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University of Texas Health Science Center at San Antonio

16 ARTICLES PUBLISHED IN JoVE

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Bioengineering

Construction of a Preclinical Multimodality Phantom Using Tissue-mimicking Materials for Quality Assurance in Tumor Size Measurement
Yongsook C. Lee 1,2, Gary D. Fullerton 2, Beth A. Goins 2
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).

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Behavior

The Attentional Set Shifting Task: A Measure of Cognitive Flexibility in Mice
Jillian M. Heisler 1, Juan Morales 1, Jennifer J. Donegan 1, Julianne D. Jett 1, Laney Redus 1, Jason C. O'Connor 1,2
1Department of Pharmacology, University of Texas Health Science Center at San Antonio, 2Audie L. Murphy VA Hospital, South Texas Veteran's Health Care System

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.

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Genetics

Methyl-binding DNA capture Sequencing for Patient Tissues
Rohit R. Jadhav 1, Yao V. Wang 1, Ya-Ting Hsu 1, Joseph Liu 1, Dawn Garcia 2, Zhao Lai 2, Tim H. M. Huang 1, Victor X. Jin 1
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.

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Immunology and Infection

Genome-wide Analysis of HDAC Inhibitor-mediated Modulation of microRNAs and mRNAs in B Cells Induced to Undergo Class-switch DNA Recombination and Plasma Cell Differentiation
Helia N. Sanchez 1, Tian Shen 1,2, Dawn Garcia 1,3, Zhao Lai 1,3, Paolo Casali 1, Hong Zan 1
1Department of Microbiology, Immunology and Molecular Genetics, University of Texas Long School of Medicine at San Antonio, 2Xiangya School of Medicine, Cental South University, 3Greehey Children’s Cancer Research Institute, University of Texas Long School of Medicine at San Antonio

Epigenetic factors can interact with genetic programs to modulate gene expression and regulate B cell function. By combining in vitro B-cell stimulation, qRT-PCR, and high-throughput microRNA-sequence and mRNA-sequence approaches, we can analyze the epigenetic modulation of miRNA and gene expression in B cells.

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Cancer Research

Fabrication of Tongue Extracellular Matrix and Reconstitution of Tongue Squamous Cell Carcinoma In Vitro
Yupeng Yao 1, Weifan Lin 1, Yan Zhang 1
1Key Laboratory of Gene Engineering of the Ministry of Education, State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-sen University

A method is shown here for the preparation of the tongue extracellular matrix (TEM) with efficient decellularization. The TEM can be used as functional scaffolds for the reconstruction of a tongue squamous cell carcinoma (TSCC) model under static or stirred culture conditions.

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Neuroscience

Adaptation of Microelectrode Array Technology for the Study of Anesthesia-induced Neurotoxicity in the Intact Piglet Brain
Emily D. Geyer *1, Prithvi A. Shetty *1, Christopher J. Suozzi *1, David Z. Allen *1,2, Pamela P. Benavidez *1,2, Joseph Liu *1,3, Charles N. Hollis 1, Greg A. Gerhardt 4, Jorge E. Quintero 4, Jason J. Burmeister 4, Emmett E. Whitaker 1,3
1Department of Anesthesiology, Ohio State University College of Medicine, 2Medical Student Research Program, Ohio State University College of Medicine, 3Department of Anesthesiology and Pain Medicine, Nationwide Children's Hospital, 4Department of Neuroscience, University of Kentucky Medical Center

This study explores the novel use of enzyme-based microelectrode array (MEA) technology to monitor in vivo neurotransmitter activity in piglets. The hypothesis was that glutamate dysregulation contributes to the mechanism of anesthetic neurotoxicity. Here, we present a protocol to adapt MEA technology to study the mechanism of anesthesia-induced neurotoxicity.

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Cancer Research

Three-Dimensional Bone Extracellular Matrix Model for Osteosarcoma
Yu Zhang 1, Yupeng Yao 1, Yan Zhang 1
1MOE Key Laboratory of Gene Function and Regulation, School of Life Sciences, Sun Yat-sen University

The bone extracellular matrix (BEM) model for osteosarcoma (OS) is well established and shown here. It can be used as a suitable scaffold for mimicking primary tumor growth in vitro and providing an ideal model for studying the histologic and cytogenic heterogeneity of OS.

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Behavior

A Chronic Immobilization Stress Protocol for Inducing Depression-Like Behavior in Mice
Hyeonwi Son 1, Ju Hwan Yang 2, Hyun Joon Kim *1, Dong Kun Lee *3
1Department of Anatomy and Convergence Medical Sciences, Institute of Health Sciences, Bio Anti-aging Medical Research Center, Gyeongsang National University Medical School, 2Department of Biological Sciences, Pusan National University, 3Department of Physiology and Convergence Medical Sciences, Institute of Health Sciences, Gyeongsang National University Medical School

This article provides a simplified and standardized protocol for induction of depressive-like behavior in chronically immobilized mice by using a restrainer. In addition, behavior and physiological techniques to verify induction of depression are explained.

