Uniformed Services University of the Health Sciences View Institution's Website 8 articles published in JoVE Developmental Biology Preparation of Drosophila Larval and Pupal Testes for Analysis of Cell Division in Live, Intact Tissue Darya Karabasheva1, Jeremy T. Smyth1 1Department of Anatomy, Physiology, and Genetics, F. Edward Hébert School of Medicine, Uniformed Services University of the Health Sciences The goal of this protocol is to analyze cell division in intact tissue by live and fixed cell microscopy using Drosophila meiotic spermatocytes. The protocol demonstrates how to isolate whole, intact testes from Drosophila larvae and early pupae, and how to process and mount them for microscopy. Immunology and Infection A Simple Flow Cytometry Based Assay to Determine In Vitro Antibody Dependent Enhancement of Dengue Virus Using Zika Virus Convalescent Serum William G. Valiant1, Joseph J. Mattapallil1 1Department of Microbiology and Immunology, Uniformed Services University We describe a simple and easy protocol for measuring antibody dependent enhancement of infection by Zika virus convalescent serum using Dengue virus Reporter Viral Particles. Immunology and Infection Measuring Local Anaphylaxis in Mice Holly Evans1, Kristin E. Killoran1, Edward Mitre1 1Department of Microbiology and Immunology, Uniformed Services University of the Health Sciences Allergic responses, characterized by the activation of mast cells and basophils, are driven by the cross-linking of IgE and release of proinflammatory mediators. A quantitative assessment of allergic responses can be achieved by using Evans Blue dye to monitor changes in vascular permeability after allergen challenge. Medicine Biomarkers in an Animal Model for Revealing Neural, Hematologic, and Behavioral Correlates of PTSD Min Jia1, Fei Meng2, Stanley E. Smerin1, Guoqiang Xing1, Lei Zhang1, David M. Su2, David Benedek1, Robert Ursano1, Yan A. Su2, He Li1 1Department of Psychiatry, Center for the Study of Traumatic Stress, Uniformed Services University of the Health Sciences, Bethesda, Maryland, 2Department of Gene and Protein Biomarkers, GenProMarkers, Inc. We describe a rat model of post traumatic stress disorder (PTSD) that reveals the persistent alterations in neuroendocrine function and the delayed long-term, exaggerated fear response, characteristic of PTSD patients. The animal model and methods described here are useful for correlating biomarkers in brain nuclei, which are mechanistic but cannot be measured in patients, with biomarkers in peripheral white blood cells, which can. Immunology and Infection Primary Microglia Isolation from Mixed Glial Cell Cultures of Neonatal Rat Brain Tissue Tami T. Tamashiro1, Clifton Lee Dalgard1,2,3, Kimberly R. Byrnes2 1Neuroscience Program, Uniformed Services University, 2Department of Anatomy, Physiology, and Genetics, Uniformed Services University, 3Molecular and Cell Biology, Uniformed Services University Isolating primary microglia from the cellular heterogeneity of the brain is essential to investigate their role in both physiological and pathological conditions. This protocol describes a mechanical isolation and mixed cell culture technique that provides high yield and high purity, viable primary microglial cells for in vitro study and downstream applications. Biology Imaging of Estrogen Receptor-α in Rat Pial Arterioles using a Digital Immunofluorescent Microscope Niloofar Rezvani1, Andrei V. Blokhin1, Emil Zeynalov1, Marguerite T. Littleton-Kearney1 1Graduate School of Nursing, Uniformed Services University of the Health Sciences The goal of this article is to demonstrate a method to optimize immunofluorescent detection of estrogen receptor-α (ERα) in rat pial arterial slices using a digital immunofluorescent microscope. Neuroscience Migratory Behavior of Cells Generated in Ganglionic Eminence Cultures Marcin Gierdalski*1, Thomas McFate*1, Joseph Abbah2, Sharon L. Juliano1,2 1Dept. of Anatomy, Physiology and Genetics, Uniformed Services University, 2Neuroscience Program, Uniformed Services University Time lapse imaging of 3D tissue culture allows studying migratory behavior of individual cells originating from ganglionic eminence in reaction to fractionated protein extract from cerebral cortex. Biology Mouse Mammary Epithelial Cells form Mammospheres During Lactogenic Differentiation Bethanie Morrison1, Mary Lou Cutler1 1Department of Pathology, F. Edward Hebert School of Medicine, Uniformed Services University of the Health Sciences, Bethesda, MD HC11 lactogenic differentiation can be characterized by the formation of domed structures referred to as mammospheres. The structures can be enumerated by phase contrast microscopy to aid in quantifying lactogenic differentiation.
