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German Center for Cardiovascular Research (DZHK), Partner Site Munich, Munich Heart Alliance (MHA)

7 ARTICLES PUBLISHED IN JoVE

Immunology and Infection

Following in Real Time the Impact of Pneumococcal Virulence Factors in an Acute Mouse Pneumonia Model Using Bioluminescent Bacteria

Malek Saleh¹, Mohammed R. Abdullah¹, Christian Schulz¹, Thomas Kohler¹, Thomas Pribyl¹, Inga Jensch¹, Sven Hammerschmidt¹

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.

Medicine

Surgical Placement of Catheters for Long-term Cardiovascular Exercise Testing in Swine

Daphne P M De Wijs-Meijler^*1, Kelly Stam^*2, Richard W B van Duin¹, Annemarie Verzijl², Irwin K Reiss³, Dirk J Duncker², Daphne Merkus²

¹Experimental Cardiology and Neonatology, Erasmus MC, ²Experimental Cardiology, Erasmus MC, ³Neonatology, Erasmus MC

Here we present a protocol to assess cardiopulmonary function in awake swine, at rest and during graded treadmill exercise. Chronic instrumentation allows for repeated hemodynamic measurements uninfluenced by cardiodepressive anesthetic agents.

Medicine

Isolation and Culture of Resident Cardiac Macrophages from the Murine Sinoatrial and Atrioventricular Node

Ruibing Xia^1,2,3, Simone Loy¹, Stefan Kääb^1,3, Anna Titova¹, 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.

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.

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 Simahendra³, Ludwig T. Weckbach^1,2,3, Stefan Brunner¹, 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.

Medicine

Preparation of Human Myocardial Tissue for Long-Term Cultivation

Jules Hamers^1,2, Payel Sen^1,2, Daphne Merkus^1,2,3, Thomas Seidel^4,5, Kun Lu^1,6, Andreas Dendorfer^1,2

¹Walter-Brendel-Centre of Experimental Medicine, University Hospital, LMU Munich, ²German Center for Cardiovascular Research (DZHK), Partner Site Munich, Munich Heart Alliance (MHA), ³Division of Experimental Cardiology, Department of Cardiology, Erasmus MC, University Medical Center Rotterdam, ⁴Institute of Cellular and Molecular Physiology, Friedrich-Alexander-Universität Erlangen-Nürnberg, ⁵Muscle Research Center Erlangen (MURCE), Friedrich-Alexander-Universität Erlangen-Nürnberg, ⁶Department of Cardiac Surgery, University Hospital, LMU Munich

We present a protocol for ex vivo cultivation of human ventricular myocardial tissue. It allows for detailed analysis of contraction force and kinetics, as well as the application of pre- and afterload to mimic the in vivo physiological environment more closely.

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.