University of Gothenburg View Institution's Website 16 articles published in JoVE Neuroscience A Neonatal Rodent Model of Retroorbital Vein Injection Eridan Rocha-Ferreira1, Syam Nair1, Owen Herrock1, Erik Axel Andersson2, Carl Joakim Ek2, Carina Mallard2, Henrik Hagberg1 1Centre of Perinatal Medicine and Health, Institute of Clinical Sciences, Department of Obstetrics and Gynecology, Sahlgrenska Academy, University of Gothenburg, 2Centre of Perinatal Medicine and Health, Institute of Neuroscience and Physiology, Sahlgrenska Academy, University of Gothenburg This protocol aims to demonstrate a reproducible venous administration route that can be used in rats and mice throughout the neonatal period. This procedure is important for preclinical rodent studies that wish to mirror drug administration in neonatal care units primarily using intravenous administration. Neuroscience Event Related Potentials (ERPs) and other EEG Based Methods for Extracting Biomarkers of Brain Dysfunction: Examples from Pediatric Attention Deficit/Hyperactivity Disorder (ADHD) Geir Ogrim1,2,3, Juri D. Kropotov4,5 1Neuropsychiatric Team, Åsebråten Outpatient Clinic, Østfold Hospital Trust, 2Institute of Psychology, Norwegian University of Science and Technology, 3Gillberg Neuropsychiatry Centre, University of Gothenburg, 4P. Bechtereva Institute of the Human Brain, Russian Academy of Sciences, 5Department of Neuropsychology, Andrzej Frycz-Modrzewski Krakow University EEG-methods are applied for extracting biomarkers of brain dysfunctions. The focus is on multi-channel event-related potentials (ERPs) recorded in a cued GO/NOGO task. Non-brain artifacts are corrected and ERPs are compared with the normative data. Examples relate to biomarkers for ADHD diagnosis and prediction of medication response. Biology Isolation and Culture of Human Mature Adipocytes Using Membrane Mature Adipocyte Aggregate Cultures (MAAC) Ida Alexandersson1, Matthew J. Harms1, Jeremie Boucher1,2,3 1Bioscience Metabolism, Research and Early Development, Cardiovascular, Renal and Metabolism (CVRM), BioPharmaceuticals R&D, AstraZeneca, 2The Lundberg Laboratory for Diabetes Research, University of Gothenburg, 3Wallenberg Centre for Molecular and Translational Medicine, University of Gothenburg Membrane mature adipocyte aggregate culture (MAAC) is a new method to culture mature human adipocytes. Here we detail how to isolate adipocytes from human adipose and how to set up MAAC. Engineering Spectral and Angle-Resolved Magneto-Optical Characterization of Photonic Nanostructures Mikko Kataja1, Rafael Cichelero1,2, Gervasi Herranz1 1Institut de Ciència de Materials de Barcelona, 2Department of Physics, University of Gothenburg Photonic band structure enables understanding how confined electromagnetic modes propagate within a photonic crystal. In photonic crystals that incorporate magnetic elements, such confined and resonant optical modes are accompanied by enhanced and modified magneto-optical activity. We describe a measurement procedure to extract the magneto-optical band structure by Fourier space microscopy. Engineering Safe Experimentation in Optical Levitation of Charged Droplets Using Remote Labs Daniel Galán1, Oscar Isaksson2, Jonas Enger2, Mats Rostedt2, Andreas Johansson2, Dag Hanstorp2, Luis de la Torre1 1Departamento de Informática y Automática, UNED, 2Department of Physics, University of Gothenburg Optical levitation is a method for levitating micrometer-sized dielectric objects using laser light. Utilizing computers and automation systems, an experiment on optical levitation can be controlled remotely. Here, we present a remotely controlled optical levitation system that is used both for educational and research purposes. Bioengineering Isolation and Decellularization of a Whole Porcine Pancreas Vijay Kumar Kuna*1, Niclas Kvarnström*2, Erik Elebring1, Suchitra Sumitran Holgersson1 1Department of Surgery, Institute of Clinical Sciences, Sahlgrenska Academy, University of Gothenburg, 2The Transplant Institute, Sahlgrenska University Hospital Tissue engineering of the whole pancreas is a challenge because of its exocrine and endocrine functions. We show a method for the dissection of an intact porcine pancreas and the process of successful decellularization by perfusion of detergents Triton X-100, sodium deoxycholate, and deoxyribonuclease. Environment The Barnacle Balanus improvisus as a Marine Model - Culturing and Gene Expression Per R. Jonsson1, Anna-Lisa Wrange3, Ulrika Lind2, Anna Abramova2, Martin Ogemark1, Anders Blomberg2 1Department of Marine Sciences, University of Gothenburg, 2Department of Chemistry and Molecular Biology, University of Gothenburg, 3IVL Swedish Environmental Research Institute The barnacle Balanus (Amphibalanus) improvisus is a model for studying osmoregulation and antifouling. However, natural seasonal spawning yields an unpredictable supply of cyprid larvae. Here, a protocol for the all-year-round culturing of B. improvisus is described, including the production of larvae. The use of cultured barnacles in gene expression studies is illustrated. Cancer Research Target Cell Pre-enrichment and Whole Genome Amplification for Single Cell Downstream Characterization Shukun Chen*1, Amin El-Heliebi*1, Julia Schmid1, Karl Kashofer2, Zbigniew T. Czyż3, Bernhard Michael Polzer3, Klaus Pantel4, Thomas Kroneis1,5, Peter Sedlmayr1 1Institute of Cell Biology, Histology and Embryology, Medical University of Graz, 2Institute of Pathology, Medical University of Graz, 3Fraunhofer Institute for Toxicology and Experimental Medicine ITEM, 4Department of Tumor Biology, University