University of Arkansas for Medical Sciences View Institution's Website 18 articles published in JoVE Neuroscience Using the Chick Embryo Brain as a Model for In Vivo and Ex Vivo Analyses of Human Glioblastoma Cell Behavior Nicole G. Pastorino1, Saori Tomatsu1, Amy Lin1, Jackson Doerr1, Zachary Waterman1, Krisztina Sershen1, Pulak Ray2, Analiz Rodriguez3, Deni S. Galileo1 1Department of Biological Sciences, University of Delaware, 2Helen F. Graham Cancer Center and Research Institute, Christiana Care, 3Department of Neurosurgery, Winthrop P. Rockefeller Cancer Institute, University of Arkansas for Medical Sciences Chick embryos are used for studying human glioblastoma (GBM) brain tumors in ovo and in ex vivo brain slice co-cultures. GBM cell behavior can be recorded by time-lapse microscopy in ex vivo co-cultures, and both preparations can be analyzed at the experimental endpoint by detailed 3D confocal analysis. Biology Image-Based Methods to Study Membrane Trafficking Events in Stomatal Lineage Cells Qin He1, Huiliang Zhang2, Xingyun Qi1 1Department of Biology, Rutgers University, 2Pharmacology and Toxicology Department, University of Arkansas for Medical Sciences Several commonly used methods are introduced here to study the membrane trafficking events of a plasma membrane receptor kinase. This manuscript describes detailed protocols including the plant material preparation, pharmacological treatment, and confocal imaging setup. Medicine Ferric Chloride-Induced Arterial Thrombosis and Sample Collection for 3D Electron Microscopy Analysis Smita Joshi*1, Alexis N. Smith1, Kanakanagavalli Shravani Prakhya1, Hammodah R. Alfar1, Joshua Lykins1, Ming Zhang1, Irina Pokrovskaya2, Maria Aronova3, Richard D. Leapman3, Brian Storrie2, Sidney W. Whiteheart*1 1Department of Molecular and Cellular Biochemistry, University of Kentucky, 2Department of Physiology and Cell Biology, University of Arkansas for Medical Sciences, 3Laboratory of Cellular Imaging and Macromolecular Biophysics, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health The present protocol describes how to use a FeCl3-mediated injury to induce arterial thrombosis, and how to collect and prepare arterial injury samples at various stages of thrombosis for electron microscopy analysis. Medicine Bridging the Technology Divide in the COVID-19 Era: Using Virtual Outreach to Expose Middle and High School Students to Imaging Technology Kevin D. Phelan1, Mohsin Syed1, Noor Akhter1, Tiffany W. Huitt1, Gregory R. Snead2, Billy R. Thomas3, Karen L. Yanowitz4 1Division of Clinical Anatomy, Department of Neurobiology and Developmental Sciences, College of Medicine, University of Arkansas for Medical Sciences, 2Department of Emergency Medicine, College of Medicine, University of Arkansas for Medical Sciences, 3Department of Pediatrics, College of Medicine, University of Arkansas for Medical Sciences, 4Department of Psychology and Counseling, Arkansas State University This article presents an overview of how synchronous web-based virtual outreach can be used to expose 6th-12th grade students to advanced imaging technologies such as ultrasound, computerized tomography, and electroencephalography. The paper discusses the methods and equipment needed to livestream integrated educational sessions for effective student engagement in STEM. Neuroscience Combined Mechanical and Enzymatic Dissociation of Mouse Brain Hippocampal Tissue Madison Trujillo1,2,3, Taylor McElroy1,2,3, Taurean Brown1,2,3, Pilar Simmons1,2, Fabio Ntagwabira1,2,3, Antiño R. Allen1,2,3 1Division of Radiation Health, University of Arkansas for Medical Sciences, 2Department of Pharmaceutical Sciences, University of Arkansas for Medical Sciences, 3Neurobiology & Developmental Sciences, University of Arkansas for Medical Sciences This neural cell dissociation protocol is intended for samples with a low amount of starting material and yields a highly viable single-cell suspension for downstream analysis, with optional fixation and staining steps. Cancer Research Isolating Human Peripheral Blood Mononuclear Cells and CD4+ T cells from Sézary Syndrome Patients for Transcriptomic Profiling Syed J. Mehdi1, Andrea