Neuroscience

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    Protocols in Current Issue
    Measurement of LRRK2 Kinase Activity by Proximity Ligation Assay
    Authors:  Matthew T. Keeney, Eric K. Hoffman, J. Timothy Greenamyre and Roberto Di Maio, date: 09/05/2021, view: 403, Q&A: 0

    Missense mutations in leucine rich-repeat kinase 2 (LRRK2) cause forms of familial Parkinson’s disease and have been linked to ‘idiopathic’ Parkinson’s disease. Assessment of LRRK2 kinase activity has been very challenging

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    Measurement of LRRK2 Kinase Activity by Proximity Ligation Assay
    Authors:  Matthew T. Keeney, Eric K. Hoffman, J. Timothy Greenamyre and Roberto Di Maio, date: 09/05/2021, view: 403, Q&A: 0
    [Abstract]

    Missense mutations in leucine rich-repeat kinase 2 (LRRK2) cause forms of familial Parkinson’s disease and have been linked to ‘idiopathic’ Parkinson’s disease. Assessment of LRRK2 kinase activity has been very challenging due to its size, complex structure, and relatively low abundance. A standard in the field to assess

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    Unraveling the Physicochemical Determinants of Protein Liquid-liquid Phase Separation by Nanoscale Infrared Vibrational Spectroscopy
    [Abstract]

    The phenomenon of reversible liquid-liquid phase separation of proteins underlies the formation of membraneless organelles, which are crucial for cellular processes such as signalling and transport. In addition, it is also of great interest to uncover the mechanisms of further irreversible maturation of the functional dense liquid phase into

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    3D-printed Recoverable Microdrive and Base Plate System for Rodent Electrophysiology
    [Abstract]

    Extracellular recordings in freely moving animals allow the monitoring of brain activity from populations of neurons at single-spike temporal resolution. While state-of-the-art electrophysiological recording devices have been developed in recent years (e.g., µLED and Neuropixels silicon probes), implantation methods for silicon probes in

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    Isolation of Nuclei from Mouse Dorsal Root Ganglia for Single-nucleus Genomics
    Authors:  Lite Yang, Ivan Tochitsky, Clifford J. Woolf and William Renthal, date: 08/05/2021, view: 661, Q&A: 0
    [Abstract]

    Primary somatosensory neurons, whose cell bodies reside in the dorsal root ganglion (DRG) and trigeminal ganglion, are specialized to transmit sensory information from the periphery to the central nervous system. Our molecular understanding of peripheral sensory neurons has been limited by both their heterogeneity and low abundance compared with

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    Optical Clearing and 3D Analysis Optimized for Mouse and Human Pancreata
    [Abstract]

    The pancreas is a heavily innervated organ, but pancreatic innervation can be challenging to comprehensively assess using conventional histological methods. However, recent advances in whole-mount tissue clearing and 3D rendering techniques have allowed detailed reconstructions of pancreatic innervation. Optical clearing is used to enhance tissue

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    CRISPR-mediated Labeling of Cells in Chick Embryos Based on Selectively Expressed Genes
    Authors:  Masahito Yamagata and Joshua R. Sanes, date: 08/05/2021, view: 1190, Q&A: 0
    [Abstract]

    The abilities to mark and manipulate specific cell types are essential for an increasing number of functional, structural, molecular, and developmental analyses in model organisms. In a few species, this can be accomplished by germline transgenesis; in other species, other methods are needed to selectively label somatic cells based on the genes

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    A Simple Spatial-independent Associative and Reversal Learning Task in Mice
    Authors:  Fulvio Magara, Benjamin Boury-Jamot and Hanna Hörnberg, date: 08/05/2021, view: 529, Q&A: 0
    [Abstract]

    The ability to adapt one's behavior in response to changing circumstances, or cognitive flexibility, is often altered in neuropsychiatric and neurodevelopmental conditions. In rodents, cognitive flexibility is frequently assessed using associative learning paradigms with a reversal component. The majority of existing protocols rely on

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    Cell-attached and Whole-cell Patch-clamp Recordings of Dopamine Neurons in the Substantia Nigra Pars Compacta of Mouse Brain Slices
    Authors:  Stefano Cattaneo, Maria Regoni, Jenny Sassone and Stefano Taverna, date: 08/05/2021, view: 745, Q&A: 0
    [Abstract]

    The Substantia Nigra pars compacta (SNc) is a midbrain dopaminergic nucleus that plays a key role in modulating motor and cognitive functions. It is crucially involved in several disorders, particularly Parkinson’s disease, which is characterized by a progressive loss of SNc dopaminergic cells. Electrophysiological studies on SNc neurons are

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    Method for Rapid Enzymatic Cleaning for Reuse of Patch Clamp Pipettes: Increasing Throughput by Eliminating Manual Pipette Replacement between Patch Clamp Attempts
    [Abstract]

    The whole-cell patch-clamp method is a gold standard for single-cell analysis of electrical activity, cellular morphology, and gene expression. Prior to our discovery that patch-clamp pipettes could be cleaned and reused, experimental throughput and automation were limited by the need to replace pipettes manually after each experiment. This

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    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: 1438, 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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