Nodular tumor-like lesions or mucoid flesh, arising from tendon sheaths, Ligaments
, or Joint capsule
, especially of the hands, wrists, or feet. They are not true cysts as they lack epithelial wall. They are distinguished from Synovial cysts
by the lack of communication with a joint cavity or the Synovial membrane
1Discipline of Biomedical Science, School of Medical Sciences, Sydney Medical School and Bosch Institute, University of Sydney, 2The MARCS Institute, University of Western Sydney, 3Discipline of Physiology, School of Medical Sciences, Sydney Medical School and Bosch Institute, University of Sydney
This video article illustrates the set-up, the procedures to patch cell bodies and how to implement dynamic clamp recordings from ganglion cells in whole-mount mouse retinae. This technique allows the investigation of the precise contribution of excitatory and inhibitory synaptic inputs, and their relative magnitude and timing to neuronal spiking.
Published May 16, 2013. Keywords: Neuroscience, Neurobiology, Biomedical Engineering, Anatomy, Physiology, Molecular Biology, Cellular Biology, Neurons, Retinal Neurons, Retinal Ganglion Cells, Eye, Retina, Neurosciences, retina, ganglion cells, synaptic conductance, artificial conductance, tetrodotoxin (TTX), patch clamp, dynamic clamp, conductance clamp, electrophysiology, mouse, animal model
1Department of Neuroscience, University of Minnesota
This article provides a description of how to dissect and record from the isolated retinal preparation in mouse. In particular, we describe how to record light responses from a fluorescently labeled ganglion cell population and subsequently identify and analyze its morphology.
Published January 26, 2011. Keywords: Neuroscience, isolated, retina, ganglion cell, electrophysiology, patch clamp, transgenic, mouse, fluorescent
1National Eye Institute, NIH, 2Ophthalmology Department, The Second Hospital of Harbin Medical University
This protocol shows how to retrogradely label retinal ganglion cells, and how to subsequently make an optic nerve crush injury in order to analyze retinal ganglion cell survival and apoptosis. It is an experimental disease model for different types of optic neuropathy, including glaucoma.
Published April 25, 2011. Keywords: Neuroscience, optic nerve crush injury, retinal ganglion cell, glaucoma, optic neuropathy, retrograde labeling
1Institute of Neurobiology, Ulm University, 2School of Computing Science & Institute of Neuroscience, Newcastle University
Here we present the methodology for fast and high resolution fluorescent voltage-sensitive dye imaging of detailed activity of neurons in the crab stomatogastric ganglion.
Published March 23, 2011. Keywords: Neuroscience, stomatogastric ganglion, voltage sensitive dye, neuron resolution imaging, central pattern generator
1Department of Biological Sciences, Purdue University
We demonstrate how to dissect and culture chick E4 statoacoustic ganglion and E6 spinal cord explants. Explants are cultured under serum-free conditions in 3D collagen gels for 24 hours. Neurite responsiveness is tested with growth factor-supplemented medium and with protein-coated beads.
Published December 20, 2011. Keywords: Neuroscience, chicken, dissection, morphogen, NT-3, neurite outgrowth, spinal cord, statoacoustic ganglion, Wnt5a
1Laboratory of Neurodevelopement and Repair, University of Science and Technology of China
We introduce an efficient method to retrograde label retinal ganglion cells (RGCs) in adult zebrafish.
Published May 3, 2014. Keywords: Neuroscience, Adult Zebrafish, Retinal Ganglion Cell, Retrograde Labeling, DiI
1Departments of Pathology and Cell Biology, and Neuroscience, Columbia University College of Physicians and Surgeons, 2Department of Ophthalmology, Columbia University College of Physicians and Surgeons
Here we present two techniques for manipulating gene expression in murine retinal ganglion cells (RGCs) by in utero and ex vivo electroporation. These techniques enable one to examine how alterations in gene expression affect RGC development, axon guidance, and functional properties.