M. Moerman-Herzog1, Henry K. Wong1 1Department of Dermatology, University of Arkansas for Medical Sciences We present a simple protocol for the isolation of peripheral blood mononuclear cells from whole blood obtained from patients diagnosed with Sézary Syndrome, followed by selection of CD4+ T cells, their stimulation with phorbol12-myristate13-acetate and A23187 ionophore, and preparation of RNA for transcriptomic profiling. Biochemistry Assessment of Cellular Oxidation using a Subcellular Compartment-Specific Redox-Sensitive Green Fluorescent Protein Alev Tascioglu Aliyev1,2, Francesca LoBianco1, Kimberly J. Krager1, Nukhet Aykin-Burns1 1Division of Radiation Health, Pharmaceutical Sciences, University of Arkansas for Medical Sciences, 2Department of Pharmaceutical Toxicology, Faculty of Pharmacy, Ege University This protocol describes the assessment of subcellular compartment-specific redox status within the cell. A redox-sensitive fluorescent probe allows convenient ratiometric analysis in intact cells. Cancer Research Preparation of Primary Acute Lymphoblastic Leukemia Cells in Different Cell Cycle Phases by Centrifugal Elutriation Magdalena Delgado*1, Anisha Kothari*1, Walter N. Hittelman2, Timothy C. Chambers1 1Department of Biochemistry and Molecular Biology, University of Arkansas for Medical Sciences, 2Department of Experimental Therapeutics, University of Texas MD Anderson Cancer Center This protocol describes the use of centrifugal elutriation to separate primary acute lymphoblastic leukemia cells into different cell cycle phases. Chemistry Extraction and Purification of Polyphenols from Freeze-dried Berry Powder for the Treatment of Vascular Smooth Muscle Cells In Vitro Rafaela G. Feresin1, Shirin Pourafshar2,3, Jingwen Huang2, Yitong Zhao2, Bahram H. Arjmandi2,3, Gloria Salazar2,3 1Department of Dietetics and Nutrition, University of Arkansas for Medical Sciences, 2Department of Nutrition, Food and Exercise Sciences, Florida State University, 3Center for Advancing Exercise and Nutrition Research on Aging (CAENRA), Florida State University This work details a step-by-step method to prepare polyphenol-rich extracts from freeze-dried berry powder. In addition, it provides a thorough description of how to use these polyphenol-rich extracts in cell culture in the presence of the peptide hormone angiotensin II (Ang II) using Vascular Smooth Muscle Cells (VSMCs). Neuroscience Assessment of Hippocampal Dendritic Complexity in Aged Mice Using the Golgi-Cox Method Thomas R. Groves1,2,3, Jing Wang1,2, Marjan Boerma1,2, Antiño R. Allen1,2,3 1Division of Radiation Health, University of Arkansas for Medical Sciences, 2Department of Pharmaceutical Sciences, University of Arkansas for Medical Sciences, 3Neurobiology & Developmental Sciences, University of Arkansas for Medical Sciences Here we present a Golgi-Cox protocol in extensive detail. This reliable tissue stain method allows for a high-quality assessment of the cytoarchitecture in the hippocampus, and throughout the entire brain, with minimal troubleshooting. Biology Clock Scan Protocol for Image Analysis: ImageJ Plugins Maxim Dobretsov1, Georg Petkau2, Abdallah Hayar3, Eugen Petkau1 1Department of Anesthesiology, University of Arkansas for Medical Sciences, 2Department of Lymphocyte Development, Max Plank Institute for Infection Biology, 3Department of Neurobiology & Developmental Sciences, University of Arkansas for Medical Sciences This paper describes two novel ImageJ plugins for 'Clock Scan' image analysis. These plugins expand the functionality of the original visual basic 6 program and, most importantly, make the program available to a large research community by bundling it with the ImageJ free image analysis software package. Genetics Detection of Inter-chromosomal Stable Aberrations by Multiple Fluorescence In Situ Hybridization (mFISH) and Spectral Karyotyping (SKY) in Irradiated Mice Rupak Pathak1, Igor Koturbash2, Martin Hauer-Jensen1,3 1Division of Radiation Health, Department of Pharmaceutical Sciences, College of Pharmacy, University of Arkansas for Medical Sciences, 2Department of Environmental Health, Fay W. Boozman School of Public