Neuroscience

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    Protocols in Current Issue
    Generation of Human iPSC-derived Neural Progenitor Cells (NPCs) as Drug Discovery Model for Neurological and Mitochondrial Disorders
    Authors:  Annika Zink, Pawel Lisowski and Alessandro Prigione, date: 03/05/2021, view: 99, Q&A: 0

    The high attrition rate in drug development processes calls for additional human-based model systems. However, in the context of brain disorders, sampling live neuronal cells for compound testing is not applicable. The use of human induced

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    Generation of Mouse Primary Hypothalamic Neuronal Cultures for Circadian Bioluminescence Assays
    Authors:  Cosima X. Schmidt, Anthony H. Tsang and Henrik Oster, date: 03/05/2021, view: 27, Q&A: 0

    An endogenous circadian clock system enables organisms to adapt to time-of-day dependent environmental changes. In consequence, most physiological processes exhibit daily rhythms of, e.g., energy metabolism, immune function, sleep, or hormone

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    Dissociating Behavior and Spatial Working Memory Demands Using an H Maze
    Authors:  Hannah S. Wirtshafter, Moqing Quan and Matthew A. Wilson, date: 03/05/2021, view: 41, Q&A: 0

    The development of mazes for animal experiments has allowed for the investigation of cognitive maps and place cells, spatial working memory, naturalistic navigation, perseverance, exploration, and choice and motivated behavior. However, many mazes,

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    Transcardiac Perfusion of the Mouse for Brain Tissue Dissection and Fixation
    Authors:  Jinyun Wu, Yuqi Cai, Xiaoyang Wu, Yue Ying, Yilin Tai and Miao He, date: 03/05/2021, view: 17, Q&A: 0
    Transcardiac perfusion with saline followed by 4% paraformaldehyde (PFA) is widely used to clear blood and preserve brain for immunostaining or in situ hybridization. PFA breaks into formaldehyde in solution, which cross-link protein and ...
    Transcardiac Perfusion of the Mouse for Brain Tissue Dissection and Fixation
    Authors:  Jinyun Wu, Yuqi Cai, Xiaoyang Wu, Yue Ying, Yilin Tai and Miao He, date: 03/05/2021, view: 17, Q&A: 0
    [Abstract] Transcardiac perfusion with saline followed by 4% paraformaldehyde (PFA) is widely used to clear blood and preserve brain for immunostaining or in situ hybridization. PFA breaks into formaldehyde in solution, which cross-link protein and DNA molecules to preserve tissue and cell structure. Here we provide a step by step guide for ...
    Dissociating Behavior and Spatial Working Memory Demands Using an H Maze
    Authors:  Hannah S. Wirtshafter, Moqing Quan and Matthew A. Wilson, date: 03/05/2021, view: 41, Q&A: 0
    [Abstract]

    The development of mazes for animal experiments has allowed for the investigation of cognitive maps and place cells, spatial working memory, naturalistic navigation, perseverance, exploration, and choice and motivated behavior. However, many mazes, such as the T maze, currently developed to test learning and memory, do not distinguish temporally

    ...
    Generation of Mouse Primary Hypothalamic Neuronal Cultures for Circadian Bioluminescence Assays
    Authors:  Cosima X. Schmidt, Anthony H. Tsang and Henrik Oster, date: 03/05/2021, view: 27, Q&A: 0
    [Abstract]

    An endogenous circadian clock system enables organisms to adapt to time-of-day dependent environmental changes. In consequence, most physiological processes exhibit daily rhythms of, e.g., energy metabolism, immune function, sleep, or hormone production. Hypothalamic circadian clocks have been identified to play a particular role in coordinating

    ...
    Generation of Human iPSC-derived Neural Progenitor Cells (NPCs) as Drug Discovery Model for Neurological and Mitochondrial Disorders
    Authors:  Annika Zink, Pawel Lisowski and Alessandro Prigione, date: 03/05/2021, view: 99, Q&A: 0
    [Abstract]

    The high attrition rate in drug development processes calls for additional human-based model systems. However, in the context of brain disorders, sampling live neuronal cells for compound testing is not applicable. The use of human induced pluripotent stem cells (iPSCs) has revolutionized the field of neuronal disease modeling and drug discovery.

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    Tracing Nitrogen Metabolism in Mouse Tissues with Gas Chromatography-Mass Spectrometry
    Authors:  Rong Xu, Yekai Wang and Jianhai Du, date: 02/20/2021, view: 643, Q&A: 0
    [Abstract]

    Nitrogen-containing metabolites including ammonia, amino acids, and nucleotides, are essential for cell metabolism, growth, and neural transmission. Nitrogen metabolism is tightly coordinated with carbon metabolism in the breakdown and biosynthesis of amino acids and nucleotides. Both nuclear magnetic resonance spectroscopy and mass spectrometry

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    In vitro Measurement of Membrane Attack Complex in RPE Cells
    Authors:  Kelly Mulfaul and Sarah L. Doyle, date: 02/20/2021, view: 575, Q&A: 0
    [Abstract]

    Initiation of the complement system results in the formation of a multiprotein pore termed the membrane attack complex (MAC, C5b-C9). MAC pores accumulate on a cell surface and can result in cell lysis. The retinal pigment epithelium (RPE) is a single monolayer of pigmented epithelial cells located at the posterior poll of the eye that forms the

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    Immuno-electrophysiology on Neuromuscular Junctions of Drosophila Third Instar Larva
    Authors:  Raffaella Klima, Giulia Romano, Monsurat Gbadamosi, Aram Megighian and Fabian Feiguin, date: 02/05/2021, view: 223, Q&A: 0
    [Abstract]

    Alterations in synaptic transmission are critical early events in neuromuscular disorders. However, reliable methodologies to analyze the functional organization of the neuromuscular synapses are still needed. This manuscript provides a detailed protocol to analyze the molecular assembly of the neuromuscular synapses through

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    Relative Quantification of NaV1.1 Protein in Mouse Brains Using a Meso Scale Discovery-Electrochemiluminescence (MSD-ECL) Method
    Authors:  Zhou Han, Anne Christiansen, Meena and Gene Liau, date: 02/05/2021, view: 230, Q&A: 0
    [Abstract]

    Densitometric analysis is often used to quantify NaV1.1 protein on immunoblots, although the sensitivity and dilution linearity of the method are usually poor. Here we present a protocol for quantification of NaV1.1 in mouse brain tissues using a Meso Scale Discovery-Electrochemiluminescence (MSD-ECL) method. MSD-ECL is based on ELISA

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    Preparation of Synaptoneurosomes to Study the Synapse in the Murine Cerebral Cortex
    Authors:  Ariel Diaz, Enrique Torre and Manuel Yepes, date: 01/20/2021, view: 587, Q&A: 1
    [Abstract]

    The synapse is a complex structure where the transmission of information takes place. Synaptic dysfunction is one of the earliest pathophysiological events in several diseases, such as traumatic brain injury, cerebral ischemia, and neurodegenerative diseases. Thus, a methodology to study synaptic structure and function is crucial for the

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    Low- and High-resolution Dynamic Analyses for Magnetic Resonance Spectroscopy Data
    Author:  Reuben Rideaux, date: 01/20/2021, view: 384, Q&A: 0
    [Abstract]

    Magnetic resonance spectroscopy (MRS) can be used to measure in vivo concentrations of neurometabolites. This information can be used to identify neurotransmitter involvement in healthy (e.g., perceptual and cognitive processes) and unhealthy brain function (e.g., neurological and psychiatric illnesses). The standard approach for analyzing MRS

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