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: 433, Q&A: 0
    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 ...
    Mechanical Tissue Compression and Whole-mount Imaging at Single Cell Resolution for Developing Murine Epididymal Tubules
    Author:  Tsuyoshi Hirashima, date: 08/05/2020, view: 433, 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 ...
    Isolation, Culture, and Differentiation of Primary Myoblasts Derived from Muscle Satellite Cells
    Authors:  Kun Ho Kim, Jiamin Qiu and Shihuan Kuang, date: 07/20/2020, view: 296, Q&A: 0
    [Abstract] The skeletal muscle is key for body mobility and motor performance, but aging and diseases often lead to progressive loss of muscle mass due to wasting or degeneration of muscle cells. Muscle satellite cells (MuSCs) represent a population of tissue stem cells residing in the skeletal muscles and are responsible for homeostatic maintenance and ...
    In vitro Induction and Detection of Acrosomal Exocytosis in Human Spermatozoa
    Authors:  Shenae L. Cafe, Amanda L. Anderson and Brett Nixon, date: 07/20/2020, view: 270, Q&A: 0
    [Abstract] The acrosome reaction is a highly regulated exocytotic event that primes spermatozoa for successful fertilization. Upon induction, acrosomal exocytosis proceeds via a wave of vesiculation that radiates across the sperm head, destabilizing the acrosomal vesicle and resulting in the release of the acrosomal contents. Having shed their acrosome, ...
    Organotypic Slice Culture of the Embryonic Mouse Brain
    Authors:  James M. Clegg and Thomas Pratt, date: 07/05/2020, view: 457, Q&A: 0
    [Abstract] Organotypic slice culture is a powerful technique for exploring the embryonic development of the mammalian brain. In this protocol we describe a basic slice culture technique we have used for two sets of experiments: axon guidance transplant assays and bead culture assays.
    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: 732, 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: 635, 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: 944, 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: 1157, 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: 763, 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 ...



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