Health, University of Arkansas for Medical Sciences, 3Surgical Service, Central Arkansas Veterans Healthcare System The present protocol describes the usefulness of multiple fluorescence in situ hybridization (mFISH) and spectral karyotyping (SKY) in identifying inter-chromosomal stable aberrations in the bone marrow cells of mice after exposure to total body irradiation. Genetics Analysis of the Ambient Particulate Matter-induced Chromosomal Aberrations Using an In Vitro System Isabelle R. Miousse1, Igor Koturbash1, Marie-Cécile Chalbot2, Martin Hauer-Jensen3, Ilias Kavouras2, Rupak Pathak3 1Department of Occupational and Environmental Health, University of Arkansas for Medical Sciences, 2Department of Environmental Health Sciences, University of Alabama at Birmingham, 3Division of Radiation Health, University of Arkansas for Medical Sciences This protocol describes techniques for the quantification and characterization of chromosomal aberrations in vitro in RAW264.7 mouse macrophages after treatment with ambient air particulate matter. Neuroscience Recording Gamma Band Oscillations in Pedunculopontine Nucleus Neurons Francisco J. Urbano1, Brennon R. Luster2, Stasia D'Onofrio2, Susan Mahaffey2, Edgar Garcia-Rill2 1IFIBYNE-CONICET, University of Buenos Aires, 2Center for Translational Neuroscience, University of Arkansas for Medical Sciences The pedunculopontine nucleus (PPN) is located in the brainstem and its neurons are maximally activated during waking and rapid eye movement (REM) sleep brain states. This work describes the experimental approach to record in vitro gamma band subthreshold membrane oscillation in PPN neurons. Medicine Image-guided Convection-enhanced Delivery into Agarose Gel Models of the Brain Karl A. Sillay1,2, S. Gray McClatchy3, Brandon A. Shepherd1, Garrett T. Venable1, Tyler S. Fuehrer4 1University of Tennessee Health Science Center, 2Semmes-Murphey Clinic, 3University of Arkansas for Medical Sciences, 4Restorative Neurosciences Foundation Convection-enhanced delivery (CED) has been proposed as a treatment option for a wide range of neurological diseases. In order to prepare health care professionals for adoption of CED, accessible training models are needed. We describe the use of agarose gel as such a model of the human brain for testing, research, and training. Biology Assessment of Mitochondrial Functions and Cell Viability in Renal Cells Overexpressing Protein Kinase C Isozymes Grażyna Nowak1, Diana Bakajsova1 1Department of Pharmaceutical Sciences, College of Pharmacy, University of Arkansas for Medical Sciences The effects of activation of protein kinase C (PKC) isozymes on mitochondrial functions associated with respiration and oxidative phosphorylation and on cell viability are described. The approach adapts adenoviral technique to selectively overexpress PKC isozymes in primary cell culture and a variety of assays to determine mitochondrial functions and energy status of the cell. Biology Application of MassSQUIRM for Quantitative Measurements of Lysine Demethylase Activity Lauren P. Blair1, Nathan L. Avaritt1, Alan J. Tackett1 1Department of Biochemistry and Molecular Biology, University of Arkansas for Medical Sciences We present a method for using MALDI mass spectrometry and reductive methylation chemistry to quantify changes in lysine methylation. Immunology and Infection Use of Interferon-γ Enzyme-linked Immunospot Assay to Characterize Novel T-cell Epitopes of Human Papillomavirus Xuelian Wang1, William W. Greenfield2, Hannah N. Coleman3, Lindsey E. James3, Mayumi Nakagawa3 1Department of Microbiology and Parasitology, College of Basic Medical Sciences, China Medical University, 2Department of Obstetrics and Gynecology, College of Medicine, University of Arkansas for Medical Sciences, 3Department of Pathology, College of Medicine, University of Arkansas for Medical Sciences Characterizing T-cell epitopes of pathogens that cause localized infections such as human papillomavirus is a challenge because of limited number of T cells in circulation. A method is described in which rare T cells were isolated and were characterized starting with a very small number of cells.