Neuroscience

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    Protocols in Current Issue
    In vitro Measurement of Membrane Attack Complex in RPE Cells
    Authors:  Kelly Mulfaul and Sarah L. Doyle, date: 02/20/2021, view: 563, Q&A: 0

    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

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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: 622, Q&A: 0

    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

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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: 563, 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

    ...
    Tracing Nitrogen Metabolism in Mouse Tissues with Gas Chromatography-Mass Spectrometry
    Authors:  Rong Xu, Yekai Wang and Jianhai Du, date: 02/20/2021, view: 622, 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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    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: 221, 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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    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: 212, 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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    Polyamine Transport Assay Using Reconstituted Yeast Membranes
    Authors:  Sarah van Veen, Shaun Martin, Marleen Schuermans and Peter Vangheluwe, date: 01/20/2021, view: 846, Q&A: 0
    [Abstract]

    ATP13A2/PARK9 is a late endo-/lysosomal P5B transport ATPase that is associated with several neurodegenerative disorders. We recently characterized ATP13A2 as a lysosomal polyamine exporter, which sheds light on the molecular identity of the unknown mammalian polyamine transport system. Here, we describe step by step a protocol to measure

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    Characterization of Hippocampal Adult-borne Granule Cells in a Transient Cerebral Ischemia Model
    Authors:  Mihai Ceanga, Madlen Guenther, Ina Ingrisch and Albrecht Kunze, date: 01/20/2021, view: 454, Q&A: 0
    [Abstract]

    Long-term consequences of stroke significantly impair the quality of life in a growing population of stroke survivors. Hippocampal adult neurogenesis has been hypothesized to play a role in the pathophysiology of cognitive and neuropsychiatric long-term sequelae of stroke. Reliable animal models of stroke are paramount to understanding their

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    Low- and High-resolution Dynamic Analyses for Magnetic Resonance Spectroscopy Data
    Author:  Reuben Rideaux, date: 01/20/2021, view: 373, 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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    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: 574, 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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    Calcium Imaging of Neuronal Activity under Gradually Changing Odor Stimulation in Caenorhabditis elegans
    Authors:  Yuki Tanimoto and Koutarou D. Kimura, date: 01/05/2021, view: 572, Q&A: 0
    [Abstract]

    Olfactory behavior is among the most fundamental animal behaviors both in the wild and in the laboratory. To elucidate the neural mechanisms underlying olfactory behavior, it is critical to measure neural responses to odorant concentration changes resembling those that animals actually sense during olfactory behavior. However, reproducing the

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    Optimized Immunostaining of Embryonic and Early Postnatal Mouse Brain Sections
    Authors:  Kawssar Harb, Michele Bertacchi and Michèle Studer, date: 01/05/2021, view: 889, Q&A: 0
    [Abstract]

    The mammalian neocortex, the outer layer of the cerebrum and most recently evolved brain region, is characterized by its unique areal and laminar organization. Distinct cortical layers and areas can be identified by the protein expression of graded transcription factors and molecular determinants that define the identity of different projection

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