Developmental Biology

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    Protocols in Current Issue
    Isolation of First-Trimester and Full-term Human Placental Hofbauer Cells
    Authors:  Anna Appios, Jake R. Thomas and Naomi McGovern, date: 06/05/2021, view: 1024, Q&A: 0
    [Abstract]

    The placenta is the crucial organ that regulates the health of both mother and fetus during pregnancy. The human placenta is composed of villous tree-like structures that embed into the maternal decidua. Within the stroma of the villi resides a population of fetally-derived macrophages, the Hofbauer cells (HBC). HBC are the only fetal immune cells

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    Analysis of Monocyte Cell Fate by Adoptive Transfer in a Murine Model of TLR7-induced Systemic Inflammation
    Authors:  Jaba Gamrekelashvili, Hermann Haller and Florian P. Limbourg, date: 05/05/2021, view: 928, Q&A: 0
    [Abstract]

    Myeloid plasticity is a hallmark of the innate immune response to Toll-like receptor (TLR) activation. Here, we provide a protocol for monocyte cell fate tracking by adoptive transfer in the context of systemic inflammation induced by TLR7 activation, the principal innate immune receptor sensing viral RNA in mice. Defined monocyte subsets are

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    Identification of R-loop-forming Sequences in Drosophila melanogaster Embryos and Tissue Culture Cells Using DRIP-seq
    Authors:  Célia Alecki and Nicole J. Francis, date: 05/05/2021, view: 979, Q&A: 0
    [Abstract]

    R-loops are non-canonical nucleic structures composed of an RNA–DNA hybrid and a displaced ssDNA. Originally identified as a source of genomic instability, R-loops have been shown over the last decade to be involved in the targeting of proteins and to be associated with different histone modifications, suggesting a regulatory function. In

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    A Workflow for High-pressure Freezing and Freeze Substitution of the Caenorhabditis elegans Embryo for Ultrastructural Analysis by Conventional and Volume Electron Microscopy
    Authors:  Mohammad M. Rahman, Irene Y. Chang, Orna Cohen-Fix and Kedar Narayan, date: 04/05/2021, view: 1087, Q&A: 0
    [Abstract]

    The free-living nematode Caenorhabditis elegans is a popular model system for studying developmental biology. Here we describe a detailed protocol to high-pressure freeze the C. elegans embryo (either ex vivo after dissection, or within the intact worm) followed by quick freeze substitution. Processed samples are suitable for ultrastructural

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    Extraction and Quantification of Sphingolipids from Hemiptera Insects by Ultra-Performance Liquid Chromatography Coupled to Tandem Mass Spectrometry
    Authors:  Ni Wang, Xiaoxiao Shi, Chao Zhang, Wenwu Zhou and Zeng-Rong Zhu, date: 02/20/2021, view: 1363, Q&A: 0
    [Abstract]

    Sphingolipids are major structural components of endomembranes and have also been described as an intracellular second messenger involved in various biological functions in all eukaryotes and a few prokaryotes. Ceramides (Cer), the central molecules of sphingolipids, have been depicted in cell growth arrest, cell differentiation, and apoptosis.

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    Induction of Epithelial-mesenchymal Transition in MDCK II Cells
    Authors:  Agnieszka Pastuła and Richard Lundmark, date: 02/05/2021, view: 671, Q&A: 0
    [Abstract]

    Epithelial-mesenchymal transition (EMT) is a reversible process of epithelial cell transdifferentiation into a mesenchymal cell, that enables initiation of cell migration. EMT plays an important role in embryonic development, tissue repair and cancer metastasis. Better understanding of cellular and molecular events during EMT will not only provide

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    Whole-mount Immunohistochemistry of Adult Zebrafish Retina for Advanced Imaging
    Authors:  Mikiko Nagashima and Peter F. Hitchcock, date: 12/20/2020, view: 1367, Q&A: 0
    [Abstract]

    Immunohistochemistry is a widely used technique to examine the expression and subcellular localization of proteins. This technique relies on the specificity of antibodies and requires adequate penetration of antibodies into tissues. The latter is especially challenging for thick specimens, such as embryos and other whole-mount preparations. Here

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    Use of Optogenetic Amyloid-β to Monitor Protein Aggregation in Drosophila melanogaster, Danio rerio and Caenorhabditis elegans
    Authors:  Prameet Kaur, Caroline Kibat, Emelyne Teo, Jan Gruber, Ajay Mathuru and Nicholas S. Tolwinski, date: 12/05/2020, view: 1570, Q&A: 0
    [Abstract]

    Alzheimer’s Disease (AD) has long been associated with accumulation of extracellular amyloid plaques (Aβ) originating from the Amyloid Precursor Protein. Plaques have, however, been discovered in healthy individuals and not all AD brains show plaques, suggesting that extracellular Aβ aggregates may play a smaller role than

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    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 ...
    DigiTAG–a RNA Sequencing Approach to Analyze Transcriptomes of Rare Cell Populations in Drosophila melanogaster
    Authors:  Lisa Landskron, Francois Bonnay, Thomas R. Burkard and Jürgen A. Knoblich, date: 11/05/2020, view: 1323, Q&A: 0
    [Abstract]

    Cell-type specific transcriptional programs underlie the development and maintenance of organs. Not only distinct cell types within a tissue, even cells with supposedly identical cell fates show a high degree of transcriptional heterogeneity. Inevitable, low cell numbers are a major hurdle to study transcriptomes of pure cell populations. Here we

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