16 articles published in JoVE
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.
Published November 27, 2013. Keywords: Neuroscience, membrane potential dye, ventromedial hypothalamus, adult neurons, glucose sensing, fluorescence imaging, arcuate nucleus
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.
Published November 12, 2013. Keywords: Neuroscience, Primary hippocampal neuron, calcium phosphate transfection, Coculture, astroglial cells, DNA
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.
Published May 17, 2013. Keywords: Neuroscience, Medicine, Neurobiology, Cellular Biology, Molecular Biology, Genetics, Developmental Biology, Cell Physiological Phenomena, neurosciences, beta-amyloid, C. elegans, Caenorhabditis elegans, apoptosis, amylopathy, amyloid beta, Alzheimer's disease, AD, neurons, cell culture, transgenic worm, animal model
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.
Published November 1, 2012. Keywords: Physiology, extensor digitorum longus, soleus, in vitro contractility, calcium signaling, muscle-tendon complex, mechanic alternans
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.
Published August 14, 2012. Keywords: Cancer Biology, Medicine, Neuroscience, Cell Culture, Tissue Engineering, 3D model, multicellular spheroids, therapeutic drugs, neuroendocrine tumor cell lines, agarose overlay platform, paraffin embedding
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.
Published February 13, 2012. Keywords: Cellular Biology, Mn quenching, 2-APB, Fura-2, Orai1, esophageal squamous cell carcinoma, skinned muscle fiber
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.
Published February 5, 2012. Keywords: Developmental Biology, Chick, Embryo, Retina, Electroporation, Injection, Egg, GFP, In Ovo, Development
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.
Published August 22, 2011. Keywords: Neuroscience, Lateral fluid percussion, hippocampus, traumatic brain injury, Morris Water Maze, mouse model of moderate injury
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.
Published August 10, 2011. Keywords: Bioengineering, tissue engineering, live imaging, axon stretch growth, nerve development, neuron, neuroscience
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.
Published June 30, 2011. Keywords: Cell Biology, mouse, cell membrane, muscle injury, tissue repair, treadmill, MG53, confocal microscopy, vesicle trafficking
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.
Published May 6, 2011. Keywords: Bioengineering, 3D, hanging drop cultures, cell sorting-out, differential adhesion
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.
Published April 8, 2011. Keywords: Bioengineering, 3D, aggregate cohesion, tissue surface tension, parallel plate compression
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.
Published March 20, 2011. Keywords: Neuroscience, Retina, in vitro, Porcine, Photoreceptor
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.
Published June 15, 2010. Keywords: Cellular Biology, translation, ribosome, polyribosome, gradient, fractionation
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.
Published December 21, 2009. Keywords: Developmental Biology, Drosophila, transmission electron microscopy, fixation, sectioning, embryonic heart tube, lumen
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.
Published September 11, 2008. Keywords: cell biology, neuron, laser, in vitro, culture dish,