Developmental Biology

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    Protocols in Current Issue
    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: 526, 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

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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: 1417, 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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    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: 1817, 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

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    Analysis of TORC1-body Formation in Budding Yeast
    Authors:  Ryan L. Wallace, Eric Lu, Arron Sullivan, James E. Hughes Hallett and Andrew P. Capaldi, date: 04/05/2021, view: 1990, Q&A: 0
    [Abstract]

    The Target of Rapamycin kinase Complex I (TORC1) is the master regulator of cell growth and metabolism in eukaryotes. In the presence of pro-growth hormones and abundant nutrients, TORC1 is active and drives protein, lipid, and nucleotide synthesis by phosphorylating a wide range of proteins. In contrast, when nitrogen and/or glucose levels fall,

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    Ligand and Carbohydrate Engagement (LACE) Assay and Fluorescence Quantification on Murine Neural Tissue
    Authors:  James M. Clegg and Thomas Pratt, date: 03/20/2021, view: 2234, Q&A: 0
    [Abstract]

    The interaction between cell surface heparan sulphate and diffusible ligands such as FGFs is of vital importance for downstream signaling, however, there are few techniques that can be used to investigate this binding event. The ligand and carbohydrate engagement (LACE) assay is a powerful tool which can be used to probe the molecular interaction

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    Molecular and Phenotypic Characterization Following RNAi Mediated Knockdown in Drosophila
    Authors:  Saurabh Jayesh Kumar Mehta, Pradyumna A. Joshi and Ram Kumar Mishra, date: 02/20/2021, view: 2068, Q&A: 0
    [Abstract]

    Loss of function studies shed significant light on the involvement of a gene or gene product in different cellular processes. Short hairpin RNA (shRNA) mediated RNA interference (RNAi) is a classical yet straightforward technique frequently used to knock down a gene for assessing its function. Similar perturbations in gene expression can be

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    DigiTAG–a RNA Sequencing Approach to Analyze Transcriptomes of Rare Cell Populations in Drosophila melanogaster
    Authors:  Lisa Landskron, Francois Bonnay, Thomas R. Burkard and Jürgen A. Knoblich, date: 11/05/2020, view: 1626, Q&A: 0
    [Abstract]

    Cell-type specific transcriptional programs underlie the development and maintenance of organs. Not only distinct cell types within a tissue, even cells with supposedly identical cell fates show a high degree of transcriptional heterogeneity. Inevitable, low cell numbers are a major hurdle to study transcriptomes of pure cell populations. Here we

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    Production and Bioassay of a Diffusible Factor that Induces Gametophyte-to-Sporophyte Developmental Reprogramming in the Brown Alga Ectocarpus
    [Abstract] The brown alga Ectocarpus has a haploid-diploid life cycle that involves alternation between two multicellular generations, the sporophyte and the gametophyte. Life cycle generation is not determined by ploidy but by a genetic system that includes two different three amino acid loop extension homeodomain transcription factors called ...
    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: 1786, 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: 1753, 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.



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