection and monitoring of cognitive dysfunction. Medicine Patient Derived Cell Culture and Isolation of CD133+ Putative Cancer Stem Cells from Melanoma Yvonne Welte1,2, Cathrin Davies1,3, Reinhold Schäfer1,4, Christian R.A. Regenbrecht1,3,4 1Institute of Pathology, Laboratory of Molecular Tumor Pathology, Charité - Universitätsmedizin Berlin, 2Institute for Chemistry and Biochemistry, Free University Berlin, 3Laboratory for Functional Genomics Charité (LFGC), Charité - Universitätsmedizin Berlin, 4Comprehensive Cancer Center Charité, Charité - Universitätsmedizin Berlin This article describes the preparation of freshly obtained melanoma tissue into primary cell cultures, and how to remove contaminations of erythrocytes and fibroblasts from the tumor cells. Finally, we describe how CD133+ putative melanoma stem cells are sorted from the CD133- bulk using Magnetic Activated Cell Sorting (MACS). Immunology and Infection IgY Technology: Extraction of Chicken Antibodies from Egg Yolk by Polyethylene Glycol (PEG) Precipitation Diana Pauly1, Pablo A. Chacana2, Esteban G. Calzado3, Björn Brembs4, Rüdiger Schade5 1Center for Biological Security, Robert Koch-Institute, 2CICVyA - INTA Castelar, Instituto de Virología, 3Center of Molecular Immunology, Ciudad de la Habana, Cuba, 4Department of Biology, Chemistry, Pharmacy, Institute of Biology-Neurobiology, Free University of Berlin, 5Institut of Pharmacology, Charité-University Medicine of Berlin This protocol describes in particular the extraction of total IgY from egg yolk by means of polyethylene glycol precipitation and gives general information about IgY technology. Neuroscience Behavioural Pharmacology in Classical Conditioning of the Proboscis Extension Response in Honeybees (Apis mellifera) Johannes Felsenberg*1, Katrin B. Gehring*1, Victoria Antemann1, Dorothea Eisenhardt1 1Fachbereich Bio/Chem/Pharm, Institut für Biologie – Neurobiologie, Freie Universität Berlin We demonstrate how to implement a behavioral pharmacology method in an appetitive olfactory conditioning paradigm in honeybees (Apis mellifera) by systemic application of drugs. This method allows investigation of the mechanisms underlying learning and memory formation in a simple and reliable way. Biology In vivo Ca2+- Imaging of Mushroom Body Neurons During Olfactory Learning in the Honey Bee Melanie Haehnel1, Anja Froese2, Randolf Menzel2 1Institut für Biologie - Neurobiologie, Freie Universität Berlin, 2Institut für Biologie - Neurobiologie, Free University Berlin - Freie Universitaet Berlin Bees can be conditioned in an appetitive olfactory learning paradigm (PER-conditioning). Using odors as stimuli, we established a method in which behavior is recorded while simultaneously Calcium Imaging is used to measure odor evoked activity in mushroom body neurons in vivo. Biology Operant Learning of Drosophila at the Torque Meter Bjoern Brembs1 1Department of Neurobiology, Free University of Berlin Measuring the yaw torque of tethered Drosophila with the torque meter allows the neuroscientist exquisite control of the stimulus situation of the experimental animal. Together with the unique genetic tools available in the fruit fly, this paradigm is used for a wide variety of neurobiological research.