JoVE Neuroscience is a multidisciplinary section devoted to investigations of the structure, function, physiology, and pathophysiology of the brain and nervous system. Included methodologies range from molecular and cellular level studies to full central and peripheral neural systems. Potential treatment platforms and surgical techniques for neurological diseases and disorders are also presented in this section.
1NIH Stem Cell Unit, National Institute of Neurological Disorders and Stroke, National Institutes of Health, 2Craniofacial and Skeletal Diseases Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health
Here, we present human pluripotent stem cell (hPSC) culture protocols, based on non-colony type monolayer (NCM) growth of dissociated single cells. This new method, utilizing Rho-associated kinase inhibitors or the laminin isoform 521 (LN-521), is suitable for producing large amounts of homogeneous hPSCs, genetic manipulation, and drug discovery.
Published July 24, 2014. Keywords: Stem Cell Biology, Pluripotent stem cells, human embryonic stem cells, induced pluripotent stem cells, cell culture, non-colony type monolayer, single cell, plating efficiency, Rho-associated kinase, Y-27632, transfection, transduction
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 Behavioral Physiology and Sociobiology (Zoology II) Biozentrum, University of Würzburg
Simultaneous extracellular long term recordings from two different brain neuropiles or two different anatomical tracts were established in honeybees. These recordings allow the investigation of temporal aspects of neuronal processing across different brain areas at the single neuron as well as at the ensemble level in a behaving animal.
Published July 21, 2014. Keywords: Neuroscience, honeybee brain, olfaction, extracellular long term recordings, double recordings, differential wire electrodes, single unit, multi-unit recordings
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
1National Heart, Lung, and Blood Institute, National Institutes of Health
This protocol details a streamlined method used to conduct live cell imaging in the context of an intact larval brain. Live cell imaging approaches are invaluable for the study of asymmetric neural stem cell divisions as well as other neurogenic and developmental processes, consistently uncovering mechanisms that were previously overlooked.
Published July 7, 2014. Keywords: Neuroscience, live imaging, Drosophila, neuroblast, stem cell, asymmetric division, centrosome, brain, cell cycle, mitosis
1Neuroscience, Genentech, Inc., 2Department of Discovery Oncology, Genentech, Inc., 3Department of Pathology, Genentech, Inc.
To obtain high-resolution images of fluorescently labeled cells within large tissues, ideally, the biological samples should be imaged without sectioning. 3DISCO is a straightforward tissue clearing procedure based on sequential incubation with organic solvents. Upon clearing, the organs become transparent allowing an end-to-end laser scan of the specimen.
Published July 7, 2014. Keywords: Neuroscience, 3D imaging, tissue clearing, transparent tissue, intact organs, optical clearing, histology, laser scanning, light-sheet microscopy, fluorescent imaging, 3DISCO, ultramicroscope
1Department of Physics, Lewis & Clark College, 2Program in Biochemistry and Molecular Biology, Lewis & Clark College, 3Jungers Center for Neuroscience Research, Oregon Health & Science University, 4Department of Chemistry, Lewis & Clark College
We describe how to implement photoactivated localization microscopy (PALM)-based studies of vesicles in fixed, cultured neurons. Key components of our protocol include labeling vesicles with photoconvertible chimeras, collecting sparsely sampled raw images with a super-resolution microscopy system, and processing the raw images to produce a super-resolution image.
Published July 2, 2014. Keywords: Neuroscience, photoactivated localization microscopy, Dendra2, dense-core vesicle, synapse, hippocampal neuron, cluster
1Psychology Department, University of Illinois Urbana-Champaign, 2Neuroscience Program, University of Illinois Urbana-Champaign, 3Beckman Institute for Advanced Science and Technology, University of Illinois Urbana-Champaign
The present work provides a comprehensive set of guidelines for manually tracing the medial temporal lobe (MTL) structures. This protocol can be applied to research involving structural and/or combined structural-functional magnetic resonance imaging (MRI) investigations of the MTL, in both healthy and clinical groups.
Published July 2, 2014. Keywords: Neuroscience, Anatomy, Segmentation, Medial Temporal Lobe, MRI, Manual Tracing, Amygdala, Hippocampus, Perirhinal Cortex, Entorhinal Cortex, Parahippocampal Cortex
1Department of Psychology, University of Pennsylvania
Informational connectivity measures the correspondence between time courses of multi-voxel information across different brain regions. Multi-voxel pattern discriminability time series are extracted from regions and compared, revealing networks that are not identified in a typical functional connectivity approach.
Published July 1, 2014. Keywords: Neuroscience, fMRI, MVPA, connectivity, informational connectivity, functional connectivity, networks, multi-voxel pattern analysis, decoding, classification, method, multivariate
1Department of Molecular Biology and Genetics, Howard Hughes Medical Institute, Johns Hopkins University School of Medicine, 2Department of Neuroscience, Howard Hughes Medical Institute, Johns Hopkins University School of Medicine, 3Department of Ophthalmology, Howard Hughes Medical Institute, Johns Hopkins University School of Medicine
Mammalian skin contains a diverse array of structures - such as hair follicles and nerve endings - that exhibit distinctive patterns of spatial organization. Analyzing skin as a flat mount takes advantage of the 2-dimensional geometry of this tissue to produce full-thickness high-resolution images of skin structures.
Published June 25, 2014. Keywords: Physiology, arrector pili, sebaceous gland, Merkel cell, cutaneous nerve, planar cell polarity, Frizzled