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The Ohio State University Wexner Medical Center

15 ARTICLES PUBLISHED IN JoVE

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Biology

Visualization of MG53-mediated Cell Membrane Repair Using in vivo and in vitro Systems
Noah Weisleder 1, Peihui Lin 1, Xiaoli Zhao 1, Matthew Orange 1, Hua Zhu 1, Jianjie Ma 1
1Department of Physiology and Biophysics, Robert Wood Johnson Medical School

Described here are protocols used to visualize the dynamic process of MG53-mediated cell membrane repair in whole animals and at the cellular level. These methods can be applied to investigate the cell biology of plasma membrane resealing and regenerative medicine.

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Biology

Ex Vivo Assessment of Contractility, Fatigability and Alternans in Isolated Skeletal Muscles
Ki Ho Park 1, Leticia Brotto 2, Oanh Lehoang 1, Marco Brotto 2, Jianjie Ma 1, Xiaoli Zhao 1,3
1Department of Physiology and Biophysics, UMDNJ-Robert Wood Johnson Medical School, 2Muscle Biology Research Group, University of Missouri-Kansas City, 3Pharmacology division, College of Pharmacy, DHLRI, Ohio State University

We describe a method to directly measure muscle force, muscle power, contractile kinetics and fatigability of isolated skeletal muscles in an in vitro system using field stimulation. Valuable information on Ca2+ handling properties and contractile machinery of the muscle can be obtained using different stimulating protocols.

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Biology

Assessment of Calcium Sparks in Intact Skeletal Muscle Fibers
Ki Ho Park 1, Noah Weisleder 2, Jingsong Zhou 3, Kristyn Gumpper 1, Xinyu Zhou 1, Pu Duann 4, Jianjie Ma 1, Pei-Hui Lin 1
1Department of Surgery, Davis Heart and Lung Research Institute, The Ohio State University Wexner Medical Center, 2Department of Physiology and Cell Biology, Davis Heart and Lung Research Institute, The Ohio State University Wexner Medical Center, 3Department of Molecular Biophysics and Physiology, Rush University Medical Center, 4Department of Internal Medicine, Davis Heart and Lung Research Institute, The Ohio State University Wexner Medical Center

Described here is a method to directly measure calcium sparks, the elementary units of Ca2+ release from sarcoplasmic reticulum in intact skeletal muscle fibers. This method utilizes osmotic-stress-mediated triggering of Ca2+ release from ryanodine receptor in isolated muscle fibers. The dynamics and homeostatic capacity of intracellular Ca2+ signaling can be employed to assess muscle function in health and disease.

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Medicine

A Murine Model of Myocardial Ischemia-reperfusion Injury through Ligation of the Left Anterior Descending Artery
Zhaobin Xu 1, Jenna Alloush 1, Eric Beck 1, Noah Weisleder 1
1Davis Heart & Lung Research Institute, Department of Physiology and Cell Biology, The Ohio State University

We introduce a surgical method to induce experimental ischemia/reperfusion (I/R) injury to simulate myocardial infarction (MI) in mouse models that allows for more clarity in positioning of the ligation on the left anterior descending artery (LAD) to increase the reproducibility of MI experiments in mice.

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Medicine

Method of Isolated Ex Vivo Lung Perfusion in a Rat Model: Lessons Learned from Developing a Rat EVLP Program
Kevin Nelson 1,2, Christopher Bobba 1,2, Emre Eren 3, Tyler Spata 4, Malak Tadres 2, Don Hayes, Jr. 5,6, Sylvester M. Black 3,7, Samir Ghadiali *1,2,3, Bryan A. Whitson *2,3,4
1Department of Biomedical Engineering, Ohio State University Wexner Medical Center, 2Davis Heart & Lung Research Institute, Ohio State University Wexner Medical Center, 3The Collaboration for Organ Perfusion, Protection, Engineering and Regeneration (COPPER) Laboratory, Ohio State University Wexner Medical Center, 4Division of Cardiac Surgery, Department of Surgery, Ohio State University Wexner Medical Center, 5Departments of Pediatrics and Internal Medicine, Ohio State University, 6Advanced Lung Disease Program, Lung and Heart-Lung Transplant Programs, Nationwide Children's Hospital, 7Division of Transplantation, Department of Surgery, Ohio State University Wexner Medical Center

Ex-Vivo Lung Perfusion (EVLP) has allowed lung transplantation in humans to become more readily available by enabling the ability to assess organs and expand the donor pool. Here, we describe the development of a rat EVLP program and refinements that allow for a reproducible model for future expansion.

