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Calcium imaging of odor-evoked responses in the Drosophila antennal lobe

Calcium imaging of odor-evoked responses in the Drosophila antennal lobe

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SILBERING, Ana, Rati BELL, Giovanni GALIZIA, Richard BENTON, 2012. Calcium imaging of odor-evoked responses in the Drosophila antennal lobe. In: Journal of Visualized Experiments(60). eISSN 1940-087X. Available under: doi: 10.3791/2976

@article{Silbering2012Calci-19023, title={Calcium imaging of odor-evoked responses in the Drosophila antennal lobe}, year={2012}, doi={10.3791/2976}, number={60}, journal={Journal of Visualized Experiments}, author={Silbering, Ana and Bell, Rati and Galizia, Giovanni and Benton, Richard} }

Silbering, Ana Galizia, Giovanni terms-of-use Bell, Rati First publ. in: Journal of Visualized Experiments : JoVE ; 61 (2012). - e2976 Benton, Richard eng The antennal lobe is the primary olfactory center in the insect brain and represents the anatomical and functional equivalent of the vertebrate olfactory bulb1-5. Olfactory information in the external world is transmitted to the antennal lobe by olfactory sensory neurons (OSNs), which segregate to distinct regions of neuropil called glomeruli according to the specific olfactory receptor they express. Here, OSN axons synapse with both local interneurons (LNs), whose processes can innervate many different glomeruli, and projection neurons (PNs), which convey olfactory information to higher olfactory brain regions. Optical imaging of the activity of OSNs, LNs and PNs in the antennal lobe - traditionally using synthetic calcium indicators (e.g. calcium green, FURA-2) or voltage-sensitive dyes (e.g. RH414) - has long been an important technique to understand how olfactory stimuli are represented as spatial and temporal patterns of glomerular activity in many species of insects6-10. Development of genetically-encoded neural activity reporters, such as the fluorescent calcium indicators G-CaMP11,12 and Cameleon13, the bioluminescent calcium indicator GFP-aequorin14,15, or a reporter of synaptic transmission, synapto-pHluorin16 has made the olfactory system of the fruitfly, Drosophila melanogaster, particularly accessible to neurophysiological imaging, complementing its comprehensively-described molecular, electrophysiological and neuroanatomical properties2,4,17. These reporters can be selectively expressed via binary transcriptional control systems (e.g. GAL4/UAS18, LexA/LexAop19,20, Q system21) in<br />defined populations of neurons within the olfactory circuitry to dissect with high spatial and temporal resolution how odor-evoked neural activity is<br />represented, modulated and transformed22-24.<br />Here we describe the preparation and analysis methods to measure odor-evoked responses in the Drosophila antennal lobe using G-CaMP25-27. The animal preparation is minimally invasive and can be adapted to imaging using wide-field fluorescence, confocal and two-photon microscopes. 2012-04-24T09:43:22Z 2012 Bell, Rati Silbering, Ana 2012-04-24T09:43:22Z Calcium imaging of odor-evoked responses in the Drosophila antennal lobe Benton, Richard Galizia, Giovanni

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