This protocol is an excerpt from&#160;Gevedon&#160;et al.,&#160;In Vivo Forward Genetic Screen to Identify Novel Neuroprotective Genes in&#160;Drosophila melanogaster,&#160;J. Vis. Exp.&#160;(2019).
1. Climbing Assay (Adapted from Ali et al., J. Vis. Exp. (2011))
<ol>
	<li>Validate the climbing assay. In this study, test the climbing assay based on previously described methodology by Ali et al., using&#160;ElavC155&#62;UAS-TauR406W&#160;(exhibiting lower climbing pass rates) and&#160;ElavC155&#62;UAS-GFP&#160;(control) and&#160;ElavC155&#62;UAS-mCherry&#160;(control) flies.</li>
	<li>Flip the flies into a testing chamber, one line at a time (do not use CO2&#160;anesthesia) and allow them to recover for 5 min. If testing is done on different days, perform the assay at the same time of the day. In this study, all climbing assays were performed in the afternoon to control for circadian effects on locomotion. 
	NOTE&#160;Avoid exposing the flies to direct sunlight when testing; this will interfere with their ability to climb.</li>
	<li>Forcibly tap the chamber (marked side down) onto a mouse pad placed on a solid surface 3 times so all flies begin the assay at the bottom. Observe fly locomotion over a period of 10 s.</li>
	<li>Record the number of flies that reach or pass the 5 cm line within this time, as well as the total number of flies tested. Repeat the protocol, waiting 1 min between each trial, for a minimum of 3 trial replicates per line. 
	NOTE&#160;In the present study, each vial contained between 2 and 19 flies for the initial screening (male flies) and between 6 and 21 flies for the mutant deficiency analysis of crossed female lines.</li>
</ol>

<table><tbody><tr><td>VWR&amp;reg; Drosophila Vial; Narrow</td><td>VWR</td><td>75813-160</td><td></td></tr><tr><td>VWR&amp;reg; General-Purpose Laboratory Labeling Tape</td><td>VWR</td><td>89097-912</td><td></td></tr><tr><td>Standard mouse pad</td><td></td><td></td><td></td></tr><tr><td>Stereoscope</td><td>Motic</td><td></td><td>Model SMZ-168</td></tr><tr><td>Drosophila Incubator</td><td>VWR</td><td>89510-750</td><td></td></tr><tr><td>R software package</td><td></td><td></td><td></td></tr></tbody></table>

fly climbing assay: a method to test general locomotion in drosophila melanogaster

Fly Climbing Assay: A Method to Test General Locomotion in Drosophila melanogaster

Source: Gevedon, O., et al. <a href="https://www.jove.com/video/59720/in-vivo-forward-genetic-screen-to-identify-novel-neuroprotective">In Vivo Forward Genetic Screen to Identify Novel Neuroprotective Genes in Drosophila melanogaster</a>. J. Vis. Exp. (2019).
This video describes a simple method to quantify fly locomotion and shows the climbing assay performed.&#160; The fly climbing assay is a general test of motor function which relies on a negative geotaxis response.

Source: Gevedon, O., et al. In Vivo Forward Genetic Screen to Identify Novel Neuroprotective Genes in Drosophila melanogaster. J. Vis. Exp. (2019).
This ...

-First, obtain a set number of adult Drosophila. The testing chamber for the climbing assay is composed of two vials taped together. One of these vials should be marked with a line. Without carbon dioxide anesthesia, which can impair climbing behavior, transfer a set number of flies to the testing chamber. Allow the flies to recover for a predetermined amount of time. With the marked side of the chamber down, firmly tap the chamber on a mouse pad placed on a solid surface. This will force the flies to the bottom of the chamber. The climbing assay takes advantage of Drosophila's natural tendency to climb upwards against gravity, a behavior called negative geotaxis. During the trial, the flies shouldn't be exposed to direct sunlight because the flies might display phototaxis behavior, or movement in response to light stimulus. Record the number of flies that reach or pass the line. Then obtain the percent pass rate by dividing this number by the total number of flies tested. Allow the flies to recover before repeating the trial. In the example, we will observe a fly climbing assay being used to assess Drosophila motor function after chemical mutagenesis. -To begin, collect homozygous flies from lines belonging to the ENU mutagenized collection of Drosophila mutants mapped to the second chromosome. Flip the flies aged 10 to 12 days into a testing chamber without anesthesia one line at a time and allow them to recover for five minutes. With the five-centimeter line marked side down, forcibly tap the chamber three times on a mouse pad placed on a solid surface so that all flies begin the assay at the bottom. Observe fly locomotion over a period of 10 seconds. Record the number of flies that reach or pass the five-centimeter line within this time as well as the total number of flies tested. Wait one minute before starting a second trial. Repeat a minimum of three trial replicates per line.

fly-climbing-assay-method-to-test-general-locomotion-drosophila

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JoVE Encyclopedia of Experiments

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Drosophila melanogaster (fruit fly)

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14.0K Views. -First, obtain a set number of adult Drosophila. The testing chamber for the climbing assay is composed 

Fly Climbing Assay: A Method to Test General Locomotion in Drosophila melanogaster

Abstract

Protocol

<table><tbody><tr><td>VWR&reg; Drosophila Vial; Narrow</td><td>VWR</td><td>75813-160</td><td></td></tr><tr><td>VWR&reg; General-Purpose Laboratory Labeling Tape</td><td>VWR</td><td>89097-912</td><td></td></tr><tr><td>Standard mouse pad</td><td></td><td></td><td></td></tr><tr><td>Stereoscope</td><td>Motic</td><td></td><td>Model SMZ-168</td></tr><tr><td>Drosophila Incubator</td><td>VWR</td><td>89510-750</td><td></td></tr><tr><td>R software package</td><td></td><td></td><td></td></tr></tbody></table>

Fly Climbing Assay: A Method to Test General Locomotion in Drosophila melanogaster

In This Article

Abstract

Protocol

Materials

Tags

Name	Company	Catalog Number	Comments
VWR® Drosophila Vial; Narrow	VWR	75813-160
VWR® General-Purpose Laboratory Labeling Tape	VWR	89097-912
Standard mouse pad
Stereoscope	Motic		Model SMZ-168
Drosophila Incubator	VWR	89510-750
R software package