DOI: 10.5061/dryad.ffbg79d5n
title: Control of odor sensation by light and cryptochrome in the Drosophila antenna

description: Olfaction is a sense employed by insects to differentiate safe from
 harmful food options, evaluate potential mates, and identify oviposition
 sites. Here, we found that the fruit fly, Drosophila melanogaster,
 responds differently to a set of repulsive odors depending on ambient
 light conditions. Ultraviolet (UV) or blue light reduces the behavioral
 aversion and electrophysiological responses of olfactory receptor neurons
 (ORNs) to certain repellent odors, such as benzaldehyde. We found that
 cryptochrome (cry) is strongly expressed in the antennal support cells
 that lie adjacent to ORNs, and mutation of cry eliminates the
 light-dependent reduction in aversion. Thus, these data indicate that
 support cells in an olfactory organ serve a sensory function as light
 responsive cells. It has been shown that light activation of Cry creates
 reactive oxygen species (ROS), and ROS activates the TRPA1 channel. We
 demonstrate that the TRPA1-C isoform is expressed and required in ORNs for
 benzaldehyde repulsion, and that TRPA1-C is activated in vitro by
 benzaldehyde. Overexpression of dual oxygenase, which generates hydrogen
 peroxide, reduced the aversion under dark conditions. Our data support the
 model that light-dependent creation of hydrogen peroxide persistently
 activates TRPA1-C. Consequently, the channel is no longer effectively
 activated by benzaldehyde. Since flies sleep much more at night, and begin
 feeding at dawn, we propose that the light-induced reduction in aversion
 to certain odors provides a mechanism to lower the barrier to feeding
 following the transition from night to day.
description: # Control of odor sensation by light and cryptochrome in the Drosophila
 antenna Dataset DOI: [10.5061/dryad.ffbg79d5n](10.5061/dryad.ffbg79d5n) ##
 Description of the data and file structure We tested the effects of
 ambient light on the olfactory behavior of Drosophila melanogaster. The
 following are video recordings, electrophysiology, immunohistochemistry
 and functional imaging that were performed to assay the olfactory behavior
 and physiology of Drosophila. ### Files and variables #### File:
 Odorant-cap.stl **Description:** 3D print file that prints the
 odorant-housing and assay-tube-capping implement used in the behavior
 assays in this work. This part is best printed using a resin-based printer
 on a fine precision setting. #### File: Imaging-Chamber.stl
 **Description:** 3D print file that prints the imaging chamber constructed
 to place one to two Drosophila antennae in place for functional imaging.
 This part is best printed using a resin-based printer in order to obtain
 smooth surface finish. Finishing the printed chamber for use requires
 piercing the narrowest point of the bottom of the chamber with a sharp
 pair of tweezers or a fine needle in order to create an aperture only
 large enough to securely hold a Drosophila antenna. A dissected antenna
 should be placed next to the bottom while creating this aperture for
 accuracy. The bottom of the chamber then is sealed with a standard clear
 coverslip held in place by vacuum grease and tape.  #### File: Funnel.stl
 **Description:** 3D print file that prints the funnel that straddles the
 lip of the glass tubes used for behavior assays in this work. This part
 can be printed using either a filament- or resin- based printer. The inner
 ridges of the funnel hold it against the tube securely while flies are
 tapped into the tube by rapidly inverting a fly vial into the open mouth
 of the funnel. #### File: Figure\_1.zip **Description:** This dataset
 contains video files with .avi extension that can be viewed in any
 standard video software and analyzed to obtain behavior metrics using the
 two Python code files included in this work - CoM_tracker.py and
 PI_tracker.py.  The raw videos are organized in sub-folders "Expt
 1" to "Expt 5", each corresponding to an experiment. Each
 experimental folder consists of 1 to 2 tubes containing 1, 2, 4, 8, 16, 32
 or 64 flies. #### File: Figure\_3.zip **Description:** This dataset
 contains electroantennograms recorded as .eag files that are readable in
 EAGPro (Ockenfels Syntech GmbH). This dataset also contains video files
 with .avi extension that can be viewed in any standard video software and
 analyzed to obtain behavior metrics using the two Python code files
 included in this work.  This dataset is organized in 4 subfolders.
