University of Texas Health Science Center, San Antonio View Institution's Website 17 articles published in JoVE Biology Microinjection of Recombinant RCAS(A) Retrovirus into Embryonic Chicken Lens Francisca M. Acosta1, Bo Ma1, Sumin Gu1, Jean X. Jiang1 1Department of Biochemistry and Structural Biology, University of Texas Health Science Center, San Antonio This protocol paper describes the methodology of embryonic chicken lens microinjection of an RCAS(A) retrovirus as a tool for studying in situ function and expression of proteins during lens development. Bioengineering Three-Dimensional Culture of Vascularized Thermogenic Adipose Tissue from Microvascular Fragments Francisca M. Acosta1, Maria A. Gonzalez Porras2,3, Katerina Stojkova2, Settimio Pacelli2, Christopher R. Rathbone2,3, Eric M. Brey2,3 1Department of Biochemistry and Structural Biology, University of Texas Health Science Center, 2Department of Biomedical Engineering and Chemical Engineering, University of Texas at San Antonio, 3Institute of Regenerative Medicine, University of Texas at San Antonio Here, we present a detailed protocol outlining the use of microvascular fragments isolated from rodent or human fat tissue as a straightforward approach to engineer functional, vascularized beige adipose tissue. Immunology and Infection Isolation of Mouse Kidney-Resident CD8+ T cells for Flow Cytometry Analysis Wei Liao1,2, Chaoyu Ma1, Nu Zhang1 1Department of Microbiology, Immunology and Molecular Genetics, School of Medicine, University of Texas Health Science Center at San Antonio, 2Department of Dermatology, Hunan Key Laboratory of Medical Epigenomics, The Second Xiangya Hospital, Central South University Following viral infection, kidney harbors a relatively large number of CD8+ T cells and offers an opportunity to study non-mucosal TRM cells. Here, we describe a protocol to isolate mouse kidney lymphocytes for flow cytometry analysis. Biology Visualization of DNA Repair Proteins Interaction by Immunofluorescence Bárbara de la Peña Avalos1,2, Eloïse Dray1,2 1Department of Biochemistry and Structural Biology, University of Texas Health Science Center at San Antonio, 2Mays Cancer Center at UT Health San Antonio MD Anderson Cancer Center Following DNA damage, human cells activate essential repair pathways to restore the integrity of their genome. Here, we describe the method of indirect immunofluorescence as a means to detect DNA repair proteins, analyze their spatial and temporal recruitment, and help interrogate protein-protein interaction at the sites of DNA damage. Immunology and Infection Identification and Characterization of Immunogenic RNA Species in HDM Allergens that Modulate Eosinophilic Lung Inflammation Hamad H. Alanazi1,2, Li She1,3, Xiao-Dong Li1 1Department of Microbiology, Immunology and Molecular Genetics, Long School of Medicine, University of Texas Health San Antonio, 2Department of Clinical Laboratory Sciences, College of Applied Medical Sciences, Jouf University, 3Department of Otolaryngology-Head and Neck Surgery, Xiangya Hospital, Central South University Environmental allergens such as house dust mites (HDM) often contain microbial substances that activate innate immune responses to regulate allergic inflammation. The protocol presented here demonstrates the identification of dsRNA species in HDM allergens and characterization of their immunogenic activities in modulating eosinophilic lung inflammation. Genetics Methyl-binding DNA capture Sequencing for Patient Tissues Rohit R. Jadhav1, Yao V. Wang1, Ya-Ting Hsu1, Joseph Liu1, Dawn Garcia2, Zhao Lai2, Tim H. M. Huang1, Victor X. Jin1 1Department of Molecular Medicine, University of Texas Health Science Center at San Antonio, 2Greehey Children's Cancer Research Institute, University of Texas Health Science Center at San Antonio Here we present a protocol to investigate genome wide DNA methylation in large scale clinical patient screening studies using the Methyl-Binding DNA Capture sequencing (MBDCap-seq or MBD-seq) technology and the subsequent bioinformatics analysis pipeline. Medicine Rose Bengal Photothrombosis by Confocal Optical Imaging In Vivo: A Model of Single Vessel Stroke Lora Talley Watts1, Wei Zheng1, R. Justin Garling2, Victoria C. Frohlich3, James Donald Lechleiter1 1Department of Cellular and Structural Biology, The University of Texas Health Science Center San Antonio, 2School of Medicine, The University of Texas Health Science Center San Antonio, 3 Here, we describe a semi-invasive optical microscopy approach for the induction of a Rose Bengal photothrombotic clot in the somatosensory cortex of a mouse in vivo. The technical aspects of the imaging procedure are described from induction of a photothrombotic event to application and data collection. Medicine Visualization of Streptococcus pneumoniae within Cardiac Microlesions and Subsequent Cardiac Remodeling Armand O. Brown1, Carlos J. Orihuela1 1Department of Microbiology and Immunology, The University of Texas Health Science Center at San Antonio Streptococcus pneumoniae forms discrete non-purulent