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: 433, Q&A: 0

    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

    An in vitro Blood-brain Barrier Model to Study the Penetration of Nanoparticles
    Authors:  Sezin Aday, Wen Li, Jeffrey M. Karp and Nitin Joshi, date: 02/20/2022, view: 2458, Q&A: 0

    The blood-brain barrier (BBB), a crucial protection mechanism in the central nervous system (CNS), is a selective barrier comprised of endothelial cells. It hampers the development of therapeutic and diagnostic tools for neurological diseases due to the poor penetration of most of these agents. Rationally engineered nanoparticles (NP) can

    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: 2847, Q&A: 0

    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

    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: 1775, 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 production. Hypothalamic circadian clocks have been identified to play a particular role in coordinating

    A Rigorous Quantitative Approach to Analyzing Phagocytosis Assays
    Authors:  Michael D. Caponegro, Kaitlyn Koenig Thompson, Maryam Tayyab and Stella E. Tsirka, date: 01/05/2021, view: 3125, Q&A: 0
    [Abstract] Studying monocytic cells in isolated systems in vitro contributes significantly to the understanding of innate immune physiology. Functional assays produce read outs which can be used to measure responses to selected stimuli, such as pathogen exposure, antigen loading, and cytokine stimulation. Integration of these results with high ...
    Identification of Socially-activated Neurons
    Authors:  Mary L. Phillips and Lucas Pozzo-Miller, date: 09/05/2020, view: 4086, Q&A: 0
    [Abstract] Determining the neuronal circuitry responsible for specific behaviors is a major focus in the field of neurobiology. Activity-dependent immediate early genes (IEGs), transcribed and translated shortly after neurons discharge action potentials, have been used extensively to either identify or gain genetic access to neurons and brain regions ...
    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 ...
    Method for Prolonged Incubation of Brain Slices
    Authors:  Orsolya Kekesi and Yossi Buskila, date: 07/20/2020, view: 2576, Q&A: 0
    [Abstract] Slices of neuronal tissue maintain a high degree of topographical and functional properties of neurons and glia and therefore are extensively used for measurements of neuronal activity at the molecular, cellular and network levels. However, the lifespan of slice preparations is narrow, averaging of 6-8 hours. Moreover, the average viability of ...
    Organotypic Slice Culture of the Embryonic Mouse Brain
    Authors:  James M. Clegg and Thomas Pratt, date: 07/05/2020, view: 2493, Q&A: 0
    [Abstract] Organotypic slice culture is a powerful technique for exploring the embryonic development of the mammalian brain. In this protocol we describe a basic slice culture technique we have used for two sets of experiments: axon guidance transplant assays and bead culture assays.
    Isolation of Tumor Cells Based on Their Distance from Blood Vessels
    Authors:  Saran Kumar, Husni Sharife, Tirzah Kreisel, Libat Bar-Lev, Myriam Grunewald and Eli Keshet, date: 05/20/2020, view: 3526, Q&A: 0
    [Abstract] Differential exposure of tumor cells to microenvironmental cues greatly impacts cell phenotypes, raising a need for position based sorting of tumor cells amenable to multiple OMICs and functional analyses. One such key determinant of tumor heterogeneity in solid tumors is its vasculature. Proximity to blood vessels (BVs) profoundly affects tumor ...

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