Neuroscience

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    Protocols in Current Issue
    Organotypic Slice Culture of the Embryonic Mouse Brain
    Authors:  James M. Clegg and Thomas Pratt, date: 07/05/2020, view: 448, Q&A: 0
    [Abstract] Organotypic slice culture is a powerful technique for exploring the embryonic development of the mammalian brain. In this protocol we describe a basic slice culture technique we have used for two sets of experiments: axon guidance transplant assays and bead culture assays.
    Rapid Generation of Human Neuronal Cell Models Enabling Inducible Expression of Proteins-of-interest for Functional Studies
    [Abstract] CRISPR-Cas9 technology has transformed the ability to edit genomic sequences and control gene expression with unprecedented ease and scale. However, precise genomic insertions of coding sequences using this technology remain time-consuming and inefficient because they require introducing adjacent single-strand cuts through Cas9 nickase action and ...
    Laser Capture Micro-dissection (LCM) of Neonatal Mouse Forebrain for RNA Isolation
    Authors:  Achira Roy, Mei Deng, Kimberly A. Aldinger, Ian A. GLass and Kathleen J. Millen, date: 01/05/2020, view: 1795, Q&A: 0
    [Abstract] Precise and reproducible isolation of desired cell types or layers from heterogeneous tissues is crucial to analyze specific gene profiles and molecular interactions in vivo. Forebrain is the core site of higher functions, like cognition and memory consolidation. It is composed of heterogeneous and distinct cell types, interconnected to ...
    Gallyas Silver Impregnation of Myelinated Nerve Fibers
    Authors:  Sabitha Joseph, Hauke B. Werner and Judith Stegmüller, date: 11/20/2019, view: 1586, Q&A: 0
    [Abstract] In the nervous system of vertebrates, nerve impulse propagation is accelerated by the ensheathment of neuronal axons with myelin. Myelin sheaths are molecularly specialized, lipid-rich plasma membrane extensions of Schwann cells in the peripheral nervous system and oligodendrocytes in the central nervous system (CNS). To visualize myelinated nerve ...
    Explant Culture of the Embryonic Mouse Spinal Cord and Gene Transfer by ex vivo Electroporation
    [Abstract] Developing axons change responsiveness to guidance cues during the journey to synapse with target cells. Axon crossing at the ventral midline serves as a model for studying how axons accomplish such a switch in their response. Although primary neuron culture has been a versatile technique for elucidating various developmental mechanisms, many in ...
    A Modified Barnes Maze for Juvenile Rats
    Authors:  Nazanin Valibeigi, Daniel G. McHail, Carmen L. Kimball and Theodore C. Dumas, date: 11/20/2018, view: 2510, Q&A: 0
    [Abstract] To better understand neural codes for spatial navigation and to address cognitive development questions, the neurobiology of postnatal hippocampal development is receiving increased attention. While the Morris Water Maze is the gold standard for assessing hippocampal integrity, potential confounds can be difficult to control for in juvenile ...
    6-hydroxydopamine (6-OHDA) Oxidative Stress Assay for Observing Dopaminergic Neuron Loss in Caenorhabditis elegans
    Authors:  Sarah-Lena Offenburger and Anton Gartner, date: 09/20/2018, view: 3323, Q&A: 0
    [Abstract] The nematode Caenorhabditis elegans is a powerful genetic model that can be used to investigate neuronal death. Research using C. elegans has been crucial to characterize cell death programmes that are conserved in mammals. Many neuronal signaling components, such as those mediating dopaminergic neurotransmission, are preserved ...
    Organotypic Explants of the Embryonic Rodent Hippocampus: An Accessible System for Transgenesis
    Authors:  Archana Iyer , Lakshmi Subramanian and Shubha Tole, date: 03/20/2018, view: 3982, Q&A: 0
    [Abstract] This protocol describes the technique of ex-vivo electroporation to target embryonic hippocampal progenitors in an organotypic slice preparation. This technique allows gene perturbation for examining developmental processes in the embryonic hippocampus while retaining the environment and connectivity of the cells. Gene perturbation can ...
    Mandibular Explant Assay for Investigating Extrinsic Stimuli on Bone and Cartilage Development
    Authors:  Jamie Jiang, Jessica W. Bertol and Walid D. Fakhouri, date: 12/05/2017, view: 4518, Q&A: 0
    [Abstract] A major issue in developmental biology is to determine how embryonic tissues respond to molecular signals in a timely manner and given the position-restricted instructions during morphogenesis, of which Meckel’s cartilage is a classical example. The ex-vivo explant model is a practical and convenient system that allows investigation of ...
    FM1-43 Photoconversion and Electron Microscopy Analysis at the Drosophila Neuromuscular Junction
    Authors:  Nadezhda S. Sabeva and Maria Bykhovskaia, date: 09/05/2017, view: 5279, Q&A: 0
    [Abstract] We developed a protocol for photoconversion of endocytic marker FM1-43 followed by electron microscopy analysis of synaptic boutons at the Drosophila neuromuscular junction. This protocol allows detection of stained synaptic vesicle even when release rates are very low, such as during the spontaneous release mode. The preparations are ...



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