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Analysis of locomotor activity rhythms in Drosophila

Abstract

The genetic, molecular and anatomical dissection of the circadian clock in Drosophila and other higher organisms relies on the quantification of rhythmic phenotypes. Here, we introduce the methods currently in use in our laboratories for the analysis of fly locomotor activity rhythms. This phenotype provides a relatively simple, automated, efficient, reliable and robust output for the circadian clock. Thus it is not surprising that it is the preferred readout for measuring rhythmicity under a variety of conditions for most fly clock laboratories. The procedure requires at least 10 days of data collection and several days for analysis. In this protocol we advise on fly maintenance and on experimental design when studying the genetics of behavioral traits. We describe the setup for studying locomotor activity rhythms in the fruit fly and we introduce the statistical methods in use in our laboratories for the analysis of periodic data.

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Figure 1: Setup for locomotor activity analysis.
Figure 2: Properties of rhythmic data.
Figure 3: Calculation of τ.
Figure 4: Drosophila activity monitors from TriKinetics: a single DAM2 monitor that will take 32 individual activity tubes.
Figure 5
Figure 6: Double plot of activity data using the 'Actogram Plotting' function in MAZ software.
Figure 7: CLEAN and autocorrelation analyses of an arrhythmic fly.
Figure 8: Activity profile of an arrhythmic fly.
Figure 9: Anchored phase-response curve for wild-type D. melanogaster

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Acknowledgements

We wish to thank M.A. Zordan for the implementation of the MAZ software. We also thank BBSRC, NERC, the Royal Society and the Wellcome Trust for funding work in our laboratories. C.P.K. acknowledges a Royal Society Wolfson Research Merit Award, and a grant from the 6th Framework Project EUCLOCK (No. 018741).

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Correspondence to Charalambos P Kyriacou.

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Rosato, E., Kyriacou, C. Analysis of locomotor activity rhythms in Drosophila. Nat Protoc 1, 559–568 (2006). https://doi.org/10.1038/nprot.2006.79

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