Neuroscience

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    Protocols in Current Issue
    Isolation of Microglia and Analysis of Protein Expression by Flow Cytometry: Avoiding the Pitfall of Microglia Background Autofluorescence
    Authors:  Jeremy C. Burns, Richard M. Ransohoff and Michaël Mingueneau, date: 07/20/2021, view: 56, Q&A: 0
    [Abstract]

    Microglia are a unique type of tissue-resident innate immune cell found within the brain, spinal cord, and retina. In the healthy nervous system, their main functions are to defend the tissue against infectious microbes, support neuronal networks through synapse remodeling, and clear extracellular debris and dying cells through phagocytosis. Many

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    A Method for Estimating the Potential Synaptic Connections Between Axons and Dendrites From 2D Neuronal Images
    Authors:  Carolina Tecuatl, Diek W. Wheeler and Giorgio A. Ascoli, date: 07/05/2021, view: 1254, Q&A: 0
    [Abstract]

    Computational neuroscience aims to model, reproduce, and predict network dynamics for different neuronal ensembles by distilling knowledge derived from electrophysiological and morphological evidence. However, analyses and simulations often remain critically limited by the sparsity of direct experimental constraints on essential parameters, such

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    Immunofluorescence of GFAP and TNF-α in the Mouse Hypothalamus
    Authors:  Prasad S. Dalvi and Denise D. Belsham, date: 07/05/2021, view: 599, Q&A: 0
    [Abstract]

    Immunofluorescence is a reliable method for identifying specific proteins in neuronal and glial cell populations of the hypothalamus. Several immunofluorescence protocols are available to detect protein markers and neuropeptides in the hypothalamus; however, published methods may vary in subtle details that can potentially impact the final outcome

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    Differential Analysis of N-glycopeptide Abundance and N-glycosylation Site Occupancy for Studying Protein N-glycosylation Dysregulation in Human Disease
    Authors:  Qi Zhang, Cheng Ma, Lian Li and Lih-Shen Chin, date: 06/20/2021, view: 845, Q&A: 0
    [Abstract]

    Protein N-glycosylation plays a vital role in diverse cellular processes, and dysregulated N-glycosylation is implicated in a variety of human diseases including neurodegenerative disorders and cancer. With recent advances in high-resolution mass spectrometry-based glycoproteomics technologies enabling large-scale N-glycoproteome profiling of

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    A Lipidomics Approach to Measure Phosphatidic Acid Species in Subcellular Membrane Fractions Obtained from Cultured Cells
    Authors:  Nawal Kassas, Laetitia Fouillen, Stéphane Gasman and Nicolas Vitale, date: 06/20/2021, view: 934, Q&A: 0
    [Abstract]

    Over the last decade, lipids have emerged as possessing an ever-increasing number of key functions, especially in membrane trafficking. For instance, phosphatidic acid (PA) has been proposed to play a critical role in different steps along the secretory pathway or during phagocytosis. To further investigate in detail the precise nature of PA

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    Dual Color, Live Imaging of Vesicular Transport in Axons of Cultured Sensory Neurons
    Authors:  Yoko Bekku and James L. Salzer, date: 06/20/2021, view: 844, Q&A: 0
    [Abstract]

    The function of neurons in afferent reception, integration, and generation of electrical activity relies on their strikingly polarized organization, characterized by distinct membrane domains. These domains have different compositions resulting from a combination of selective targeting and retention of membrane proteins. In neurons, most proteins

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    In vivo Fluorescence Imaging of Extracellular ATP in the Mouse Cerebral Cortex with a Hybrid-type Optical Sensor
    [Abstract]

    Adenosine 5’-triphosphate (ATP) works as an extracellular signaling molecule for cells in the brain, such as neurons and glia. Cellular communication via release of ATP is involved in a range of processes required for normal brain functions, and aberrant communication is associated with brain disorders. To investigate the mechanisms

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    A New Method for Studying RNA-binding Proteins on Specific RNAs
    Authors:  Weiping Sun, Ziheng Zhang, Ji-Long Liu and Min Zhuang, date: 05/20/2021, view: 4076, Q&A: 0
    [Abstract]

    Proximity-based protein labeling has been developed to identify protein-nucleic acid interactions. We have reported a novel method termed CRUIS (CRISPR-based RNA-United Interacting System), which captures RNA-protein interactions in living cells by combining the RNA-binding capacity of CRISPR/Cas13 and the proximity-tagging activity of PUP-IT.

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    Imaging Microtubules in vitro at High Resolution while Preserving their Structure
    Authors:  Camille Cuveillier, Yasmina Saoudi, Isabelle Arnal and Christian Delphin, date: 04/05/2021, view: 718, Q&A: 0
    [Abstract]

    Microtubules (MT) are the most rigid component of the cytoskeleton. Nevertheless, they often appear highly curved in the cellular context and the mechanisms governing their overall shape are poorly understood. Currently, in vitro microtubule analysis relies primarily on electron microscopy for its high resolution and Total Internal Reflection

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    Investigate Synaptic Vesicles Mobility in Neuronal Culture Axons by FRAP Imaging
    Authors:  Xiao Min Zhang, Fabrice P. Cordelieres and Etienne Herzog, date: 03/20/2021, view: 1992, Q&A: 0
    [Abstract]

    Synaptic vesicles (SVs) are clustered in the presynaptic terminals and consistently trafficking along axons. Based on their release features, SVs are classified into different “pools”. Imaging of SVs that are traveling among multiple presynaptic terminals has helped define a new pool named “SV super-pool”. Here we describe

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