University of Medicine and Dentistry of New Jersey View Institution's Website 16 articles published in JoVE Neuroscience Membrane Potential Dye Imaging of Ventromedial Hypothalamus Neurons From Adult Mice to Study Glucose Sensing Reema P. Vazirani1, Xavier Fioramonti2, Vanessa H. Routh1 1Department of Pharmacology and Physiology, Rutgers New Jersey Medical School, 2Center for Taste and Feeding Behavior, Universite de Bourgogne The activity of single neurons from adult-aged mice can be studied by dissociating neurons from specific brain regions and using fluorescent membrane potential dye imaging. By testing responses to changes in glucose, this technique can be used to study the glucose sensitivity of adult ventromedial hypothalamic neurons. Neuroscience Calcium Phosphate Transfection of Primary Hippocampal Neurons Miao Sun*1, Laura P. Bernard*1, Victoria L. DiBona1, Qian Wu1, Huaye Zhang1 1Department of Neuroscience and Cell Biology, Robert Wood Johnson Medical School, Rutgers University Calcium phosphate precipitation is a convenient and economical method for transfection of cultured cells. With optimization, it is possible to use this method on hard-to-transfect cells like primary neurons. Here we describe our detailed protocol for calcium phosphate transfection of hippocampal neurons cocultured with astroglial cells. Neuroscience A Caenorhabditis elegans Model System for Amylopathy Study Zhibing Duan1, Federico Sesti1 1Department of Neuroscience and Cell Biology, Robert Wood Johnson Medical School, University of Medicine and Dentistry of New Jersey We describe methods to study aspects of amylopathies in the worm C. elegans. We show how to construct worms expressing human Aβ42 in neurons and how to test their function in behavioral assays. We further show how to obtain primary neuronal cultures that can be used for pharmacological testing. Biology Ex Vivo Assessment of Contractility, Fatigability and Alternans in Isolated Skeletal Muscles Ki Ho Park1, Leticia Brotto2, Oanh Lehoang1, Marco Brotto2, Jianjie Ma1, Xiaoli Zhao1,3 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. Medicine Human Neuroendocrine Tumor Cell Lines as a Three-Dimensional Model for the Study of Human Neuroendocrine Tumor Therapy Chung Wong1, Evan Vosburgh1,2, Arnold J. Levine2,3, Lei Cong2, Eugenia Y. Xu1,2 1Raymond and Beverly Sackler Foundation, 2The Cancer Institute of New Jersey, University of Medicine and Dentistry of New Jersey, 3School of Natural Sciences, Institute for Advanced Study, Princeton, New Jersey We present a simple agarose overlay platform to grow 3D multicellular spheroids using neuroendocrine cancer cell lines. This method provides a very convenient way to examine the effect of therapeutic drugs on the neuroendocrine tumor cells. It could also help us establish human neuroendocrine tumor spheroids for cancer therapy. Biology Fluorescence-based Measurement of Store-operated Calcium Entry in Live Cells: from Cultured Cancer Cell to Skeletal Muscle Fiber Zui Pan1, Xiaoli Zhao2, Marco Brotto3 1Department of Physiology and Biophysics, Confocal Microscopy and Cell Imaging Core, Robert Wood Johnson Medical School, 2Department of Physiology and Biophysics, Robert Wood Johnson Medical School, 3Muscle Biology Research Group-MUBIG Schools of Nursing & Medicine, University of Missouri-Kansas City The extent of store-operated Ca2+ entry (SOCE) can be monitored using fluorescent Ca2+ indicators. Mn2+ quenching of such indicators assays SOCE in cultured cells and skeletal muscle fibers. A technique allowing spatial and temporal resolution of SOCE by confocal imaging of mechanically skinned muscle fibers is also described. Biology In Ovo Electroporation in Embryonic Chick Retina Mohammed M. Islam1, Sung Tae Doh2, Li Cai1,2 1Department of Pharmacology, University of Medicine and Dentistry of New Jersey-Robert Wood Johnson Medical School, 2Department of Biomedical Engineering, Rutgers University The overall goal of this video is to show how to perform targeted retinal injection and in ovo electroporation of DNA/RNA constructs into the chick embryonic retina at the Hamburger and Hamilton stage 22-23, which is about embryonic day 4 (E4). This technique is very useful to study gene expression, gene regulation, and morphological change in developing chick retina. Neuroscience Lateral Fluid Percussion: Model of Traumatic Brain Injury in Mice Janet Alder1, Wendy Fujioka1, Jonathan Lifshitz2,3, David P. Crockett1, Smita Thakker-Varia1 1Department of Neuroscience and Cell Biology, University of Medicine and Dentistry of New Jersey-Robert Wood Johnson