East Carolina University View Institution's Website 11 articles published in JoVE Immunology and Infection Quantification of Autoreactive Antibodies in Mice upon Experimental Autoimmune Encephalomyelitis Jonathan J. Carver1, Alessandro Didonna1 1Department of Anatomy and Cell Biology, Brody School of Medicine, East Carolina University Experimental autoimmune encephalomyelitis (EAE) is an animal model of multiple sclerosis (MS), which shares with the human disease a robust humoral autoimmune response. Here, we report a simple and flexible ELISA protocol to quantify autoantibodies in the serum of EAE immunized mice. Cancer Research Three-Dimensional Culture of Murine Colonic Crypts to Study Intestinal Stem Cell Function Ex Vivo Amna N. Naser1, Qun Lu1, Yan-Hua Chen1 1Department of Anatomy and Cell Biology, Brody School of Medicine, East Carolina University The present protocol describes establishing a murine colonic organoid system to study the activity and functioning of colonic stem cells in a claudin-7 knockout model. Behavior Drosophila Passive Avoidance Behavior as a New Paradigm to Study Associative Aversive Learning Elena S. Pak1, Alexander K. Murashov1 1Department of Physiology, East Carolina University This work describes a simple behavioral paradigm that allows the analysis of aversive associative learning in adult fruit flies. The method is based on suppressing the innate negative geotaxis behavior due to the association formed between a specific environmental context and an electric shock. Biochemistry Measuring Mitochondrial Electron Transfer Complexes in Previously Frozen Cardiac Tissue from the Offspring of Sow: A Model to Assess Exercise-Induced Mitochondrial Bioenergetics Changes Daniel Barrera1, Sierra Upton2, Megan Rauch3, Tara Notarianni4, Ki Suk Eum5, Megan Liberty6, Sarmila Venkoba Sah7, Robert Liu8, Sean Newcomer9, Linda E. May10, Emre Agbas11, Jessica Sage12, Edina Kosa7, Abdulbaki Agbas7,13 1AdventHelath, 2University of Illinois Chicago School of Public Health, 3Lincoln Memorial University, 4Cincinnati Children’s Hospital Medical Center, 7Kansas City University, 8Roblex Rex Veterans Affairs Medical Center, 9California State University San Marcos, 10East Carolina University, 11Massachusetts Institute of Technology, 12Boehringer Ingelheim Norway KS, 13Heartland Center for Mitochondrial Medicine Preparation of mitochondria-enriched samples from previously frozen archived solid tissues allowed the investigators to perform both functional and analytical assessments of mitochondria in various experimental modalities. This study demonstrates how to prepare mitochondria-enriched preparations from frozen heart tissue and perform analytical assessments of mitochondria. Developmental Biology Using Chicken Embryo as a Powerful Tool in Assessment of Developmental Cardiotoxicities Qixiao Jiang1, Xiaohui Xu1, Jamie C. DeWitt2, Yuxin Zheng1 1School of Public Health, Qingdao University, 2Department of Pharmacology & Toxicology, Brody School of Medicine, East Carolina University Chicken embryos, as a classical developmental model, are used in our lab to assess developmental cardiotoxicities following exposure to various environmental contaminants. Exposure methods and morphological/functional assessment methods established are described in this manuscript. Biology In vivo Measurement of Knee Extensor Muscle Function in Mice Camille R. Brightwell1,2, Ted G. Graber3, Benjamin D. Brightwell4,5, Matthew Borkowski6, Brian Noehren5,7, Christopher S. Fry1,2 1Department of Athletic Training and Clinical Nutrition, University of Kentucky, 2Center for Muscle Biology, University of Kentucky, 3Department of Physical Therapy, East Carolina University, 4Kinesiology and Health Promotion Graduate Program, University of Kentucky, 5Biomotion Lab, College of Health Sciences, University of Kentucky, 6Aurora Scientific, 7Department of Physical Therapy, College of Health Sciences, University of Kentucky Quantification of knee extensor maximal strength is imperative to understand functional adaptations to aging, disease, injury, and rehabilitation. We present a novel method to repeatedly measure in vivo knee extension isometric peak tetanic torque. Immunology and Infection In Vivo Assessment of Alveolar Macrophage Efferocytosis Following Ozone Exposure Myles X. Hodge1, Sky W. Reece1, Jennifer H. Madenspacher2, Kymberly M. Gowdy1 1Department of Pharmacology and Toxicology, East Carolina University, 2Research Triangle Park, National Institute of Environmental Health and Sciences This manuscript describes a protocol for determining whether exposure to ozone, a criteria air pollutant, impairs alveolar macrophage efferocytosis in vivo. This protocol utilizes commonly used reagents and techniques and can be adapted to multiple models of pulmonary injury to determine effects on alveolar macrophage efferocytosis. Neuroscience The Use of Trace Eyeblink Classical Conditioning to Assess Hippocampal Dysfunction in a Rat Model of Fetal Alcohol Spectrum Disorders Tuan D. Tran1,2, Aenia Amin1,2, Keith G. Jones2, Ellen M. Sheffer1, Lidia Ortega1, Keith Dolman2 1Department of Psychology, East Carolina University, 2Multidisciplinary Studies Program in Neuroscience, East Carolina University Trace eyeblink classical conditioning (ECC) was used to assess hippocampal-dependent associative learning in adult rats that were administered a high concentration (11.9% v/v) of alcohol during early neonatal brain development. In general, ECC procedures are sound diagnostic tools for detecting brain dysfunction across many psychological and biomedical settings. Biology In vitro Cell Migration and Invasion Assays Calvin R. Justus1, Nancy Leffler1, Maria Ruiz-Echevarria1, Li V. Yang1 1Division of Hematology/Oncology, Department of Oncology, East Carolina University Commonly used, highly accessible methods for examining cell migration and invasion in vitro are described. The first method is the cell wound closure assay that measures cell motility. The second method is the transwell migration and invasion assay that assesses the chemotactic and invasive capacity of cells. Bioengineering Graphene Coatings for Biomedical Implants Ramakrishna Podila1,2, Thomas Moore3, Frank Alexis3, Apparao Rao1,4 1Department of Physics, Clemson University, 2Department of Pharmacology and Toxicology, East Carolina University, 3Department of Bioengineering, Clemson University, 4Center for Optical Materials Science and Engineering Technologies, Clemson University Graphene offers potential as a coating material for biomedical implants. In this study we demonstrate a method for coating nitinol alloys with nanometer thick layers of graphene and determine how graphene may influence implant response. Medicine Coronary Artery Ligation and Intramyocardial Injection in a Murine Model of Infarction Jitka A.I. Virag1, Robert M. Lust1 1Department of Physiology, Brody School of Medicine, East Carolina University Numerous genetic manipulations and/or intramyocardial injections of genes, proteins, cells, and/or biomaterials are superimposed upon the dimension of time in studies of acute ischemia/ reperfusion injury and chronic remodeling in mice. This video illustrates the microsurgical procedures for ischemia/reperfusion, permanent coronary artery ligation, and intramyocardial injection studies.