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JoVE Core

Electrophysiological Motor Unit Number Estimation (MUNE) Measuring Compound Muscle Action Potential (CMAP) in Mouse Hindlimb Muscles
W. David Arnold 1,2,3, Kajri A. Sheth 1, Christopher G. Wier 4, John T. Kissel 1,3, Arthur H. Burghes 1,3,4, Stephen J. Kolb 1,3,4
1Department of Neurology, The Ohio State University Wexner Medical Center, 2Department of Physical Medicine and Rehabilitation, The Ohio State University, 3Department of Neuroscience, The Ohio State University Wexner Medical Center, 4Department of Biochemistry and Pharmacology, The Ohio State University Wexner Medical Center

We present refined protocols that allow in vivo monitoring of motor unit function in the mouse. Techniques to measure compound muscle action potential (CMAP) and motor unit number estimation (MUNE) in the mouse hind limb muscles innervated by the sciatic nerve are described.

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Medicine

Method of Direct Segmental Intra-hepatic Delivery Using a Rat Liver Hilar Clamp Model
Eliza W. Beal 1, Curtis Dumond 1, Jung-Lye Kim 1, Khalid Mumtaz 1, Don Hayes Jr. 1, Ken Washburn 1, Bryan A. Whitson 1, Sylvester M. Black 1
1Collaboration for Organ Perfusion, Protection, Engineering and Regeneration (COPPER) Lab, Division of Transplant, Department of Surgery, Comprehensive Transplant Center, The Ohio State University Wexner Medical Center

A unique rat liver hilar clamp model was developed for studying the impact of pharmacologic molecules in ameliorating ischemia-reperfusion injury. This model includes direct cannulation of the portal supply to the ischemic liver segment via a branch of the portal vein, allowing for direct hepatic delivery.

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Medicine

A Small Animal Model of Ex Vivo Normothermic Liver Perfusion
Eliza W. Beal 1,2, Curtis Dumond 1, Jung-Lye Kim 1,2, Clifford Akateh 1,2, Emre Eren 1, Katelyn Maynard 1, Chandan K. Sen 3, Jay L. Zweier 4, Kenneth Washburn 2, Bryan A. Whitson 1,3, Sylvester M. Black 1,2
1Collaboration for Organ Perfusion, Protection, Engineering and Regeneration (COPPER) Lab, Division of Transplant, Department of Surgery, Comprehensive Transplant Center, Ohio State University Wexner Medical Center, 2Department of Surgery, Division of Transplant, Ohio State University Wexner Medical Center, 3Department of Surgery, Division of CardioThoracic Surgery, Ohio State University Wexner Medical Center, 4Department of Medicine, Ohio State University Wexner Medical Center

There is a significant liver donor shortage, and criteria for liver donors have been expanded. Normothermic ex vivo liver perfusion (NEVLP) has been developed to evaluate and modify organ function. This study demonstrates a rat model of NEVLP and tests the ability of pegylated-catalase, to mitigate liver preservation injury.

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Neuroscience

3D Kinematic Analysis for the Functional Evaluation in the Rat Model of Sciatic Nerve Crush Injury
Tianshu Wang 1, Akira Ito 2, Junichi Tajino 1,3, Hiroshi Kuroki 2, Tomoki Aoyama 1
1Department of Development and Rehabilitation of Motor Function, Human Health Sciences, Graduate School of Medicine, Kyoto University, 2Department of Motor Function Analysis, Human Health Sciences, Graduate School of Medicine, Kyoto University, 3Department of Otolaryngology, The Ohio State University Wexner Medical Center

We introduce a kinematic analysis method that uses a three-dimensional motion capture apparatus containing four cameras and data processing software for performing functional evaluations during fundamental research involving rodent models.

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Developmental Biology

A Protocol for Immunohistochemistry and RNA In-situ Distribution within Early Drosophila Embryo
Wei Zhang *1, Xinjuan Lei *1, Xin Zhou *2,3, Boling He 1, Liqin Xiao 1, Huimin Yue 1, Shulin Wang 1, Yuting Sun 1, Yajun Wu 1, Liyang Wang 1,4, George Ghartey-Kwansah 1, Odell D. Jones 5, Joseph L. Bryant 6, MengMeng Xu 7, Jianjie Ma 3, Xuehon Xu 1
1National Engineering Laboratory for Resource Development of Endangered Crude Drugs in Northwest of China/CGDB, Shaanxi Normal University College of Life Sciences, 2Shaanxi Key Laboratory of Ischemic Cardiovascular Disease, Shaanxi Key Laboratory of Brain disorders, Institute of Basic & Translational Medicine, Xi'an Medical University, 3Ohio State University College of Medicine, 4Hematology-Oncology Division, Department of Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, 5University of Pennsylvania ULAR, 6University of Maryland School of Medicine, 7Columbia University Medical Center

Here, we describe a protocol for detection and localization of Drosophila embryo protein and RNA from collection to pre-embedding and embedding, immunostaining, and mRNA in situ hybridization.

