Developmental Biology

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    Protocols in Current Issue
    Primed Track: Reliable Volumetric Single-cell Tracking and Lineage Tracing of Living Specimen with Dual-labeling Approaches
    Authors:  Maaike Welling, Konstantinos Kalyviotis and Periklis Pantazis, date: 06/05/2020, view: 550, Q&A: 0
    [Abstract] Mammalian embryonic development starts with a single fertilized zygote that develops into a blastocyst embryo consisting of three cell types that evolve into either embryonic or extra-embryonic tissues. Lineage tracing of these cells can provide important information about the molecular and cellular dynamics contributing to fate allocation during ...
    Ex vivo Culture Assay Using Human Hair Follicles to Study Circadian Characteristics
    Authors:  Atsuhiro Nishida, Yoshiki Miyawaki, Koichi Node and Makoto Akashi, date: 06/05/2020, view: 469, Q&A: 0
    [Abstract] Ex vivo culture assays of biopsy specimens are advantageous for the experimental evaluation of human circadian characteristics. We developed a simple and non-invasive experimental evaluation method for monitoring the expression of circadian clock genes in an ex vivo culture assay using human hair follicles. This method imposes ...
    FRET Reporter Assays for cAMP and Calcium in a 96-well Format Using Genetically Encoded Biosensors Expressed in Living Cells
    Authors:  Brandon T. Milliken, Robert P. Doyle, George G. Holz and Oleg G. Chepurny, date: 06/05/2020, view: 643, Q&A: 0
    [Abstract] Stimulation of G protein-coupled receptors (GPCR) by hormones and neurotransmitters elicits cellular responses, many of which result from alterations in the concentrations of cytosolic cAMP and Ca2+. Here, we describe a microplate reader fluorescence resonance energy transfer (FRET) assay that uses the genetically encoded biosensors ...
    Direct Reprogramming of Mouse Embryonic Fibroblasts to Conventional Type 1 Dendritic Cells by Enforced Expression of Transcription Factors
    Authors:  Fábio F. Rosa, Cristiana F. Pires, Olga Zimmermannova and Carlos-Filipe Pereira, date: 05/20/2020, view: 808, Q&A: 0
    [Abstract] Ectopic expression of transcription factor combinations has been recently demonstrated to reprogram differentiated somatic cells towards the dendritic cell (DC) lineage without reversion to a multipotent state. DCs have the ability to induce potent and long-lasting adaptive immune responses. In particular, conventional type 1 DCs (cDC1s) excel on ...
    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: 589, 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 ...
    A Modified Semisolid Clonal Culture for Identification of B-1 and B-2 Progenitor Colony Forming Ability of Mouse Embryonic Hemogenic Endothelial Cells
    Authors:  Michihiro Kobayashi and Momoko Yoshimoto, date: 05/05/2020, view: 659, Q&A: 0
    [Abstract] The search for the origin of the first hematopoietic stem cells (HSCs) in the mouse embryo has been a hot topic in the field of developmental hematopoiesis. Detecting lymphoid potential is one of the supportive evidence to show the definitive hematopoietic activity of HSCs. However, the first B-lymphoid potential in the mouse embryos are reported ...
    Auxin-mediated Protein Degradation in Caenorhabditis elegans
    Authors:  Michael A. Q. Martinez and David Q. Matus, date: 04/20/2020, view: 931, 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: 547, 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 ...
    Self-organization Assay for Min Proteins of Escherichia coli in Micro-droplets Covered with Lipids
    Authors:  Shunshi Kohyama, Kei Fujiwara, Natsuhiko Yoshinaga and Nobuhide Doi, date: 03/20/2020, view: 697, Q&A: 0
    [Abstract] The Min system determines the cell division plane of bacteria. As a cue of spatiotemporal regulation, the Min system uses wave propagation of MinD protein (Min wave). Therefore, the reconstitution of the Min wave in cell-sized closed space will lead to the creation of artificial cells capable of cell division. The Min waves emerge via coupling ...



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