Neuroscience

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    Protocols in Current Issue
    Axon-seq for in Depth Analysis of the RNA Content of Neuronal Processes
    Authors:  Jik Nijssen, Julio Aguila and Eva Hedlund, date: 07/20/2019, view: 590, Q&A: 0
    [Abstract] Neuronal processes have an RNA composition that is distinct from the cell body. Therefore, to fully understand neuronal biology in health and disease we need to study both somas, dendrites and axons. Here we describe a detailed protocol of a newly refined method, Axon-seq, for RNA sequencing of axons (and dendrites) grown in isolation using single ...
    Live-cell Migration Assays to Study Motility of Neural and Glial (Oligodendrocyte) Progenitor Cells
    Authors:  Chu-Yen Chen, Fu-Sheng Chou and Pei-Shan Wang, date: 06/20/2019, view: 540, Q&A: 0
    [Abstract] Cell motility has been extensively studied in in vitro models using fibroblasts and keratocytes, but the cell type-specific mechanisms underlying migration of lineage- or disease-specific cells, such as neural and glial progenitor cells, remain an active field for investigation. The migrating neural and glial progenitor cells contribute ...
    Protocol to Quantitatively Assess the Structural Integrity of Perineuronal Nets ex vivo
    Authors:  Bhanu P. Tewari and Harald Sontheimer, date: 05/20/2019, view: 713, Q&A: 0
    [Abstract] Perineuronal nets (PNNs) are extracellular matrix assemblies of highly negatively charged proteoglycans that wrap around fast-spiking parvalbumin (PV) expressing interneurons in the cerebral cortex. PNNs play important roles in neuronal plasticity and modulate biophysical properties of the enclosed interneurons. Various central nervous system ...
    Preparation and Manipulation of Olfactory Epithelium Explant Cultures for Measurement of the Mechanical Tension of Individual Axons Using the Biomembrane Force Probe
    Authors:  Coralie Fouquet and Alain Trembleau, date: 04/20/2019, view: 564, Q&A: 0
    [Abstract] In this paper, we describe a protocol allowing measurement of the mechanical tension of individual axons grown ex vivo from neural tissue explants. This protocol was developed with primary cultures of olfactory epithelium explants from embryonic (E13.5) mice. It includes a detailed description of explant dissection and culture, as well as ...
    Time-lapse Whole-field Fluorescence Imaging of Microglia Processes Motility in Acute Mouse Hippocampal Slices and Analysis
    [Abstract] Microglia are the resident immune cells of the central nervous system (CNS). In the last year, the improvements in the transgenic mouse technologies and imaging techniques have shed light on microglia functions under physiological conditions. Microglia continuously scan the brain parenchyma with their highly motile processes, maintaining tissue ...
    Primary Embryonic Rat Cortical Neuronal Culture and Chronic Rotenone Treatment in Microfluidic Culture Devices
    Authors:  Victor S. Van Laar, Beth Arnold and Sarah B Berman, date: 03/20/2019, view: 1029, Q&A: 0
    [Abstract] In the study of neurodegenerative diseases, it is imperative to study the cellular and molecular changes associated with pathogenesis in the relevant cell type, central nervous system neurons. The unique compartmentalized morphology and bioenergetic needs of primary neurons present complications for their study in culture. Recent microculture ...
    Differentiation of Human Induced Pluripotent Stem Cells (iPSCs) into an Effective Model of Forebrain Neural Progenitor Cells and Mature Neurons
    [Abstract] Induced Pluripotent Stem Cells (iPSCs) are pluripotent stem cells that can be generated from somatic cells, and provide a way to model the development of neural tissues in vitro. One particularly interesting application of iPSCs is the development of neurons analogous to those found in the human forebrain. Forebrain neurons play a central ...
    Isolation of Neural Stem Cells from the Embryonic Mouse Hippocampus for in vitro Growth or Engraftment into a Host Tissue
    Authors:  Oksana Rybachuk, Olga Kopach, Tetyana Pivneva and Vitaliy Kyryk, date: 02/20/2019, view: 1254, Q&A: 0
    [Abstract] For both stem cell research and treatment of the central nervous system disorders, neural stem/progenitor cells (NSPCs) represent an important breakthrough tool. In the expanded stem cell-based therapy use, NSPCs not only provide a powerful cell source for neural cell replacement but a useful model for developmental biology research. Despite ...
    Heterochronic Phenotype Analysis of Hypodermal Seam Cells in Caenorhabditis elegans
    Authors:  Yon Ju Ji and Jiou Wang, date: 01/05/2019, view: 1369, Q&A: 0
    [Abstract] Heterochrony refers to changes in the timing of developmental events, and it is precisely regulated in the organisms by the heterochronic genes such as C. elegans lin-4 and let-7. Mutations in these genes cause precocious or retarded development of certain cell lineages. With well-defined cell lineages, C. elegans ...
    Estimation of the Readily Releasable Synaptic Vesicle Pool at the Drosophila Larval Neuromuscular Junction
    Authors:  Pragya Goel, Xiling Li and Dion Dickman, date: 01/05/2019, view: 1664, Q&A: 0
    [Abstract] Presynaptic boutons at nerve terminals are densely packed with synaptic vesicles, specialized organelles for rapid and regulated neurotransmitter secretion. Upon depolarization of the nerve terminal, synaptic vesicles fuse at specializations called active zones that are localized at discrete compartments in the plasma membrane to initiate synaptic ...



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