Neuroscience

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    Protocols in Current Issue
    A Step-by-step Protocol for Obtaining Mature Microglia from Mice
    Authors:  Min-Jung Yoo and Min-Soo Kwon, date: 08/05/2022, view: 794, Q&A: 0
    [Abstract]

    In mice, microglial precursors in the yolk sac migrate to the brain parenchyma through the head neuroepithelial layer between embryonic days 8.5 (E8.5)–E16.5 and acquire their unique identity with a ramified form. Based on the microglial developmental process, we dissected the neuroepithelial layer (NEL) of E13.5 mice, which is composed

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    Generation of a Human Conditionally Immortalized Cell-based Multicellular Spheroidal Blood-Brain Barrier Model for Permeability Evaluation of Macromolecules
    Authors:  Ryuto Isogai, Hanae Morio, Ayaka Okamoto, Keita Kitamura and Tomomi Furihata, date: 08/05/2022, view: 573, Q&A: 0
    [Abstract]

    There is an urgent need for the development of brain drug delivery carriers based on middle-sized or macromolecules, to which in vitro blood-brain barrier (BBB) models are expected to contribute significantly through evaluation of BBB permeability. As part of efforts to develop such models, we have been working on human conditionally

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    Gene Expression Analysis in Stem Cell-derived Cortical Neuronal Cultures Using Multi-well SYBR Green Quantitative PCR Arrays
    Authors:  Vasavi Nallur Srinivasaraghavan, Faria Zafar and Birgitt Schüle, date: 07/20/2022, view: 844, Q&A: 0
    [Abstract]

    To optimize differentiation protocols for stem cell-based in vitro modeling applications, it is essential to assess the change in gene expression during the differentiation process. This allows controlling its differentiation efficiency into the target cell types. While RNA transcriptomics provides detail at a larger scale, timing and cost are

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    Identification and Monitoring of Nucleotide Repeat Expansions Using Southern Blotting in Drosophila Models of C9orf72 Motor Neuron Disease and Frontotemporal Dementia
    Authors:  Joanne L. Sharpe, Nikki S. Harper and Ryan J. H. West, date: 05/20/2022, view: 710, Q&A: 0
    [Abstract]

    Repeat expansion diseases, including fragile X syndrome, Huntington’s disease, and C9orf72-related motor neuron disease and frontotemporal dementia, are a group of disorders associated with polymorphic expansions of tandem repeat nucleotide sequences. These expansions are highly repetitive and often hundreds to thousands of repeats in

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    High Throughput Blood-brain Barrier Organoid Generation and Assessment of Receptor-Mediated Antibody Transcytosis
    Authors:  Elena Kassianidou, Claire Simonneau, Alina Gavrilov and Roberto Villaseñor, date: 04/20/2022, view: 1045, Q&A: 0
    [Abstract]

    Targeting receptor-mediated transcytosis (RMT) is a successful strategy for drug delivery of biologic agents across the blood-brain barrier (BBB). The recent development of human BBB organoid models is a major advancement to help characterize the mechanisms of RMT and thus accelerate the design of brain delivery technologies. BBB organoids exhibit

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    Operant Self-medication for Assessment of Spontaneous Pain Relief and Drug Abuse Liability in Mouse Models of Chronic Pain
    [Abstract]

    The search for safe and efficient chronic pain treatments is dampened by the lack of reliable models that faithfully reproduce current pharmacological treatments for chronic spontaneous pain in humans. Preclinical models often assess the antinociceptive efficacy of non-contingent pharmacological treatments evaluated in the short-term. Here, we

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    A Novel Standardized Peripheral Nerve Transection Method and a Novel Digital Pressure Sensor Device Construction for Peripheral Nerve Crush Injury
    [Abstract]

    Peripheral nerve injury (PNI) is common in all walks of life, and the most common PNIs are nerve crush and nerve transection. While optimal functional recovery after crush injury occurs over weeks, functional recovery after nerve transection with microsurgical repair and grafting is poor, and associated with permanent disability. The gold-standard

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    Immunoprecipation Assay to Quantify the Amount of tRNAs associated with Their Interacting Proteins in Tissue and Cell Culture
    Authors:  Sarada Das, Amila Zuko, Robin Thompson, Erik Storkebaum and Zoya Ignatova, date: 02/20/2022, view: 1630, Q&A: 0
    [Abstract]

    Transfer RNAs (tRNAs) are highly abundant species and, along their biosynthetic and functional path, they establish interactions with a plethora of proteins. The high number of nucleobase modifications in tRNAs renders conventional RNA quantification approaches unsuitable to study protein-tRNA interactions and their associated functional roles in

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    Induction of Repeated Social Defeat Stress in Rats
    Authors:  Soumyabrata Munshi, Alexandra Ritger and J. Amiel Rosenkranz, date: 02/05/2022, view: 1108, Q&A: 0
    [Abstract]

    Repeated social defeat stress (RSDS) is a model of chronic stress in rodents. There are several variants of social defeat procedures that exert robust effects in mice, but few published detailed protocols to produce a robust stress and altered immunological profile in rats. In this article, we describe the protocol for the induction of RSDS in

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    Immunoprecipitation for Protein-Protein Interactions and for RNA Enrichment in Drosophila melanogaster
    Authors:  Giulia Romano, Raffaella Klima and Fabian Feiguin, date: 12/05/2021, view: 1256, Q&A: 0
    [Abstract]

    To determine the molecular and functional interactions between RNA-binding proteins (RBPs) and their targets RNAs, is of fundamental importance to understand the dynamic organization of the nervous system in health and disease. Nevertheless, this task has remained elusive due to the lack of specific protocols and experimental systems that would

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