Medical Center Hamburg-Eppendorf, 5Sahlgrenska Cancer Center, University of Gothenburg This protocol is to recover and prepare rare target cells from a mixture with non-target background cells for molecular genetic characterization at the single-cell level. DNA quality is equal to non-treated single cells and allows for single-cell application (both screening based and targeted analysis). Neuroscience Monitoring the Effect of Osmotic Stress on Secretory Vesicles and Exocytosis Hoda Fathali1, Johan Dunevall1, Soodabeh Majdi2, Ann-Sofie Cans1 1Department of Chemistry and Chemical Engineering, Chalmers University of Technology, 2Department of Chemistry and Molecular Biology, University of Gothenburg Osmotic stress affects exocytosis and the amount of neurotransmitter released during this process. We demonstrate how combining electrochemical methods together with transmission electron microscopy can be used to study the effect of extracellular osmotic pressure on exocytosis activity, vesicle quantal size, and the amount of neurotransmitter released during exocytosis. Biology Simultaneous Mapping and Quantitation of Ribonucleotides in Human Mitochondrial DNA Katrin Kreisel1, Martin K.M. Engqvist1,2, Anders R. Clausen1 1Department for Medical Biochemistry and Cell Biology, University of Gothenburg, 2Department of Biology and Biological Engineering, Chalmers University of Technology Here we describe a method amenable to simultaneously quantitate and genome-wide map ribonucleotides in highly intact DNA at single-nucleotide resolution, combining enzymatic cleavage of genomic DNA with its alkaline hydrolysis and subsequent 5´-end sequencing. Genetics Comprehensive DNA Methylation Analysis Using a Methyl-CpG-binding Domain Capture-based Method in Chronic Lymphocytic Leukemia Patients Santhilal Subhash1, Meena Kanduri2 1Department of Medical Genetics, Institute of Biomedicine, Gothenburg University, 2Department of Clinical Chemistry and Transfusion Medicine, Institute of Biomedicine, Gothenburg University This work describes an optimized methyl-CpG-binding domain (MBD) sequencing protocol and a computational pipeline to identify differentially methylated CpG-rich regions in chronic lymphocytic leukemia (CLL) patients. Medicine A High Throughput, Multiplexed and Targeted Proteomic CSF Assay to Quantify Neurodegenerative Biomarkers and Apolipoprotein E Isoforms Status Wendy E. Heywood*1, Anna Baud*1, Emily Bliss1, Ernestas Sirka1, Jonathan M. Schott2, Henrik Zetterberg3, Daniela Galimberti4, Neil J. Sebire5, Kevin Mills1 1Centre for Translational Omics, Genetics and Genomic Medicine Deptartment, Great Ormond Street Institute of Child Health, University College London, 2Dementia Research Centre, Institute of Neurology, University College London, 3Clinical Neurochemistry Laboratory, Institute of Neuroscience and Physiology, Department of Psychiatry and Neurochemistry, The Sahlgrenska Academy, University of Gothenburg, 4Neurology Unit, Department of Pathophysiology and Transplantation, University of Milan, 5Great Ormond Street Hospital for Children, University College London We describe a high-throughput, multiplex, and targeted proteomic cerebrospinal fluid (CSF) assay developed with potential for clinical translation. The test can quantitate potential markers and risk factors for neurodegeneration, such as the apolipoprotein E variants (E2, E3 and E4), and measure their allelic expression. Immunology and Infection Non-Invasive Model of Neuropathogenic Escherichia coli Infection in the Neonatal Rat Fatma Dalgakiran1, Luci A. Witcomb1, Alex J. McCarthy1, George M. H. Birchenough2, Peter W. Taylor1 1School of Pharmacy, University College London, 2Mucin Biology Group, University of Gothenburg Here, a procedure is described for the establishment of systemic infection in the neonatal rat with cultures of Escherichia coli K1. This non-invasive procedure permits colonization of the gastrointestinal tract, translocation of the pathogen to the systemic circulation, and invasion of the central nervous system at the choroid plexus. Medicine Modified Technique for Coronary Artery Ligation in Mice Yangzhen Shao1, Björn Redfors1, Elmir Omerovic1 1Wallenberg Laboratory, Sahlgrenska Academy, University of Gothenburg The surgical procedure used to induce experimental myocardial infarction in mice begins with left thoracotomy between the third and the fourth ribs in order to visualize the anterior surface of the heart and left lung. The left coronary artery is ligated, the chest is closed and the mouse is allowed to recover spontaneously. Biology Isolation and Characterization of RNA-Containing Exosomes Cecilia Lässer*1, Maria Eldh*1, Jan Lötvall1 1Krefting Research Centre, Department of Internal Medicine, Sahlgrenska Academy, University of Gothenburg This paper demonstrates methods for the isolation, purification and detection of exosomes, as well as techniques for analysis of their molecular content. These methods are adaptable for exosome isolation from both cell culture media and biological fluids, and can beyond analysis of molecular content also be useful in functional studies. Biology Rat Mesentery Angiogenesis Assay Klas C. Norrby1 1Department of Pathology, Institute of Biomedicine, University of Gothenburg Normal adult vascularized mammalian tissue that lacks physiologic angiogenesis and that has not been exposed to surgical intervention is used to study: (i) the initiation and development of angiogenesis following intraperitoneal administration of test agents; and (ii) modification of angiogenesis following systemic administration of selected test agents.