Developmental Biology

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    Protocols in Current Issue
    Analysis of Caenorhabditis elegans Sperm Number, Size, Activation, and Mitochondrial Content
    Authors:  Amy M. Hammerquist, Chia-An Yen and Sean P. Curran, date: 06/05/2021, view: 170, Q&A: 0

    Infertility is a widespread and often unexplained issue. Studying reproduction using C. elegans males offers insight into the influence of individual factors on male fertility in humans. We have created a collection of protocols to assess several

    ...
    Rice Root Hair Phenotypes Imaged by Cryo-SEM

    Cryo-scanning electron microscopy (cryo-SEM) was first introduced for scientific use in the 1980s. Since then, cryo-SEM has become a routine technique for studying the surfaces and internal structures of biological samples with a high water content.

    ...
    Quantitation of Secretory Granule Size in Drosophila Larval Salivary Glands
    Authors:  Cheng-I J. Ma and Julie A. Brill, date: 06/05/2021, view: 161, Q&A: 0

    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

    ...
    Visualization and Quantitation of Wg trafficking in the Drosophila Wing Imaginal Epithelium

    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

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

    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

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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: 578, Q&A: 0

    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

    ...
    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: 578, 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

    ...
    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

    ...
    Visualization and Quantitation of Wg trafficking in the Drosophila Wing Imaginal Epithelium
    [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

    ...
    Quantitation of Secretory Granule Size in Drosophila Larval Salivary Glands
    Authors:  Cheng-I J. Ma and Julie A. Brill, date: 06/05/2021, view: 161, 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

    ...
    Rice Root Hair Phenotypes Imaged by Cryo-SEM
    [Abstract]

    Cryo-scanning electron microscopy (cryo-SEM) was first introduced for scientific use in the 1980s. Since then, cryo-SEM has become a routine technique for studying the surfaces and internal structures of biological samples with a high water content. In contrast to traditional SEM, cryo-SEM requires no sample pretreatment processes; thus, we can

    ...
    Analysis of Caenorhabditis elegans Sperm Number, Size, Activation, and Mitochondrial Content
    Authors:  Amy M. Hammerquist, Chia-An Yen and Sean P. Curran, date: 06/05/2021, view: 170, Q&A: 0
    [Abstract]

    Infertility is a widespread and often unexplained issue. Studying reproduction using C. elegans males offers insight into the influence of individual factors on male fertility in humans. We have created a collection of protocols to assess several aspects of C. elegans sperm quality, including number, size, rate of activation, and mitochondrial

    ...
    Imaging and Fluorescence Quantification in Caenorhabditis elegans with Flow Vermimetry and Automated Microscopy
    Authors:  Elissa Tjahjono, Alexey V. Revtovich and Natalia V. Kirienko, date: 05/20/2021, view: 611, Q&A: 0
    [Abstract]

    Gene activation and cellular biomarkers are commonly monitored using fluorescent signals from transgenic reporters or dyes. These quantifiable markers are critical for biological research and serve as an incredibly powerful tool, even more so when combined with high-throughput screening. Caenorhabditis elegans is a particularly useful model in

    ...
    Identification of R-loop-forming Sequences in Drosophila melanogaster Embryos and Tissue Culture Cells Using DRIP-seq
    Authors:  Célia Alecki and Nicole J. Francis, date: 05/05/2021, view: 517, Q&A: 0
    [Abstract]

    R-loops are non-canonical nucleic structures composed of an RNA–DNA hybrid and a displaced ssDNA. Originally identified as a source of genomic instability, R-loops have been shown over the last decade to be involved in the targeting of proteins and to be associated with different histone modifications, suggesting a regulatory function. In

    ...
    Analysis of Monocyte Cell Fate by Adoptive Transfer in a Murine Model of TLR7-induced Systemic Inflammation
    Authors:  Jaba Gamrekelashvili, Hermann Haller and Florian P. Limbourg, date: 05/05/2021, view: 505, Q&A: 0
    [Abstract]

    Myeloid plasticity is a hallmark of the innate immune response to Toll-like receptor (TLR) activation. Here, we provide a protocol for monocyte cell fate tracking by adoptive transfer in the context of systemic inflammation induced by TLR7 activation, the principal innate immune receptor sensing viral RNA in mice. Defined monocyte subsets are

    ...
    Development of a Chemical Reproductive Aging Model in Female Rats
    [Abstract]

    Women are born with an abundant but finite pool of ovarian follicles, which naturally and progressively decreased during their reproductive years until menstrual periods stop permanently (menopause). Perimenopause represents the transition from reproductive to non-reproductive life. It is usually characterized by neuroendocrine, metabolic and

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