microscopic lesions in the heart. Outlined is the protocol for a murine model of cardiac microlesion formation. Instruction is provided on microlesion visualization using microscopy, discrimination between early and late microlesions, and methods to detect cardiac remodeling in hearts of convalescent animals. Behavior The Attentional Set Shifting Task: A Measure of Cognitive Flexibility in Mice Jillian M. Heisler1, Juan Morales1, Jennifer J. Donegan1, Julianne D. Jett1, Laney Redus1, Jason C. O'Connor1,2 1Department of Pharmacology, University of Texas Health Science Center at San Antonio, 2 The goal of this protocol is to perform a behavioral assay such as the attentional set shifting task (AST) to assess prefrontal cortex-mediated cognitive flexibility in mice. Bioengineering Construction of a Preclinical Multimodality Phantom Using Tissue-mimicking Materials for Quality Assurance in Tumor Size Measurement Yongsook C. Lee1,2, Gary D. Fullerton2, Beth A. Goins2 1Department of Radiation Oncology, University of Kansas School of Medicine, 2Department of Radiology, University of Texas Health Science Center at San Antonio This paper describes in-house procedures of constructing a preclinical multimodality phantom made of tissue-mimicking (TM) materials for quality assurance (QA) of tumor size measurement in animal imaging modalities such as ultrasound (US), computed tomography (CT) and magnetic resonance imaging (MRI). Medicine In vitro Measurements of Tracheal Constriction Using Mice Iurii Semenov1, Jeremiah T. Herlihy1, Robert Brenner1 1Department of Physiology, UT Health Science Center, San Antonio Transgenic mice have been extremely useful in ascribing physiological function to genes. As such, research in general, and functional studies of airway, in particular, have undergone a remarkable shift toward murine models. Here we provide protocols for in vitro trachea constriction studies to evaluate smooth muscle function in murine airway. Neuroscience Dissection of a Mouse Eye for a Whole Mount of the Retinal Pigment Epithelium Alison Claybon1, Alexander J. R. Bishop1 1UT Health Science Center at San Antonio, Greehey Children's Cancer Research Institute and Department of Cellular and Structural Biology A formal demonstration of the dissection of a mouse eye, resulting in a whole mount of the retinal pigment epithelium. Medicine Contrast Enhanced Vessel Imaging using MicroCT Suresh I. Prajapati1,2, Charles Keller1,3,4 1Greehey Children's Cancer Research Institute, University of Texas Health Science Center at San Antonio, 2Small Animal Imaging Resources facility, University of Texas Health Science Center at San Antonio, 3Department of Cellular and Structural Biology, University of Texas Health Science Center at San Antonio, 4Department of Pediatrics, University of Texas Health Science Center at San Antonio Contrast enhanced small animal vessel imaging by microCT is a rapid, cost-effective and high-throughput technique for serial in situ examination for tumor development, for analyzing the network of blood vessels that nourish them, and for following the response of tumors to preclinical therapeutic intervention(s). Biology Proper Care and Cleaning of the Microscope Victoria Centonze Frohlich1 1Core Optical Imaging Facility, University of Texas Health Science Center at San Antonio (UTHSCSA) Keeping the microscope optics clean is important for high-quality imaging. Dust, fingerprints, excess immersion oil, or mounting medium on or in a microscope causes reduction in contrast and resolution. DIC is especially sensitive to contamination and scratches on the lens surfaces. This protocol details the procedure for keeping the microscope clean. Biology Phase Contrast and Differential Interference Contrast (DIC) Microscopy Victoria Centonze Frohlich1 1Core Optical Imaging Facility, University of Texas Health Science Center at San Antonio (UTHSCSA) This protocol highlights the principles and practical applications of Phase and Differential Interference Contrast (DIC) Microscopy Biology Major Components of the Light Microscope Victoria Centonze Frohlich1 1Core Optical Imaging Facility, University of Texas Health Science Center at San Antonio (UTHSCSA) The light microscope is a basic tool for the cell biologist, who should have a thorough understanding of how it works, how it should be aligned for different applications, and how it should be maintained as required to obtain maximum image-forming capacity and resolution. The components of the microscope are described in detail here. Biology Loading Drosophila Nerve Terminals with Calcium Indicators Adam J. Rossano1, Gregory T. Macleod1 1Department of Physiology, University of Texas Health Science Center at San Antonio (UTHSCSA) Calcium is a ubiquitous messenger in the nervous system, essential for triggering neurotransmitter release and changes in synaptic strength. Here we demonstrate a technique for loading Ca2+-indicators into Drosophila nerve terminals. We also demonstrate fabrication of the required apparatus and emphasize points critical for the technique's success.
