Developmental Biology

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    Protocols in Current Issue
    Isolation and in vitro Culture of Mouse Oocytes
    Authors:  Jessica Greaney, Goutham Narayanan Subramanian, Yunan Ye and Hayden Homer, date: 08/05/2021, view: 23, Q&A: 0

    Females are endowed at birth with a fixed reserve of oocytes, which declines both in quantity and quality with advancing age. Understanding the molecular mechanisms regulating oocyte quality is crucial for improving the chances of pregnancy success

    ...
    Mouse Periosteal Cell Culture, in vitro Differentiation, and in vivo Transplantation in Tibial Fractures

    The periosteum covering the outer surface of bone contains skeletal stem/progenitor cells that can efficiently form cartilage and bone during bone repair. Several methods have been described to isolate periosteal cells based on bone scraping and/or

    ...
    Fe-NTA Microcolumn Purification of Phosphopeptides from Immunoprecipitation (IP) Eluates for Mass Spectrometry Analysis
    Authors:  Ethan J. Sanford and Marcus B. Smolka, date: 08/05/2021, view: 14, Q&A: 0

    Protein phosphorylation is a nearly universal signaling mechanism. To date, a number of proteomics tools have been developed to analyze phosphorylation. Phosphoproteome-wide analyses using whole cell extracts suffer from incomplete coverage, often

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    A Genetically Engineered Mouse Model of Venous Anomaly and Retinal Angioma-like Vascular Malformation
    Authors:  Xudong Cao, Beibei Xu, Xiao Li, Taotao Li and Yulong He, date: 08/05/2021, view: 16, Q&A: 0

    Characterization of key regulators in vein development will advance our understanding of mechanisms underlying venous anomalies and provide therapeutic targets for the treatment of vascular malformations. Here, we provide a detailed protocol for the

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    A Genetically Engineered Mouse Model of Venous Anomaly and Retinal Angioma-like Vascular Malformation
    Authors:  Xudong Cao, Beibei Xu, Xiao Li, Taotao Li and Yulong He, date: 08/05/2021, view: 16, Q&A: 0
    [Abstract]

    Characterization of key regulators in vein development will advance our understanding of mechanisms underlying venous anomalies and provide therapeutic targets for the treatment of vascular malformations. Here, we provide a detailed protocol for the generation of genetically engineered mouse models targeting the Tek gene for the analysis of vein

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    Fe-NTA Microcolumn Purification of Phosphopeptides from Immunoprecipitation (IP) Eluates for Mass Spectrometry Analysis
    Authors:  Ethan J. Sanford and Marcus B. Smolka, date: 08/05/2021, view: 14, Q&A: 0
    [Abstract]

    Protein phosphorylation is a nearly universal signaling mechanism. To date, a number of proteomics tools have been developed to analyze phosphorylation. Phosphoproteome-wide analyses using whole cell extracts suffer from incomplete coverage, often missing phosphorylation events from low-abundance proteins. In order to increase coverage of

    ...
    Mouse Periosteal Cell Culture, in vitro Differentiation, and in vivo Transplantation in Tibial Fractures
    [Abstract]

    The periosteum covering the outer surface of bone contains skeletal stem/progenitor cells that can efficiently form cartilage and bone during bone repair. Several methods have been described to isolate periosteal cells based on bone scraping and/or enzymatic digestion. Here, we describe an explant culture method to isolate periosteum-derived

    ...
    Isolation and in vitro Culture of Mouse Oocytes
    Authors:  Jessica Greaney, Goutham Narayanan Subramanian, Yunan Ye and Hayden Homer, date: 08/05/2021, view: 23, Q&A: 0
    [Abstract]

    Females are endowed at birth with a fixed reserve of oocytes, which declines both in quantity and quality with advancing age. Understanding the molecular mechanisms regulating oocyte quality is crucial for improving the chances of pregnancy success in fertility clinics. In vitro culture systems enable researchers to analyse important molecular and

    ...
    GeneWeld: Efficient Targeted Integration Directed by Short Homology in Zebrafish
    [Abstract]

    Efficient precision genome engineering requires high frequency and specificity of integration at the genomic target site. Multiple design strategies for zebrafish gene targeting have previously been reported with widely varying frequencies for germline recovery of integration alleles. The GeneWeld protocol and pGTag (plasmids for Gene Tagging)

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    An Improved Method for Individual Tracking of Voluntary Wheel Running in Pair-housed Juvenile Mice
    Authors:  David A. Valientes, Anthony M. Raus and Autumn S. lvy, date: 07/05/2021, view: 697, Q&A: 0
    [Abstract]

    Rodent cages equipped with access to a voluntary running wheel are commonly used to study the effects of aerobic physical activity on physiology and behavior. Notable discoveries in exercise neurobiology, including the key role of brain-derived neurotrophic factor (BDNF) in neural plasticity and cognition, have been made using rodents housed with

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    Isolation of First-Trimester and Full-term Human Placental Hofbauer Cells
    Authors:  Anna Appios, Jake R. Thomas and Naomi McGovern, date: 06/05/2021, view: 1573, Q&A: 0
    [Abstract]

    The placenta is the crucial organ that regulates the health of both mother and fetus during pregnancy. The human placenta is composed of villous tree-like structures that embed into the maternal decidua. Within the stroma of the villi resides a population of fetally-derived macrophages, the Hofbauer cells (HBC). HBC are the only fetal immune cells

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    Generation of Mouse Pluripotent Stem Cell-derived Trunk-like Structures: An in vitro Model of Post-implantation Embryogenesis
    [Abstract]

    Post-implantation mammalian embryogenesis involves profound molecular, cellular, and morphogenetic changes. The study of these highly dynamic processes is complicated by the limited accessibility of in utero development. In recent years, several complementary in vitro systems comprising self-organized assemblies of mouse embryonic stem cells, such

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    Visualization and Quantitation of Wg trafficking in the Drosophila Wing Imaginal Epithelium
    Authors:  Leonie Witte, Karen Linnemannstöns, Mona Honemann-Capito and Julia Christina Gross, date: 06/05/2021, view: 1193, Q&A: 0
    [Abstract]

    Secretory Wnt trafficking can be studied in the polarized epithelial monolayer of Drosophila wing imaginal discs (WID). In this tissue, Wg (Drosophila Wnt-I) is presented on the apical surface of its source cells before being internalized into the endosomal pathway. Long-range Wg secretion and spread depend on secondary secretion from endosomal

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    Quantitation of Secretory Granule Size in Drosophila Larval Salivary Glands
    Authors:  Cheng-I J. Ma and Julie A. Brill, date: 06/05/2021, view: 1149, Q&A: 0
    [Abstract]

    Maturation of secretory granules is a crucial process that ensures the bioactivity of cargo proteins undergoing regulated secretion. In Drosophila melanogaster, the larval salivary glands produce secretory granules that are up to four-fold larger in cross-sectional area after maturation. Therefore, we developed a live imaging microscopy approach

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