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University of Texas at Austin

26 ARTICLES PUBLISHED IN JoVE

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Neuroscience

Novel Apparatus and Method for Drug Reinforcement
Allison A. Feduccia 1, Christine L. Duvauchelle 1
1College of Pharmacy, Division of Pharmacology and Toxicology, University of Texas at Austin

Operant drug self-administration and conditioned place preference (CPP) procedures are expansively used in research to model various components of drug reinforcement, consumption, and addiction in humans. In this report, we combined traditional CPP and self-administration methods as a novel approach to studying drug reinforcement and addiction in rats.

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Neuroscience

How to Build a Laser Speckle Contrast Imaging (LSCI) System to Monitor Blood Flow
Adrien Ponticorvo 1, Andrew K. Dunn 1
1Biomedical Engineering Department, University of Texas at Austin

This video demonstrates how to build a Laser Speckle Contrast Imaging (LSCI) system that can easily be used to monitor blood flow.

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Neuroscience

Assessment of Ultrasonic Vocalizations During Drug Self-administration in Rats
Esther Y. Maier 1,2, Sean T. Ma 3, Allison Ahrens 2,4, Timothy J. Schallert 2,4,5, Christine L. Duvauchelle 1,2,4
1College of Pharmacy, Division of Pharmacology and Toxicology, University of Texas at Austin, 2The Waggoner Center of Addiction and Alcohol Research, University of Texas at Austin, 3Department of Psychology, University of Michigan, 4Institute for Neuroscience, University of Texas at Austin, 5Department of Psychology, University of Texas at Austin

Drug self-administration and ultrasonic vocalizations (USV) are used as behavioral assessments in animal research, but rarely in combination. The purpose of this article is to describe the advantages of recording USVs during drug self-administration procedures to assess affective responses to drug experience.

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Neuroscience

The Vermicelli and Capellini Handling Tests: Simple quantitative measures of dexterous forepaw function in rats and mice
Kelly A. Tennant 1, Aaron L. Asay 2, Rachel P. Allred 3, Angela R. Ozburn 4, Jeffrey A. Kleim 5, Theresa A. Jones 1,2
1Institute for Neuroscience, University of Texas at Austin, 2Department of Psychology, University of Texas at Austin, 3Department of Neurology, University of Florida, 4Department of Psychiatry, University of Texas Southwestern Medical Center, 5Department of Neuroscience, McKnight Brain Institute, University of Florida

The Vermicelli and Capellini Handling Tests of forepaw dexterity take advantage of the natural inclination of rodents to manipulate food items using skillful forepaw and digit movements. Animals are videotaped while handling short strands of uncooked dry pasta. Slow motion video playback allows for the quantification of forepaw adjustments.

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Immunology and Infection

'Bioluminescent' Reporter Phage for the Detection of Category A Bacterial Pathogens
David A. Schofield 1, Ian J. Molineux 2, Caroline Westwater 3
1BioSciences Division, Guild Associates, Inc., 2Department of Molecular Genetics and Microbiology, University of Texas at Austin, 3Department of Craniofacial Biology, Medical University of South Carolina

A simple method for the identification of priority bacterial pathogens is to use genetically engineered reporter phage. These reporter phage, which are specific to their particular host species, are capable of rapidly transducing a bioluminescent signal response to host cells. Herein, we describe the use of reporter phage for the detection of Yersinia pestis.

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Bioengineering

Plasma Lithography Surface Patterning for Creation of Cell Networks
Michael Junkin 1, Siu Ling Leung 1, Yongliang Yang 1, Yi Lu 1, Justin Volmering 1, Pak Kin Wong 1,2
1Aerospace and Mechanical Engineering, University of Arizona , 2Biomedical Engineering IDP and BIO5 Institute, University of Arizona

A versatile plasma lithography technique has been developed to generate stable surface patterns for guiding cellular attachment. This technique can be applied to create cell networks including those that mimic natural tissues and has been used for studying several, distinct cell types.

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Immunology and Infection

Determination of Tolerable Fatty Acids and Cholera Toxin Concentrations Using Human Intestinal Epithelial Cells and BALB/c Mouse Macrophages
Farshad Tamari 1, Joanna Tychowski 2, Laura Lorentzen 3
1Department of Biological Sciences, Kingsborough Community College, 2Institute for Cellular and Molecular Biology, University of Texas at Austin, 3New Jersey Center for Science, Technology and Mathematics, Kean University

We set out to determine tolerable concentrations of three fatty acids (oleic, linoleic and linolenic acids) and cholera toxin that did not significantly and adversely affect cell survival by solubilizing the fatty acids and the toxin and using them in cell survival assays.

