Martin Luther University Halle-Wittenberg 8 articles published in JoVE Immunology and Infection Analyzing the Permeability of the Blood-Brain Barrier by Microbial Traversal through Microvascular Endothelial Cells Veronika Weber1, Kaya Bork1, Rüdiger Horstkorte1, Heidi Olzscha1 1Institute of Physiological Chemistry, Martin Luther University Halle-Wittenberg The human blood-brain barrier selectively prevents penetration of hydrophilic molecules and pathogens into the brain. Several pathologies, including meningitis and postoperative delirium, are associated with an increased permeability of the blood-brain barrier. Here, we describe an endothelial cell culture model to test the barrier permeability by microbial traversal. Biochemistry Affinity Purification of Chloroplast Translocon Protein Complexes Using the TAP Tag Venkatasalam Shanmugabalaji1, Véronique Douet1, Birgit Agne2, Felix Kessler1 1Laboratory of Plant Physiology, University of Neuchatel, 2Institut fur Biochemie und Biotechnologie, Martin-Luther-Universitat Halle-Wittenberg We here present a proven and tested protocol for the purification of chloroplast protein import complexes (TOC-TIC complex) using the TAP-tag. The one-step affinity-isolation protocol can potentially be applied to any protein and be used to identify new interaction partners by mass spectrometry. Neuroscience Optimized Management of Endovascular Treatment for Acute Ischemic Stroke Katharina Schregel1,2, Daniel Behme1, Ioannis Tsogkas1, Michael Knauth1, Ilko Maier3, André Karch4, Rafael Mikolajczyk4,5, Mathias Bähr3, Jörn Schäper6, José Hinz6, Jan Liman3, Marios-Nikos Psychogios1 1Institute of Neuroradiology, University Medical Center Goettingen, 2 The outcome of patients with acute ischemic stroke depends on swift restoration of cerebral blood flow. This protocol aims at optimizing the management of such patients by minimizing peri-procedural timings and rendering the time from hospital admission to reperfusion as short as possible. Biology Combining Chemical Cross-linking and Mass Spectrometry of Intact Protein Complexes to Study the Architecture of Multi-subunit Protein Assemblies Caroline Haupt*1, Tommy Hofmann*1, Sabine Wittig*1, Susann Kostmann1, Argyris Politis2, Carla Schmidt1 1Interdisciplinary research center HALOmem, Martin Luther University Halle-Wittenberg, 2Department of Chemistry, Kings College London The architecture of protein complexes is essential for their function. Combining various mass spectrometric techniques proved powerful to study their assembly. We provide protocols for chemical cross-linking and native mass spectrometry and show how these complementary techniques help to elucidate the architecture of multi-subunit protein assemblies. Bioengineering Metabolic Glycoengineering of Sialic Acid Using N-acyl-modified Mannosamines Paul R. Wratil1,2, Rüdiger Horstkorte3 1Max von Pettenkofer-Institut & Genzentrum, Virologie, Nationales Referenzzentrum für Retroviren, Medizinische Fakultät, LMU München, 2Institut für Laboratoriumsmedizin, klinische Chemie und Pathobiochemie, Charité - Universitätsmedizin Berlin, 3Institut für Physiologische Chemie, Martin-Luther-Universität Halle-Wittenberg Sialic acid is a typical monosaccharide-unit found in glycoconjugates. It is involved in a plethora of molecular and cellular interactions. Here we present a method to modify cell surface sialic acid expression using metabolic glycoengineering with N-acetylmannosamine derivatives. Neuroscience Organotypic Hippocampal Slice Cultures As a Model to Study Neuroprotection and Invasiveness of Tumor Cells Urszula Grabiec*1, Tim Hohmann*1, Niels Hammer2, Faramarz Dehghani1 1Department of Anatomy and Cell Biology, Martin Luther University Halle-Wittenberg, 2Department of Anatomy, University of Otago Organotypic hippocampal slice cultures (OHSC) represent an in vitro model that simulates the in vivo situation very well. Here we describe a vibratome-based improved slicing protocol to obtain high quality slices for use in assessing the neuroprotective potential of novel substances or the biological behavior of tumor cells. Biology Relating Stomatal Conductance to Leaf Functional Traits Wenzel Kröber1, Isa Plath1, Heike Heklau1, Helge Bruelheide1,2 1Institute of Biology / Geobotany and Botanical Garden, Martin-Luther-University Halle-Wittenberg, 2German Centre for Integrative Biodiversity Research Unraveling how physiology and morphology are linked allows a deeper understanding of mechanistic functioning of plant leaves. We present both a procedure to derive parameters of stomatal regulation from stomatal conductance measurements and correlations with traditional functional leaf traits. Medicine Coculture System with an Organotypic Brain Slice and 3D Spheroid of Carcinoma Cells Han-Ning Chuang1, Raphaela Lohaus1, Uwe-Karsten Hanisch2, Claudia Binder1, Faramarz Dehghani*3, Tobias Pukrop*1 1Department of Hematology and Oncology, University of Göttingen, 2Institute of Neuropathology, University of Göttingen, 3Institute of Anatomy and Cellbiology, University of Halle The organotypic brain slice coculture with carcinoma cells enables visualizing morphological changes by fluorescence as well as bright field (video) microscopy during the process of carcinoma cell invasion of brain tissue. This model system also allows for cell exchange and replenishment approaches and offers a wide variety of manipulations and analyses.