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Medicine

A Rat Lung Transplantation Model of Warm Ischemia/Reperfusion Injury: Optimizations to Improve Outcomes
Yong Gyu Lee 1,2, Jung-Lye Kim 1,2, Andre F. Palmer 3, Brenda F. Reader 4, Jianjie Ma 1, Sylvester M. Black 2,4, Bryan A. Whitson 1,2
1Department of Surgery Division of Cardiac Surgery, The Ohio State University Wexner Medical Center, 2The Collaboration for Organ Perfusion, Protection, Engineering and Regeneration (COPPER) Laboratory, The Ohio State University College of Medicine, 3William G. Lowrie Department of Chemical and Biomolecular Engineering, College of Engineering, The Ohio State University, 4Comprehensive Transplant Center, The Ohio State University Wexner Medical Center

Here, we present optimizations to a rat lung transplantation model that serve to improve outcomes. We provide a size guide for cuffs based on body weight, a measurement strategy to ascertain the 4th intercostal space, and methods of wound closure and BAL (bronchoalveolar lavage) fluid and tissue collection.

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Neuroscience

Comprehensive Understanding of Inactivity-Induced Gait Alteration in Rodents
Junichi Tajino 1,2, Tomoki Aoyama 1, Hiroshi Kuroki 1, Akira Ito 1
1Department of Motor Function Analysis, Human Health Sciences, Graduate School of Medicine, Kyoto University, 2Otolaryngology - Head & Neck Surgery, Ohio State Wexner Medical Center

The present protocol describes three-dimensional motion tracking/evaluation to depict gait motion alteration of rats after exposure to a simulated disuse environment.

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Cancer Research

Magnetic Fluorescent Bead-Based Dual-Reporter Flow Analysis of PDL1-Vaxx Peptide Vaccine-Induced Antibody Blockade of the PD-1/PD-L1 Interaction
Jay Overholser *1, Linlin Guo *1, Pravin T. P. Kaumaya 1,2
1Department of Obstetrics & Gynecology, The Ohio State University Wexner Medical Center, 2The James Comprehensive Cancer Center

Checkpoint inhibitors are important targets in developing therapies for the battle against cancer. This report introduces a novel PDL1 peptide-based cancer vaccine, PDL1-Vaxx, which induces neutralizing polyclonal antibody production that blocks PD-1/PDL1 complex formation. This work also details the development and testing of a fluorescent bead-based assay for analyzing this activity.

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Medicine

A Randomized, Sham-Controlled Trial of Cranial Electrical Stimulation for Fibromyalgia Pain and Physical Function, Using Brain Imaging Biomarkers
Anna Ree 1, Benjamin Rapsas 2, Chanse Denmon 1, Mark Vernon 1, Sheila AM Rauch 1,2, Ying Guo 3, Xiangqin Cui 1,3, Jennifer S Stevens 1,2, Venkatagiri Krishnamurthy 1,2, Vitaly Napadow 4, Jessica A Turner 5, Anna Woodbury 1,2
1Atlanta Veterans Affairs Healthcare System, 2Emory University School of Medicine, 3Rollins School of Public Health, 4Spaulding Rehabilitation Hospital, Harvard Medical School, 5The Ohio State University Wexner Medical Center

The current study is a randomized, placebo-controlled trial to determine the efficacy of cranial electrical stimulation (CES) for improving pain and function in fibromyalgia and further develop resting functional connectivity magnetic resonance imaging (rs-fcMRI) as a clinical tool to assess the neural correlates and mechanisms of chronic pain and analgesic response.

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Medicine

Exploring Alternative Perfusion Solutions Using Next-Generation Polymerized Hemoglobin-Based Oxygen Carriers in a Model of Rat Ex Vivo Lung Perfusion
Doug A. Gouchoe *1,2, Yong Gyu Lee *1,2, Alisyn Greenfield 3, Clayton Cuddington 3, Jung-Lye Kim 1,2, Sylvester M. Black 1, Andre F. Palmer 3, Bryan A. Whitson 1,2
1COPPER Laboratory, Department of Surgery, The Ohio State University Wexner Medical Center, 2Division of Cardiac Surgery, Department of Surgery, The Ohio State University Wexner Medical Center, 3William G. Lowrie Department of Chemical and Biomolecular Engineering, College of Engineering, The Ohio State University

Here, we describe the application of a polymerized human hemoglobin (PolyhHb)-based oxygen carrier as a perfusate and the protocol in which this perfusion solution can be tested in a model of rat ex vivo lung perfusion.

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