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German Center for Neurodegenerative Diseases (DZNE)

12 ARTICLES PUBLISHED IN JoVE

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Medicine

High Content Screening in Neurodegenerative Diseases
Shushant Jain 1, Ronald E. van Kesteren 2, Peter Heutink 1
1Department of Clinical Genetics, VU University Medical Center, 2Center for Neurogenomics and Cognitive Research, Neuroscience Campus Amsterdam

We describe a methodology combining automated cell culturing with high-content imaging to visualize and quantify multiple cellular processes and structures, in a high-throughput manner. Such methods can aid in the further functional annotation of genomes as well as identify disease gene networks and potential drug targets.

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Medicine

In vivo 19F MRI for Cell Tracking
Mangala Srinivas 1, Philipp Boehm-Sturm 2, Markus Aswendt 2, Eberhard D. Pracht 2,3, Carl G. Figdor 1, I. Jolanda de Vries 1, Mathias Hoehn 2
1Department of Tumor Immunology, Nijmegen Center for Molecular Life Sciences, Radboud University Medical Center, 2In-Vivo-NMR Laboratory, Max Planck Institute for Neurological Research, 3German Center for Neurodegenerative Diseases (DZNE)

We describe a general protocol for in vivo cell tracking using MRI in a mouse model with ex vivo labeled cells. A typical protocol for cell labeling, image acquisition processing and quantification is included.

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Neuroscience

Non-restraining EEG Radiotelemetry: Epidural and Deep Intracerebral Stereotaxic EEG Electrode Placement
Anna Papazoglou 1, Andreas Lundt 1, Carola Wormuth 1, Dan Ehninger 2, Christina Henseler 1, Julien Soós 1, Karl Broich 1, Marco Weiergräber 1
1Department of Neuropsychopharmacology, Federal Institute for Drugs and Medical Devices (Bundesinstitut für Arzneimittel und Medizinprodukte, BfArM), 2Molecular and Cellular Cognition Lab, German Center for Neurodegenerative Diseases (Deutsches Zentrum für Neurodegenerative Erkrankungen, DZNE)

Non-restraining EEG radiotelemetry is a valuable methodological approach to record in vivo long-term electroencephalograms from freely moving rodents. This detailed protocol describes stereotaxic epidural and deep intracerebral electrode placement in different brain regions in order to obtain reliable recordings of CNS rhythmicity and CNS related behavioral stages.

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Behavior

Automatic Detection of Highly Organized Theta Oscillations in the Murine EEG
Ralf Müller 1, Anna Papazoglou 2, Julien Soos 2, Andreas Lundt 2, Carola Wormuth 2, Christina Henseler 2, Dan Ehninger 3, Karl Broich 4, Marco Weiergräber 2
1Department of Psychiatry and Psychotherapy, University of Cologne, 2Department of Neuropsychopharmacology, Federal Institute for Drugs and Medical Devices, 3Molecular and Cellular Cognition Lab, German Center for Neurodegenerative Diseases, 4Federal Institute for Drugs and Medical Devices

Theta activity in the hippocampus is related to specific cognitive and behavioral stages. Here, we describe an analytical method to detect highly-organized theta oscillations within the hippocampus using a time-frequency (i.e., wavelet analysis)-based approach.

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Neuroscience

Analyzing Synaptic Modulation of Drosophila melanogaster Photoreceptors after Exposure to Prolonged Light
Atsushi Sugie 1,2,5, Christoph Möhl 3, Satoko Hakeda-Suzuki 4, Hideaki Matsui 1,2, Takashi Suzuki *4, Gaia Tavosanis *5
1Department of Neuroscience of Disease, Center for Transdisciplinary Research, Niigata University, 2Brain Research Institute, Niigata University, 3Image and Data Analysis Facility, German Center for Neurodegenerative Diseases (DZNE), 4Graduate School of Life Science and Technology, Tokyo Institute of Technology (Titech), 5Dendrite Differentiation, German Center for Neurodegenerative Diseases (DZNE)

Here we show how to quantify the number and spatial distribution of synaptic active zones in Drosophila melanogaster photoreceptors, highlighted with a genetically encoded molecular marker, and their modulation after prolonged exposure to light.

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Neuroscience

Bioluminescence Imaging of Neuroinflammation in Transgenic Mice After Peripheral Inoculation of Alpha-Synuclein Fibrils
Sara Breid 1, Maria E. Bernis 1, Julius B. Tachu 1, Maria C. Garza 2, Holger Wille 2, Gültekin Tamgüney 1
1German Center for Neurodegenerative Diseases (DZNE), 2Centre for Prions and Protein Folding Diseases & Department of Biochemistry, University of Alberta

Peripheral injection of alpha-synuclein fibrils into the peritoneum or tongue of Tg(M83+/-:Gfap-luc+/-) mice, which express human alpha-synuclein with the familial A53T mutation and firefly luciferase, can induce neuropathology, including neuroinflammation, in their central nervous system.

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Biology

Methods to Study Changes in Inherent Protein Aggregation with Age in Caenorhabditis elegans
Nicole Groh 1,2, Ivan Gallotta 1, Marie C. Lechler 1,2, Chaolie Huang 1, Raimund Jung 1, Della C. David 1
1Protein Aggregation and Aging, German Center for Neurodegenerative Diseases (DZNE), 2Graduate School of Cellular and Molecular Neuroscience, German Center for Neurodegenerative Diseases (DZNE)

The goal of the method presented here is to explore protein aggregation during normal aging in the model organism C. elegans. The protocol represents a powerful tool to study the highly insoluble large aggregates that form with age and to determine how changes in proteostasis impact protein aggregation.