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Chemistry

Cercosporin-Photocatalyzed [4+1]- and [4+2]-Annulations of Azoalkenes Under Mild Conditions
Philippe Icyishaka *1, Chenxing Li *1, Liushen Lu 1, Wenhao Bao 1, Jia Li 1, Yan Zhang 2, Yijian Rao 1
1Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 2School of Pharmaceutical Science, Jiangnan University

New routes for the synthesis of nitrogen-containing heterocycles utilizing cercosporin as a metal-free photocatalyst were developed.

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Biology

Visualization of DNA Repair Proteins Interaction by Immunofluorescence
Bárbara de la Peña Avalos 1,2, Eloïse Dray 1,2
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.

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Immunology and Infection

Isolation of Mouse Kidney-Resident CD8+ T cells for Flow Cytometry Analysis
Wei Liao 1,2, Chaoyu Ma 1, Nu Zhang 1
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.

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Biology

Three-Dimensional, Serum-Free Culture System for Lacrimal Gland Stem Cells
Hongzi Chen 1, Pan Huang 1, Yan Zhang 1
1MOE Key Laboratory of Gene Function and Regulation, School of Life Sciences, Sun Yat-sen University

The three-dimensional, serum-free culture method for adult lacrimal gland (LG) stem cells is well established for the induction of LG organoid formation and differentiation into acinar or ductal-like cells.

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Developmental Biology

Fixation of Embryonic Mouse Tissue for Cytoneme Analysis
Eric T. Hall 1, Christina A. Daly 1,2, Yan Zhang 1, Miriam E. Dillard 1, Stacey K. Ogden 1
1Department of Cell and Molecular Biology, St. Jude Children’s Research Hospital, 2St. Jude Graduate School of Biomedical Sciences

Here, an optimized step-by-step protocol is provided for fixation, immunostaining, and sectioning of embryos to detect specialized signaling filopodia called cytonemes in developing mouse tissues.

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Biology

Protocols for CRISPR/Cas9 Mutagenesis of the Oriental Fruit Fly Bactrocera dorsalis
Jinxi Yuan 1, Jie Zhang 1, Yan Zhang 1, WuYun QiQiGe 1, Wei Liu 2, Shanchun Yan 1, Guirong Wang 2
1Key Laboratory of Sustainable Forest Ecosystem Management - Ministry of Education, Northeast Forestry University, 2Shenzhen Branch, Guangdong Laboratory of Lingnan Modern Agriculture, Genome Analysis Laboratory of the Ministry of Agriculture and Rural Affairs, Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences

This paper presents the step-by-step protocols for CRISPR/Cas9 mutagenesis of the Oriental fruit fly Bactrocera dorsalis. Detailed steps provided by this standardized protocol will serve as a useful guide for generating mutant flies for functional gene studies in B. dorsalis.

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Cancer Research

Measurement of the Compressibility of Cell and Nucleus Based on Acoustofluidic Microdevice
Qibin Fu 1, Yan Zhang 1, Tuchen Huang 1, Yang Liu 1
1Sino-French Institute of Nuclear Engineering and Technology, Sun Yat-sen University

Here a protocol is presented to build a fast and non-destructive system for measuring cell or nucleus compressibility based on acoustofluidic microdevice. Changes in mechanical properties of tumor cells after epithelial-mesenchymal transition or ionizing radiation were investigated, demonstrating the application prospect of this method in scientific research and clinical practice.

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Neuroscience

In Vivo Calcium Imaging of Neuronal Ensembles in Networks of Primary Sensory Neurons in Intact Dorsal Root Ganglia
John Shannonhouse 1, Ruben Gomez 1, Hyeonwi Son 1, Yan Zhang 1, Yu Shin Kim 1,2
1Department of Oral & Maxillofacial Surgery, School of Dentistry, University of Texas Health Science Center at San Antonio, 2Programs in Integrated Biomedical Sciences, Translational Sciences, Biomedical Engineering, Radiological Sciences, University of Texas Health Science Center at San Antonio

This protocol describes the surgical exposure of the dorsal root ganglion (DRG) followed by GCaMP3 (genetically-encoded Ca2+ indicator; Green Fluorescent Protein-Calmodulin-M13 Protein 3) Ca2+ imaging of the neuronal ensembles using Pirt-GCaMP3 mice while applying a variety of stimuli to the ipsilateral hind paw.

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