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Heinrich Heine University Düsseldorf

9 ARTICLES PUBLISHED IN JoVE

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Neuroscience

Multi-photon Intracellular Sodium Imaging Combined with UV-mediated Focal Uncaging of Glutamate in CA1 Pyramidal Neurons
Christian Kleinhans *1, Karl W. Kafitz *1, Christine R. Rose 1
1Institute of Neurobiology, Heinrich Heine University Düsseldorf

We describe the combination of focal UV-induced photo-activation of neuro-active compounds with whole-cell patch-clamp and multi-photon imaging of intracellular sodium transients in dendrites and spines of hippocampal neurons in acute tissue slices of the mouse brain.

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Neuroscience

Double-barreled and Concentric Microelectrodes for Measurement of Extracellular Ion Signals in Brain Tissue
Nicole Haack 1, Simone Durry 1, Karl W. Kafitz 1, Mitchell Chesler 2, Christine R. Rose 1
1Institute of Neurobiology, Heinrich Heine University Düsseldorf, 2Departments of Physiology and Neuroscience, New York University School of Medicine

We demonstrate the fabrication, calibration and properties of two types of ion-selective microelectrodes (double-barreled and concentric) for measurement of ion concentrations in brain tissue. These are then used in the mouse hippocampal slice preparation to show that excitatory activity changes both extracellular potassium and sodium concentrations.

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Immunology and Infection

Intraperitoneal Glucose Tolerance Test, Measurement of Lung Function, and Fixation of the Lung to Study the Impact of Obesity and Impaired Metabolism on Pulmonary Outcomes
Katharina Dinger 1, Jasmine Mohr 1, Christina Vohlen 1,2, Dharmesh Hirani 1, Eva Hucklenbruch-Rother 2, Regina Ensenauer 3, Jörg Dötsch 2, Miguel A. Alejandre Alcazar 1,2
1Translational Experimental Pediatrics, Experimental Pulmonology, Department of Pediatrics and Adolescent Medicine, University of Cologne, 2Department of Pediatrics and Adolescent Medicine, University of Cologne, 3Division of Experimental Pediatrics and Metabolism, University Children's Hospital, Heinrich Heine University Düsseldorf

The incidence of obesity is rising and increases the risk of chronic lung diseases. To establish the underlying mechanisms and preventive strategies, well-defined animal models are needed. Here, we provide three methods (glucose-tolerance-test, body plethysmography, and lung fixation) to study the effect of obesity on pulmonary outcomes in mice.

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Neuroscience

Optogenetic Entrainment of Hippocampal Theta Oscillations in Behaving Mice
Franziska Bender 1,2, Tatiana Korotkova 2,3, Alexey Ponomarenko 1,2
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.

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Medicine

Impact of Intracardiac Neurons on Cardiac Electrophysiology and Arrhythmogenesis in an Ex Vivo Langendorff System
Christiane Jungen 1,2, Katharina Scherschel 1,2, Nadja I. Bork 2,3, Pawel Kuklik 1, Christian Eickholt 1, Helge Kniep 1, Niklas Klatt 1,2, Stephan Willems 1,2, Viacheslav O. Nikolaev 2,3, Christian Meyer 1,2
1Department of Cardiology-Electrophysiology, cNEP (cardiac Neuro- and Electrophysiology research group), University Heart Center, University Hospital Hamburg-Eppendorf, 2DZHK (German Center for Cardiovascular Research), 3Institute of Experimental Cardiovascular Research, University Medical Center Hamburg-Eppendorf

Here, we present a protocol for the modulation of the intracardiac autonomic nervous system and the assessment of its influence on basic electrophysiology, arrhythmogenesis, and cAMP dynamics using an ex vivo Langendorff setup.

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Neuroscience

Imaging of Intracellular ATP in Organotypic Tissue Slices of the Mouse Brain using the FRET-based Sensor ATeam1.03YEMK
Rodrigo Lerchundi *1, Karl W. Kafitz *1, Marcel Färfers 1, Felix Beyer 2, Na Huang 1, Christine R. Rose 1
1Institute of Neurobiology, Heinrich Heine University Düsseldorf, 2Department of Neurology, Düsseldorf University Hospital

We describe a protocol for cell-type specific expression of the genetically encoded FRET-based sensor ATeam1.03YEMK in organotypic slice cultures of the mouse forebrain. Furthermore, we show how to use this sensor for dynamic imaging of cellular ATP levels in neurons and astrocytes.

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Medicine

Location, Dissection, and Analysis of the Murine Stellate Ganglion
Katharina Scherschel 1,2,3, Hanna Bräuninger 2,4, Klara Glufke 2, Christiane Jungen 2,4,5, Nikolaj Klöcker 3, Christian Meyer 1,2,3
1Division of Cardiology, EVK Düsseldorf, cNEP, cardiac Neuro- and Electrophysiology Research Consortium, 2DZHK (German Centre for Cardiovascular Research), 3Institute of Neural and Sensory Physiology, Medical Faculty, Heinrich Heine University Düsseldorf, 4Clinic for Cardiology, University Heart & Vascular Centre, University Hospital Hamburg-Eppendorf, 5Department of Cardiology, Leiden University Medical Center

Pathophysiological changes in the cardiac autonomic nervous system, especially in its sympathetic branch, contribute to the onset and maintenance of ventricular arrhythmias. In the present protocol, we show how to characterize murine stellate ganglia to improve the understanding of the underlying molecular and cellular processes.

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Neuroscience

Generation of Human Brain Organoids for Mitochondrial Disease Modeling
Stephanie Le 1, Laura Petersilie 2, Gizem Inak 1,4, Carmen Menacho-Pando 1, Karl W. Kafitz 2, Agnieszka Rybak-Wolf 3, Nikolaus Rajewsky 3, Christine R. Rose 2, Alessandro Prigione 1,5
1Department of General Pediatrics, Neonatology and Pediatric Cardiology, Duesseldorf University Hospital, Medical Faculty, Heinrich Heine University, 2Institute of Neurobiology, Heinrich Heine University, 3Berlin Institute for Medical Systems Biology (BIMSB), Max Delbrueck Center for Molecular Medicine (MDC), 4Seaver Autism Center for Research and Treatment, Icahn School of Medicine at Mount Sinai, 5Max Delbrueck Center for Molecular Medicine (MDC)

We describe a detailed protocol for the generation of human induced pluripotent stem cell-derived brain organoids and their use in modeling mitochondrial diseases.

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Biochemistry

NMR 15N Relaxation Experiments for the Investigation of Picosecond to Nanoseconds Structural Dynamics of Proteins
Tobias Stief 1,2, Katharina Vormann 1,2, Nils-Alexander Lakomek 1,2
1Institute of Biological Information Processing (IBI-7), Forschungszentrum Jülich, 2Institute of Physical Biology, Faculty of Mathematics and Natural Sciences, Heinrich Heine University Düsseldorf

Nuclear magnetic resonance (NMR) spectroscopy can characterize structural protein dynamics in a residue-specific manner. We provide a hands-on protocol for recording NMR 15N R1 and R2 relaxation and {1H}-15N heteronuclear Overhauser effect (hetNOE) experiments, sensitive to the picoseconds to nanoseconds timescale.

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