Wake Forest University

8 ARTICLES PUBLISHED IN JoVE

Medicine

Martin K. Childers¹, Robert W. Grange², Joe N. Kornegay³

¹Department of Neurology and Wake Forest Institute for Regenerative Medicine, Wake Forest University, ²Department of Human Nutrition, Foods and Exercise, Virginia Polytechnic Institute and State University, ³Departments of Pathology and Laboratory Medicine and Neurology and the Gene Therapy Center , University of North Carolina-Chapel Hill

We describe a minimally-invasive and painless method to measure canine hindlimb muscle strength and muscle response to repeated eccentric contractions.

Biology

Tissue Triage and Freezing for Models of Skeletal Muscle Disease

Hui Meng¹, Paul M.L. Janssen², Robert W. Grange³, Lin Yang⁴, Alan H. Beggs⁵, Lindsay C. Swanson⁵, Stacy A. Cossette^1,6, Alison Frase⁷, Martin K. Childers⁸, Henk Granzier⁹, Emanuela Gussoni⁵, Michael W. Lawlor¹

¹Division of Pediatric Pathology, Department of Pathology and Laboratory Medicine, Medical College of Wisconsin, ²Department of Physiology and Cell Biology, The Ohio State University, ³Department of Human Nutrition, Foods and Exercise, Virginia Tech, ⁴Division of Biomedical Informatics, Department of Biostatistics, Department of Computer Science, University of Kentucky, ⁵Division of Genetics and Genomics, The Manton Center for Orphan Disease Research, Boston Children's Hospital, Harvard Medical School, ⁶Cure Congenital Muscular Dystrophy, ⁷Joshua Frase Foundation, ⁸Department of Rehabilitation Medicine, University of Washington, ⁹Department of Physiology, University of Arizona

The analysis of skeletal muscle tissues to determine structural, functional, and biochemical properties is greatly facilitated by appropriate preparation. This protocol describes appropriate methods to prepare skeletal muscle tissue for a broad range of phenotyping studies.

Engineering

High Temperature Fabrication of Nanostructured Yttria-Stabilized-Zirconia (YSZ) Scaffolds by In Situ Carbon Templating Xerogels

Sixbert P. Muhoza¹, Matthew A. Cottam¹, Michael D. Gross^1,2

¹Department of Chemistry, Wake Forest University, ²Center for Energy, Environment, and Sustainability, Wake Forest University

A protocol for fabricating porous, nanostructured yttria-stabilized-zirconia (YSZ) scaffolds at temperatures between 1,000 °C and 1,400 °C is presented.

Medicine

Evaluating the Procedure for Performing Awake Cystometry in a Mouse Model

Travis K. Mann-Gow^*1, Troy R. Larson^*1, Chrissie T. Wøien², Thomas M. Andersen², Karl-Erik Andersson^3,4, Peter Zvara^1,2

¹Department of Surgery, University of Vermont, ²Department of Urology and Biomedical Laboratory, University of Southern Denmark, ³Wake Forest Institute for Regenerative Medicine, Wake Forest University, ⁴Institute for Clinical Medicine, Aarhus University

This study describes the surgical procedures and experimental techniques for performing awake cystometry in a freely moving mouse. In addition, it provides experimental evidence to support its optimization and standardization.

Genetics

A Deep-sequencing-assisted, Spontaneous Suppressor Screen in the Fission Yeast Schizosaccharomyces pombe

Bahjat F. Marayati^1,2, James B. Pease¹, Ke Zhang^1,2

¹Department of Biology, Wake Forest University, ²Center for Molecular Communication and Signaling, Wake Forest University

We present a simple suppressor screen protocol in fission yeast. This method is efficient, mutagen-free, and selective for mutations that often occur at a single genomic locus. The protocol is suitable for isolating suppressors that alleviate growth defects in liquid culture that are caused by a mutation or a drug.

Biology

In Vivo Measurement of Hindlimb Dorsiflexor Isometric Torque from Pig

Benjamin T. Corona¹, Jarrod A. Call^2,3, Matthew Borkowski⁴, Sarah M. Greising⁵

¹School of Medicine, Wake Forest University, ²Department of Kinesiology, University of Georgia, ³Regenerative Bioscience Center, University of Georgia, ⁴Aurora Scientific Inc., ⁵School of Kinesiology, University of Minnesota

The present protocol describes concise experimental details on the evaluation and interpretation of in vivo torque data obtained via electrical stimulation of the common peroneal nerve in anesthetized pigs.

Immunology and Infection

A TNBS-Induced Rodent Model to Study the Pathogenic Role of Mechanical Stress in Crohn's Disease

Ramasatyaveni Geesala^*1, You-Min Lin^*1, Ke Zhang^*1, Suimin Qiu², Xuan-Zheng Shi¹

¹Department of Internal Medicine, the University of Texas Medical Branch, ²Department of Pathology, the University of Texas Medical Branch

The present protocol describes the development of a Crohn's-like colitis model in rodents. Transmural inflammation leads to stenosis at the TNBS instillation site, and mechanical enlargement is observed in the segment proximal to the stenosis. These changes allow studying mechanical stress in colitis.

Biochemistry

Advancing High-Resolution Imaging of Virus Assemblies in Liquid and Ice

Liza-Anastasia DiCecco^*1,2, Samantha Berry^*1, G. M. Jonaid^*1,3, Maria J. Solares^1,4, Liam Kaylor^1,4, Jennifer L. Gray⁵, Carol Bator⁶, William J. Dearnaley¹, Michael Spilman⁷, Madeline J. Dressel-Dukes⁸, Kathryn Grandfield², Sarah M. McDonald Esstman⁹, Deborah F. Kelly^1,5,6

¹Department of Biomedical Engineering, Pennsylvania State University, ²Department of Materials Science and Engineering, McMaster University, ³Bioinformatics and Genomics Graduate Program, Huck Institutes of the Life Sciences, Pennsylvania State University, ⁴Molecular, Cellular, and Integrative Biosciences Graduate Program, Huck Institutes of the Life Sciences, Pennsylvania State University, ⁵Materials Research Institute, Pennsylvania State University, ⁶Huck Institutes of the Life Sciences, Pennsylvania State University, ⁷Applications team, Direct Electron, ⁸Application Scientist, Protochips, Inc., ⁹Department of Biology, Wake Forest University

Here protocols are described to prepare virus assemblies suitable for liquid-EM and cryo-EM analysis at the nanoscale using transmission electron microscopy.