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October, 2006
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Ganglion Cysts: 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.
 JoVE Neuroscience

Implementing Dynamic Clamp with Synaptic and Artificial Conductances in Mouse Retinal Ganglion Cells

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

JoVE 50400

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.

 JoVE Neuroscience

An Isolated Retinal Preparation to Record Light Response from Genetically Labeled Retinal Ganglion Cells

1Department of Neuroscience, University of Minnesota

JoVE 2367

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.

 JoVE Neuroscience

An Optic Nerve Crush Injury Murine Model to Study Retinal Ganglion Cell Survival

1National Eye Institute, NIH, 2Ophthalmology Department, The Second Hospital of Harbin Medical University

JoVE 2685

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.

 JoVE Neuroscience

Optical Imaging of Neurons in the Crab Stomatogastric Ganglion with Voltage-sensitive Dyes

1Institute of Neurobiology, Ulm University, 2School of Computing Science & Institute of Neuroscience, Newcastle University

JoVE 2567

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.

 JoVE Neuroscience

Dissection and Culture of Chick Statoacoustic Ganglion and Spinal Cord Explants in Collagen Gels for Neurite Outgrowth Assays

1Department of Biological Sciences, Purdue University

JoVE 3600

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.

 JoVE Biology

In utero and ex vivo Electroporation for Gene Expression in Mouse Retinal Ganglion Cells

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

JoVE 1333

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.

 JoVE Neuroscience

Genetic Study of Axon Regeneration with Cultured Adult Dorsal Root Ganglion Neurons

1Department of Orthopaedic Surgery, Johns Hopkins University School of Medicine, 2Solomon H. Snyder Department of Neuroscience, Johns Hopkins University School of Medicine

JoVE 4141

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.

 JoVE Neuroscience

Transfection of Mouse Retinal Ganglion Cells by in vivo Electroporation

1Department of Neurobiology, Yale University, 2Program in Developmental Biology, Baylor College of Medicine

JoVE 2678

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.

 JoVE Biology

Retrograde Labeling of Retinal Ganglion Cells by Application of Fluoro-Gold on the Surface of Superior Colliculus

1The University of Hong Kong - HKU

JoVE 819

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.

 JoVE Neuroscience

Extracellularly Identifying Motor Neurons for a Muscle Motor Pool in Aplysia californica

1Department of Biology, Case Western Reserve University, 2Department of Neurosciences, Case Western Reserve University, 3Department of Biomedical Engineering, Case Western Reserve University

JoVE 50189

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.

 JoVE Neuroscience

The Swimmeret System of Crayfish: A Practical Guide for the Dissection of the Nerve Cord and Extracellular Recordings of the Motor Pattern

1Emmy Noether Group, Institute of Zoology, University of Cologne

JoVE 52109

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.

 JoVE Neuroscience

An In Vitro Preparation for Eliciting and Recording Feeding Motor Programs with Physiological Movements in Aplysia californica

1Department of Biology, Case Western Reserve University, 2Department of Neurosciences, Case Western Reserve University, 3Department of Biomedical Engineering, Case Western Reserve University

JoVE 4320

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.

 JoVE Neuroscience

Intracellular Recording, Sensory Field Mapping, and Culturing Identified Neurons in the Leech, Hirudo medicinalis

1Department of Biology, University of Kentucky, 2Department of Biology, College of Science, University of Salahaddin, Iraq, 3Department of Neurobiology and Cognitive Neuroscience, SISSA, Italy

JoVE 50631

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.

 JoVE Neuroscience

Whole Cell Patch Clamp for Investigating the Mechanisms of Infrared Neural Stimulation

1Biotactical Engineering, Faculty of Engineering and Industrial Science, Swinburne University of Technology, 2Department of Otolaryngology, The University of Melbourne

JoVE 50444

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.

 JoVE Neuroscience

Immunohistochemical and Calcium Imaging Methods in Wholemount Rat Retina

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

JoVE 51396

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.

