Protocols in Current Issue
    A Parkinson’s Disease-relevant Mitochondrial and Neuronal Morphology High-throughput Screening Assay in LUHMES Cells

    Parkinson’s disease is a devastating neurodegenerative disorder affecting 2-3% of the population over 65 years of age. There is currently no disease-modifying treatment. One of the predominant pathological features of Parkinson’s disease is mitochondrial dysfunction, and much work has aimed to identify therapeutic compounds which can

    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: 2670, 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 ...
    FM1-43 Photoconversion and Electron Microscopy Analysis at the Drosophila Neuromuscular Junction
    Authors:  Nadezhda S. Sabeva and Maria Bykhovskaia, date: 09/05/2017, view: 5648, 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 ...
    Microinjection of Virus into Lumbar Enlargement of Spinal Dorsal Horn in Mice
    Authors:  Zhi-Jun Zhang, Peng-Bo Jing and Yong-Jing Gao, date: 11/20/2016, view: 11017, Q&A: 0
    [Abstract] In order to explore the role of a specific gene/protein in the specific segment of the spinal cord, the technique of intraspinal injection is particularly used to deliver viral vectors targeting the specific gene/protein. These viral vectors can knockdown or overexpress the specific gene/protein in specific cells (glial cells or neurons). In this ...
    In utero Electroporation of the Embryonic Mouse Retina
    Authors:  Francisco Nieto-Lopez and Luisa Sanchez-Arrones, date: 10/05/2014, view: 12141, Q&A: 0
    [Abstract] This protocol is useful to manipulate gene expression in the embryonic retina and compare the result with the contralateral non electroporated retina. In addition, the electroporation of a membrane or cytoplasmic tagged GFP allows to determine the effects of gene manipulation on the outgrowth of retinal ganglion cell axons (Garcia-Frigola et al. ...
    Open-book Preparations from Chick Embryos and DiI Labeling of Commissural Axons
    Authors:  Nicole H. Wilson and Esther T. Stoeckli, date: 07/05/2014, view: 10249, Q&A: 0
    [Abstract] Successful neural circuit formation relies on the accurate navigation of axons towards their targets during development. Axons are guided by a combination of short-range and long-range, attractive and repulsive cues. The commissural axons of the developing spinal cord have provided an informative in vivo model for the identification of ...
    Neural Stem Cell Differentiation and Prion Infection
    Authors:  Yoshifumi Iwamaru, Takato Takenouchi and Hiroshi Kitani, date: 03/20/2014, view: 9120, Q&A: 0
    [Abstract] Prion diseases are transmissible, fatal, neurodegenerative diseases in human and animals. The molecular basis of neurodegeneration in prion diseases is largely unclear. Developing a cellular model capable of monitoring prion-induced cytotoxicity would be a promising approach for better understanding the prion pathogenesis. One candidate cellular ...
    Isolation and Culture of Neurospheres for the Study of Pathogenesis of Prion Disease
    Authors:  Yoshifumi Iwamaru, Takato Takenouchi and Hiroshi Kitani, date: 03/20/2014, view: 18786, Q&A: 0
    [Abstract] Neurosphere contains neural stem cells that are capable of self-renewal and multilineage differentiation including neurons, astrocytes, and oligodendrocytes (Gage, 2000). Cell culture model using differentiated neurosphere cultures are suggested to be a valuable tool for studying the pathogenesis of prion disease at the cellular level (Iwamaru et ...
    Neuron Culture from Mouse Superior Cervical Ganglion
    Authors:  Marisa Jackson and Warren Tourtellotte, date: 01/20/2014, view: 11438, Q&A: 0
    [Abstract] The rodent superior cervical ganglion (SCG) is a useful and readily accessible source of neurons for studying the mechanisms of sympathetic nervous system (SNS) development and growth in vitro. The sympathetic nervous system (SNS) of early postnatal animals undergoes a great deal of remodeling and development; thus, neurons taken from ...

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