 Subfolder "Figure 3AB" consists of electroantennograms recorded
 as .eag files corresponding to Figure 3A and 3B. Subfolder "Figure
 3C" consists of subfolders corresponding to the genotype of the flies
 tested: "Control", "ninaEI17", "norpAP24",
 "Rh3-2", "Rh4-1", "Rh7-1",
 "Rh7-L". Each subfolder corresponding to a genotype contains two
 subfolders - one for experiments performed in the dark (named
 "Dark") and the other for those performed under UV illumination
 (named "UV"), each of which contains raw video files in .avi
 format. Subfolder "Figure 3D" consists of subfolders
 corresponding to the genotype of the flies and the wavelength of light
 that they were tested in (for e.g. "cry01 365 nm" corresponds to
 cry01 mutant flies tested under 365 nm light illumination), each of which
 contains raw video files in .avi format. Subfolder "Figure 3EF"
 consists of electroantennograms recorded as .eag files corresponding to
 Figure 3E and 3F. #### File: Figure\_S2.zip **Description:** This folder
 contains a confocal imaging file in .czi format that can be viewed in
 Zeiss Zen software or ImageJ. The file is labeled according to the
 transgenes expressed by the flies. #### File: Figure\_S1.zip
 **Description:** This dataset contains video files with .avi extension
 that can be viewed in any standard video software and analyzed to obtain
 behavior metrics using the two Python code files included in this work.
 The dataset is arranged in subfolders corresponding to individual figures.
 Subfolder "Figure S1AC" contains raw video files in .avi format.
 Subfolder "Figure S1D" contains subfolders for each of the
 concentrations of benzaldehyde that were tested. For example, "1
 percent" contains raw video files in .avi format for which the flies
 were tested against 1 % benzaldehyde. "pt 1
 percent" contains raw video files in .avi format for which the flies
 were tested against 0.1 % benzaldehyde. Each subfolder corresponding to a
 concentration contains subfolders corresponding to different experiments
 conducted for the same concentration of benzaldehyde. Subfolder
 "Figure S1E" contains subfolders corresponding to different
 experiments conducted for either only male, only female or mixed
 populations of flies.  #### File: Figure\_S5.zip **Description:** This
 folder contains confocal imaging files in .czi format that can be viewed
 in Zeiss Zen software or ImageJ. The files are labeled according to the
 genotype of the flies and whether they were baseline recordings or after
 UV stimulation. For example "cry01 UV.czi" indicates a sample
 taken from cry01 mutant flies tested after UV stimulation. #### File:
 Figure\_2.zip **Description:** This dataset contains video files with .avi
 extension that can be viewed in any standard video software and analyzed
 to obtain behavior metrics using the two Python code files included in
 this work. The dataset is arranged in subfolders corresponding to
 individual figures. Subfolder "Figure 2A" contains subfolders
 which contain raw video files in .avi format. "Expt 1" and
 "Expt 2" subfolders contain .avi files corresponding to flies
 tested in the dark. "Expt 3" and "Expt 4" subfolders
 contain .avi files corresponding to flies tested under white light
 illumination. Subfolder "Figure 2B" consists of subfolders
 corresponding to each color of light that was used for illumination. For
 example subfolder "Blue" consists of two subfolders "Expt
 1" and "Expt 2", both of which contain raw video files in
 .avi format corresponding to experiments in which flies were exposed to
 blue light.  Subfolder "Figure 2C" consists of subfolders
 corresponding to each intensity of light that was used for illumination.
