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Heinrich Heine University

5 ARTICLES PUBLISHED IN JoVE

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. Rose¹

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.

Neuroscience

Double-barreled and Concentric Microelectrodes for Measurement of Extracellular Ion Signals in Brain Tissue

Nicole Haack¹, Simone Durry¹, Karl W. Kafitz¹, Mitchell Chesler², Christine R. Rose¹

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.

Medicine

The Inverted Heart Model for Interstitial Transudate Collection from the Isolated Rat Heart

Kezhe Tan^*1,2, Zhaoping Ding^*1,2, Bodo Steckel², Sonja Hartwig³, Stefan Lehr³, Xiaoming Deng¹, Jürgen Schrader²

¹Department of Anesthesiology, Changhai Hospital, Second Military Medical University, ²Department of Molecular Cardiology, Heinrich Heine University, ³Department of Clinical Biochemistry and Pathobiochemistry, German Diabetes Center, Heinrich Heine University

This protocol describes a method to collect cardiac interstitial fluid from the isolated, perfused rat heart. To physically separate interstitial transudate from coronary venous effluent perfusate, the Langendorff perfused heart is inverted, and the transudate (interstitial fluid) formed on the cardiac surface is collected using a soft latex cap.

Neuroscience

Imaging of Intracellular ATP in Organotypic Tissue Slices of the Mouse Brain using the FRET-based Sensor ATeam1.03^YEMK

Rodrigo Lerchundi^*1, Karl W. Kafitz^*1, Marcel Färfers¹, Felix Beyer², Na Huang¹, Christine R. Rose¹

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 ATeam1.03^YEMK 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.

Neuroscience

Generation of Human Brain Organoids for Mitochondrial Disease Modeling

Stephanie Le¹, Laura Petersilie², Gizem Inak^1,4, Carmen Menacho-Pando¹, Karl W. Kafitz², Agnieszka Rybak-Wolf³, Nikolaus Rajewsky³, Christine R. Rose², 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.