The tomato/GFP-FLP/FRT method for live imaging of mosaic adult Drosophila photoreceptor cells

Pierre Dourlen, Clemence Levet, Alexandre Mejat, Alexis Gambis, Bertrand Mollereau

    Research output: Contribution to journalArticle

    Abstract

    The Drosophila eye is widely used as a model for studies of development and neuronal degeneration. With the powerful mitotic recombination technique, elegant genetic screens based on clonal analysis have led to the identification of signaling pathways involved in eye development and photoreceptor (PR) differentiation at larval stages. We describe here the Tomato/GFP-FLP/FRT method, which can be used for rapid clonal analysis in the eye of living adult Drosophila. Fluorescent photoreceptor cells are imaged with the cornea neutralization technique, on retinas with mosaic clones generated by flipase-mediated recombination. This method has several major advantages over classical histological sectioning of the retina: it can be used for high-throughput screening and has proved an effective method for identifying the factors regulating PR survival and function. It can be used for kinetic analyses of PR degeneration in the same living animal over several weeks, to demonstrate the requirement for specific genes for PR survival or function in the adult fly. This method is also useful for addressing cell autonomy issues in developmental mutants, such as those in which the establishment of planar cell polarity is affected.

    Original languageEnglish (US)
    Article numbere50610
    JournalJournal of Visualized Experiments
    Issue number79
    DOIs
    StatePublished - Sep 20 2013

    Fingerprint

    Photoreceptor Cells
    Lycopersicon esculentum
    Drosophila
    Imaging techniques
    Genetic Recombination
    Retina
    Screening
    Animals
    Genes
    Throughput
    Cell Polarity
    Kinetics
    Diptera
    Cornea
    Clone Cells

    Keywords

    • Cornea neutralization
    • Degeneration
    • Development
    • Developmental
    • Developmental Biology
    • Drosophila
    • Eye
    • Genes
    • Issue 79
    • Live imaging
    • Mitotic recombination
    • Neuron
    • Photoreceptor
    • Photoreceptor Cells
    • Visualization

    ASJC Scopus subject areas

    • Neuroscience(all)
    • Chemical Engineering(all)
    • Immunology and Microbiology(all)
    • Biochemistry, Genetics and Molecular Biology(all)

    Cite this

    The tomato/GFP-FLP/FRT method for live imaging of mosaic adult Drosophila photoreceptor cells. / Dourlen, Pierre; Levet, Clemence; Mejat, Alexandre; Gambis, Alexis; Mollereau, Bertrand.

    In: Journal of Visualized Experiments, No. 79, e50610, 20.09.2013.

    Research output: Contribution to journalArticle

    Dourlen, Pierre ; Levet, Clemence ; Mejat, Alexandre ; Gambis, Alexis ; Mollereau, Bertrand. / The tomato/GFP-FLP/FRT method for live imaging of mosaic adult Drosophila photoreceptor cells. In: Journal of Visualized Experiments. 2013 ; No. 79.
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