Protocols in Current Issue
    Establishing an Adult Mouse Brain Hippocampal Organotypic Slice Culture System that Allows for Tracing and Pharmacological Manipulation of ex vivo Neurogenesis
    Authors:  Steffen Mayerl and Charles ffrench-Constant, date: 01/05/2021, view: 399, Q&A: 0

    The function of the hippocampus depends on the process of adult hippocampal neurogenesis which underpins the exceptional neural plasticity of this structure, and is also frequently affected in CNS pathologies. Thus, manipulation of this process represents an important therapeutic goal. To identify potential strategies, organotypic adult brain

    A Quantitative Assay to Measure Stress Granule Association of Proteins and Peptides in Semi-permeabilized Human Cells
    Authors:  Saskia Hutten and Dorothee Dormann, date: 12/20/2020, view: 828, Q&A: 0

    Stress granules (SGs) are membrane-less organelles that form in the cytoplasm through phase separation, in response to diverse stressors. SGs contain translationally stalled mRNAs, proteins involved in translation, and various RNA-binding proteins (RBPs). Due to the high local concentration of aggregation-prone RBPs, SGs might act as condensation

    In vitro Time-lapse Imaging of Primary Cilium in Migrating Neuroblasts
    [Abstract] Neuronal migration is a critical step for the development of neuronal circuits in the brain. Immature new neurons (neuroblasts) generated in the postnatal ventricular-subventricular zone (V-SVZ) show a remarkable potential to migrate for a long distance at a high speed in the postnatal mammalian brain, and are thus a powerful model to analyze the ...
    In vitro AMPylation/Adenylylation of Alpha-synuclein by HYPE/FICD
    Authors:  Ali Camara, Anwesha Sanyal, Sayan Dutta, Jean-Christophe Rochet and Seema Mattoo, date: 09/20/2020, view: 1287, Q&A: 0
    [Abstract] One of the major histopathological hallmarks of Parkinson’s disease are Lewy bodies (LBs) –cytoplasmic inclusions, enriched with fibrillar forms of the presynaptic protein alpha-synuclein (α-syn). Progressive deposition of α-syn into LBs is enabled by its propensity to fibrillize into insoluble aggregates. We recently described a marked reduction ...
    Assessing Gαq/15-signaling with IP-One: Single Plate Transfection and Assay Protocol for Cell-Based High-Throughput Assay
    [Abstract] Cell-based functional assays are an important part of compound screening and drug lead optimization, and they can also play a crucial role in the determination of the residues involved in ligand binding and signaling for a particular G-protein-coupled receptor. Conventional methods used for Gαq/15-coupled receptors rely on the use of ...
    Co-culture of Murine Neurons Using a Microfluidic Device for The Study of Tau Misfolding Propagation
    [Abstract] The deposition of misfolded, aggregated tau protein is a hallmark of several neurodegenerative diseases, collectively termed “tauopathies”. Tau pathology spreads throughout the brain along connected pathways in a prion-like manner. The process of tau pathology propagation across circuits is a focus of intense research and has been investigated in ...
    Generation of Functional Mouse Hippocampal Neurons
    Authors:  Francesco Tomassoni-Ardori, Zhenyi Hong, Gianluca Fulgenzi and Lino Tessarollo, date: 08/05/2020, view: 1661, Q&A: 0
    [Abstract] Primary culture of mouse hippocampal neurons is a very useful in vitro model for studying neuronal development, axonal and dendritic morphology, synaptic functions, and many other neuronal features. Here we describe a step-by-step process of generating primary neurons from mouse embryonic hippocampi (E17.5/E18.5). Hippocampal neurons ...
    An ex vivo Approach to Assess Mitochondrial ROS by Flow Cytometry in AAV-tagged Astrocytes in Adult Mice
    Authors:  Carlos Vicente-Gutierrez and Juan P. Bolaños, date: 03/20/2020, view: 1740, Q&A: 0
    [Abstract] Mitochondrial reactive oxygen species (mROS) are naturally produced signalling molecules extremely relevant for understanding both health- and disease-associated biological processes. The study of mROS in the brain is currently underway to decipher their physiopathological roles and contributions in neurological diseases. Recent advances in this ...
    Shared Pheromonal Communication of Specific Fear Between Adult Sprague Dawley Rats
    Authors:  Samantha J. Carew and Abhinaba Ghosh, date: 03/20/2020, view: 1066, Q&A: 0
    [Abstract] Rats are highly social animals, and mainly communicate with one another in two ways: through ultrasonic vocalizations and pheromones. Most research on pheromones has been dedicated those regarding sexual behavior, but more recently pheromones which signal danger to conspecifics have been identified in rodents. In fact, rats are capable of ...
    Induction of Temporal Lobe Epilepsy in Mice with Pilocarpine
    Authors:  Muhammad Nauman Arshad and Janice R Naegele, date: 02/20/2020, view: 2068, Q&A: 0
    [Abstract] In the pilocarpine model of temporal lobe epilepsy (TLE) in rodents, systemic injections of pilocarpine induce continuous, prolonged limbic seizures, a condition termed “Status Epilepticus” (SE). With appropriate doses, many inbred strains of mice show behavioral seizures within an hour after pilocarpine is injected. With the behavioral scoring ...

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