Leibniz Institute of Molecular Pharmacology (FMP-Berlin) 5 articles published in JoVE Biology In Vivo Quantification of Protein Turnover in Aging C. Elegans using Photoconvertible Dendra2 Maria Lucia Pigazzini1,2, Janine Kirstein1,3 1Leibniz Research Institute for Molecular Pharmacology im Forschungsverbund, 2NeuroCure Cluster of Excellence, Charité - Universitätsmedizin Berlin, 3Department of Biology and Chemistry, University of Bremen Presented here is a protocol to monitor degradation of the protein huntingtin fused to the photoconvertible fluorophore Dendra2. Biochemistry Characterization of Amyloid Structures in Aging C. Elegans Using Fluorescence Lifetime Imaging Maria Lucia Pigazzini*1,2, Christian Gallrein*1, Manuel Iburg*1, Gabriele Kaminski Schierle3, Janine Kirstein1,4 1Leibniz Research Institute for Molecular Pharmacology im Forschungsverbund Berlin, 2NeuroCure Cluster of Excellence, Charité - Universitätsmedizin Berlin, 3Molecular Neuroscience Group, Department of Chemical Engineering and Biotechnology, University of Cambridge, 4Cell Biology, University of Bremen Fluorescence lifetime imaging monitors, quantifies and distinguishes the aggregation tendencies of proteins in living, aging, and stressed C. elegans disease models. Neuroscience Optogenetic Entrainment of Hippocampal Theta Oscillations in Behaving Mice Franziska Bender1,2, Tatiana Korotkova2,3, Alexey Ponomarenko1,2 1Systems Neurophysiology Research Group, Institute of Clinical Neuroscience and Medical Psychology, Medical Faculty, Heinrich Heine University Düsseldorf, 2Behavioural Neurodynamics Group, Leibniz Institute for Molecular Pharmacology (FMP)/ NeuroCure Cluster of Excellence, 3Neuronal Circuits and Behavior Research Group, Max Planck Institute for Metabolism Research We describe the use of optogenetics and electrophysiological recordings for selective manipulations of hippocampal theta oscillations (5-10 Hz) in behaving mice. The efficacy of the rhythm entrainment is monitored using local field potential. A combination of opto- and pharmacogenetic inhibition addresses the efferent readout of hippocampal synchronization. Biochemistry Reconstitution of Nucleosomes with Differentially Isotope-labeled Sister Histones Stamatios Liokatis1 1Department of Structural Biology, Leibniz Institute of Molecular Pharmacology (FMP-Berlin) This protocol describes the reconstitution of nucleosomes containing differentially isotope-labeled sister histones. At the same time, asymmetrically post-translationally modified nucleosomes can be generated after using a premodified histone copy. These preparations can be readily used to study modification crosstalk mechanisms, simultaneously on both sister histones, by using high-resolution NMR spectroscopy. Biology Culturing Primary Rat Inner Medullary Collecting Duct Cells Dörte Faust1, Andrea Geelhaar1, Beate Eisermann1, Jenny Eichhorst2, Burkhard Wiesner2, Walter Rosenthal1,3, Enno Klussmann1 1Anchored Signalling, Max-Delbrück-Center for Molecular Medicine, 2Leibniz Institute for Molecular Pharmacology (FMP), 3Charité University Medicine Berlin Arginine-vasopressin (AVP) controls fine-tuning of body water homeostasis through facilitating water reabsorption by renal principal cells. Here, we present a protocol for the cultivation of primary rat inner medullary collecting duct cells suitable for the elucidation of molecular mechanisms underlying AVP-mediated water reabsorption.