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Neuroscience

Data Acquisition and Analysis In Brainstem Evoked Response Audiometry In Mice
Andreas Lundt 1,2, Julien Soos 1, Christina Henseler 1, Muhammad Imran Arshaad 1, Ralf Müller 3, Dan Ehninger 4, Jürgen Hescheler 5, Agapios Sachinidis 5, Karl Broich 6, Carola Wormuth 1,7, Anna Papazoglou 1, Marco Weiergräber 1
1Experimental Neuropsychopharmacology, Federal Institute for Drugs and Medical Devices, 2KBRwyle GmbH, 3Cognitive Neurophysiology, Department of Psychiatry and Psychotherapy, Faculty of Medicine, University of Cologne, 4Molecular and Cellular Cognition, German Center for Neurodegenerative Diseases (DZNE), 5Institute of Neurophysiology, Faculty of Medicine, University of Cologne, 6Federal Institute for Drugs and Medical Devices (BfArM), 7Thescon GmbH

Brainstem evoked response audiometry is an important tool in clinical neurophysiology. Nowadays, brainstem evoked response audiometry is also applied in the basic science and preclinical studies involving both pharmacological and genetic animal models. Here we provide a detailed description of how auditory brainstem responses can be successfully recorded and analyzed in mice.

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Cancer Research

Generation of Orthotopic Pancreatic Tumors and Ex vivo Characterization of Tumor-Infiltrating T Cell Cytotoxicity
Sarah Spear 1,2, Iain A McNeish 1, Melania Capasso 2,3
1Division of Cancer, Department of Surgery and Cancer, Imperial College London, 2Centre for Cancer and Inflammation, Barts Cancer Institute, Queen Mary University of London, 3German Center for Neurodegenerative Diseases (DZNE)

This protocol describes the surgical generation of orthotopic pancreatic tumors and the rapid digestion of freshly isolated murine pancreatic tumors. Following digestion, viable immune cell populations can be used for further downstream analysis, including ex vivo stimulation of T cells for intracellular cytokine detection by flow cytometry.

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Biology

Automated Production of Human Induced Pluripotent Stem Cell-Derived Cortical and Dopaminergic Neurons with Integrated Live-Cell Monitoring
Ashutosh Dhingra *1, Joachim Täger *1, Elisangela Bressan *1, Salvador Rodriguez-Nieto 1, Manmeet-Sakshi Bedi 1, Stefanie Bröer 1, Eldem Sadikoglou 1, Noémia Fernandes 2, Melissa Castillo-Lizardo 1, Patrizia Rizzu 2, Peter Heutink 1,3
1Genome Biology of Neurodegenerative Diseases, German Center for Neurodegenerative Diseases (DZNE), 2Applied Genomics for Neurodegenerative Diseases, German Center for Neurodegenerative Diseases (DZNE), 3Department for Neurodegenerative Diseases, Hertie Institute for Clinical Brain Research, University of Tübingen

We show the automation of human induced pluripotent stem cell (hiPSC) cultures and neuronal differentiations compatible with automated imaging and analysis.

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Neuroscience

Manual Segmentation of the Human Choroid Plexus Using Brain MRI
Deepthi Bannai *1,2, Yuan Cao *3,4, Matcheri Keshavan 1,2, Martin Reuter 5,6,7, Paulo Lizano 1,2,8
1Department of Psychiatry, Beth Israel Deaconess Medical Center, 2Division of Translational Neuroscience, Beth Israel Deaconess Medical Center, 3Huaxi MR Research Center (HMRRC), Department of Radiology, West China Hospital of Sichuan University, 4Department of Psychiatry and Psychotherapy, Jena University Hospital, 5AI in Medical Imaging, German Center for Neurodegenerative Diseases (DZNE), 6A. A. Martinos Center for Biomedical Imaging, Massachusetts General Hospital, 7Department of Radiology, Harvard Medical School, 8Department of Psychiatry, Harvard Medical School

Despite the crucial role of the choroid plexus in the brain, neuroimaging studies of this structure are scarce due to the lack of reliable automated segmentation tools. The present protocol aims to ensure gold-standard manual segmentation of the choroid plexus that can inform future neuroimaging studies.

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Neuroscience

Recording and Analyzing Multimodal Large-Scale Neuronal Ensemble Dynamics on CMOS-Integrated High-Density Microelectrode Array
Brett Addison Emery *1, Shahrukh Khanzada *1, Xin Hu *1, Diana Klütsch 1, Hayder Amin 1,2,3
1Group of "Biohybrid Neuroelectronics (BIONICS)", German Center for Neurodegenerative Diseases (DZNE), 2Faculty of Medicine Carl Gustav Carus, Technical University Dresden, 3Dresden Center for Intelligent Materials (DCIM), Technical University Dresden

Here, we employ HD-MEA to delve into computational dynamics of large-scale neuronal ensembles, particularly in hippocampal, olfactory bulb circuits, and human neuronal networks. Capturing spatiotemporal activity, combined with computational tools, provides insights into neuronal ensemble complexity. The method enhances understanding of brain functions, potentially identifying biomarkers and treatments for neurological disorders.

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