Cell Biology

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    Protocols in Current Issue
    Spatiotemporal Quantification of Cytosolic pH in Arabidopsis Pollen Tubes
    Authors:  Maria Teresa Portes, Daniel S.C. Damineli and José A. Feijó, date: 07/20/2021, view: 440, Q&A: 0
    [Abstract]

    Ion-specific probes and fluorescent indicators have been key in establishing the role of ion signaling, namely calcium, protons, and anions, in plant development, providing a robust approach for monitoring spatiotemporal changes in intracellular ion dynamics. The integration of protons/pH in signaling mechanisms is especially important as reports

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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: 788, Q&A: 0
    [Abstract]

    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 technique employs a pair of complementary intrabodies loaded into living cells that co-translationally bind

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    A Method for Estimating the Potential Synaptic Connections Between Axons and Dendrites From 2D Neuronal Images
    Authors:  Carolina Tecuatl, Diek W. Wheeler and Giorgio A. Ascoli, date: 07/05/2021, view: 1306, Q&A: 0
    [Abstract]

    Computational neuroscience aims to model, reproduce, and predict network dynamics for different neuronal ensembles by distilling knowledge derived from electrophysiological and morphological evidence. However, analyses and simulations often remain critically limited by the sparsity of direct experimental constraints on essential parameters, such

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    Fluorescence-based Single-cell Analysis of Whole-mount-stained and Cleared Microtissues and Organoids for High Throughput Screening
    Authors:  Henrik Renner, Mandy Otto, Martha Grabos, Hans R. Schöler and Jan M. Bruder, date: 06/20/2021, view: 1115, Q&A: 1
    [Abstract]

    Three-dimensional (3D) cell culture, especially in the form of organ-like microtissues (“organoids”), has emerged as a novel tool potentially mimicking human tissue biology more closely than standard two-dimensional culture. Typically, tissue sectioning is the standard method for immunohistochemical analysis. However, it removes cells

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    In vivo Optical Access to Olfactory Sensory Neurons in the Mouse Olfactory Epithelium
    Authors:  Shigenori Inagaki, Ryo Iwata and Takeshi Imai, date: 06/20/2021, view: 1043, Q&A: 0
    [Abstract]

    In neuroscience, it is fundamental to understand how sensory stimuli are translated into neural activity at the entry point of sensory systems. In the olfactory system, odorants inhaled into the nasal cavity are detected by ~1,000 types of odorant receptors (ORs) that are expressed by olfactory sensory neurons (OSNs). Since each OSN expresses only

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    Dual Color, Live Imaging of Vesicular Transport in Axons of Cultured Sensory Neurons
    Authors:  Yoko Bekku and James L. Salzer, date: 06/20/2021, view: 877, Q&A: 0
    [Abstract]

    The function of neurons in afferent reception, integration, and generation of electrical activity relies on their strikingly polarized organization, characterized by distinct membrane domains. These domains have different compositions resulting from a combination of selective targeting and retention of membrane proteins. In neurons, most proteins

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    Analysis of Caenorhabditis elegans Sperm Number, Size, Activation, and Mitochondrial Content
    Authors:  Amy M. Hammerquist, Chia-An Yen and Sean P. Curran, date: 06/05/2021, view: 1167, Q&A: 0
    [Abstract]

    Infertility is a widespread and often unexplained issue. Studying reproduction using C. elegans males offers insight into the influence of individual factors on male fertility in humans. We have created a collection of protocols to assess several aspects of C. elegans sperm quality, including number, size, rate of activation, and mitochondrial

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    Quantitation of Secretory Granule Size in Drosophila Larval Salivary Glands
    Authors:  Cheng-I J. Ma and Julie A. Brill, date: 06/05/2021, view: 1130, Q&A: 0
    [Abstract]

    Maturation of secretory granules is a crucial process that ensures the bioactivity of cargo proteins undergoing regulated secretion. In Drosophila melanogaster, the larval salivary glands produce secretory granules that are up to four-fold larger in cross-sectional area after maturation. Therefore, we developed a live imaging microscopy approach

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    Visualization and Quantitation of Wg trafficking in the Drosophila Wing Imaginal Epithelium
    Authors:  Leonie Witte, Karen Linnemannstöns, Mona Honemann-Capito and Julia Christina Gross, date: 06/05/2021, view: 1169, Q&A: 0
    [Abstract]

    Secretory Wnt trafficking can be studied in the polarized epithelial monolayer of Drosophila wing imaginal discs (WID). In this tissue, Wg (Drosophila Wnt-I) is presented on the apical surface of its source cells before being internalized into the endosomal pathway. Long-range Wg secretion and spread depend on secondary secretion from endosomal

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    In vivo Fluorescence Imaging of Extracellular ATP in the Mouse Cerebral Cortex with a Hybrid-type Optical Sensor
    [Abstract]

    Adenosine 5’-triphosphate (ATP) works as an extracellular signaling molecule for cells in the brain, such as neurons and glia. Cellular communication via release of ATP is involved in a range of processes required for normal brain functions, and aberrant communication is associated with brain disorders. To investigate the mechanisms

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