German Center for Diabetes Research (DZD)

4 ARTICLES PUBLISHED IN JoVE

Neuroscience

Analysis of Immune Cells in Single Sciatic Nerves and Dorsal Root Ganglion from a Single Mouse Using Flow Cytometry

Asa S. Hidmark¹, Peter P. Nawroth^1,2,3, Thomas Fleming^1,3

¹Department of Medicine I and Clinical Chemistry, University Hospital of Heidelberg, ²Institute for Diabetes and Cancer IDC Helmholtz Center Munich, Germany & Joint Heidelberg-IDC Translational Diabetes Program, ³German Center for Diabetes Research (DZD)

Quantitative analysis of cell content within the murine sciatic nerve is difficult due to the scarcity of the tissue. This protocol describes a method for tissue digestion and preparation that provides sufficient cells for flow cytometry analysis of immune cell populations from nerves of individual mice.

Biochemistry

Plate-based Large-scale Cultivation of Caenorhabditis elegans: Sample Preparation for the Study of Metabolic Alterations in Diabetes

Katharina Kohl¹, Thomas Fleming^2,3, Kübra Acunman¹, Hans-Peter Hammes^1,4, Michael Morcos^*1, Andrea Schlotterer^*1

¹5th Medical Department, Medical Faculty Mannheim, Heidelberg University, ²Department of Internal Medicine, Heidelberg University, ³German Center for Diabetes Research (DZD), ⁴European Center for Angioscience, Medical Faculty Mannheim, Heidelberg University

This protocol describes a method for the large-scale cultivation of Caenorhabditis elegans on solid media. As an alternative to liquid culture, this protocol allows obtaining parameters of different scales under plate-based cultivation. This increases the comparability of results by omitting the morphological and metabolic differences between liquid and solid media culture.

Biochemistry

Measuring and Interpreting Oxygen Consumption Rates in Whole Fly Head Segments

Louisa Jutta Dietz¹, Anuroop Venkateswaran Venkatasubramani¹, Annika Müller-Eigner², Martin Hrabe de Angelis^3,4,5, Axel Imhof¹, Lore Becker³, Shahaf Peleg^2,6,7

¹Munich Center of Integrated Protein Science and Biomedical Center, Ludwig-Maximilians University of Munich, ²Laboratory for Metabolism and Epigenetics in Aging, Leibniz Institute for Farm Animal Biology (FBN), ³German Mouse Clinic, Helmholtz Zentrum Munich, German Research Center for Environment and Health (GmbH), ⁴German Center for Diabetes Research (DZD), ⁵Chair of Experimental Genetics, School of Life Science Weihenstephan, Technische Universität München, ⁶Laboratory for Metabolism and Epigenetics in Brain Aging, Institute of Neuroregeneration & Neurorehabilitation of Qingdao University, ⁷Molecular Biology Division, Biomedical Center, Faculty of Medicine, Ludwig-Maximilians University of Munich

Measuring alterations in metabolic rates is central to understanding the progression of various diseases and aging. Here, we present a novel technique to measure whole head oxygen consumption that more closely resembles the physiological state and may aid in revealing novel drugs that modify mitochondrial activity.

Immunology and Infection

Observing Islet Function and Islet-Immune Cell Interactions in Live Pancreatic Tissue Slices

Mollie K. Huber¹, Denise M. Drotar^2,3,4, Helmut Hiller¹, Maria L. Beery¹, Paul Joseph¹, Irina Kusmartseva¹, Stephan Speier^2,3,4, Mark A. Atkinson¹, Clayton E. Mathews¹, Edward A. Phelps⁵

¹Department of Pathology, Immunology, and Laboratory Medicine, University of Florida, ²Paul Langerhans Institute Dresden (PLID) of the Helmholtz Zentrum München at the University Clinic Carl Gustav Carus of Technische Universität Dresden, ³Institute of Physiology, Faculty of Medicine, Technische Universität Dresden, ⁴German Center for Diabetes Research (DZD), ⁵J. Crayton Pruitt Family Department of Biomedical Engineering, University of Florida

This study presents the application of live pancreatic tissue slices to the study of islet physiology and islet-immune cell interactions.