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LMU Munich

14 ARTICLES PUBLISHED IN JoVE

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Immunology and Infection

Following in Real Time the Impact of Pneumococcal Virulence Factors in an Acute Mouse Pneumonia Model Using Bioluminescent Bacteria
Malek Saleh 1, Mohammed R. Abdullah 1, Christian Schulz 1, Thomas Kohler 1, Thomas Pribyl 1, Inga Jensch 1, Sven Hammerschmidt 1
1Department Genetics of Microorganisms, Interfaculty Institute for Genetics and Functional Genomics, University of Greifswald

Streptococcus pneumoniae is the leading pathogen causing severe community-acquired pneumonia and responsible for over 2 million deaths worldwide. The impact of bacterial factors implicated in fitness or virulence can be monitored in real-time in an acute mouse pneumonia or bacteremia model using bioluminescent bacteria.

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Medicine

Modeling Stroke in Mice: Permanent Coagulation of the Distal Middle Cerebral Artery
Gemma Llovera *1,2, Stefan Roth *1,2, Nikolaus Plesnila 1,2, Roland Veltkamp 3,4, Arthur Liesz 1,2
1Institute for Stroke and Dementia Research, University Hospital Munich, 2Munich Cluster for Systems Neurology (SyNergy), 3Department of Neurology, University Heidelberg, 4Imperial College, Charing Cross Hospital

Various murine models of middle cerebral artery occlusion (MCAO) are widely used in experimental brain research. Here, we demonstrate the model of transcranial permanent distal MCAO which produces consistent cortical infarction of a size corresponding to damage imposed by the majority of human ischemic strokes.

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Medicine

In Vivo Morphometric Analysis of Human Cranial Nerves Using Magnetic Resonance Imaging in Menière's Disease Ears and Normal Hearing Ears
Wilhelm H. Flatz 1, Annika Henneberger 2, Maximilian F. Reiser 1, Robert Gürkov 3, Birgit Ertl-Wagner 1
1Department of Radiology, University Hospital, LMU Munich, 2Department of Otorhinolaryngology - Head and Neck Surgery, SLK-Kliniken Heilbronn GmbH, 3Department of Otorhinolaryngology Head and Neck Surgery, Ludwig-Maximilians-University Hospital Munich, German Centre for Vertigo and Balance Disorder

To evaluate morphological changes of cranial nerves such as loss of neural structures or swelling of cranial nerves in Menière's Disease (MD) or in healthy persons in vivo, a protocol of evaluation has been developed using magnetic resonance imaging (MRI). Additional MRI-based confirmation of MD was performed.

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Medicine

Sampling Strategies and Processing of Biobank Tissue Samples from Porcine Biomedical Models
Andreas Blutke 1, Rüdiger Wanke 1
1Institute of Veterinary Pathology at the Centre for Clinical Veterinary Medicine, LMU Munich

The practical application and performance of methods for generation of representative tissue samples of porcine animal models for a broad spectrum of downstream analyses in biobank projects are demonstrated, including volumetry, systematic random sampling, and differential processing of tissue samples for qualitative and quantitative morphologic and molecular analyses types.

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Medicine

Multiplex Therapeutic Drug Monitoring by Isotope-dilution HPLC-MS/MS of Antibiotics in Critical Illnesses
Carina Schuster 1, Sebastian Sterz 1, Daniel Teupser 1, Mathias Brügel 1, Michael Vogeser 1, Michael Paal 1
1Institute of Laboratory Medicine, University Hospital, LMU Munich

Here we present a tandem mass spectrometry-based protocol for the quantification of frequently used antibiotics in intensive care units, namely cefepime, meropenem, ciprofloxacin, moxifloxacin, linezolid, and piperacillin.

