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Abstract

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Behavior

An Automated Method to Determine the Performance of Drosophila in Response to Temperature Changes in Space and Time

Published: October 12th, 2018

DOI:

10.3791/58350

1Groningen Institute for Evolutionary Life Sciences, University of Groningen, 2Department of Cell Biology, University of Groningen, University Medical Center Groningen, 3Ruijsink Dynamic Engineering, 4Department of Psychology, University of Groningen
* These authors contributed equally

Here we present a protocol to automatically determine the locomotor performance of Drosophila at changing temperatures using a programmable temperature-controlled arena that produces fast and accurate temperature changes in time and space.

Temperature is a ubiquitous environmental factor that affects how species distribute and behave. Different species of Drosophila fruit flies have specific responses to changing temperatures according to their physiological tolerance and adaptability. Drosophila flies also possess a temperature sensing system that has become fundamental to understanding the neural basis of temperature processing in ectotherms. We present here a temperature-controlled arena that permits fast and precise temperature changes with temporal and spatial control to explore the response of individual flies to changing temperatures. Individual flies are placed in the arena and exposed to pre-programmed temperature challenges, such as uniform gradual increases in temperature to determine reaction norms or spatially distributed temperatures at the same time to determine preferences. Individuals are automatically tracked, allowing the quantification of speed or location preference. This method can be used to rapidly quantify the response over a large range of temperatures to determine temperature performance curves in Drosophila or other insects of similar size. In addition, it can be used for genetic studies to quantify temperature preferences and reactions of mutants or wild-type flies. This method can help uncover the basis of thermal speciation and adaptation, as well as the neural mechanisms behind temperature processing.

Temperature is a constant environmental factor that affects how organisms function and behave1. Differences in latitude and altitude lead to differences in the type of climates organism are exposed to, which results in evolutionary selection for their responses to temperature2,3. Organisms respond to different temperatures through morphological, physiological, and behavioral adaptations that maximize performance under their particular environments4. For instance, in the fruit fly Drosophila melanogaster, populations from different regions have different ....

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1. Preparation of Fly Food Medium

  1. Pour 1 L of tap water into a 2 L glass beaker and add a magnetic stir bar. Put the beaker on a magnetic hot plate at 300 °C until boiling temperature is reached.
  2. Stir at 500 rounds/min and add the following: 10 g of agar, 30 g of glucose, 15 g of sucrose, 15 g of cornmeal, 10 g of wheat germ, 10 g of soy flour, 30 g of molasses, and 35 g of active dry yeast.
  3. When the mix foams vigorously, turn down the hot plate temperature to 120 °C while contin.......

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The temperature-controlled arena (Figure 1A) consists of three copper tiles whose temperature can be individually controlled through a programmable circuit. Each copper tile possesses a temperature sensor that gives feedback to the programmable circuit. The circuit activates a power supply to increase the temperature of each tile. Passive thermoelectric elements act as constant heating elements to maintain the desired temperature, while a heat sink cooled by .......

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Here we have presented an automated temperature-controlled arena (Figure 1) that produces precise temperature changes in time and space. This method allows exposure of individual Drosophila not only to pre-programmed gradual increases of temperature (Figure 2 and Figure 3), but also to dynamic temperature challenges in which each tile of the fly arena was heated independently to a different temperature (

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This work was supported in part by a scholarship from the Behavioural and Cognitive Neuroscience Program of the University of Groningen and a graduate scholarship from the Consejo Nacional de Ciencia y Tecnología (CONACyT) from Mexico, granted to Andrea Soto-Padilla, and a grant from the John Templeton Foundation for the study of time awarded to Hedderik van Rijn and Jean-Christophe Billeter. We are also thankful to Peter Gerrit Bosma for his participation in developing the FlySteps tracker.

Scripts TemperaturePhases,FlySteps, and FlyStepAnalysis can be found as supplementary information and in the following ....

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Name Company Catalog Number Comments
Arduino Due Arduino A000062 Software RUG
Electronics Board Ruijsink Dynamic Engineering FF-Main-02-2014
Power supply Boost XP-Power 48. V 65 W ECS65US48 Set to 53 Volt
Power supply Tile Heating XP-Power 15. V 80 W VFT80US15
Power supply Cooling XP-Power 15. V 130 W ECS130U515
Peltier elements Marlow Industries RC12-4 2 Elements, controlled DC feed
Heat sink Fisher Technik LA 9/150-230V Decoupled for vibration
Temperature sensors Measurement Specialties MCD_10K3MCD1 Micro Thermistor Probe
Copper block/tiles Ruijsink Dynamic Engineering FF-CB-01-2014
Auminum ring Ruijsink Dynamic Engineering FF-RoF-02-2015
Tesa 4104 white tape 25 x 66 mm RS Components 111-2300  White conductive tape
Red LEDs Lucky Ligt ll-583vc2c-v1-4da Wavelength between 625 nm, 20 mAmp and 6 V
Warm white LED strip Ledstripkoning HQ-3528-SMD 60 LEDs per meter
Switch Power Supply Generic T-36-12
Logitech c920 Logitech Europe S.A PN960-001055
QuickTime Player Apple Computer Recording program
Tracking analysis software R Packages: pacman
Tracking analysis software MATLAB
Thermal Imaging FLIR T400sc
Graphs and Statisticts Software Graph Pad Prism
Sigmacote Sigma-Aldrich SL2-100ML Siliconising agent
Fly rearing bottles Flystuff 32-130 6oz Drosophila stock bottle
Flypad Flystuff 59-114
Fly rearing vials Dominique Dutscher 789008 Drosophila tubes narrow 25x95 mm
Incubator Sanyo MIR-154
Magnetic hot plate Heidolph 505-20000-00 MR Hei-Standard
Agar Caldic Ingredients B.V. 010001.26.0
Glucose Gezond&wel 1019155 Dextrose/Druivensuiker
Sucrose Van Gilse Granulated sugar
Cornmeal Flystuff 62-100
Wheat germ Gezond&wel 1017683
Soy flour Flystuff 62-115
Molasses Flystuff 62-117
Active dry yeast Red Star
Tegosept Flystuff 20-258 100%

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