Neuroscience

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    Protocols in Current Issue
    Fluorescence Screens for Identifying Central Nervous System–Acting Drug–Biosensor Pairs for Subcellular and Supracellular Pharmacokinetics

    Subcellular pharmacokinetic measurements have informed the study of central nervous system (CNS)–acting drug mechanisms. Recent investigations have been enhanced by the use of genetically encoded fluorescent biosensors for drugs of interest at the

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    In vitro Preparation of Homogenous Actin Filaments for Dynamic and Electrophoretic Light Scattering Measurements
    Authors:  Ernesto Alva, Annitta George, Lorenzo Brancaleon and Marcelo Marucho, date: 11/20/2022, view: 350, Q&A: 0

    Actin filaments are essential for various biological activities in eukaryotic cellular processes. Available in vitro experimental data on these systems often lack details and information on sample preparation protocols and experimental techniques,

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    Fluorescence Screens for Identifying Central Nervous System–Acting Drug–Biosensor Pairs for Subcellular and Supracellular Pharmacokinetics
    [Abstract]

    Subcellular pharmacokinetic measurements have informed the study of central nervous system (CNS)–acting drug mechanisms. Recent investigations have been enhanced by the use of genetically encoded fluorescent biosensors for drugs of interest at the plasma membrane and in organelles. We describe screening and validation protocols for identifying hit

    ...
    In vitro Preparation of Homogenous Actin Filaments for Dynamic and Electrophoretic Light Scattering Measurements
    Authors:  Ernesto Alva, Annitta George, Lorenzo Brancaleon and Marcelo Marucho, date: 11/20/2022, view: 350, Q&A: 0
    [Abstract]

    Actin filaments are essential for various biological activities in eukaryotic cellular processes. Available in vitro experimental data on these systems often lack details and information on sample preparation protocols and experimental techniques, leading to unreproducible results. Additionally, different experimental techniques and polymerization

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    MiniSOG2-mediated Specific Photoablation of Motor Neurons in Ascidian Embryos
    Authors:  Madoka K. Utsumi, Taichi Akahoshi, Kotaro Oka and Kohji Hotta, date: 10/20/2022, view: 749, Q&A: 0
    [Abstract]

    When understanding the neuronal function of a specific neural circuit, single-cell level photoablation of a targeted cell is one of the useful experimental approaches. This protocol describes a method to photoablate specific motor neurons via the mini singlet oxygen generator (miniSOG2), a light–oxygen–voltage (LOV)-based optogenetic

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    A Simplified Paradigm of Late Gestation Transient Prenatal Hypoxia to Investigate Functional and Structural Outcomes from a Developmental Hypoxic Insult
    Authors:  Elyse C. Gadra and Ana G. Cristancho, date: 10/05/2022, view: 512, Q&A: 0
    [Abstract]

    Late-gestation transient intrauterine hypoxia is a common cause of birth injury. It can lead to long-term neurodevelopmental disabilities even in the absence of gross anatomic injury. Currently, postnatal models of hypoxia–ischemia are most commonly used to study the effect of oxygen deprivation in the fetal brain. These models, however, are

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    In situ Dephosphorylation Assay with Recombinant Nil Phosphatase
    Authors:  Nilay Nandi, Charles Tracy and Helmut Krämer, date: 09/20/2022, view: 944, Q&A: 0
    [Abstract]

    The activity of numerous autophagy-related proteins depends on their phosphorylation status, which places importance on understanding the responsible kinases and phosphatases. Great progress has been made in identifying kinases regulating autophagy, but much less is known about the phosphatases counteracting their function. Genetic screens and

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    Characterizing Hedonic Responses to Flavors Paired with Internal Pain and Nausea through the Taste Reactivity Test in Rats
    Authors:  Matías López, Dominic M. Dwyer, Patricia Gasalla, Claudia Jove and Azucena Begega, date: 09/20/2022, view: 824, Q&A: 0
    [Abstract]

    Feeding behavior is a complex experience that involves not only sensory (i.e., visual, odor, taste, or texture) but also affective or emotional aspects (i.e., pleasure, palatability, or hedonic value) of foods. As such, behavioral tests that assess the hedonic impact of foods are necessary to fully understand the factors involved in ingestive

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    Evaluation of Mitochondrial Turnover Using Fluorescence Microscopy in Drosophila
    Author:  Felipe Martelli, date: 09/05/2022, view: 745, Q&A: 0
    [Abstract]

    Mitochondrial dysfunction is associated with perturbations in the cellular oxidative status, changes in energy production and metabolic rate, and the onset of pathological processes. Classic methods of assessing mitochondrial dysfunction rely on indirect measures, such as evaluating mitochondrial DNA copy numbers, or direct but more costly and

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    Aerotaxis Assay in Caenorhabditis elegans to Study Behavioral Plasticity
    Authors:  Qiaochu Li, Daniel-Cosmin Marcu, Paul H. Dear and Karl Emanuel Busch, date: 08/20/2022, view: 966, Q&A: 0
    [Abstract]

    C. elegans shows robust and reproducible behavioral responses to oxygen. Specifically, worms prefer O2 levels of 5–10% and avoid too high or too low O2. Their O2 preference is not fixed but shows plasticity depending on experience, context, or genetic background. We recently showed that this experience-dependent plasticity declines with age,

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    A Step-by-step Protocol for Obtaining Mature Microglia from Mice
    Authors:  Min-Jung Yoo and Min-Soo Kwon, date: 08/05/2022, view: 1097, 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: 914, 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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