Developmental Biology

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    Protocols in Current Issue
    Mechanical Tissue Compression and Whole-mount Imaging at Single Cell Resolution for Developing Murine Epididymal Tubules
    Author:  Tsuyoshi Hirashima, date: 08/05/2020, view: 460, 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: 770, 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: 1348, 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: 1118, 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: 1450, 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: 2240, 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 ...
    Reconstituting Breast Tissue with Organotypic Three-dimensional Co-culture of Epithelial and Stromal Cells in Discontinuous Extracellular Matrices
    Authors:  Gang Ren, Vandana Sharma, Joshua Letson, Yashna Walia, Veani Fernando and Saori Furuta, date: 10/05/2019, view: 1676, Q&A: 0
    [Abstract] Co-culture systems utilizing reconstituted or synthetic extracellular matrix (ECM) and micropatterning techniques have enabled the reconstruction of surface epithelial tissues. This technique has been utilized in the regeneration, disease modeling and drug screening of the surface epithelia, such as the skin and esophagus. On the other hand, the ...
    A Widely Applicable Urea-based Fluorescent/Colorimetric mRNA in situ Hybridization Protocol
    Author:  Chiara Sinigaglia, date: 09/05/2019, view: 2178, Q&A: 0
    [Abstract] In situ hybridization methods are routinely employed to detect nucleic acid sequences, allowing to localize gene expression or to study chromosomal organization in their native context. These methods rely on the pairwise binding of a labeled probe to the target endogenous nucleic acid sequence–the hybridization step, followed by detection ...
    Detection of mRNA by Whole Mount in situ Hybridization and DNA Extraction for Genotyping of Zebrafish Embryos
    Authors:  Rachna Narayanan and Andrew C. Oates, date: 03/20/2019, view: 6523, Q&A: 0
    [Abstract] In situ hybridization is used to visualize the spatial distribution of gene transcripts in tissues and in embryos, providing important information about disease and development. Current methods involve the use of complementary riboprobes incorporating non-radioactive labels that can be detected by immunohistochemistry and coupled to ...



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