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Freie Universität Berlin

13 ARTICLES PUBLISHED IN JoVE

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Biology

In vivo Ca2+- Imaging of Mushroom Body Neurons During Olfactory Learning in the Honey Bee
Melanie Haehnel 1, Anja Froese 2, Randolf Menzel 2
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.

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Neuroscience

Behavioural Pharmacology in Classical Conditioning of the Proboscis Extension Response in Honeybees (Apis mellifera)
Johannes Felsenberg *1, Katrin B. Gehring *1, Victoria Antemann 1, Dorothea Eisenhardt 1
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.

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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 Weigand 1,2,3, Yan Fan 1,3, Matti Gärtner 1,3, Melanie Feeser 1,3, Malek Bajbouj 1,2,3
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.

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Biology

Who is Who? Non-invasive Methods to Individually Sex and Mark Altricial Chicks
Iris Adam 1, Constance Scharff 1, Mariam Honarmand 1
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.

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JoVE Core

Proton Transfer and Protein Conformation Dynamics in Photosensitive Proteins by Time-resolved Step-scan Fourier-transform Infrared Spectroscopy
Víctor A. Lórenz-Fonfría 1, Joachim Heberle 1
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.

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Behavior

Introducing Clicker Training as a Cognitive Enrichment for Laboratory Mice
Charlotte Leidinger 1,2, Felix Herrmann 1, 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.

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Behavior

Systematic Assessment of Well-Being in Mice for Procedures Using General Anesthesia
Katharina Hohlbaum 1,2, Bettina Bert 2,3, Silke Dietze 2, Rupert Palme 4, Heidrun Fink 2, Christa Thöne-Reineke 1
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.

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Neuroscience

Noninvasive EEG Recordings from Freely Moving Piglets
Nora V. de Camp 1,2, Silke Dietze 1, Markus Klaßen 3, Jürgen Bergeler 1
1Institute of Animal Welfare, Animal Behavior and Laboratory Animal Science, Veterinary Medicine, Free University Berlin, 2Department of Biology, Behavioral Physiology, Humboldt University Berlin, 3Lehr- und Versuchsanstalt für Viehhaltung Hofgut Neumühle

Here, we present a protocol to record telemetric electroencephalograms (EEGs) from freely moving piglets directly in the pigpen without the use of a sedative, making it possible to record typical EEG patterns during non-REM sleep, like spindle bursts.

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Biochemistry

Workflow and Tools for Crystallographic Fragment Screening at the Helmholtz-Zentrum Berlin
Jan Wollenhaupt 1, Tatjana Barthel 1,2, Gustavo M. A. Lima 3, Alexander Metz 4, Dirk Wallacher 5, Elmir Jagudin 3, Franziska U. Huschmann 1,4, Thomas Hauß 1, Christian G. Feiler 1, Martin Gerlach 1, Michael Hellmig 1, Ronald Förster 1, Michael Steffien 1, Andreas Heine 4, Gerhard Klebe 4, Uwe Mueller 1, Manfred S. Weiss 1
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.

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Biochemistry

Visualization and Quantification of TGFβ/BMP/SMAD Signaling under Different Fluid Shear Stress Conditions using Proximity-Ligation-Assay
Paul-Lennard Mendez 1,2, Leon Obendorf 1, Petra Knaus 1
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.

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Neuroscience

A Model for Epilepsy of Infectious Etiology using Theiler's Murine Encephalomyelitis Virus
Gaelle Batot 1, Cameron S. Metcalf 1, Laura A. Bell 1,2, Alberto Pauletti 3, Karen S. Wilcox 1,2, Sonja Bröer 3
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.

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Immunology and Infection

NAD(P)H Fluorescence Lifetime Imaging for the Metabolic Analysis of the Murine Intestine and Parasites During Nematode Infection
Wjatscheslaw Liublin 1,2, Sebastian Rausch 3, Ruth Leben 1,2,3, Juliane Liebeskind 3, Anja E. Hauser 4,5, Susanne Hartmann 3, Raluca A. Niesner 2,1
1Biophysical Analytics, Deutsches Rheuma-Forschungszentrum, Berlin - A Leibniz Institute, 2Dynamic and Functional In Vivo Imaging, Department of Veterinary Medicine, Institute for Veterinary Physiology, Freie Universität Berlin, 3Institute of Immunology, Department of Veterinary Medicine, Freie Universität Berlin, 4Laboratory for Immune Dynamics, Deutsches Rheuma-Forschungszentrum, Berlin - A Leibniz Institute, 5Intravital Microscopy and Immune Dynamics, Department of Rheumatology and Clinical Immunology, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt Universität zu Berlin

The present protocol describes the NAD(P)H fluorescence lifetime imaging of an explanted murine intestine infected with the natural parasite Heligmosomoides polygyrus, which allows one to investigate metabolic processes both in host and parasite tissues in a spatially resolved manner.

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Biochemistry

Monitoring Leucine-Rich Repeat Containing 8 Channel (LRRC8/VRAC) Activity Using Sensitized-Emission F
Malte Klüssendorf 1, Sumaira Pervaiz 2, Tobias Stauber 1,2
1Institute for Molecular Medicine, MSH Medical School Hamburg, 2Institute of Chemistry and Biochemistry, Freie Universität Berlin

Electrophysiology is the gold standard for investigating ion channel activity. However, there are plenty of alternative approaches, including optical methods. Here, we describe a method to monitor the activity of the leucine-rich repeat containing 8 channel (LRRC8)-formed anion channels using an inter-subunit Förster resonance energy transfer (FRET)-based method.

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