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Methods for quantifying simple gravity sensing in Drosophila melanogaster

Abstract

Perception of gravity is essential for animals: most animals possess specific sense organs to detect the direction of the gravitational force. Little is known, however, about the molecular and neural mechanisms underlying their behavioral responses to gravity. Drosophila melanogaster, having a rather simple nervous system and a large variety of molecular genetic tools available, serves as an ideal model for analyzing the mechanisms underlying gravity sensing. Here we describe an assay to measure simple gravity responses of flies behaviorally. This method can be applied for screening genetic mutants of gravity perception. Furthermore, in combination with recent genetic techniques to silence or activate selective sets of neurons, it serves as a powerful tool to systematically identify neural substrates required for the proper behavioral responses to gravity. The assay requires 10 min to perform, and two experiments can be performed simultaneously, enabling 12 experiments per hour.

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Figure 1: Setup of the countercurrent apparatus.
Figure 2: Manipulation of the countercurrent apparatus.
Figure 3: Negative gravitaxis of wild-type flies with intact and aristae-ablated antennae.

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Acknowledgements

We thank Y. Hotta for the original countercurrent apparatus. This study was supported by the Alexander von Humboldt Foundation and the Japan Society for the Promotion of Science (to A.K.) and by the Human Frontier Science Program Organization, BIRD/Japan Science and Technology Agency and the Japan Society for the Promotion of Science (to K.I.). This paper is dedicated to the late Seymour Benzer.

Author information

Authors and Affiliations

Authors

Contributions

H.K.I., A.K. and K.I. designed experiments; H.K.I. performed experiments; H.K.I., A.K. and K.I. wrote the paper; and K.I. supervised the work. All authors discussed the concepts and results and commented on the paper.

Corresponding author

Correspondence to Kei Ito.

Supplementary information

Supplementary Video 1

Handling of the countercurrent apparatus. Actual experiments are performed in pitch darkness. Under bright light condition, flies show both negative-gravitaxis and phototaxis, leading to a high partition coefficient Cf. Note that some parts of the movie are played at 10x speed (QuickTime; 4.2 MB). (MOV 4315 kb)

Supplementary Audio 1

Audio file to be used as timer signals at pitch darkness. (WAV 1492 kb)

Supplementary Fig. 1

Drawing of the countercurrent apparatus. (PDF 52 kb)

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Inagaki, H., Kamikouchi, A. & Ito, K. Methods for quantifying simple gravity sensing in Drosophila melanogaster. Nat Protoc 5, 20–25 (2010). https://doi.org/10.1038/nprot.2009.196

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