Published September 24, 2009. Keywords: Neuroscience, Developmental Biology, retinal ganglion cells, electroporation, retinal explants, gene transfection, border assays, in utero, ex vivo
1Department of Orthopaedic Surgery, Johns Hopkins University School of Medicine, 2Solomon H. Snyder Department of Neuroscience, Johns Hopkins University School of Medicine
An in vitro model for genetic study of axon regeneration using cultured adult mouse dorsal root ganglion neurons is described. The method includes a re-suspension/re-plating step to allow axon re-growth from neurons undergoing genetic manipulation. This approach is especially useful for loss-of-function studies of axon regeneration using RNAi-based protein knockdown.…
Published August 17, 2012. Keywords: Neuroscience, Physiology, Developmental Biology, cell culture, axon regeneration, axon growth, dorsal root ganglion, spinal cord injury
1Department of Neurobiology, Yale University, 2Program in Developmental Biology, Baylor College of Medicine
We demonstrate an in vivo electroporation protocol for transfecting single or small clusters of retinal ganglion cells (RGCs) and other retinal cell types in postnatal mice over a wide range of ages. The ability to label and genetically manipulate postnatal RGCs in vivo is a powerful tool for developmental studies.
Published April 17, 2011. Keywords: Neuroscience, Retinotopy, Eye Segregation, Superior Colliculus, Lateral Geniculate Nucleus, Visual Development, Retinal Ganglion Cell, Retina, Electroporation
1The University of Hong Kong - HKU
This video describes the method of retrograde labeling of RGC by applying fluoro-gold (FG) on the surface of superior colliculus (SC). Technique involves drilling the skull, aspirating the cortex, and applying gelatin sponge over entire dorsal surface of SC.
Published June 17, 2008. Keywords: Neuroscience, Retrograde labeling, retinal ganglion cells, ophthalmology research, superior colliculus, experimental glaucoma
1Department of Biology, Case Western Reserve University, 2Department of Neurosciences, Case Western Reserve University, 3Department of Biomedical Engineering, Case Western Reserve University
In animals with large identified neurons (e.g. mollusks), analysis of motor pools is done using intracellular techniques1,2,3,4. Recently, we developed a technique to extracellularly stimulate and record individual neurons in Aplysia californica5. We now describe a protocol for using this technique to uniquely identify and characterize motor neurons within a motor pool.…
Published March 25, 2013. Keywords: Neuroscience, Physiology, Biomedical Engineering, Anatomy, Behavior, Neurobiology, Animal, Neurosciences, Neurophysiology, Electrophysiology, Aplysia, Aplysia californica, California sea slug, invertebrate, feeding, buccal mass, ganglia, motor neurons, neurons, extracellular stimulation and recordings, extracellular electrodes, animal model
1Emmy Noether Group, Institute of Zoology, University of Cologne
Here we describe the dissection of the crayfish abdominal nerve cord. We also demonstrate an electrophysiological technique to record fictive locomotion from swimmeret motor neurons.
Published November 25, 2014. Keywords: Neurobiology, crustacean, dissection, extracellular recording, fictive locomotion, motor neurons, locomotion
1Department of Biology, Case Western Reserve University, 2Department of Neurosciences, Case Western Reserve University, 3Department of Biomedical Engineering, Case Western Reserve University
We describe a technique to extracellularly record and stimulate from nerves, muscles, and individual identified neurons in vitro while eliciting and observing different types of feeding behaviors in the feeding apparatus of Aplysia.
Published December 5, 2012. Keywords: Neuroscience, Physiology, Biomedical Engineering, Anatomy, Marine Biology, Aplysia, Aplysia californica, California sea slug, invertebrate, feeding, neurobiology, buccal mass, semi-intact preparation, extracellular electrodes, extracellular recording, neurons, animal model
1Department of Biology, University of Kentucky, 2Department of Biology, College of Science, University of Salahaddin, Iraq, 3Department of Neurobiology and Cognitive Neuroscience, SISSA, Italy
This article describes three nervous system preparations using leeches: intracellular recording from neurons in ventral ganglia, culturing neurons from ventral ganglia, and recording from a patch of innervated skin to map sensory fields.
Published November 4, 2013. Keywords: Neuroscience, leech, Neurobiology, culture, neurons, electrophysiology, synapse, neurophysiology, neuroethology, developmental biology, ganglion, central nervous system (CNS)
1Biotactical Engineering, Faculty of Engineering and Industrial Science, Swinburne University of Technology, 2Department of Otolaryngology, The University of Melbourne
Infrared nerve stimulation has been proposed as an alternative to electrical stimulation in a range of nerve types, including those associated with the auditory system. This protocol describes a patch clamp method for studying the mechanism of infrared nerve stimulation in a culture of primary auditory neurons.