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Behavior

Brain Infarct Segmentation and Registration on MRI or CT for Lesion-symptom Mapping
J. Matthijs Biesbroek 1, Hugo J. Kuijf 2, Nick A. Weaver 1, Lei Zhao 3, Marco Duering 4, , Geert Jan Biessels 1
1Department of Neurology and Neurosurgery, UMC Utrecht Brain Center, University Medical Center Utrecht, Utrecht University, 2Image Sciences Institute, University Medical Center Utrecht, 3BrainNow Research Institute, 4Institute for Stroke and Dementia Research, University Hospital, LMU Munich

Provided here is a practical tutorial for an open-access, standardized image processing pipeline for the purpose of lesion-symptom mapping. A step-by-step walkthrough is provided for each processing step, from manual infarct segmentation on CT/MRI to subsequent registration to standard space, along with practical recommendations and illustrations with exemplary cases.

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Chemistry

18F-Labeling of Radiotracers Functionalized with a Silicon Fluoride Acceptor (SiFA) for Positron Emission Tomography
David Connolly *1, Justin J. Bailey *1, Harun Ilhan 2,3, Peter Bartenstein 2,3, Carmen Wängler 4, Björn Wängler 5, Melinda Wuest 1, Frank Wuest 1, Ralf Schirrmacher 1
1Department of Oncology, University of Alberta, 2Department of Nuclear Medicine, University Hospital of Munich, 3ENETS Centre of Excellence, Interdisciplinary Center of Neuroendocrine Tumors of the GastroEnteroPancreatic System, LMU Munich, 4Biomedical Chemistry, Department of Clinical Radiology and Nuclear Medicine, Medical Faculty Mannheim of Heidelberg University, 5Molecular Imaging and Radiochemistry, Department of Clinical Radiology and Nuclear Medicine, Medical Faculty Mannheim of Heidelberg University

The synthesis of fluorine-18 (18F) labeled radiopharmaceuticals for positron emission tomography typically requires months of experience. When incorporated into a radiotracer, the silicon-fluoride acceptor (SiFA) motif enables a simple 18F-labeling protocol that is independent of costly equipment and preparatory training, while reducing precursor quantity needed and utilizing milder reaction conditions.

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Medicine

Isolation and Culture of Resident Cardiac Macrophages from the Murine Sinoatrial and Atrioventricular Node
Ruibing Xia 1,2,3, Simone Loy 1, Stefan Kääb 1,3, Anna Titova 1, Christian Schulz 1,2,3, Steffen Massberg 1,2,3, Sebastian Clauss 1,2,3
1University Hospital Munich, Department of Medicine I, Ludwig-Maximilian-Unversity Munich (LMU), 2Insitute of Surgical Research at the Walter Brendel Center of Experimental Medicine, University Hospital Munich, Ludwig-Maximilians-University (LMU), 3German Center for Cardiovascular Research (DZHK), Partner Site Munich, Munich Heart Alliance (MHA)

The protocol presented here provides a step-by-step approach for the isolation of cardiac resident macrophages from the sinoatrial node (SAN) and atrioventricular node (AVN) region of mouse hearts.

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Medicine

Analyzing Long-Term Electrocardiography Recordings to Detect Arrhythmias in Mice
Philipp Tomsits 1,2,3, Kavi Raj Chataut 1,2, Aparna Sharma Chivukula 1,2, Li Mo 1,2, Ruibing Xia 1,2, Dominik Schüttler 1,2,3, Sebastian Clauss 1,2,3
1Department of Medicine I, University Hospital Munich, Campus Großhadern, Ludwig-Maximilians University Munich (LMU), 2DZHK (German Centre for Cardiovascular Research), Partner Site Munich, Munich Heart Alliance (MHA), 3Walter Brendel Centre of Experimental Medicine, Ludwig-Maximilians University Munich (LMU)

Here we present a step-by-step protocol for a semiautomated approach to analyze murine long-term electrocardiography (ECG) data for basic ECG parameters and common arrhythmias. Data are obtained by implantable telemetry transmitters in living and awake mice and analyzed using Ponemah and its analysis modules.

