Cell Biology

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    Cell-attached and Whole-cell Patch-clamp Recordings of Dopamine Neurons in the Substantia Nigra Pars Compacta of Mouse Brain Slices
    Authors:  Stefano Cattaneo, Maria Regoni, Jenny Sassone and Stefano Taverna, date: 08/05/2021, view: 17, Q&A: 0

    The Substantia Nigra pars compacta (SNc) is a midbrain dopaminergic nucleus that plays a key role in modulating motor and cognitive functions. It is crucially involved in several disorders, particularly Parkinson’s disease, which is

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    Mouse Periosteal Cell Culture, in vitro Differentiation, and in vivo Transplantation in Tibial Fractures

    The periosteum covering the outer surface of bone contains skeletal stem/progenitor cells that can efficiently form cartilage and bone during bone repair. Several methods have been described to isolate periosteal cells based on bone scraping and/or

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    CRISPR-mediated Labeling of Cells in Chick Embryos Based on Selectively Expressed Genes
    Authors:  Masahito Yamagata and Joshua R. Sanes, date: 08/05/2021, view: 62, Q&A: 0

    The abilities to mark and manipulate specific cell types are essential for an increasing number of functional, structural, molecular, and developmental analyses in model organisms. In a few species, this can be accomplished by germline transgenesis;

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    Isolation and in vitro Culture of Mouse Oocytes
    Authors:  Jessica Greaney, Goutham Narayanan Subramanian, Yunan Ye and Hayden Homer, date: 08/05/2021, view: 23, Q&A: 0

    Females are endowed at birth with a fixed reserve of oocytes, which declines both in quantity and quality with advancing age. Understanding the molecular mechanisms regulating oocyte quality is crucial for improving the chances of pregnancy success

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    Optical Clearing and 3D Analysis Optimized for Mouse and Human Pancreata

    The pancreas is a heavily innervated organ, but pancreatic innervation can be challenging to comprehensively assess using conventional histological methods. However, recent advances in whole-mount tissue clearing and 3D rendering techniques have

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    A Multi-color Bicistronic Biosensor to Compare the Translation Dynamics of Different Open Reading Frames at Single-molecule Resolution in Live Cells
    Authors:  Amanda L. Koch, Tatsuya Morisaki and Timothy J. Stasevich, date: 07/20/2021, view: 859, Q&A: 0

    Here, we describe how to image and quantitate the translation dynamics of a bicistronic biosensor that we recently created to fairly compare cap-dependent and IRES-mediated translation at single-molecule resolution in live human cells. This

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    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: 1056, 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

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    Modeling the Nonlinear Dynamics of Intracellular Signaling Networks
    Authors:  Oleksii S. Rukhlenko and Boris N. Kholodenko, date: 07/20/2021, view: 948, Q&A: 0

    This protocol illustrates a pipeline for modeling the nonlinear behavior of intracellular signaling pathways. At fixed spatial points, nonlinear signaling dynamics are described by ordinary differential equations (ODEs). At constant parameters,

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    Analysis of the Effects of Hexokinase 2 Detachment From Mitochondria-Associated Membranes with the Highly Selective Peptide HK2pep
    Authors:  Francesco Ciscato, Federica Chiara, Riccardo Filadi and Andrea Rasola, date: 07/20/2021, view: 818, Q&A: 0

    The crucial role of hexokinase 2 (HK2) in the metabolic rewiring of tumors is now well established, which makes it a suitable target for the design of novel therapies. However, hexokinase activity is central to glucose utilization in all tissues;

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    Preparation of Human Chondrocytes for Profiling Using Cytometry by Time-of-flight (cyTOF)
    Authors:  Fiorella Carla Grandi and Nidhi Bhutani, date: 07/20/2021, view: 810, Q&A: 0

    Single-cell technologies have allowed high-resolution profiling of tissues and thus a deeper understanding of tissue homeostasis and disease heterogeneity. Understanding this heterogeneity can be especially important for tailoring treatments in a

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