Developmental Biology


    Protocols in Current Issue
    Optimized Immunostaining of Embryonic and Early Postnatal Mouse Brain Sections
    Authors:  Kawssar Harb, Michele Bertacchi and Michèle Studer, date: 01/05/2021, view: 1116, Q&A: 0

    The mammalian neocortex, the outer layer of the cerebrum and most recently evolved brain region, is characterized by its unique areal and laminar organization. Distinct cortical layers and areas can be identified by the protein expression of graded transcription factors and molecular determinants that define the identity of different projection

    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: 1727, 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 ...
    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: 1463, 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 ...
    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: 2221, 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 ...
    Electron Microscopic Detection of Proteins and Protein Complexes in Mammalian Cells Using APEX-tagged, Conditionally Stable Nanobodies
    Authors:  Thomas E. Hall, James Rae, Nicholas Ariotti and Robert G. Parton, date: 11/20/2018, view: 3156, Q&A: 0
    [Abstract] The recent development of 3D electron microscopic techniques for cells and tissues has necessitated the development of new methods for the detection of proteins and protein-complexes in situ. The development of new genetic tags, such as the ascorbate peroxidase, APEX, for electron microscopic detection of tagged proteins has expanded the ...
    Transmission Electron Microscopy for Analysis of Mitochondria in Mouse Skeletal Muscle
    Authors:  Joseph D. McMillan and Michael A. Eisenback, date: 05/20/2018, view: 12351, Q&A: 0
    [Abstract] Skeletal muscle is the most abundant tissue in the human body and regulates a variety of functions including locomotion and whole-body metabolism. Skeletal muscle has a plethora of mitochondria, the organelles that are essential for aerobic generation of ATP which provides the chemical energy to fuel vital functions such as contraction. The number ...
    Construction and Cloning of Minigenes for in vivo Analysis of Potential Splice Mutations
    Authors:  Lisa Maria Riedmayr, Sybille Böhm, Stylianos Michalakis and Elvir Becirovic, date: 03/05/2018, view: 8283, Q&A: 0
    [Abstract] Disease-associated mutations influencing mRNA splicing are referred to as splice mutations. The majority of splice mutations are found on exon-intron boundaries defining canonical donor and acceptor splice sites. However, mutations in the coding region (exonic mutations) can also affect mRNA splicing. Exact knowledge of the disease mechanism of ...
    Preparation of Precisely Oriented Cryosections of Undistorted Drosophila Wing Imaginal Discs for High Resolution Confocal Imaging
    Authors:  Samuel Petshow and Marcel Wehrli, date: 02/05/2018, view: 6302, Q&A: 0
    [Abstract] The combination of immunofluorescence and laser scanning confocal microscopy (LSM) is essential to high-resolution detection of molecular distribution in biological specimens. A frequent limitation is the need to image deep inside a tissue or in a specific plane, which may be inaccessible due to tissue size or shape. Recreating high-resolution 3D ...
    Rapid IFM Dissection for Visualizing Fluorescently Tagged Sarcomeric Proteins
    Authors:  Yu Shu Xiao, Frieder Schöck and Nicanor González-Morales, date: 11/20/2017, view: 7191, Q&A: 0
    [Abstract] Sarcomeres, the smallest contractile unit of muscles, are arguably the most impressive actomyosin structure. Yet a complete understanding of sarcomere formation and maintenance is missing. The Drosophila indirect flight muscle (IFM) has proven to be a very valuable model to study sarcomeres. Here, we present a protocol for the rapid ...

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