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Drexel University College of Medicine

15 ARTICLES PUBLISHED IN JoVE

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Biology

Combining Peripheral Nerve Grafting and Matrix Modulation to Repair the Injured Rat Spinal Cord
John D. Houle 1, Arthi Amin 1, Marie-Pascale Cote 1, Michel Lemay 1, Kassi Miller 1, Harra Sandrow 1, Lauren Santi 1, Jed Shumsky 1, Veronica Tom 1
1Department of Neurobiology and Anatomy, Drexel University College of Medicine

Traumatic injury to the spinal cord disrupts communication with the brain. To restore lost connectivity we utilize a peripheral nerve graft to provide a substratum for regenerating fibers in combination with neurotrophic factors and matrix-modulating enzymes to remove inhibitory molecules to promote long distance growth.

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Neuroscience

Live Imaging of Dorsal Root Axons after Rhizotomy
Andrew Skuba 1, B. Timothy Himes 2,3, Young-Jin Son 4
1Temple University, Shriners Hospitals Pediatric Research Center and Department of Anatomy and Cell Biology, 2Medical Research Service, Department of Veterans Affairs Hospital, 3Department of Neurobiology and Anatomy, Drexel University College of Medicine, 4Shriners Hospitals Pediatric Research Center and Department of Anatomy and Cell Biology, Temple University School of Medicine

An in vivo imaging protocol to monitor primary sensory axons following dorsal root crush is described. The procedures utilize wide-field fluorescence microscopy and thy1-YFP transgenic mice, and permit repeated imaging of axon regeneration over 4 cm in the PNS and axon interactions with the interface of the CNS.

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Medicine

Acute and Chronic Tactile Sensory Testing after Spinal Cord Injury in Rats
Megan Ryan Detloff 1,2, Lesley C. Fisher 1, Rochelle J. Deibert 1, D. Michele Basso 1
1Center for Brain and Spinal Cord Repair, School of Allied Medical Professions, The Ohio State University, 2Spinal Cord Research Center, Drexel University College of Medicine

We describe two tactile sensory testing methods for acute or chronic periods of spinal cord injury in rats. These validated procedures can detect the development and maintenance of allodynia-like sensations.

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Neuroscience

Bilaminar Co-culture of Primary Rat Cortical Neurons and Glia
Saori Shimizu *1, Anna Abt *1, Olimpia Meucci 1
1Department of Pharmacology and Physiology, Drexel University College of Medicine

Here we provide a protocol for culturing rat cortical neurons in the presence of a glial feeder layer. The cultured neurons establish polarity and create synapses, and can be separated from the glia for use in various applications, such as electrophysiology, calcium imaging, cell survival assays, immunocytochemistry, and RNA/DNA/protein isolation.

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Neuroscience

Ex Vivo Organotypic Corneal Model of Acute Epithelial Herpes Simplex Virus Type I Infection
Oleg Alekseev 1, Anh H. Tran 1, Jane Azizkhan-Clifford 1
1Department of Biochemistry and Molecular Biology, Drexel University College of Medicine

In this video article we describe the use of a new ex vivo model of acute herpes simplex virus type I corneal epithelial infection.

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Biology

Chemically-blocked Antibody Microarray for Multiplexed High-throughput Profiling of Specific Protein Glycosylation in Complex Samples
Chen Lu 1, Joshua L. Wonsidler 1, Jianwei Li 2, Yanming Du 1, Timothy Block 3, Brian Haab 4, Songming Chen 1
1Institute for Hepatitis and Virus Research, 2Department of Microbiology and Immunology, Thomas Jefferson University, 3Drexel University College of Medicine, 4Van Andel Research Institute

In this study, we describe an improved protocol for a multiplexed high-throughput antibody microarray with lectin detection method that can be used in glycosylation profiling of specific proteins. This protocol features new reliable reagents and significantly reduces the time, cost, and lab equipment requirements as compared to the previous procedure.

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Biology

Purification and microRNA Profiling of Exosomes Derived from Blood and Culture Media
Marguerite K. McDonald 1, Kathryn E. Capasso 1, Seena K. Ajit 1
1Department of Pharmacology & Physiology, Drexel University College of Medicine

The presence of stable microRNAs (miRNAs) in exosomes has generated immense interest as a novel mode of intercellular communication, for their potential utility as biomarkers and as a route for therapeutic intervention. Here we demonstrate exosome purification from blood and culture media followed by quantitative PCR to identify miRNAs being transported.

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Identification of Cells with Markers of Cellular Senescence in Human Formalin-fixed, Paraffin-embedded Brain Tissue Sections
Elizabeth P. Crowe *1, Alessandro Bitto *1, Rekha Bhat 1, Frederick Bradley Johnson 2, John Q. Trojanowski 2, Christian Sell 1, Claudio Torres 1
1Department of Pathology and Laboratory Medicine, Drexel University College of Medicine, 2Department of Pathology and Laboratory Medicine, University of Pennsylvania School of Medicine

We describe an immunofluorescence procedure to detect markers of cellular senescence in human formalin-fixed, paraffin-embedded brain sections. In contrast to immunohistochemistry, immunofluorescence permits semi-quantitative measurements and multiple staining on the same slide, which allows detecting different cell types more rigorously and assessing the abundance of senescence markers more objectively.

