Eberhard Karls University of Tubingen 9 articles published in JoVE Neuroscience Organotypic Retinal Explant Cultures from Macaque Monkey Wenrong Xu1, Yujie Dong1, Yan Li1, Zhulin Hu1, François Paquet-Durand2, Kangwei Jiao1 1Yunnan Eye Institute & Key Laboratory of Yunnan Province, Yunnan Eye Disease Clinical Medical Center, Affiliated Hospital of Yunnan University, Yunnan University, 2Institute for Ophthalmic Research, Eberhard-Karls-Universität Tübingen Retinal explants obtained from wild-type macaques were cultured in vitro. Retinal degeneration and the cGMP-PKG signaling pathway was induced using the PDE6 inhibitor zaprinast. cGMP accumulation in the explants at different zaprinast concentrations was verified using immunofluorescence. Neuroscience Automated Gait Analysis to Assess Functional Recovery in Rodents with Peripheral Nerve or Spinal Cord Contusion Injury Johannes Heinzel1,2,3, Nicole Swiadek2,3, Mohamed Ashmwe2,3, Alexander Rührnößl2,3, Viola Oberhauser2,3, Jonas Kolbenschlag1, David Hercher2,3 1Department of Hand-, Plastic, Reconstructive and Burn Surgery, BG Trauma Center Tubingen, Eberhard Karls University, 2Ludwig Boltzmann Institute for Experimental and Clinical Traumatology, 3Austrian Cluster for Tissue Regeneration Automated gait analysis is a feasible tool to evaluate functional recovery in rodent models of peripheral nerve injury and spinal cord contusion injury. While it requires only one setup to assess locomotor function in various experimental models, meticulous hard- and soft-ware adjustment and training of the animals is highly important. Cancer Research A Chromatin Immunoprecipitation Assay to Identify Novel NFAT2 Target Genes in Chronic Lymphocytic Leukemia Alexander R. Fuchs1, Melanie Märklin1, Jonas S. Heitmann1, Stefan Futterknecht1, Michael Haap2, Stefan Wirths1, Hans-Georg Kopp1, Clemens Hinterleitner1, Daniela Dörfel1, Martin R. Müller1 1Dept. of Hematology, Oncology and Immunology, University of Tübingen, 2Dept. of Endocrinology, Diabetology, Clinical Pathology and Metabolism, University of Tübingen Chronic lymphocytic leukemia (CLL) is the most common leukemia in the western world. NFAT transcription factors are important regulators of development and activation in numerous cell types. Here, we present a protocol for the use of chromatin immunoprecipitation (ChIP) in human CLL cells to identify novel target genes of NFAT2. Medicine Induction of Nephrotic Syndrome in Mice by Retrobulbar Injection of Doxorubicin and Prevention of Volume Retention by Sustained Release Aprotinin Bernhard N. Bohnert1,2,3, Ferruh Artunc1,2,3 1Department of Internal Medicine, Division of Endocrinology, Diabetology, Vascular Disease, Nephrology and Clinical Chemistry, University Hospital Tübingen, 2Institute of Diabetes Research and Metabolic Diseases (IDM) of the Helmholtz Center Munich, University Tübingen, 3German Center for Diabetes Research (DZD), University Tübingen Here, we describe the induction of experimental nephrotic syndrome in 129S1/SvImJ mice by rapid retrobulbar injection of doxorubicin. We also treat nephrotic mice with sustained release pellets containing aprotinin to inhibit urinary serine protease activity and prevent sodium retention. Immunology and Infection Non-invasive In Vivo Fluorescence Optical Imaging of Inflammatory MMP Activity Using an Activatable Fluorescent Imaging Agent Johannes Schwenck1,2, Florian C. Maier1, Manfred Kneilling1,3, Stefan Wiehr1, Kerstin Fuchs1 1Werner Siemens Imaging Center, Department of Preclinical Imaging and Radiopharmacy, Eberhard Karls University of Tübingen, 2Department of Nuclear Medicine, Eberhard Karls University of Tübingen, 3Department of Dermatology, Eberhard Karls University of Tübingen This paper explains the application of fluorescent imaging using an activatable optical imaging probe to visualize the in vivo activity of key matrix metalloproteinases in two different experimental models of inflammation. Immunology and Infection Murine Lymphocyte Labeling by 64Cu-Antibody Receptor Targeting for In Vivo Cell Trafficking by PET/CT Sabrina H. L. Hoffmann1, Andreas Maurer1, Dorothea I. Reck1, Gerald Reischl1, Bernd J. Pichler1, Manfred Kneilling1,2, Christoph M. Griessinger1 1Werner Siemens Imaging Center, Department of Preclinical Imaging and Radiopharmacy, Eberhard Karls University Tübingen, 2Department of Dermatology, Eberhard Karls University Tübingen Following the preparation of a 64Cu-modified monoclonal antibody binding to a transgenic murine T cell receptor, T cells are radiolabeled in vivo, analyzed for viability, functionality, labeling stability and apoptosis, and adoptively transferred into mice with an airway delayed-type hypersensitivity reaction for non-invasive imaging by positron emission tomography/computed tomography (PET/CT). Chemistry The Synthesis of [Sn10(Si(SiMe3)3)4]2- Using a Metastable Sn(I) Halide Solution Synthesized via a Co-condensation Technique Mareike Binder1, Claudio Schrenk1, Andreas Schnepf1 1Chemistry Department, Institute of Inorganic Chemistry, University of Tübingen The disproportionation reaction of a metastable Sn(I) chloride solution, obtained via the preparative co-condensation technique, is used for the synthesis of a metalloid tin cluster compound. Immunology and Infection Retroviral Transduction of Helper T Cells as a Genetic Approach to Study Mechanisms Controlling their Differentiation and Function Yogesh Singh1,2, Oliver A. Garden3, Florian Lang2, Bradley S. Cobb1 1Department of Comparative Biomedical Sciences, The Royal Veterinary College, 2Institute of Physiology I, Cardiology & Vascular Medicine, Eberhard Karls University of Tuebingen, 3Department of Clinical Science and Services, The Royal Veterinary College Many experimental systems have been utilized to understand the mechanisms regulating T cell development and function in an immune response. Here a genetic approach using retroviral transduction is described, which is economic, time efficient, and most importantly, highly informative in identifying regulatory pathways. Bioengineering Imaging Cell Viability on Non-transparent Scaffolds — Using the Example of a Novel Knitted Titanium Implant Gauri Tendulkar1, Phillip Grau1, Patrick Ziegler1,2, Alfred Buck, Sr.3, Alfred Buck, Jr.3, Andreas Badke1,2, Hans-Peter Kaps1,2, Sabrina Ehnert1, Andreas K. Nussler1 1Siegfried Weller Institute for Trauma Research at the BG Trauma Center, Eberhard Karls Universität Tübingen, 2Department of Orthopaedics, BG Trauma-Center, 3Buck GmbH and Co.KG Here we present a fluorophore based imaging technique to detect cell viability on a non-transparent titanium scaffold as well as to detect glimpses of the scaffold impurities. This protocol troubleshoots the drawback of imaging cell-cell or cell-metal interactions on non-transparent scaffolds.
