Probing Drosophila visual behaviors in high throughput
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1
Howard Hughes Medical Institute, Janelia Farm Research Campus, United States
Janelia’s Fly Olympiad Project Team has developed several quantitative systems for measuring Drosophila behavior. The first and most complex behavioral apparatus we implemented, that we call “the box”, efficiently tests groups of flies in nearly-flat acrylic tubes, through a series of assays for basic locomotion, response to a mechanical startle, motion vision, photototaxis, and wavelength discrimination. The flies are filmed from above while the tubes are suspended above an infrared backlight. The experimental protocol is controlled by microprocessor controller that delivers stimuli with millisecond precision. The box is a light-tight enclosure in which the temperature can be rapidly switched. Our typical protocol is initially run at 24°C (the permissive temperature), and then at 34°C (the restrictive temperature for UAS-Shits1) and the same sequences are run on the same population of flies. The primary screen consists of GAL4 driver lines crossed to UAS-Shits1, with the goal of measuring the behavior consequence of the inactivation of small subsets of neurons. To date we have screened over 2200 lines from the Rubin lab’s GAL4 collection through the box. The assay is supported by a robust pipeline for video compression, fly tracking, data analysis, and database insertion that has been adapted to the other fly olympiad assays. The Fly Olympiad team has been reliably running over 80 lines per week through the box. We will present an analysis of the consistency of control behaviors and strategies we employ to compensate for the issues that limit this consistency over the several year screen duration.
The box was primarily designed to reliably measure robust features of fly visual behavior. While none of the implemented assays is novel, the integration of several well-established visual-motor behaviors into a single experimental apparatus readily enables comparative studies. Motion vision is presented via green LEDs along both sides of the tubes. By presenting grating patterns translating along the length of the tube at a range of speeds, we measure a following response from wild-type flies that is similar to the optomotor response of Drosophila measured from tethered flying flies. The behaviors even show a strong correspondence of the temporal frequency optimum of the motion response measured in these very different settings. Phototaxis and color preference are tested by presenting narrow-band light from single LEDs (one green and one UV) at each end of the tubes. Flies reliably walk towards either LED when illuminated, and in the UV versus green tests, show a preference for green light over dim UV that gives way to strong UV preference for increasingly brighter UV settings. For all visual behaviors, we designed the experimental protocol to produce tuning curves, so that the screen can reveal different categories of behavioral deficits. We will present a summary of all of the behavioral categories we have defined as well as specific examples of lines with behavioral deficits related to motion detection, UV-green preference, and phototaxis. The behavioral data, technology developed, and analysis methods will soon be made publicly available.
Acknowledgements
We wish to acknowledge the invaluable contributions of Nan Chen, Mary Phillips, Tanya Tabachnik, Magnus Karlsson, Chris Werner, Gus Lott, Lowell Umayam, Frank Midgley and Janelia’s Fly Core.
Keywords:
Color Vision,
Drosophila,
fly,
motion vision,
Neurogenetics,
walking behaviors
Conference:
Tenth International Congress of Neuroethology, College Park. Maryland USA, United States, 5 Aug - 10 Aug, 2012.
Presentation Type:
Poster (but consider for Participant Symposium)
Topic:
Sensory: Vision
Citation:
Reiser
MB,
Rowell
W,
Lee
A and
Korff
W
(2012). Probing Drosophila visual behaviors in high throughput.
Conference Abstract:
Tenth International Congress of Neuroethology.
doi: 10.3389/conf.fnbeh.2012.27.00420
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Received:
03 May 2012;
Published Online:
07 Jul 2012.
*
Correspondence:
Dr. Michael B Reiser, Howard Hughes Medical Institute, Janelia Farm Research Campus, Ashburn, VA, United States, reiserm@janelia.hhmi.org