Published July 31, 2013. Keywords: Neuroscience, Biomedical Engineering, Neurobiology, Molecular Biology, Cellular Biology, Physiology, Primary Cell Culture, Biophysics, Electrophysiology, fiber optics, infrared neural stimulation, patch clamp, in vitro models, spiral ganglion neurons, neurons, patch clamp recordings, cell culture
1Department of Neurobiology, University of California, Los Angeles, 2Veterans Administration Greater Los Angeles Healthcare System, 3Departments of Physiology & Biophysics and Ophthalmology & Visual Sciences, Dalhousie University, 4Departments of Neurobiology and Medicine, Jules Stein Eye Institute, CURE-Digestive Diseases Research Center, David Geffen School of Medicine, University of California, Los Angeles
Immunohistochemistry protocols are used to study the localization of a specific protein in the retina. Calcium imaging techniques are employed to study calcium dynamics in retinal ganglion cells and their axons.
Published October 13, 2014. Keywords: Neuroscience, immunohistochemistry, antibody, fluo-4, calcium imaging, ganglion cells, retina, rat
1Division of Marine Biology and Fisheries, Rosenstiel School of Marine and Atmospheric Sciences, University of Miami
We describe the dissection of the nervous system of the marine sea hare Aplysia after anesthesia, the isolation of neurons for short term-tissue culture, and recordings of single cell ion currents via the patch clamp technique.
Published July 10, 2013. Keywords: Neuroscience, Neurobiology, Anatomy, Physiology, Cellular Biology, Molecular Biology, Environmental Sciences, Marine Biology, Receptors, Neurophysiology, Neurotransmitter, Neurotransmitter Agents, Patch Clamp Recordings, Primary Cell Culture, Electrophysiology, L-Glutamate, NMDA, D-Aspartate, dissection, ganglia, buccal ganglion, neurons, invertebrate, Aplysia californica, california sea slug, mollusk, animal model
1Department of Molecular Physiology and Biophysics, Baylor College of Medicine, 2Department of Neuroscience, Baylor College of Medicine, 3Department of Neuroscience, University of California at San Diego, 4National Center for Macromolecular Imaging, Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine
To preserve neuronal processes for ultrastructural analysis, we describe a protocol for plating of primary neurons on electron microscopy grids followed by flash freezing, yielding samples suspended in a layer of vitreous ice. These samples can be examined with a cryo-electron microscope to visualize structures at the nanometer scale.
Published February 12, 2014. Keywords: Neuroscience, Neurons, Cryo-electron Microscopy, Electron Microscope Tomography, Brain, rat, primary neuron culture, morphological assay
1Department of Neurobiology and Behavior, University of California, Irvine
Here we describe a microdissection technique followed by fluorescent dye injection into the acoustic ganglion of early chick embryos for selective tracing of auditory axon fibers in the nerve and hindbrain.
Published March 18, 2013. Keywords: Neurobiology, Neuroscience, Behavior, Developmental Biology, Anatomy, Biomedical Engineering, Surgery, Development, Inner Ear, Cochlea, Auditory, Chick, Axon Tracing, VIIIth Cranial Nerve, nerve, ganglion, fiber, cochlear duct, basilar papilla, embryo, microinjection, animal model
1Department of Biology, University of Kentucky, 2Department of Physiology, University of Toronto
The experiments demonstrate an easy approach for students to gain experience in examining muscle structure, synaptic responses, the effects of ion gradients and permeability on membrane potentials. Also, a sensory-CNS-motor-muscle circuit is presented to show a means to test effects of compounds on a neuronal circuit.
Published January 18, 2011. Keywords: Neuroscience, Invertebrate, Crayfish, neurophysiology, muscle, anatomy, electrophysiology
1Department of Surgery, University of Toronto
Optic Nerve transection is a widely used model of adult CNS injury. Ninety percent of retinal ganglion cells (RGCs) whose axons are completely transected (axotomy) die within 14 days after axotomy. This model is easily amenable to experimental manipulations and highly reproducible.
Published May 12, 2011. Keywords: Neuroscience, issue 51, Central Nervous System, Retina, Apoptosis, Retinal Ganglion Cell, Axotomy, Optic Nerve Transection, Rat, Retrograde Labeling, Rat Model
1Volen Center for Complex Systems, Brandeis
We describe the fine dissection of the stomatogastric nervous system from the stomach of the American lobster (Homarus americanus).
