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Humboldt-Universität zu Berlin

6 ARTICLES PUBLISHED IN JoVE

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Neuroscience

Whole-cell Patch-clamp Recordings for Electrophysiological Determination of Ion Selectivity in Channelrhodopsins
Christiane Grimm *1, Johannes Vierock *1, Peter Hegemann 1, Jonas Wietek 1
1Experimental Biophysics, Institute of Biology, Humboldt-Universität zu Berlin

This article describes how the ion selectivity of channelrhodopsin is determined with electrophysiological whole-cell patch-clamp recordings using HEK293 cells. Here, the experimental procedure for investigating chloride selectivity of an anion-selective channelrhodopsin is demonstrated. However, the procedure is transferable to other channelrhodopsins of distinct selectivity.

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Biochemistry

Fatty Acid 13C Isotopologue Profiling Provides Insight into Trophic Carbon Transfer and Lipid Metabolism of Invertebrate Consumers
Ralph Menzel 1, Rainer Nehring 1, Dilara Simsek 1, Liliane Ruess 1
1Ecology, Institute of Biology, Humboldt-Universität zu Berlin

The fatty acid trophic marker approach, i.e., the assimilation of fatty acids as entire molecule and transfer into consumer tissue with no or minor modification, is hampered by knowledge gaps in fatty acid metabolism of small soil invertebrates. Isotopologue profiling is provided as a valuable tool to disentangle trophic interactions.

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Behavior

Eye Tracking During Visually Situated Language Comprehension: Flexibility and Limitations in Uncovering Visual Context Effects
Camilo Rodriguez Ronderos 1, Katja Münster 1, Ernesto Guerra 2, Helene Kreysa 3, Alba Rodríguez 1,4, Julia Kröger 4, Thomas Kluth 4, Michele Burigo 4, Dato Abashidze 5, Eva Nunnemann 4, Pia Knoeferle 1,6,7
1Institute of German Studies and Linguistics, Humboldt-Universität zu Berlin, 2Center for Advanced Research in Education – CIAE, Universidad de Chile, 3Institute of Psychology, Friedrich Schiller University Jena, 4Cognitive Interaction Technology Excellence Cluster (CITEC), Faculty of Linguistics and Literature, Bielefeld University, 5Department of Education, Concordia University, 6Berlin School of Mind and Brain, 7Einstein Center for Neurosciences Berlin

The present article reviews an eye-tracking methodology for studies on language comprehension. To obtain reliable data, key steps of the protocol must be followed. Among these are the correct set-up of the eye tracker (e.g., ensuring good quality of the eye and head images) and accurate calibration.

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Immunology and Infection

An Advanced Murine Model for Nonalcoholic Steatohepatitis in Association with Type 2 Diabetes
Julia Sbierski-Kind 1,2,3,4, Katharina Schmidt-Bleek 3,6, Mathias Streitz 3,5, Jonas Kath 3,5, Joachim Spranger *1,2,4, Hans-Dieter Volk *2,3,5
1Department of Endocrinology & Metabolism Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, 2Berlin Institute of Health (BIH), 3Berlin-Brandenburg Center for Regenerative Therapies (BCRT), Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, 4DZHK (German Centre for Cardiovascular Research), 5Institute of Medical Immunology, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, 6Julius Wolff Institute (JWI) and Center for Musculoskeletal Surgery, Charité - Universitätsmedizin Berlin, Berlin, Germany, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin

A simple and reliable diet-induced rodent animal model for nonalcoholic steatohepatitis (NASH) is described, achieved through non-SPF housing of the animals and administration of a specific high-fat diet. We describe identification of hepatic and adipose immune cell subsets to recapitulate human immunological conditions by exposing mice to environmental germs.

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Medicine

Magnetic Resonance Imaging of Multiple Sclerosis at 7.0 Tesla
Sonia Waiczies *1, Antje Els *1, Joseph Kuchling *2,3,4, Karin Markenroth Bloch 5, Anna Pankowska 6,7, Helmar Waiczies 8, Carl Herrmann 1, Claudia Chien 2,3, Carsten Finke 4,9, Friedemann Paul 2,3,4, Thoralf Niendorf 1,2,8
1Berlin Ultrahigh Field Facility (B.U.F.F.), Max Delbrück Center for Molecular Medicine in the Helmholtz Association, 2Experimental and Clinical Research Center, Max Delbrueck Center for Molecular Medicine and Charité – Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, 3NeuroCure Clinical Research Center, Charité – Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, 4Department of Neurology, Charité – Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, 5The Swedish National 7T Facility, Lund University Bioimaging Center, Lund University, 6Department of Radiography, Medical University of Lublin, 7ECOTECH-COMPLEX, Maria Curie-Skłodowska University, 8MRI.TOOLS GmbH, 9Berlin School of Mind and Brain, Humboldt-Universität zu Berlin

Here, we present a protocol to acquire magnetic resonance (MR) images of multiple sclerosis (MS) patient brains at 7.0 Tesla. The protocol includes preparation of the setup including the radio-frequency coils, standardized interview procedures with MS patients, subject positioning in the MR scanner and MR data acquisition.

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Developmental Biology

Generation and Downstream Analysis of Single-Cell and Single-Nuclei Transcriptomes in Brain Organoids
Miriam Wandres *1,2, Denise Aigner *1,2, Nicolai Kastelic 1,3, Anastasiya Boltengagen 1, Agnieszka Rybak-Wolf 1,3, Nikolaus Rajewsky 1,2
1Berlin Institute for Medical Systems Biology (BIMSB), Max Delbrück Center for Molecular Medicine (MDC), 2Institut für Biologie, Humboldt-Universität zu Berlin, 3Organoid platform, Berlin Institute for Medical Systems Biology (BIMSB), Max Delbrück Center for Molecular Medicine (MDC)

Here, we introduce a comprehensive protocol for the generation and downstream analysis of human brain organoids using single-cell and single-nucleus RNA sequencing.

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