Neuroscience

Categories

    Protocols in Current Issue
    Translating Ribosome Affinity Purification (TRAP) of Cell Type-specific mRNA from Mouse Brain Lysates
    Authors:  Catherine L. Salussolia, Kellen D. Winden and Mustafa Sahin, date: 05/05/2022, view: 390, Q&A: 0
    [Abstract]

    Mammalian tissues are highly heterogenous and complex, posing a challenge in understanding the molecular mechanisms regulating protein expression within various tissues. Recent studies have shown that translation at the level of the ribosome is highly regulated, and can vary independently of gene expression observed at a transcriptome level, as

    ...
    Reconstitution of Membrane-associated Components of a G-protein Signaling Pathway on Membrane-coated Nanoparticles (Lipobeads)
    Authors:  Michael J. Irwin, Xin Wang and Rick H. Cote, date: 01/20/2022, view: 752, Q&A: 0
    [Abstract]

    G-protein coupled signaling pathways are organized into multi-protein complexes called signalosomes that are located within and on cellular membranes. We describe the use of silica nanoparticles coated with a unilamellar phospholipid bilayer (lipobeads) to reconstitute the activated photoreceptor G-protein α-subunit (Gtα*) with its

    ...
    Label-free Quantification of Direct Protein-protein Interactions with Backscattering Interferometry
    Authors:  Seher Abbas and Karl-Wilhelm Koch, date: 12/20/2021, view: 1276, Q&A: 0
    [Abstract]

    The functional performance of a cell depends on how macromolecules, in particular proteins, come together in a precise orientation, how they assemble into protein complexes and interact with each other. In order to study protein-protein interactions at a molecular level, a variety of methods to investigate these binding processes yield affinity

    ...
    Analysis of Protein Stability by Synthesis Shutoff
    Authors:  Ira Buntenbroich, Tânia Simões and Mafalda Escobar-Henriques, date: 11/20/2021, view: 1191, Q&A: 0
    [Abstract]

    In this protocol, we describe the analysis of protein stability over time, using synthesis shutoff. As an example, we express HA-tagged yeast mitofusin Fzo1 in Saccharomyces cerevisiae and inhibit translation via cycloheximide (CHX). Proteasomal inhibition with MG132 is performed, as an optional step, before the addition of CHX. Proteins are

    ...
    Knockoff: Druggable Cleavage of Membrane Proteins
    Authors:  John H. Rinald, Jason D. Vevea and Edwin R. Chapman, date: 11/20/2021, view: 1143, Q&A: 0
    [Abstract]

    Comparative cell biology relies on methods that disrupt protein function. Traditional approaches target the gene that encodes the protein of interest via conventional knockout (KO) methodology, conditional Cre-lox system, or recently, flexible protocols based on CRISPR/Cas9. However, these technologies lack precise temporal control (hours),

    ...
    Isolation and Phospholipid Enrichment of Muscle Mitochondria and Mitoplasts
    [Abstract]

    The efficient ATP production in mitochondria relies on the highly specific organization of its double membrane. Notably, the inner mitochondrial membrane (IMM) displays a massive surface extension through its folding into cristae, along which concentrate respiratory complexes and oligomers of the ATP synthase. Evidence has accumulated to highlight

    ...
    Simultaneous Monitoring Cytoplasmic Calcium Ion and Cell Surface Phosphatidylserine in the Necrotic Touch Neurons of Caenorhabditis elegans
    Authors:  Yoshitaka Furuta and Zheng Zhou, date: 10/20/2021, view: 823, Q&A: 0
    [Abstract]

    Calcium ions trigger many cellular events, including the release of neurotransmitters at the synaptic terminal and excitotoxic cell death. Recently, we have discovered that a transient increase in the level of cytoplasmic Ca2+ triggers the exposure of phosphatidylserine (PS) on the surfaces of necrotic cells in the nematode Caenorhabditis elegans.

    ...
    A Co-purification Method for Efficient Production and Src Kinase-mediated Phosphorylation of Aplysia Cortactin
    Authors:  Sherlene L. Brown, Yuan Ren, Daniel M. Suter and Seema Mattoo, date: 09/20/2021, view: 1230, Q&A: 0
    [Abstract]

    Cortactin is an actin-binding protein that regulates processes like cell migration, endocytosis, and tumor cell metastasis. Although cortactin is associated with actin-cytoskeletal dynamics in non-neuronal cells and cell-free systems, the exact mechanisms underlying its fundamental roles in neuronal growth cones are not fully explored. Recent

    ...
    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: 1521, 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

    ...
    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: 2838, 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

    ...



    We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.