Published May 28, 2009. Keywords: Neuroscience, lobster, stomach, neural network, dissection, central pattern generator
1Department of Genetics, Development and Cell Biology, Neuroscience Program, Iowa State University
A method for the isolation of single retinal cells and subsequent amplification of their cDNAs is described. Single-cell transcriptomics reveals the degree of cellular heterogeneity present in a tissue and uncovers new marker genes for rare cell populations. The accompanying protocol can be adjusted to suit many different cell types.
Published April 19, 2012. Keywords: Neuroscience, Single-cells, transcriptomics, gene expression, cell-type markers, retina, neurons, genetics
1Department of Neuroscience, The Scripps Research Institute, Florida
Marine snail Aplysia californica has been widely used as a neurobiology model for the studies on cellular and molecular basis of behavior. Here a methodology is described for exploring the nervous system of Aplysia for the electrophysiological and molecular analyses of single neurons of identified neural circuitry.
Published January 13, 2014. Keywords: Neurobiology, intracellular recording, identified neuron, neural circuitry, gene expression, action potential, CREB, Aplysia californica, genomics
JoVE Developmental Biology
1Department of Anatomy and Neuroscience, The University of Melbourne, 2The Florey Institute of Neuroscience and Mental Health Research, The University of Melbourne
Here we present protocols that offer a flexible and strategic foundation for virally manipulating oligodendrocyte precursor cells to overexpress proteins of interest in order to specifically interrogate their role in oligodendrocytes via the in vitro model of central nervous system myelination.
Published January 12, 2015. Keywords: Developmental Biology, lentivirus, cocultures, oligodendrocyte, myelination, oligodendrocyte precursor cells, dorsal root ganglion neurons
1Department of Surgery, University of Toronto
Optic Nerve transection is a widely used model of adult CNS injury. This model is ideal for performing a number of experimental manipulations that target the retina globally or directly target the injured neuronal population of retinal ganglion cells.
Published May 12, 2011. Keywords: Neuroscience, Central Nervous System, Retinal Ganglion Cell, Axotomy, Optic Nerve Transection, Intraocular Injection, Nerve Stump Transfection, Viral Vector, Short Interfering RNA
1Department of Neurobiology & Anatomy, University of Utah, 2Department of Ophthalmology & Visual Sciences, University of Utah
Microglia activation and microgliosis are key responses to chronic neurodegeneration. Here, we present methods for in vivo, long-term visualization of retinal CX3CR1-GFP+ microglial cells by confocal ophthalmoscopy, and for threshold and morphometric analyses to identify and quantify their activation. We monitor microglial changes during early stages of age-related glaucoma.…
Published May 11, 2015. Keywords: Medicine, Neuroscience, microglia, neurodegeneration, glaucoma, retina, optic nerve head, confocal scanning laser ophthalmoscopy, live image analysis, segmentation by thresholding, cell morphometry CX3CR1, DBA/2J
1Bionics Institute, 2Department of Anatomical Pathology, St Vincent's Hospital Melbourne, 3Department of Pathology, University of Melbourne, 4Medical Bionics Department, University of Melbourne
Techniques for visualizing retinal cytoarchitecture directly adjacent to individual electrodes within a retinal stimulator.
Published August 2, 2013. Keywords: Medicine, Anatomy, Physiology, Biomedical Engineering, Bioengineering, Surgery, Ophthalmology, Pathology, Tissue Engineering, Prosthesis Implantation, Implantable Neurostimulators, Implants, Experimental, Histology, bionics, Retina, Prosthesis, Bionic Eye, Retinal, Implant, Suprachoroidal, Fixation, Localization, Safety, Preclinical, dissection, embedding, staining, tissue, surgical techniques, clinical techniques
1Center for Narcolepsy, Sleep and Health Research, University of Illinois at Chicago, 2Department of Pharmacology, University of Illinois at Chicago, 3Department of Biobehavioral Health Science, University of Illinois at Chicago
Afferent vagal signaling transmits important information to central nervous system from receptors located in organs of the abdomen and thorax. The nodose ganglia of vagus nerves contain many types of receptors that modulate vagal activity. This protocol describes a method of local injections of neurochemicals into the nodose ganglia.
