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Visualizing olfactory receptor expression and localization in Drosophila.
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Methods in Molecular Biology
Odor detection and discrimination by olfactory systems in vertebrates and invertebrates depend both on the selective expression of individual olfactory receptor genes in subpopulations of olfactory sensory neurons, and on the targeting of the encoded proteins to the exposed, ciliated endings of sensory dendrites. Techniques to visualize the expression and localization of olfactory receptor gene products in vivo have been essential to reveal the molecular logic of peripheral odor coding and to permit investigation of the developmental and cellular neurobiology of this sensory system. Here, we describe methods for detection of olfactory receptor transcripts and proteins in the antennal olfactory organ of the fruit fly, Drosophila melanogaster, an important genetic model organism. We include protocols both for antennal cryosections and whole-mount antennae. These methods can be adapted for detection of receptor expression in other olfactory and gustatory tissues in Drosophila, as well as in the chemosensory systems of other insects.
Animals, Cryoultramicrotomy, Drosophila melanogaster/cytology, Drosophila melanogaster/metabolism, Gene Expression Regulation, Immunohistochemistry, In Situ Hybridization, Fluorescence, Insect Proteins/genetics, Insect Proteins/metabolism, Molecular Imaging/methods, Protein Transport, RNA Probes/genetics, RNA Probes/metabolism, Receptors, Odorant/genetics, Receptors, Odorant/metabolism, Tissue Fixation
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