 For example subfolder "1 uWpercm2" consists of two subfolders
 "Expt 1" and "Expt 2", both of which contain raw video
 files in .avi format corresponding to experiments in which flies were
 exposed to 1 micro-watt/cm^2 of light intensity.  Subfolder "Figure
 2D" consists of subfolders corresponding to each intensity of light
 that was used for illumination. For example subfolder "18
 uWpercm2" consists of two subfolders "Expt 1" and
 "Expt 2", both of which contain raw video files in .avi format
 corresponding to experiments in which flies were exposed to 18
 micro-watt/cm^2 of light intensity.  Subfolder "Figure 2E"
 consists of subfolders corresponding to each concentration of benzaldehyde
 that the flies were exposed to. For example subfolder "1
 percent" consists of two subfolders "Dark" and
 "UV", both of which contain raw video files in .avi format
 corresponding to experiments in which flies were exposed to 1%
 benzaldehyde either to no light or to UV.  Subfolder "Figure 2F"
 consists of subfolders corresponding to the odor that the flies were
 exposed to. For example subfolder "AITC" consists of two
 subfolders "Dark" and "UV", both of which contain raw
 video files in .avi format corresponding to experiments in which flies
 were exposed to AITC either to no light or to UV.  #### File:
 Figure\_5.zip **Description:** This dataset contains video files with .avi
 extension that can be viewed in any standard video software and analyzed
 to obtain behavior metrics using the two Python code files included in
 this work. The dataset is arranged into subfolders corresponding to
 individual figures. Subfolder "Figure 5A" contains raw video
 files in .avi format organized by the genotype of the flies used. For
 example subfolder "Lush-GAL4-Cry01" contains video files of
 flies carrying lush-GAL4 and the cry01 mutant allele. Subfolder
 "Figure 5B" contains subfolders which contains raw imaging data
 in .czi format for control flies - "wt" or flies carrying the
 cry01 mutant allele - "cry01". Subfolder "Figure 5C"
 contains raw video files in .avi format organized by the genotype of the
 flies used. For example subfolder "Lush-GAL4 X UAS-Duox"
 contains video files of flies carrying the lush-GAL4 and UAS-Duox
 transgenes.  #### File: Figure\_7.zip **Description:** This dataset
 contains video files with .avi extension that can be viewed in any
 standard video software and analyzed to obtain behavior metrics using the
 two Python code files included in this work. This dataset also includes
 electrophysiological data recorded in .abf format that can be viewed in
 Clampex or Clampfit (Molecular Devices). The dataset is arranged into
 subfolders corresponding to individual figures. Subfolder "Figure
 7A" contains raw video files in .avi format organized by the genotype
 of the flies used. For example subfolder "trpA1-KO" contains
 video files of flies carrying the trpA1-KO mutant allele.  Subfolder
 "Figure 7B" contains raw video files in .avi format organized by
 the genotype of the flies used. For example subfolder "TRPA1-CD-GAL4
 X UAS-TRPA1C-TRPA1-1" contains video files of flies carrying the
 trpA1-GAL4 and UAS-trpA1-C and the trpa1-1 mutant allele.  Subfolder
 "Figure 7C" contains raw video files in .avi format organized by
 the genotype of the flies used. For example subfolder "Orco-GAL4 X
 Kir" contains video files of flies carrying the orco-GAL4 and UAS-Kir
 transgenes. Subfolders "Figure 7D-F" and "Figure 7G-I"
 each contain electrophysiological data recorded in .abf format. #### File:
 Figure\_S4.zip **Description:**  This folder contains confocal imaging
 files in .czi format that can be viewed in Zeiss Zen software or
 ImageJ. The files are labeled according to the transgenes expressed by the
 flies. For example, "ase-GAL4 x GFP-Cry-UASmcd8Dsred.czi" was
 the data collected from flies that expressed ase-GAL4, GFP-Cry and
 UASmcd8-Dsred. #### File: Figure\_6.zip **Description:**  This folder
 contains confocal imaging files in .lsm format that can be viewed in Zeiss
 Zen software or ImageJ. The files are labeled according to the transgenes
 expressed by the flies. For example, "TRPA1-CD-GAL4 X
 UAS-GFP.lsm" was the data collected from antennal lobes dissected
 from flies that expressed TRPA1-CD-GAl4 and UAS-GFP. #### File:
 Figure\_S6.zip **Description:**  This folder contains a confocal imaging
 file in .czi format that can be viewed in Zeiss Zen software or
 ImageJ. The file is labeled according to the transgenes expressed by the
 flies. #### File: Figure\_S7.zip **Description:** This dataset contains
 video files with .avi extension that can be viewed in any standard video
 software and analyzed to obtain behavior metrics using the two Python code
 files included in this work. The data are organized in subfolders
 corresponding to the odor that the flies were exposed to. Subfolder
 "Figure S7A" - corresponding to experiments in which flies were
 exposed to propionic acid either in the dark or under UV - consists of two
 subfolders "Dark" and "UV", both of which contain
 subfolders corresponding to experiments performed with propionic acid.