 JoVE Neuroscience

Isolation of Sensory Neurons of Aplysia californica for Patch Clamp Recordings of Glutamatergic Currents

1Division of Marine Biology and Fisheries, Rosenstiel School of Marine and Atmospheric Sciences, University of Miami

JoVE 50543

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.

 JoVE Neuroscience

Preparation of Primary Neurons for Visualizing Neurites in a Frozen-hydrated State Using Cryo-Electron Tomography

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

JoVE 50783

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.

 JoVE Neuroscience

Selective Tracing of Auditory Fibers in the Avian Embryonic Vestibulocochlear Nerve

1Department of Neurobiology and Behavior, University of California, Irvine

JoVE 50305

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.

 JoVE Neuroscience

Membrane Potentials, Synaptic Responses, Neuronal Circuitry, Neuromodulation and Muscle Histology Using the Crayfish: Student Laboratory Exercises

1Department of Biology, University of Kentucky, 2Department of Physiology, University of Toronto

JoVE 2322

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.

 JoVE Neuroscience

Optic Nerve Transection: A Model of Adult Neuron Apoptosis in the Central Nervous System

1Department of Surgery, University of Toronto

JoVE 2241

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.

 JoVE Neuroscience

Single-cell Profiling of Developing and Mature Retinal Neurons

1Department of Genetics, Development and Cell Biology, Neuroscience Program, Iowa State University

JoVE 3824

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.

 JoVE Bioengineering

Aplysia Ganglia Preparation for Electrophysiological and Molecular Analyses of Single Neurons

1Department of Neuroscience, The Scripps Research Institute, Florida

JoVE 51075

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.

 JoVE Developmental Biology

Production and Use of Lentivirus to Selectively Transduce Primary Oligodendrocyte Precursor Cells for In Vitro Myelination Assays

1Department of Anatomy and Neuroscience, The University of Melbourne, 2The Florey Institute of Neuroscience and Mental Health Research, The University of Melbourne

JoVE 52179

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.

 JoVE Neuroscience

Methods for Experimental Manipulations after Optic Nerve Transection in the Mammalian CNS

1Department of Surgery, University of Toronto

JoVE 2261

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.

 JoVE Medicine

In Vivo Dynamics of Retinal Microglial Activation During Neurodegeneration: Confocal Ophthalmoscopic Imaging and Cell Morphometry in Mouse Glaucoma

1Department of Neurobiology & Anatomy, University of Utah, 2Department of Ophthalmology & Visual Sciences, University of Utah

JoVE 52731

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.

 JoVE Medicine

Techniques for Processing Eyes Implanted With a Retinal Prosthesis for Localized Histopathological Analysis

1Bionics Institute, 2Department of Anatomical Pathology, St Vincent's Hospital Melbourne, 3Department of Pathology, University of Melbourne, 4Medical Bionics Department, University of Melbourne

JoVE 50411

Techniques for visualizing retinal cytoarchitecture directly adjacent to individual electrodes within a retinal stimulator.

 JoVE Neuroscience

A Method of Nodose Ganglia Injection in Sprague-Dawley Rat

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

JoVE 52233

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.

 JoVE Biology

Preparation of Aplysia Sensory-motor Neuronal Cell Cultures

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

JoVE 1355

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.

 JoVE Neuroscience

Acute Dissociation of Lamprey Reticulospinal Axons to Enable Recording from the Release Face Membrane of Individual Functional Presynaptic Terminals

1Biological Sciences, University of Illinois at Chicago

JoVE 51925

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.

 JoVE Neuroscience

In Situ Ca2+ Imaging of the Enteric Nervous System

1Department of Physiology, Michigan State University

JoVE 52506

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.

 JoVE Neuroscience

In vivo Imaging of Optic Nerve Fiber Integrity by Contrast-Enhanced MRI in Mice

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

JoVE 51274

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).