Published November 25, 2014. Keywords: Neuroscience, neuroscience, nodose ganglia, vagus nerve, EMG, serotonin, apnea, genioglossus, cannabinoids
1Dept. of Psychiatry and Biobehavioral Sciences, University of California, Los Angeles, 2Dept. of Biological Chemistry, University of California, Los Angeles, 3Semel Institute for Neuroscience and Human Behavior, University of California, Los Angeles
Primary cultures of Aplysia sensory-motor neurons provide a model preparation for studying synapse formation and synaptic plasticity in vitro. This video demonstrates the identification and microdissection of sensory and motor neurons from Aplysia ganglia as well as the methods for establishing and maintaining sensory-motor neurons in culture.…
Published June 8, 2009. Keywords: Neuroscience, Aplysia Californica, Synaptic Plasticity, Sensory Motor Neuronal Cultures, Invertebrates, Short-Term Facilitation, Monosynaptic, Intermediate-Term Facilitation, Ganglia, Long-Term Depression, Autapses, Sirnas, Glutamatergic Synapses, Somata
1Biological Sciences, University of Illinois at Chicago
Recording Ca2+ currents at the presynaptic release face membrane is key to a precise understanding of Ca2+ entry and neurotransmitter release. We present an acute dissociation of the lamprey spinal cord that yields functional isolated reticulospinal axons, permitting recording directly from the release face membrane of individual presynaptic terminals. …
Published October 1, 2014. Keywords: Neuroscience, reticulospinal synapse, reticulospinal axons, presynaptic terminal, presynaptic calcium, voltage-gated calcium channels, vesicle fusion, synaptic transmission, neurotransmitter release, spinal cord, lamprey, synaptic vesicles, acute dissociation
1Department of Physiology, Michigan State University
The enteric nervous system (ENS) is a network of neurons and glia located in the gut wall that controls intestinal reflexes. This protocol describes methods for recording the activity of enteric neurons and glia in live preparations of ENS using Ca2+ imaging.
Published January 29, 2015. Keywords: Neuroscience, enteric nervous system, enteric glia, gut, intestine, intestinal nervous system, autonomic, calcium (Ca2+) imaging, peripheral glia, fluorescent, live cell
1Hans Berger Department of Neurology, Jena University Hospital, 2Immunology, Leibniz Institute for Age Research, Fritz Lipmann Institute, Jena, 3Institute of Diagnostic and Interventional Radiology, Medical Physics Group, Jena University Hospital
This video illustrates a method, using a clinical 3 T scanner, for contrast-enhanced MR imaging of the naïve mouse visual projection and for repetitive and longitudinal in vivo studies of optic nerve degeneration associated with acute optic nerve crush injury and chronic optic nerve degeneration in knock-out mice (p50KO).
Published July 22, 2014. Keywords: Neuroscience, manganese-enhanced MRI, mouse retino-tectal projection, visual system, neurodegeneration, optic nerve injury, NF-κB
1Department of Biology, University of Kentucky, 2Department of Biology, University of Salahaddin, 3Oregon Institute of Marine Biology, University of Oregon
This article describes the cockroach ventral nerve cord dissection and extracellular recordings from the cercal nerve and connectives. Evoked responses are generated by electrical stimulation of the cercal nerve or direct mechanical stimulation of the cerci.
Published November 4, 2013. Keywords: Neuroscience, Life Sciences (General), electrophysiology, neural circuit, cockroach, neuroethology, neural network modeling, P. americana, action potentials (APs)
1Department of Neurobiology, Duke University, 2Department of Cell Biology, Duke University
This protocol details how to quantify synapse number both in dissociated neuronal culture and in brain sections using immunocytochemistry. Using compartment-specific antibodies, we label presynaptic terminals as well as sites of postsynaptic specialization. We define synapses as points of colocalization between the signals generated by these markers.
Published November 16, 2010. Keywords: Neuroscience, synapse, immunocytochemistry, brain, neuron, astrocyte
1Department of Genetics, UMR_S 968, Institut de la Vision, 2Department of Visual Information, UMR_S 968, Institut de la Vision, 3Exploratory Team, UMR_S 968, Institut de la Vision, 4Sorbonne Universités, Paris 06, UMR_S 968, Institut de la Vision, 5INSERM, U968, Institut de la Vision, 6CNRS, UMR_7210, Institut de la Vision
Neural retina of a mouse aged 8 days is on top of a 4% gelatin block. After isolation of the photoreceptor layer (200 µm) by vibratome, the photoreceptors are seeded after mechanical and enzymatic dissociation for culture. The photoreceptor layer can be used for molecular, biochemical analyses or transplantation.
