The Tomato/GFP-FLP/FRT Method for Live Imaging of Mosaic Adult Drosophila Photoreceptor Cells |
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Authors: | Pierre Dourlen Clemence Levet Alexandre Mejat Alexis Gambis Bertrand Mollereau |
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Institution: | 1.Laboratory of Molecular Biology of the Cell, Ecole Normale Supérieure de Lyon;2.INSERM U744, Institut Pasteur de Lille, Université Lille-Nord de France;3.Howard Hughes Medical Institute, Laboratory of Apoptosis and Cancer, The Rockefeller University |
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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. |
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Keywords: | Developmental Biology Issue 79 Eye Photoreceptor Cells Genes Developmental neuron visualization degeneration development live imaging Drosophila photoreceptor cornea neutralization mitotic recombination |
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