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Bioengineering

Cell Squeezing as a Robust, Microfluidic Intracellular Delivery Platform
Armon Sharei 1, Nahyun Cho 1, Shirley Mao 1, Emily Jackson 1, Roberta Poceviciute 1, Andrea Adamo 1, Janet Zoldan 2, Robert Langer 1,2, Klavs F Jensen 1
1Department of Chemical Engineering, Massachusetts Institute of Technology, 2David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology

Rapid mechanical deformation of cells has emerged as a promising, vector-free method for intracellular delivery of macromolecules and nanomaterials. This protocol provides detailed steps on how to use the system for a broad range of applications.

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Behavior

Compensatory Limb Use and Behavioral Assessment of Motor Skill Learning Following Sensorimotor Cortex Injury in a Mouse Model of Ischemic Stroke
Abigail L. Kerr 1, Kelly A. Tennant 2
1Psychology Department, Illinois Wesleyan University, 2Division of Medical Sciences, University of Victoria

Mouse models have become increasingly popular in studies of behavioral neuroscience. As models advance, it is important to develop sensitive behavioral measures specific to the mouse. This protocol describes the Pasta Matrix Reaching Task, which is a skilled motor task for use in mouse models of stroke.

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Chemistry

Synthesis of Immunotargeted Magneto-plasmonic Nanoclusters
Chun-Hsien Wu 1,2, Konstantin Sokolov 1,2
1Department of Biomedical Engineering, University of Texas at Austin, 2Department of Imaging Physics, University of Texas M.D. Anderson Cancer Center

Here, we describe a protocol for synthesis of magneto-plasmonic nanoparticles with a strong magnetic moment and a strong near-infrared (NIR) absorbance. The protocol also includes antibody conjugation to the nanoparticles through the Fc moiety for various biomedical applications which require molecular specific targeting.

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Medicine

Murine Model of Femoral Artery Wire Injury with Implantation of a Perivascular Drug Delivery Patch
Victoria Le 1, Collin G. Johnson 1, Jonathan D. Lee 1, Aaron B. Baker 1
1Department of Biomedical Engineering, University of Texas at Austin

We describe a surgical technique that produces wire injury in the femoral artery of mice to induce neointimal hyperplasia to serve as a model testing system for the perivascular delivery of therapeutic compounds for the inhibition of restenosis.

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Environment

A CO2 Concentration Gradient Facility for Testing CO2 Enrichment and Soil Effects on Grassland Ecosystem Function
Philip A. Fay 1, Lara G. Reichmann 2, Michael J. Aspinwall 2, Albina R. Khasanova 1, H. Wayne Polley 1
1USDA-ARS, Grassland Soil and Water Research Laboratory, 2Department of Integrative Biology, University of Texas at Austin

The Lysimeter Carbon Dioxide Gradient Facility creates a 250 to 500 µl L-1 linear carbon dioxide gradient in temperature-controlled chambers housing grassland plant communities on clay, silty clay, and sandy soil monoliths. The facility is used to determine how past and future carbon dioxide levels affect grassland carbon cycling.

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Medicine

Ultrasound Assessment of Flow-Mediated Dilation of the Brachial and Superficial Femoral Arteries in Rats
Daniel R. Machin 1, Miriam E. Leary 2, Yuxia He 1,3, Yan-Ting Shiu 1,3, Hirofumi Tanaka 2, Anthony J. Donato 1,4,5,6
1Department of Internal Medicine, University of Utah, 2Department of Kinesiology and Health Education, University of Texas at Austin, 3Division of Nephrology and Hypertension, University of Utah, 4Department of Biochemistry, University of Utah, 5Department of Exercise and Sport Science, University of Utah, 6Geriatric Research Education and Clinical Center, Department of Veterans Affairs

Non-invasive assessment of endothelial function in humans can be determined by the flow-mediated dilation technique. Although thousands of studies have used this technique, no study has performed this technique non-invasively in rats. The following article describes non-invasive measurement of flow-mediated dilation in the brachial and superficial femoral arteries of rats.