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Neuroscience

Modeling Stroke in Mice: Focal Cortical Lesions by Photothrombosis
Gemma Llovera 1, Kelsey Pinkham 1, Arthur Liesz 1,2
1Institute for Stroke and Dementia Research, LMU Munich, 2Munich Cluster for Systems Neurology (SyNergy)

Described here is the photothrombotic stroke model, where a stroke is produced through the intact skull by inducing permanent microvascular occlusion using laser illumination after administration of a photosensitive dye.

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Neuroscience

Modeling Stroke in Mice: Transient Middle Cerebral Artery Occlusion via the External Carotid Artery
Gemma Llovera 1, Alba Simats 1, Arthur Liesz 1,2
1Institute for Stroke and Dementia Research, LMU Munich, 2Munich Cluster for Systems Neurology (SyNergy)

Different models of middle cerebral artery occlusion (MCAo) are used in experimental stroke research. Here, an experimental stroke model of transient MCAo via the external carotid artery (ECA) is described, which aims to mimic human stroke, in which the cerebrovascular thrombus is removed due to spontaneous clot lysis or therapy.

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Medicine

Real-Time Electrocardiogram Monitoring During Treadmill Training in Mice
Philipp Tomsits *1,2,4, Aparna Sharma Chivukula *1,2, Kavi Raj Chataut *1,2, Agus Simahendra 3, Ludwig T. Weckbach 1,2,3, Stefan Brunner 1, Sebastian Clauss 1,2,4
1Department of Medicine I, University Hospital Munich, Ludwig-Maximilians University Munich (LMU), 2DZHK (German Centre for Cardiovascular Research), Partner Site Munich, Munich Heart Alliance (MHA), 3Institute of Cardiovascular Physiology and Pathophysiology, Biomedical Center, Ludwig-Maximilians-University, 4Institute of Surgical Research at the Walter-Brendel-Centre of Experimental Medicine, University Hospital Munich, Ludwig-Maximilians University Munich (LMU)

Electrocardiogram (ECG) is the key variable to understanding cardiac electrophysiology. Physical exercise has beneficial effects but may also be harmful in the context of cardiovascular diseases. This manuscript provides a method of recording real-time ECG during exercise, which can serve to investigate its effects on cardiac electrophysiology in mice.

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An Innovative Toolkit to Investigate the Complex Mechanisms of Cardiac Arrhythmias
Sebastian Clauss 1,2,3,4
1Department of Medicine I, University Hospital Munich (LMU), Ludwig-Maximilians-University Munich (LMU), 2Institute of Surgical Research at the Walter-Brendel-Centre of Experimental Medicine, University Hospital, LMU Munich, 3DZHK (German Centre for Cardiovascular Research) Partner Site Munich, Munich Heart Alliance (MHA), 4Interfaculty Center for Endocrine and Cardiovascular Disease Network Modelling and Clinical Transfer (ICONLMU), LMU Munich

An Innovative Toolkit to Investigate the Complex Mechanisms of Cardiac Arrhythmias

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Medicine

Microdissection and Immunofluorescence Staining of Myocardial Sleeves in Murine Pulmonary Veins
Hannes E. Villgrater *1,2,3, Ruibing Xia *1,2,3, Aparna Sharma Chivukula 1,2,3, Philipp Tomsits *1,2,3, Sebastian Clauss *1,2,3,4
1University Hospital Munich, Department of Medicine I, Ludwig-Maximilians-University Munich (LMU), 2Walter Brendel Center of Experimental Medicine (WBex), LMU Munich, 3German Center for Cardiovascular Research (DZHK), Partner Site Munich, Munich Heart Alliance (MHA), 4Interfaculty Center for Endocrine and Cardiovascular Disease Network Modelling and Clinical Transfer (ICONLMU), LMU Munich

This protocol demonstrates microscopy-guided isolation and immunofluorescence staining of murine pulmonary veins. We prepare tissue samples containing the left atrium, pulmonary veins, and the corresponding lungs and stain them for cardiac Troponin T and Connexin 43.

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