Developmental Biology


    Protocols in Current Issue
    Ex vivo Trophoblast-specific Genetic Manipulation Using Lentiviral Delivery
    Authors:  Damayanti Chakraborty, Masanaga Muto and Michael J. Soares, date: 12/20/2017, view: 257, Q&A: 0
    [Abstract] In this protocol report, we describe a lentiviral gene delivery technique for genetic modification of the rat trophoblast cell lineage. Lentiviral packaged gene constructs can be efficiently and specifically delivered to the trophoblast cell lineage of the blastocyst. The consequences of ‘gain-of-function’ and ‘loss-of-function’ blastocyst ...
    Biochemical Isolation of Myonuclei from Mouse Skeletal Muscle Tissue
    Authors:  Alicia A Cutler, Anita H Corbett and Grace K Pavlath, date: 12/20/2017, view: 387, Q&A: 0
    [Abstract] Skeletal muscle provides the contractile force necessary for movement, swallowing, and breathing and, consequently, is necessary for survival. Skeletal muscle cells are unique in that they are extremely large cells containing thousands of nuclei. These nuclei must all work in concert to maintain skeletal muscle function and thereby maintain life. ...
    Notch Ligand Binding Assay Using Flow Cytometry
    Authors:  Shweta Varshney and Pamela Stanley, date: 12/05/2017, view: 515, Q&A: 0
    [Abstract] Notch signaling is an evolutionarily conserved signaling pathway that plays an indispensable role during development, and in the maintenance of homeostatic processes, in a wide variety of tissues (Kopan, 2012; Hori et al., 2013). The multifaceted roles of Notch signaling are stringently regulated at different levels. One of the most ...
    Mandibular Explant Assay for Investigating Extrinsic Stimuli on Bone and Cartilage Development
    Authors:  Jamie Jiang, Jessica W. Bertol and Walid D. Fakhouri, date: 12/05/2017, view: 367, Q&A: 0
    [Abstract] A major issue in developmental biology is to determine how embryonic tissues respond to molecular signals in a timely manner and given the position-restricted instructions during morphogenesis, of which Meckel’s cartilage is a classical example. The ex-vivo explant model is a practical and convenient system that allows investigation of ...
    Rapid IFM Dissection for Visualizing Fluorescently Tagged Sarcomeric Proteins
    Authors:  Yu Shu Xiao, Frieder Schöck and Nicanor González-Morales, date: 11/20/2017, view: 516, 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 ...
    Protocol for Notch-ligand Binding Assays Using Dynabeads
    Authors:  Shogo Sawaguchi, Mitsutaka Ogawa and Tetsuya Okajima, date: 10/20/2017, view: 933, Q&A: 0
    [Abstract] This protocol describes how to measure interaction between Notch receptors and their ligands by cell-based assay using Dynabeads. We have used the protocol to determine binding capacity between Notch1-transfected HEK293T cells and ligand-coated Dynabeads. Expression of Eogt in Notch1-expressing cells promoted binding toward DLL4-coated ...
    In vitro Co-culture of Mesenchymal Stem Cells and Endothelial Colony Forming Cells
    Authors:  Abbas Shafiee and Kiarash Khosrotehrani, date: 10/20/2017, view: 1090, Q&A: 0
    [Abstract] The discovery of endothelial colony forming cells (ECFCs) with robust self-renewal and de novo vessel formation potentials suggests that ECFCs can be an excellent cell source for cardiovascular diseases treatment through improving neovascularization in the ischemic tissues. However, their engraftment after transplantation resulted to be low. ...
    Labelling HaloTag Fusion Proteins with HaloTag Ligand in Living Cells
    Authors:  Huy Nguyen Duc and Xiaojun Ren , date: 09/05/2017, view: 999, Q&A: 0
    [Abstract] HaloTag has been widely used to label proteins in vitro and in vivo (Los et al., 2008). In this protocol, we describe labelling HaloTag-Cbx fusion proteins by HaloTag ligands for live-cell single-molecule imaging (Zhen et al., 2016).
    Isolation of Mouse Cardiac Neural Crest Cells and Their Differentiation into Smooth Muscle Cells
    Authors:  Xia Wang and Sophie Astrof, date: 09/05/2017, view: 934, Q&A: 0
    [Abstract] Cardiac neural crest cells (CNCCs) originate at the dorsal edge of the neural tube between the otic pit and the caudal edge of the 3rd somite, and migrate into the pharyngeal arches and the heart. We have shown that fibronectin (Fn1) plays an important role in the development of the CNCC by regulating the differentiation of CNCCs into ...
    Ex vivo Ooplasmic Extract from Developing Drosophila Oocytes for Quantitative TIRF Microscopy Analysis
    Authors:  Imre Gáspár and Anne Ephrussi, date: 07/05/2017, view: 1445, Q&A: 0
    [Abstract] Understanding the dynamic behavior and the continuously changing composition of macromolecular complexes, subcellular structures and organelles is one of areas of active research in both cell and developmental biology, as these changes directly relate to function and subsequently to the development and homeostasis of the organism. Here, we ...