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11 ARTICLES PUBLISHED IN JoVE

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Medicine

Coronary Artery Ligation and Intramyocardial Injection in a Murine Model of Infarction
Jitka A.I. Virag 1, Robert M. Lust 1
1Department of Physiology, Brody School of Medicine, East Carolina University

Numerous genetic manipulations and/or intramyocardial injections of genes, proteins, cells, and/or biomaterials are superimposed upon the dimension of time in studies of acute ischemia/ reperfusion injury and chronic remodeling in mice. This video illustrates the microsurgical procedures for ischemia/reperfusion, permanent coronary artery ligation, and intramyocardial injection studies.

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Bioengineering

Graphene Coatings for Biomedical Implants
Ramakrishna Podila 1,2, Thomas Moore 3, Frank Alexis 3, Apparao Rao 1,4
1Department of Physics, Clemson University, 2Department of Pharmacology and Toxicology, East Carolina University, 3Department of Bioengineering, Clemson University, 4Center for Optical Materials Science and Engineering Technologies, Clemson University

Graphene offers potential as a coating material for biomedical implants. In this study we demonstrate a method for coating nitinol alloys with nanometer thick layers of graphene and determine how graphene may influence implant response.

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JoVE Journal

In vitro Cell Migration and Invasion Assays
Calvin R. Justus 1, Nancy Leffler 1, Maria Ruiz-Echevarria 1, Li V. Yang 1
1Division of Hematology/Oncology, Department of Oncology, East Carolina University

Commonly used, highly accessible methods for examining cell migration and invasion in vitro are described. The first method is the cell wound closure assay that measures cell motility. The second method is the transwell migration and invasion assay that assesses the chemotactic and invasive capacity of cells.

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Neuroscience

The Use of Trace Eyeblink Classical Conditioning to Assess Hippocampal Dysfunction in a Rat Model of Fetal Alcohol Spectrum Disorders
Tuan D. Tran 1,2, Aenia Amin 1,2, Keith G. Jones 2, Ellen M. Sheffer 1, Lidia Ortega 1, Keith Dolman 2
1Department of Psychology, East Carolina University, 2Multidisciplinary Studies Program in Neuroscience, East Carolina University

Trace eyeblink classical conditioning (ECC) was used to assess hippocampal-dependent associative learning in adult rats that were administered a high concentration (11.9% v/v) of alcohol during early neonatal brain development. In general, ECC procedures are sound diagnostic tools for detecting brain dysfunction across many psychological and biomedical settings.

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Immunology and Infection

In Vivo Assessment of Alveolar Macrophage Efferocytosis Following Ozone Exposure
Myles X. Hodge 1, Sky W. Reece 1, Jennifer H. Madenspacher 2, Kymberly M. Gowdy 1
1Department of Pharmacology and Toxicology, East Carolina University, 2Research Triangle Park, National Institute of Environmental Health and Sciences

This manuscript describes a protocol for determining whether exposure to ozone, a criteria air pollutant, impairs alveolar macrophage efferocytosis in vivo. This protocol utilizes commonly used reagents and techniques and can be adapted to multiple models of pulmonary injury to determine effects on alveolar macrophage efferocytosis.

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Developmental Biology

Using Chicken Embryo as a Powerful Tool in Assessment of Developmental Cardiotoxicities
Qixiao Jiang 1, Xiaohui Xu 1, Jamie C. DeWitt 2, Yuxin Zheng 1
1School of Public Health, Qingdao University, 2Department of Pharmacology & Toxicology, Brody School of Medicine, East Carolina University

Chicken embryos, as a classical developmental model, are used in our lab to assess developmental cardiotoxicities following exposure to various environmental contaminants. Exposure methods and morphological/functional assessment methods established are described in this manuscript.

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Biology

In vivo Measurement of Knee Extensor Muscle Function in Mice
Camille R. Brightwell 1,2, Ted G. Graber 3, Benjamin D. Brightwell 4,5, Matthew Borkowski 6, Brian Noehren 5,7, Christopher S. Fry 1,2
1Department of Athletic Training and Clinical Nutrition, University of Kentucky, 2Center for Muscle Biology, University of Kentucky, 3Department of Physical Therapy, East Carolina University, 4Kinesiology and Health Promotion Graduate Program, University of Kentucky, 5Biomotion Lab, College of Health Sciences, University of Kentucky, 6Aurora Scientific, 7Department of Physical Therapy, College of Health Sciences, University of Kentucky

Quantification of knee extensor maximal strength is imperative to understand functional adaptations to aging, disease, injury, and rehabilitation. We present a novel method to repeatedly measure in vivo knee extension isometric peak tetanic torque.

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Biochemistry

Measuring Mitochondrial Electron Transfer Complexes in Previously Frozen Cardiac Tissue from the Offspring of Sow: A Model to Assess Exercise-Induced Mitochondrial Bioenergetics Changes
Daniel Barrera 1, Sierra Upton 2, Megan Rauch 3, Tara Notarianni 4, Ki Suk Eum 5, Megan Liberty 6, Sarmila Venkoba Sah 7, Robert Liu 8, Sean Newcomer 9, Linda E. May 10, Emre Agbas 11, Jessica Sage 12, Edina Kosa 7, Abdulbaki Agbas 7,13
1AdventHelath, 2University of Illinois Chicago School of Public Health, 3Lincoln Memorial University, 4Children’s Mercy Hospital, 5Scripps Clinic, 6Cincinnati Children’s Hospital Medical Center, 7Kansas City University, 8Roblex Rex Veterans Affairs Medical Center, 9California State University San Marcos, 10East Carolina University, 11Massachusetts Institute of Technology, 12Boehringer Ingelheim Norway KS, 13Heartland Center for Mitochondrial Medicine

Preparation of mitochondria-enriched samples from previously frozen archived solid tissues allowed the investigators to perform both functional and analytical assessments of mitochondria in various experimental modalities. This study demonstrates how to prepare mitochondria-enriched preparations from frozen heart tissue and perform analytical assessments of mitochondria.

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JoVE Core

Drosophila Passive Avoidance Behavior as a New Paradigm to Study Associative Aversive Learning
Elena S. Pak 1, Alexander K. Murashov 1
1Department of Physiology, East Carolina University

This work describes a simple behavioral paradigm that allows the analysis of aversive associative learning in adult fruit flies. The method is based on suppressing the innate negative geotaxis behavior due to the association formed between a specific environmental context and an electric shock.

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Cancer Research

Three-Dimensional Culture of Murine Colonic Crypts to Study Intestinal Stem Cell Function Ex Vivo
Amna N. Naser 1, Qun Lu 1, Yan-Hua Chen 1
1Department of Anatomy and Cell Biology, Brody School of Medicine, East Carolina University

The present protocol describes establishing a murine colonic organoid system to study the activity and functioning of colonic stem cells in a claudin-7 knockout model.

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Immunology and Infection

Quantification of Autoreactive Antibodies in Mice upon Experimental Autoimmune Encephalomyelitis
Jonathan J. Carver 1, Alessandro Didonna 1
1Department of Anatomy and Cell Biology, Brody School of Medicine, East Carolina University

Experimental autoimmune encephalomyelitis (EAE) is an animal model of multiple sclerosis (MS), which shares with the human disease a robust humoral autoimmune response. Here, we report a simple and flexible ELISA protocol to quantify autoantibodies in the serum of EAE immunized mice.

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