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Environment

The Calibration and Use of Capacitance Sensors to Monitor Stem Water Content in Trees
Ashley M. Matheny 1, Steven R. Garrity 2, Gil Bohrer 3
1Department of Geological Sciences, Jackson School of Geosciences, University of Texas at Austin, 2METER Group, Inc., USA, 3Department of Civil, Environmental and Geodetic Engineering, Ohio State University

The hydraulic capacitance of biomass is a key component of the vegetation water budget, which serves as a buffer against short and long-term drought stresses. Here, we present a protocol for the calibration and use of soil moisture capacitance sensors to monitor water content in the stems of large trees.

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Medicine

Intraoperative Ultrasound in Spinal Surgery
Melissa M.J. Chua 1, Viren S. Vasudeva 1, Yi Lu 1
1Brigham and Women's Hospital, Harvard Medical School

Here, we present a protocol on the use of intraoperative ultrasound in spinal surgery, particularly in cases of intradural lesions and lesions in the ventral spinal canal when using a posterior approach.

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Immunology and Infection

In Vitro Differentiation of Mouse Granulocyte-macrophage-colony-stimulating Factor (GM-CSF)-producing T Helper (THGM) Cells
Yi Lu 1,2,3, Xin-Yuan Fu 3, Yongliang Zhang 1,2
1Department of Microbiology and Immunology, Yong Loo Lin School of Medicine, National University of Singapore, 2Immunology Programme, Life Sciences Institute, National University of Singapore, 3Cancer Science Institute of Singapore, Yong Loo Lin School of Medicine, National University of Singapore

Here, we present a protocol to differentiate murine granulocyte-macrophage-colony-stimulating-factor-producing T helper (THGM) cells from naive CD4+ T cells, including isolation of naive CD4+ T cells, differentiation of THGM, and analysis of differentiated THGM cells. This method can be applied to studies of the regulation and function of THGM cells.

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Immunology and Infection

Quantitative PCR of T7 Bacteriophage from Biopanning
Xiujuan Peng 1, Jasmim Leal 1, Rashmi Mohanty 1, Melissa Soto 1, Debadyuti Ghosh 1
1Division of Molecular Pharmaceutics and Drug Delivery, College of Pharmacy, University of Texas at Austin

A reproducible, accurate, and time efficient quantitative PCR (qPCR) method to enumerate T7 bacteriophage is described here. The protocol clearly describes phage genomic DNA preparation, PCR reaction preparation, qPCR cycling conditions, and qPCR data analysis.

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Medicine

Preclinical Model of Hind Limb Ischemia in Diabetic Rabbits
Andrew D. Sligar 1, Gretchen Howe 2, Julia Goldman 3, Patricia Felli 2, Varsha Karanam 1, Richard W. Smalling 2,4, Aaron B. Baker 1,5,6,7
1Department of Biomedical Engineering, University of Texas at Austin, 2Division of Cardiology, University of Texas McGovern Medical School, 3Center for Laboratory Animal Medicine and Care, UT Health Science Center at Houston, 4Memorial Herman Heart and Vascular Center, Texas Medical Center, 5Institute for Cellular and Molecular Biology, University of Texas at Austin, 6The Institute for Computational Engineering and Sciences, University of Texas at Austin, 7Institute for Biomaterials, Drug Delivery and Regenerative Medicine, University of Texas at Austin

We describe a surgical procedure used to induce peripheral ischemia in rabbits with hyperlipidemia and diabetes. This surgery acts as a preclinical model for conditions experienced in peripheral artery disease in patients. Angiography is also described as a means to measure the extent of introduced ischemia and recovery of perfusion.

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Developmental Biology

An In Vitro 3D Model and Computational Pipeline to Quantify the Vasculogenic Potential of iPSC-Derived Endothelial Progenitors
Cody O. Crosby 1, Janet Zoldan 1
1Department of Biomedical Engineering, University of Texas at Austin

Endothelial progenitors derived from induced pluripotent stem cells (iPSC-EPs) have the potential to revolutionize cardiovascular disease treatments and to enable the creation of more faithful cardiovascular disease models. Herein, the encapsulation of iPSC-EPs in three-dimensional (3D) collagen microenvironments and a quantitative analysis of these cells’ vasculogenic potential are described.