Medical School, 2Spinal Cord and Brain Injury Research Center, 3Department of Anatomy and Neurobiology, Department of Physical Medicine and Rehabilitation, University of Kentucky Chandler Medical Center Lateral fluid percussion (LFP), an established model of traumatic brain injury in mice, is demonstrated. LFP fulfills three major criteria for animal models: validity, reliability and clinical relevance. The procedure, consisting of surgical craniotomy, fixation of hub followed by induction of injury, resulting in focal and diffuse injuries, is described. Bioengineering Axon Stretch Growth: The Mechanotransduction of Neuronal Growth Joseph R. Loverde1,2, Rosa E. Tolentino1,2, Bryan J. Pfister1 1Departments of Biomedical Engineering, New Jersey Institute of Technology, 2Graduate School of Biomedical Sciences, University of Medicine and Dentistry of New Jersey A unique tissue engineering method was developed to elongate numerous nerve fibers in culture by recapitulating axon stretch growth; a form of nervous system growth whereby nerves elongate in conjunction with growth of the enlarging body. Biology Visualization of MG53-mediated Cell Membrane Repair Using in vivo and in vitro Systems Noah Weisleder1, Peihui Lin1, Xiaoli Zhao1, Matthew Orange1, Hua Zhu1, Jianjie Ma1 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. Bioengineering A Simple Hanging Drop Cell Culture Protocol for Generation of 3D Spheroids Ramsey Foty1 1Department of Surgery, UMDNJ-Robert Wood Johnson Medical School We describe a simple, rapid method of generating 3D tissue-like spheroids and their potential application to quantify differences in cell-cell interactions. Bioengineering Measurement of Aggregate Cohesion by Tissue Surface Tensiometry Christine M. Butler1, Ramsey A. Foty1 1Department of Surgery, UMDNJ-Robert Wood Johnson Medical School We describe a method of measuring binding energy, expressible as tissue surface tension, between cells within 3D tissue-like aggregates. Differences in tissue surface tension have been demonstrated to correlate with invasiveness of lung, muscle, and brain tumors, and are fundamental determinants of establishing spatial relationships between different cell types. Neuroscience Organotypic Culture of Full-thickness Adult Porcine Retina Jianfeng Wang1, Anton M. Kolomeyer2, Marco A. Zarbin2, Ellen Townes-Anderson1 1Neurology and Neurosciences, University of Medicine and Dentistry of New Jersey - UMDNJ, 2Institute of Ophthalmology and Visual Science, University of Medicine and Dentistry of New Jersey - UMDNJ Here we describe a cost-effective technique for organotypic culture of adult porcine retina for seven days. Briefly, a sterile filter paper was used to lift the neural retina off from the RPE and place photoreceptor side up on an insert raised by a custom-made stand. Biology Eukaryotic Polyribosome Profile Analysis Anthony M. Esposito1, Maria Mateyak1, Dongming He1, Marcus Lewis1, Arjun N. Sasikumar1, Jenna Hutton1, Paul R. Copeland1, Terri G. Kinzy1 1Department of Molecular Genetics, Microbiology, and Immunology, University of Medicine and Dentistry of New Jersey, Robert Wood Johnson Medical School This article describes a protocol for the extraction of translating ribosomes from eukaryotic cells. Once extracted, ribosomes are separated into monosomes and polyribosomes by sucrose gradient fractionation to allow different ribosomal populations to be analyzed. As such, this method is the gold standard for examining the regulation of translation. Biology Preparation of embryos for Electron Microscopy of the Drosophila embryonic heart tube Nadine H. Soplop1,2, Rajesh Patel1, Sunita G. Kramer1,2 1Joint Graduate Program in Cell and Developmental Biology, UMDNJ-Graduate School of Biomedical Sciences and Rutgers: The State University of New Jersey, 2Department of Pathology and Laboratory Medicine, Robert Wood Johnson Medical School, University of Medicine and Dentistry of New Jersey We describe a process for fixation, embedding, sectioning, and imaging of late stage Drosophila embryos for Trasmission Electron Microscopy of the embryonic heart tube. This technique allows for the visualization of the heart tube lumen as well as the basement membrane, which lines the lumen of the heart. Biology Using Laser Tweezers For Manipulating Isolated Neurons In Vitro Robert Clarke1, Jianfeng Wang1, Ellen Townes-Anderson1 1Department of Neurology and Neuroscience, School of Medicine, University of Medicine and Dentistry of New Jersey - UMDNJ This video describes the manipulation of cultured neurons using laser tweezers in vitro.