Arthropods

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    Protocols in Current Issue
    Identification and Monitoring of Nucleotide Repeat Expansions Using Southern Blotting in Drosophila Models of C9orf72 Motor Neuron Disease and Frontotemporal Dementia
    Authors:  Joanne L. Sharpe, Nikki S. Harper and Ryan J. H. West, date: 05/20/2022, view: 18, Q&A: 0
    [Abstract]

    Repeat expansion diseases, including fragile X syndrome, Huntington’s disease, and C9orf72-related motor neuron disease and frontotemporal dementia, are a group of disorders associated with polymorphic expansions of tandem repeat nucleotide sequences. These expansions are highly repetitive and often hundreds to thousands of repeats in

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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: 378, 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: 1300, 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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    Combination of Immunofluorescence and Quantitative Fluorescence In-situ Hybridization for Analysing Differential Gene Expression in the Niche Cells of the Drosophila Lymph Gland
    Authors:  Parvathy Ramesh, Sushmit Ghosh and Lolitika Mandal, date: 01/20/2022, view: 1194, Q&A: 0
    [Abstract]

    The Drosophila larval haematopoietic organ or lymph gland consists of multiple cell types arranged in zones. The smallest stem cell compartment consists of 40-45 cells that constitute the haematopoietic niche. In order to analyse the haematopoietic niche, it needs to be labelled with a specific antibody to differentiate it from the other cell

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    Ex-vivo Microtubule Stability Assay Using Drosophila Wing Disc
    Authors:  Jung-Wan Mok and Kwang-Wook Choi, date: 12/05/2021, view: 1020, Q&A: 0
    [Abstract]

    Regulation of microtubule stability is crucial for diverse biological processes, including cell division, morphogenesis, and signaling. Various in vitro assays for microtubule stability have been developed to identify and characterize proteins involved in controlling microtubule stability. Here, we introduce a simple ex-vivo assay for identifying

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    Immunoprecipitation for Protein-Protein Interactions and for RNA Enrichment in Drosophila melanogaster
    Authors:  Giulia Romano, Raffaella Klima and Fabian Feiguin, date: 12/05/2021, view: 871, Q&A: 0
    [Abstract]

    To determine the molecular and functional interactions between RNA-binding proteins (RBPs) and their targets RNAs, is of fundamental importance to understand the dynamic organization of the nervous system in health and disease. Nevertheless, this task has remained elusive due to the lack of specific protocols and experimental systems that would

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    A low-cost Portable Device to Deliver Smoke, Volatile or Vaporized Substances to Drosophila melanogaster , Useful for Research and/or Educational Assays
    Authors:  Manuela Santalla, Ivana M. Gómez, Carlos A. Valverde and Paola Ferrero, date: 12/05/2021, view: 1340, Q&A: 0
    [Abstract]

    Drosophila melanogaster has been used to test drugs of abuse, substances with potential benefits for medical purposes, as well as contaminants and hazardous volatile compounds. This model has also been used for the characterization of behavioral changes, physiopathological consequences, and subcellular mechanisms of the use of cocaine,

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    Preparation of Drosophila Larval Blood Cells for Single-cell RNA Sequencing
    Authors:  Sudhir Gopal Tattikota and Norbert Perrimon, date: 08/20/2021, view: 1436, Q&A: 0
    [Abstract]

    Recent advances in single-cell RNA-sequencing (scRNA-seq) technologies provide unprecedented opportunities to identify new cell types and characterize cell states. One of the most important requirements for performing scRNA-seq is to obtain high-quality single cells in suspension. Recently, we used this approach to characterize Drosophila blood

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    Efficient and Rapid Analysis of Polysomes and Ribosomal Subunits in Cells and Tissues Using Ribo Mega-SEC
    [Abstract]

    Polysome profile analysis is a popular method for separating polysomes and ribosomal subunits and is typically achieved using a sucrose density gradient (SDG). This has remained the gold standard method since ribosomes were first discovered; however, this method is time-consuming and requires multiple steps from making the gradient and long

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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: 1982, 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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