 JoVE Neuroscience

Neural Circuit Recording from an Intact Cockroach Nervous System

1Department of Biology, University of Kentucky, 2Department of Biology, University of Salahaddin, 3Oregon Institute of Marine Biology, University of Oregon

JoVE 50584

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.

 JoVE Neuroscience

Quantifying Synapses: an Immunocytochemistry-based Assay to Quantify Synapse Number

1Department of Neurobiology, Duke University, 2Department of Cell Biology, Duke University

JoVE 2270

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.

 JoVE Neuroscience

Vibratome Sectioning Mouse Retina to Prepare Photoreceptor Cultures

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

JoVE 51954

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.

 JoVE Biology

Neuronal Cell Cultures from Aplysia for High-Resolution Imaging of Growth Cones

1Department of Biological Sciences, Purdue University

JoVE 662

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.

 JoVE Biology

Optimized Protocol for Retinal Wholemount Preparation for Imaging and Immunohistochemistry

1Ophthalmology, Weill Medical College of Cornell University

JoVE 51018

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.

 JoVE Neuroscience

Recording Light-evoked Postsynaptic Responses in Neurons in Dark-adapted, Mouse Retinal Slice Preparations Using Patch Clamp Techniques

1Anatomy and Cell Biology, Wayne State University School of Medicine, 2Ophthalmology, Wayne State University School of Medicine

JoVE 52422

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.

 JoVE Neuroscience

Analysis of Developing Tooth Germ Innervation Using Microfluidic Co-culture Devices

1Institute of Oral Biology, Unit of Orofacial Development and Regeneration, University of Zurich

JoVE 53114

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.

 JoVE Neuroscience

Production and Isolation of Axons from Sensory Neurons for Biochemical Analysis Using Porous Filters

1Department of Neurology and Neurosurgery, Montreal Neurological Institute, McGill University

JoVE 51795

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.

 JoVE Neuroscience

Monitoring Changes in the Intracellular Calcium Concentration and Synaptic Efficacy in the Mollusc Aplysia

1Fishberg Department of Neuroscience and Friedman Brain Institute, Mt. Sinai School of Medicine, 2Phase Five Communications Inc.

JoVE 3907

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.

 JoVE Neuroscience

Oral Administration of Rotenone using a Gavage and Image Analysis of Alpha-synuclein Inclusions in the Enteric Nervous System

1Institute of Anatomy, Technische Universität Dresden

JoVE 2123

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.

 JoVE Neuroscience

Patch-clamp Capacitance Measurements and Ca2+ Imaging at Single Nerve Terminals in Retinal Slices

1The Vollum Institute, Oregon Health and Science University

JoVE 3345

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.

 JoVE Neuroscience

Analysis of Neural Crest Migration and Differentiation by Cross-species Transplantation

1Department of Biochemistry and Cell Biology, Rice University

JoVE 3622

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.

 JoVE Neuroscience

Efficient Gene Delivery into Multiple CNS Territories Using In Utero Electroporation

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

JoVE 2957

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.

 JoVE Neuroscience

Isolation and Culture of Dissociated Sensory Neurons From Chick Embryos

1Department of Natural Sciences, Assumption College

JoVE 51991

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.

 JoVE Medicine

A Laser-induced Mouse Model of Chronic Ocular Hypertension to Characterize Visual Defects

1Department of Ophthalmology, Northwestern University, 2Department of Neurobiology, Northwestern University

JoVE 50440

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.

 JoVE Medicine

Using Adeno-associated Virus as a Tool to Study Retinal Barriers in Disease

1Department of Therapeutics, Institut de la Vision, Sorbonne Universtés, UPMC Univ Paris 06, UMR_S 968, 2INSERM, U968, 3CNRS, UMR_7210

JoVE 52451

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.

 JoVE Neuroscience

Dissection, Culture, and Analysis of Xenopus laevis Embryonic Retinal Tissue

1Department of Biology, College of William and Mary

JoVE 4377

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.

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