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Medicine

A Radio-telemetric System to Monitor Cardiovascular Function in Rats with Spinal Cord Transection and Embryonic Neural Stem Cell Grafts
Shaoping Hou 1, Armin Blesch 2, Paul Lu 3,4
1Spinal Cord Research Center, Department of Neurobiology & Anatomy, Drexel University College of Medicine, 2Spinal Cord Injury Center, Heidelberg University Hospital, 3Veterans Administration Medical Center, San Diego, CA, 4Department of Neurosciences, University of California, San Diego

We present a protocol for using a radio-telemetric system to record cardiovascular parameters in T4 spinal cord transected rats eight weeks after embryonic brainstem neural stem cell grafting into the lesion site. Telemetry is an advanced technique to accurately evaluate cardiovascular function in conscious freely moving spinal cord injured rats.

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JoVE Journal

Preparation of Meiotic Chromosome Spreads from Mouse Spermatocytes
Ferdusy Dia 1, Tierra Strange 1, Jenny Liang 1, Jacob Hamilton 1, Karen M. Berkowitz 1,2
1Department of Biochemistry & Molecular Biology, Drexel University College of Medicine, 2Department of Obstetrics & Gynecology, Drexel University College of Medicine

Meiosis is the developmental process by which gametes are formed through a single round of DNA replication and two successive rounds of chromosome segregation. Mammalian meiosis can be examined by utilizing a technique to prepare meiotic chromosome spreads. Here, we demonstrate a method of preparing surface-spread nuclei from mouse spermatocytes.

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Biochemistry

Identification of Novel CK2 Kinase Substrates Using a Versatile Biochemical Approach
John E. Chojnowski 1, Emily A. McMillan 1, Todd I. Strochlic 1
1Department of Biochemistry and Molecular Biology, Drexel University College of Medicine

The objective of this protocol is to label, enrich, and identify substrates of protein kinase CK2 from a complex biological sample such as a cell lysate or tissue homogenate. This method leverages unique aspects of CK2 biology for this purpose.

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Genetics

Combining Laser Capture Microdissection and Microfluidic qPCR to Analyze Transcriptional Profiles of Single Cells: A Systems Biology Approach to Opioid Dependence
Sean J. O'Sullivan 1,2, Beverly A.S. Reyes 3, Rajanikanth Vadigepalli 1, Elisabeth J. Van Bockstaele 3, James S. Schwaber 1
1Daniel Baugh Institute for Functional Genomics and Computational Biology, Department of Pathology, Anatomy, and Cell Biology, Thomas Jefferson University, 2Sidney Kimmel Medical College, Thomas Jefferson University, 3Department of Pharmacology & Physiology, Drexel University College of Medicine

This protocol explains how to collect single neurons, microglia, and astrocytes from the central nucleus of the amygdala with high accuracy and anatomic specificity using laser capture microdissection. Additionally, we explain our use of microfluidic RT-qPCR to measure a subset of the transcriptome of these cells.

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Neuroscience

Uptake of Fluorescent Labeled Small Extracellular Vesicles In Vitro and in Spinal Cord
Richa Gupta 1, Xuan Luo 1, Zhucheng Lin 1, Yuzhen Tian 1, Seena K. Ajit 1
1Department of Pharmacology & Physiology, Drexel University College of Medicine

We describe a protocol to label macrophage-derived small extracellular vesicles with PKH dyes and observe their uptake in vitro and in the spinal cord after intrathecal delivery.

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Cancer Research

An Ex Vivo Brain Slice Model to Study and Target Breast Cancer Brain Metastatic Tumor Growth
Lorela Ciraku 1, Rebecca A. Moeller 1, Emily M. Esquea 1, Wiktoria A. Gocal 1, Edward J. Hartsough 2,4, Nicole L. Simone 3,4, Joshua G. Jackson 2, Mauricio J. Reginato 1,4
1Department of Biochemistry and Molecular Biology, Drexel University College of Medicine, 2Department of Pharmacology and Physiology, Drexel University College of Medicine, 3Department of Radiation Oncology, Thomas Jefferson University, 4Sidney Kimmel Cancer Center, Thomas Jefferson University

We introduce a protocol for measuring real-time drug and radiation response of breast cancer brain metastatic cells in an organotypic brain slice model. The methods provide a quantitative assay to investigate the therapeutic effects of various treatments on brain metastases from breast cancer in an ex vivo manner within the brain microenvironment interface.

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Bioengineering

Investigating Stress-relaxation and Failure Responses in the Trachea
Anita Singh 1, Tanmay Majmudar 2,3, Adi Iyer 4, Diya Iyer 4, Sriram Balasubramanian 3
1Department of Biomedical Engineering, Widener University, 2Drexel University College of Medicine, 3School of Biomedical Engineering, Science and Health Systems, Drexel University, 4Rosetree Media School District

The present protocol determines the tensile stress-relaxation and failure properties of porcine tracheae. Results from such methods can help improve the understanding of the viscoelastic and failure thresholds of the trachea and help advance the capabilities of computational models of the pulmonary system.

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