Developmental Biology

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    Protocols in Current Issue
    Quantitative Live Confocal Imaging in Aquilegia Floral Meristems
    Authors:  Ya Min, Stephanie J. Conway and Elena M. Kramer, date: 06/20/2022, view: 928, Q&A: 0
    [Abstract]

    In this study, we present a detailed protocol for live imaging and quantitative analysis of floral meristem development in Aquilegia coerulea, a member of the buttercup family (Ranunculaceae). Using confocal microscopy and the image analysis software MorphoGraphX, we were able to examine the cellular growth dynamics during floral organ primordia

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    RNA Interference Method for Gene Function Analysis in the Japanese Rhinoceros Beetle Trypoxylus dichotomus
    Authors:  Kazuki Sakura, Shinichi Morita and Teruyuki Niimi, date: 04/20/2022, view: 814, Q&A: 0
    [Abstract]

    In the Japanese rhinoceros beetle Trypoxylus dichotomus, various candidate genes required for a specific phenotype of interest are listed by next-generation sequencing analysis. Their functions were investigated using RNA interference (RNAi) method, the only gene function analysis tool for T. dichotomus developed so far. The summarized procedure

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    Measurement of Contractile Ring Tension Using Two-photon Laser Ablation during Drosophila Cellularization
    Authors:  Swati Sharma and Richa Rikhy, date: 03/20/2022, view: 476, Q&A: 0
    [Abstract]

    Cytokinesis occurs at the final step of cell division and leads to the separation of daughter cells. It requires assembly and constriction of the actomyosin contractile ring. The phases of assembly and constriction of the contractile ring show an increase in tension in the actomyosin complex. The measurement of tension in the contractile ring is

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    Labeling and Tracking Mitochondria with Photoactivation in Drosophila Embryos
    Authors:  Sayali Chowdhary and Richa Rikhy, date: 03/05/2022, view: 1475, Q&A: 0
    [Abstract]

    Mitochondria are relatively small, fragmented, and abundant in the large embryos of Drosophila, Xenopus and zebrafish. It is essential to study their distribution and dynamics in these embryos to understand the mechanistic role of mitochondrial function in early morphogenesis events. Photoactivation of mitochondrially tagged GFP (mito-PA-GFP) is

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    Total Nucleic Acid Extraction from Single Zebrafish Embryos for Genotyping and RNA-seq
    Authors:  Neha Wali, Munise Merteroglu, Richard J. White and Elisabeth M. Busch-Nentwich, date: 01/05/2022, view: 1700, Q&A: 0
    [Abstract]

    RNA sequencing allows for the quantification of the transcriptome of embryos to investigate transcriptional responses to various perturbations (e.g., mutations, infections, drug treatments). Previous protocols either lack the option to genotype individual samples, or are laborious and therefore difficult to do at a large scale. We have developed a

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    Whole-mount Immunohistochemistry to Visualize Mouse Embryonic Dermal Lymphatic Vasculature
    Authors:  Xin Geng and R. Sathish Srinivasan, date: 10/20/2021, view: 1060, Q&A: 0
    [Abstract]

    Lymphatic vessels are abundant in the skin where they regulate interstitial fluid uptake and immune surveillance. Defects in dermal lymphatic vessels, such as fewer vessels and abnormal lymphatic vessel coverage with mural cells, are frequently associated with lymphedema and other lymphatic disorders. Whole-mount immunohistochemistry allows the

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    Phalloidin Staining of Actin Filaments for Visualization of Muscle Fibers in Caenorhabditis elegans
    Authors:  Mario Romani and Johan Auwerx, date: 10/05/2021, view: 2077, Q&A: 0
    [Abstract]

    Advances in C. elegans research have allowed scientists to recapitulate different human disorders, from neurodegenerative diseases to muscle dysfunction, in these nematodes. Concomitantly, the interest in visualizing organs affected by these conditions has grown, leading to the establishment of different antibody- and dye-based staining protocols

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    En masse DNA Electroporation for in vivo Transcriptional Assay in Ascidian Embryos
    Author:  Sébastien Darras, date: 09/20/2021, view: 1584, Q&A: 0
    [Abstract]

    Ascidian embryos are powerful models for functional genomics, in particular, due to the ease of generating a large number of transgenic embryos by electroporation. In addition, the small size of their genome makes them an attractive model for studying cis-regulatory elements that control gene expression during embryonic development. Here, I

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    Inter-species Transplantation of Blastocysts between Medaka and Zebrafish
    Authors:  Jana F. Fuhrmann, Jasmin Onistschenko and Lázaro Centanin, date: 09/20/2021, view: 1382, Q&A: 0
    [Abstract]

    Transplantation of blastocysts from a donor to a host blastula constitutes a powerful experimental tool to tackle major developmental biology questions. The technique is widely implemented in diverse biological models including teleost fish, where it is typically used for intra-species blastula transplantations – i.e., labeled blastocysts

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    Direct-TRI: High-throughput RNA-extracting Method for all Stages of Zebrafish Development
    Authors:  Kota Ujibe, Kanako Nishimura, Makoto Kashima and Hiromi Hirata, date: 09/05/2021, view: 1798, Q&A: 0
    [Abstract]

    Recent popularization of next-generation sequencing enables conducting easy transcriptome analysis. Nevertheless, substantial RNA isolation work prior to RNA sequencing, as well as the high cost involved, still makes the routine use of large-scale transcriptome analysis difficult. For example, conventional phenol-chloroform RNA extraction cannot

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