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Measuring Interleukin-6 in Human Serum from Children with a Pediatric Concussion using a Single Molecule Counting Platform
Nicole D. Osier 1,2, David Hutto 3, Claire Slote 1, Adam S. Venable 4
1School of Nursing, University of Texas at Austin, 2Department of Neurology, University of Texas at Austin, 3Cockrell School of Engineering, University of Texas at Austin, 4MilliporeSigma

Methods are described for ultrasensitive quantification of biomarker proteins using an immunoassay system capable of single molecule counting. The platform can be used to quantify a wide array of proteins in a variety of tissues from several species. This exemplar assays interleukin-6 in human serum from pediatric concussion patients.

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Genetics

Antibody-Free Assay for RNA Methyltransferase Activity Analysis
Samantha B. Shelton 1, Blerta Xhemalce 1
1Department of Molecular Biosciences, University of Texas at Austin

Here, an antibody-free in vitro assay for direct analysis of methyltransferase activity on synthetic or in vitro transcribed RNA is described.

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Immunology and Infection

Use of Viral Entry Assays and Molecular Docking Analysis for the Identification of Antiviral Candidates against Coxsackievirus A16
Jonathan Y. Wang 1, Chien-Ju Lin 2, Ching-Hsuan Liu 3,4, Liang-Tzung Lin 3,5
1Department of Molecular Biosciences, University of Texas at Austin, 2School of Pharmacy, College of Pharmacy, Kaohsiung Medical University, 3Graduate Institute of Medical Sciences, College of Medicine, Taipei Medical University, 4Department of Microbiology and Immunology, Dalhousie University, 5Department of Microbiology and Immunology, School of Medicine, College of Medicine, Taipei Medical University

The goal of the protocol is to illustrate the different assays relating to viral entry that can be used to identify candidate viral entry inhibitors.

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Engineering

Evaluation of an Exclusive Spur Dike U-Turn Design with Radar-Collected Data and Simulation
Yang Shao 1, Hongtao Yu 1, Huan Wu 2, Xueyan Han 1, Xizhen Zhou 1, Christian G. Claudel 3, Hualing Zhang 4, Chen Yang 5
1Highway Academy, Chang'an University, 2Human Resource Department, Xi'an Shiyou University, 3Cockrell School of Engineering, University of Texas at Austin, 4Department of Earth and Atmospheric Science, University of Houston, 5School of Foreign Languages, Xi'an Shiyou University

This protocol describes the process of solving a microscopic traffic problem with simulation. The whole process contains a detailed description of data collection, data analysis, simulation model build, simulation calibration, and sensitive analysis. Modifications and troubleshooting of the method are also discussed.

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Neuroscience

Three-dimensional Navigation-guided, Prone, Single-position, Lateral Lumbar Interbody Fusion Technique
Samantha E. Hoffman 1, Saksham Gupta 1, Kevin Huang 1, Neil Klinger 1, Yi Lu 1
1Department of Neurosurgery, Brigham and Women’s Hospital, Harvard Medical School

The single-position, prone, lateral approach allows for both lateral lumbar interbody placement and direct posterior decompression with pedicle screw placement in one position.

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Biology

Preparing and Rearing Axenic Insects with Tissue Cultured Seedlings for Host-Gut Microbiota Interaction Studies of the Leaf Beetle
Meiqi Ma 1, Pei Liu 1, Jingya Yu 2, Runhua Han 3, Letian Xu 1
1State Key Laboratory of Biocatalysis and Enzyme Engineering, School of Life Sciences, Hubei University, 2Institute of Plant Protection, Wuhan Institute of Landscape Architecture, 3McKetta Department of Chemical Engineering, University of Texas at Austin

To obtain an axenic insect, its egg surface is sterilized, and the hatched larva is subsequently reared using axenic leaves. This method provides an efficient way for axenic insect preparation without administering antibiotics or developing an artificial diet, which can also be applied to other leaf-eating insects.

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Bioengineering

In Vitro Selection of Aptamers to Differentiate Infectious from Non-Infectious Viruses
Marcos Ezequiel Gramajo *1, Ryan J. Lake *2, Yi Lu 3, Ana Sol Peinetti 1
1INQUIMAE (CONICET), Departamento de Química Inorgánica, Analítica y Química Física, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, 2Department of Chemistry, University of Illinois at Urbana-Champaign, 3Department of Chemistry, University of Texas at Austin

We provide a protocol that can be generally applied to select aptamers that bind to infectious viruses only and not to viruses that have been rendered non-infectious by a disinfection method or to any other similar viruses. This opens the possibility of determining infectivity status in portable and rapid tests.

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