Published December 22, 2014. Keywords: Neuroscience, Primary photoreceptor cell culture, Inherited retinal degeneration, Rod-derived Cone Viability Factor, S-Antigen, Flat-mounted mouse retina, Transplantation.
1Department of Biological Sciences, Purdue University
Aplysia californica neurons develop large growth cones in culture that are excellent for high-resolution imaging of growth cone motility and guidance. Here, we present a protocol for dissection and plating of Aplysia bag cell neurons as well as for setting up a chamber for live cell imaging.
Published February 20, 2008. Keywords: Neuroscience, Aplysia californica, abdominal ganglion, nervous system, bag cell neuron, neuronal growth cone, neuronal cell culture, live cell imaging, cytoskeletal dynamics, growth cone motility and guidance
1Ophthalmology, Weill Medical College of Cornell University
The protocol described here is for structural assessment of a wholemount retinal preparation. This includes descriptions of tissue dissection, mounting onto a hydrophilized polytetrafluoroethylene (PTFE) membrane insert, bolus loading with fluorescent markers, and a comparison of fixation with carbodiimide and paraformaldehyde for immunohistochemical analysis of cellular and synaptic components.…
Published December 13, 2013. Keywords: Basic Protocol, hydrophilized PTFE membrane, retina, bolus loading, carbodiimide fixation, immunohistochemistry, antibody staining, microscopy
1Anatomy and Cell Biology, Wayne State University School of Medicine, 2Ophthalmology, Wayne State University School of Medicine
We will demonstrate how to prepare retinal slices from the mouse eye and record light responses in retinal neurons. The entire procedure is conducted in dark-adapted conditions.
Published February 11, 2015. Keywords: Neuroscience, Retina, Patch clamp recording, light response, mouse, dark adaptation, infrared
1Institute of Oral Biology, Unit of Orofacial Development and Regeneration, University of Zurich
Co-cultures represent a valuable method to study the interactions between nerves and target tissues and organs. Microfluidic systems allow co-culturing ganglia and whole developing organs or tissues in different culture media, thus representing a valuable tool for the in vitro study of the crosstalk between neurons and their targets.
Published August 14, 2015. Keywords: Neuroscience, Developmental biology, orofacial development, tooth, innervation, trigeminal ganglion, microfluidics, co-culture systems
1Department of Neurology and Neurosurgery, Montreal Neurological Institute, McGill University
This article describes a neuronal culture system that can be used to obtain large quantities of pure axonal samples for biochemical and immunocytochemical analyses. This technique will allow an improved understanding of normal axonal physiology and signaling pathways leading to neurodegeneration.
Published July 8, 2014. Keywords: Neuroscience, neuron, axon, filter inserts, culture system, dorsal root ganglion, axonal degeneration
1Fishberg Department of Neuroscience and Friedman Brain Institute, Mt. Sinai School of Medicine, 2Phase Five Communications Inc.
We demonstrate how changes in the intracellular free calcium concentration and synaptic efficacy can be simultaneously monitored in a ganglion preparation of Aplysia. We image intracellular calcium using a fluorescent dye, Calcium Orange, and induce and monitor synaptic transmission with sharp (intracellular) electrodes.
Published July 15, 2012. Keywords: Neuroscience, Molecular Biology, Marine Biology, calcium imaging, intracellular recording, invertebrate, mollusc, Aplysia, Calcium Orange, facilitation, synaptic plasticity
1Institute of Anatomy, Technische Universität Dresden
Parkinson's disease has been related to the exposure to pesticides. Here we show a method to deliver pesticides using a gastric tube at the desired concentration and a method to analyze their effect in alpha-synuclein accumulation in the enteric nervous system.
Published October 26, 2010. Keywords: Neuroscience, neurogical disorders, Parkinson's disease, animal model, mouse, rotenone, gavage, image analysis
1The Vollum Institute, Oregon Health and Science University
Here we describe a protocol for the preparation of agar-embedded retinal slices that are suitable for electrophysiology and Ca2+ imaging. This method allows one to study ribbon-type synapses in retinal microcircuits using direct patch-clamp recordings of single presynaptic nerve terminals.