 These subfolders contain raw video files in .avi format. Subfolder
 "Figure S7B" - corresponding to experiments in which flies were
 exposed to R-limonene either in the dark or under UV - consists of two
 subfolders "Dark" and "UV", both of which contain
 subfolders corresponding to experiments performed with R-limonene. These
 subfolders contain raw video files in .avi format. Subfolder "Figure
 S7C" - corresponding to experiments in which flies were exposed to
 AITC either in the dark or under UV - consists of two subfolders
 "Dark" and "UV", both of which contain subfolders
 corresponding to experiments performed with AITC. These subfolders contain
 raw video files in .avi format. Subfolder "Figure S7D" -
 corresponding to experiments in which flies were exposed to citronellal
 either in the dark or under UV - consists of two subfolders
 "Dark" and "UV", both of which contain subfolders
 corresponding to experiments performed with citronellal. These subfolders
 contain raw video files in .avi format. Subfolder "Figure S7E" -
 corresponding to experiments in which flies were exposed to isovaleric
 acid either in the dark or under UV - consists of two subfolders
 "Dark" and "UV", both of which contain subfolders
 corresponding to experiments performed with isovaleric acid. These
 subfolders contain raw video files in .avi format. Subfolder "Figure
 S7F" - corresponding to experiments in which flies were exposed to
 lemongrass either in the dark or under UV - consists of two subfolders
 "Dark" and "UV", both of which contain subfolders
 corresponding to experiments performed with lemongrass. These subfolders
 contain raw video files in .avi format. #### File: Figure\_4.zip
 **Description:** This folder contains a confocal imaging file in .czi
 format that can be viewed in Zeiss Zen software or ImageJ. The files are
 labeled according to the transgenes expressed by the flies and the tissue
 that was imaged. For example, "LushGAL4 X GFP-Cry-UASmcd8dsred
 antenna" indicates flies expressing Lush-GAL4, GFP-Cry and
 UASmcd8-Dsred and the tissue that was imaged was the antenna.  #### File:
 Figure\_S3.zip **Description:** This folder contains confocal imaging
 files in .czi format that can be viewed in Zeiss Zen software or ImageJ.
 The files are labeled according to the transgenes expressed by the flies
 and the tissue that was imaged. For example, "OrcoRFP-UAS-GFP X
 Cry-GAL4 brain" indicates flies expressing Orco-RFP, UAS-GFP and
 Cry-GAL4 and the tissue that was imaged was the brain.  ## Code/software
 The two included python scripts - CoM_tracker and PI_tracker can be used
 to analyze the behavior data .avi video files. For this work, the scripts
 were run in Phycharm Community edition (JetBrains). The following python
 libraries are required: matplotlib, numpy, opencv, pandas and tkinter.
 Upon running either script, a dialog box appears, asking the user to
 specify the video files to be analyzed. A single folder in which all video
 files that need to be analyzed should be selected. The user is required to
 write the destination of the analysis files within the script. The output
 is in the form of .csv files saved in the destination provided by the
 user. The .csv files contain center of mass or preference index values
 that can be analyzed in standard spreadsheets and graphing software.  ##
 Access information Data was derived from the following source: * Data used
 in Supplementary information Figure S4 A-F was derived
 from [https://flycellatlas.org/](https://flycellatlas.org/)