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The present protocol assesses the locomotor activity of Drosophila by tracking and analyzing the movement of flies in a hand-made arena using open-source software Fiji, compatible with plugins to segment pixels of each frame based on high-definition video recording to calculate parameters of speed, distance, etc.
Drosophila melanogaster is an ideal model organism for studying various diseases due to its abundance of advanced genetic manipulation techniques and diverse behavioral features. Identifying behavioral deficiency in animal models is a crucial measure of disease severity, for example, in neurodegenerative diseases where patients often experience impairments in motor function. However, with the availability of various systems to track and assess motor deficits in fly models, such as drug-treated or transgenic individuals, an economical and user-friendly system for precise evaluation from multiple angles is still lacking. A method based on the AnimalTracker application programming interface (API) is developed here, which is compatible with the Fiji image processing program, to systematically evaluate the movement activities of both adult and larval individuals from recorded video, thus allowing for the analysis of their tracking behavior. This method requires only a high-definition camera and a computer peripheral hardware integration to record and analyze behavior, making it an affordable and effective approach for screening fly models with transgenic or environmental behavioral deficiencies. Examples of behavioral tests using pharmacologically treated flies are given to show how the techniques can detect behavioral changes in both adult flies and larvae in a highly repeatable manner.
Drosophila melanogaster provides an excellent model organism for investigating cellular and molecular functions in neuronal disease models created by gene modification1, drug treatment2, and senescence3. The high conservation of biological pathways, physical properties, and disease-associated homolog genes between humans and Drosophila makes the fruit fly an ideal mimic from the molecular to the behavioral level4. In many disease models, behavioral deficiency is an important index, providing a helpful model for various human neuropathies5....
W1118 adult flies and third instar larvae were used for the present study.
1. Experimental preparation
NOTE: An open-field arena for Drosophila locomotion tracking is made withacolorless and odorless silica gel.
In the present study, locomotor deficits in adult flies and third instar larvae treated with rotenone were examined and compared in their motor activity to that of a control fly fed with the drug solvent dimethyl sulfoxide (DMSO). Treatment with rotenone in Drosophila has been shown to cause dopaminergic neuron loss in the brain22 and lead to significant locomotor deficits23. As shown in Figure 11 and Figure 12
We have designed a method, based on the open-source material AnimalTracker API compatible with the Fiji image processing program, that can enable researchers to systematically evaluate locomotor activity by tracking both adult and individual larval flies. AnimalTracke is a tool written in Java that can be easily integrated into existing databases or other tools to facilitate the analysis of application-designed animal-tracking behavior24. Upon a frame-by-frame analysis by a softw.......
This work was supported by a special launch fund from Soochow University and the National Science Foundation of China (NSFC) (82171414). We thank Prof. Chunfeng Liu's lab members for their discussion and comments.
....Name | Company | Catalog Number | Comments |
Animal tracker | Hungarian Brain Research Program | version: 1.7 | pfficial website: http://animaltracker.elte.hu/main/downloads |
Camera software | Microsoft | version: 2021.105.10.0 | built-in windows 10 system |
Computer | DELL | Vostro-14-5480 | a comupter running win 10 system is available |
Drosophila carbon dioxide anesthesia workstation | Wu han Yihong technology | #YHDFPCO2-018 | official website: http://www.yhkjwh.com/ |
Fiji software | Fiji team | version: 1.53v | official website: https://fiji.sc/ |
Format factory software | Pcfreetime | version: X64 5.4.5 | official website: http://www.pcfreetime.com/formatfactory/CN/index.html |
Graph pad prism | GraphPad Software | version: 8.0.2 | official website: https://www.graphpad-prism.cn |
Hight definition camera | TTQ | Jingwang2 (HD1080P F1.6 6-60mm) | official website: http://www.ttq100.com/product_show.php?id=35 |
Office software | Microsoft | version: office 2019 | official website: https://www.microsoftstore.com.cn/software/office |
Petri dish | Bkman | 110301003 | size: 60 mm |
Silica gel | DOW | SYLGARD 184 Silicone Elastomer Kit | Mix well according to the instructions |
Sodium bicarbonate | Macklin | #144-55-8 | Mix well with silica gel |
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