Published January 19, 2012. Keywords: Neuroscience, synaptic physiology, axon terminal, synaptic ribbon, retina microcircuits, goldfish, zebrafish, brain slices, patch-clamp, membrane capacitance measurements, calcium-imaging, exocytosis, transmitter release
1Department of Biochemistry and Cell Biology, Rice University
An approach for analyzing migration and eventual fate of avian neural crest cells in quail-chick chimeric embryos is described. This method is a simple and straightforward technique for tracing neural crest cells during migration and differentiation that are otherwise difficult to distinguish within an unmanipulated chick embryo.
Published February 7, 2012. Keywords: Neuroscience, Neural crest, chick, quail, chimera, fate map, cell migration, cell differentiation
1Department of Biochemistry and Molecular Biology, Hotchkiss Brain Institute, Alberta Children’s Hospital Research Institute, University of Calgary, 2Department of Medical Genetics, Alberta Children’s Hospital Research Institute, Hotchkiss Brain Institute, University of Calgary
In utero electroporation allows for rapid gene delivery in a spatially- and temporally-controlled manner in the developing central nervous system (CNS). Here we describe a highly adaptable in utero electroporation protocol that can be used to deliver expression constructs into multiple embryonic CNS domains, including the telencephalon, diencephalon and retina.
Published June 23, 2011. Keywords: Neuroscience, In utero electroporation, embryonic central nervous system, telencephalon, diencephalon, retina, gene delivery, mouse, gain-of-function, loss-of-function
1Department of Natural Sciences, Assumption College
Cell culture models provide detailed control over environmental conditions and thus provide a powerful platform to elucidate numerous aspects of neuronal cell biology. We describe a rapid, inexpensive, and reliable method to isolate, dissociate, and culture sensory neurons from chick embryos. Details of substrata preparation and immunocytochemistry are also provided.
Published September 24, 2014. Keywords: Neuroscience, dorsal root gangia, DRG, chicken, in vitro, avian, laminin-1, embryonic, primary
1Department of Ophthalmology, Northwestern University, 2Department of Neurobiology, Northwestern University
Chronic ocular hypertension is induced using laser photocoagulation of the trabecular meshwork in mouse eyes. The intraocular pressure (IOP) is elevated for several months after laser treatment. The decrease of visual acuity and contrast sensitivity of experimental animals are monitored using the optomotor test.
Published August 14, 2013. Keywords: Medicine, Biomedical Engineering, Neurobiology, Anatomy, Physiology, Neuroscience, Cellular Biology, Molecular Biology, Ophthalmology, Retinal Neurons, Retinal Neurons, Retinal Ganglion Cells, Neurodegenerative Diseases, Ocular Hypertension, Retinal Degeneration, Vision Tests, Visual Acuity, Eye Diseases, Retinal Ganglion Cell (RGC), Ocular Hypertension, Laser Photocoagulation, Intraocular pressure (IOP), Tonometer; Visual Acuity, Contrast Sensitivity, Optomotor, animal model
1Department of Therapeutics, Institut de la Vision, Sorbonne Universtés, UPMC Univ Paris 06, UMR_S 968, 2INSERM, U968, 3CNRS, UMR_7210
To investigate the blood-retinal barrier permeability and the inner limiting membrane integrity in animal models of retinal disease, we used several adeno-associated virus (AAV) variants as tools to label retinal neurons and glia. Virus mediated reporter gene expression is then used as an indicator of retinal barrier permeability.
Published April 19, 2015. Keywords: Medicine, Blood-Retinal Barrier (BRB), Inner Limiting Membrane (ILM), Adeno-Associated Virus (AAV)
1Department of Biology, College of William and Mary
Xenopus laevis provides an ideal model system for studying cell fate specification and physiological function of individual retinal cells in primary cell culture. Here we present a technique for dissecting retinal tissues and generating primary cell cultures that are imaged for calcium activity and analyzed by in situ hybridization.
Published December 23, 2012. Keywords: Developmental Biology, Neuroscience, Cellular Biology, Surgery, Anatomy, Physiology, Ophthalmology, retina, primary cell culture, dissection, confocal microscopy, calcium imaging, fluorescent in situ hybridization, FISH, Xenopus laevis, animal model