Developmental Biology

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    Protocols in Current Issue
    Advances in Proximity Ligation in situ Hybridization (PLISH)
    Authors:  Monica Nagendran, Adam M Andruska, Pehr B Harbury and Tushar J Desai, date: 11/05/2020, view: 683, Q&A: 0
    [Abstract] Understanding tissues in the context of development, maintenance and disease requires determining the molecular profiles of individual cells within their native in vivo spatial context. We developed a Proximity Ligation in situ Hybridization technology (PLISH) that enables quantitative measurement of single cell gene expression ...
    Multitarget Immunohistochemistry for Confocal and Super-resolution Imaging of Plant Cell Wall Polysaccharides
    Authors:  Kalina T. Haas, Methieu Rivière, Raymond Wightman and Alexis Peaucelle, date: 10/05/2020, view: 811, Q&A: 0
    [Abstract] The plant cell wall (PCW) is a pecto-cellulosic extracellular matrix that envelopes the plant cell. By integrating extra-and intra-cellular cues, PCW mediates a plethora of essential physiological functions. Notably, it permits controlled and oriented tissue growth by tuning its local mechano-chemical properties. To refine our knowledge of these ...
    A Reproducible Protocol to Measure the Critical Swimming Speed of Adult Zebrafish
    Authors:  Yuma Wakamatsu, Makoto Kashima and Hiromi Hirata, date: 08/20/2020, view: 849, Q&A: 0
    [Abstract] The quantitative measurement of water flow-induced swimming of fish species using a swimmill is a powerful method to evaluate motor ability of individual fish. Zebrafish is a commonly used vertebrate that enables the study of morphological, physiological and behavioral characteristics associated with genes. We here established a reproducible ...
    Mechanical Tissue Compression and Whole-mount Imaging at Single Cell Resolution for Developing Murine Epididymal Tubules
    Author:  Tsuyoshi Hirashima, date: 08/05/2020, view: 1024, Q&A: 0
    [Abstract] Cells inside the body are subjected to various mechanical stress, such as stretch or compression provided by surrounding cells, shear stresses by blood or lymph flows, and normal stresses by luminal liquids. Force loading to the biological tissues is a fundamental method to better understand cellular responses to such mechanical stimuli. There ...
    Application of Mechanical Forces on Drosophila Embryos by Manipulation of Microinjected Magnetic Particles
    Authors:  Arturo D’Angelo and Jérôme Solon, date: 05/05/2020, view: 1056, Q&A: 0
    [Abstract] Cells generate mechanical forces to shape tissues during morphogenesis. These forces can activate several biochemical pathways and trigger diverse cellular responses by mechano-sensation, such as differentiation, division, migration and apoptosis. Assessing the mechano-responses of cells in living organisms requires tools to apply controlled local ...
    Live Cell Imaging of Male Meiosis in Arabidopsis by a Landmark-based System
    [Abstract] Live cell imaging has tremendously promoted our understanding of cellular and subcellular processes such as cell division. Here, we present a step-by-step protocol for a robust and easy-to-use live cell imaging approach to study male meiosis in the plant Arabidopsis thaliana as recently established. Our method relies on the concomitant ...
    Auxin-mediated Protein Degradation in Caenorhabditis elegans
    Authors:  Michael A. Q. Martinez and David Q. Matus, date: 04/20/2020, view: 1832, Q&A: 0
    [Abstract] The auxin-inducible degron (AID) technology was recently adapted for use in the nematode Caenorhabditis elegans. Rapid degradation of C. elegans proteins tagged with an AID is mediated by a plant-specific F-box protein, transport inhibitor response 1 (TIR1), and occurs only in the presence of the phytohormone auxin. The first ...
    Bimolecular Fluorescence Complementation (BiFC) for Studying Sarcomeric Protein Interactions in Drosophila
    Authors:  Océane Marescal, Frieder Schöck and Nicanor González-Morales, date: 04/05/2020, view: 1530, Q&A: 0
    [Abstract] Protein-protein interactions in Drosophila myofibrils are essential for their function and formation. Bimolecular Fluorescence Complementation (BiFC) is an effective method for studying protein interactions and localization. BiFC relies on the reconstitution of a monomeric fluorescent protein from two half-fragments when in proximity. Two ...
    Mouse Embryonic Tooth Germ Dissection and Ex vivo Culture Protocol
    Authors:  Xue Han, Keigo Yoshizaki, Tian Tian, Kanako Miyazaki, Ichiro Takahashi and Satoshi Fukumoto, date: 02/05/2020, view: 1889, Q&A: 1
    [Abstract] A tooth germ ex vivo organ culture allows visualization of its development in different stages, thus enabling investigation of the molecular mechanisms of regulatory factors. Tooth germs can be rapidly dissected from E13 mouse embryos and placed on cell culture inserts for observation of subsequent tooth germ development in a ...
    3D Organoid Formation from the Murine Salivary Gland Cell Line SIMS
    Authors:  Harleen K. Athwal and Isabelle M. A. Lombaert, date: 10/05/2019, view: 2700, Q&A: 0
    [Abstract] Salivary glands consist of multiple phenotypically and functionally unique cell populations, such as the acinar, ductal, and myoepithelial cells that help produce, modify, and secrete saliva (Lombaert et al., 2011). Identification of mechanisms and factors that regulate these populations has been of key interest, as salivary gland-related ...



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