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Simultaneous measurement of sleep and feeding in individual Drosophila

Abstract

Drosophila is widely used for the dissection of genetic and neuronal mechanisms of behavior. Recently, flies have emerged as a model for investigating the regulation of feeding and sleep. Although typically studied in isolation, increasing evidence points to a fundamental connection between these behaviors. Thus, a system for measuring sleep and feeding simultaneously in a single integrated system is important for interpreting behavioral shifts of either state. Here, we describe the construction and use of the Activity Recording Capillary Feeder or CAFE (ARC), a machine-vision (automated image tracking)-based system for the integrated measurement of sleep and feeding in individual Drosophila. Flies feed on liquid food from a microcapillary, and consumption is measured by tracking the liquid meniscus over time. Sleep measurements are obtained from positional tracking of individual animals, and arousal threshold can be determined by vibrational stimulus response. Using this system, a single computer and experimenter can track diverse behaviors from up to 60 individual flies in a single integrated system. The ARC is efficiently assembled with minimal training, and each experiment can be run for up to 7 d, with a total setup and breakdown time of 2 h.

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Figure 1: ARC schematic.
Figure 2: Arousal threshold measurement components.
Figure 3: ARC data capture.
Figure 4: Flow chart of ARC implementation of the JavaGrinders library.
Figure 5: Expected data from the ARC.

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Acknowledgements

We thank J. Jacobs and A. Sehgal for comments on the manuscript. This work was funded by the National Institutes of Health (R21DK092735 to W.W.J.).

Author information

Authors and Affiliations

Authors

Contributions

K.R.M., J.H.P., R.H., and W.W.J. contributed to the conception, development, and testing of the ARC; K.R.M., R.H., and W.W.J. wrote the manuscript; K.R.M., J.H.P., R.H., and W.W.J. revised the manuscript.

Corresponding authors

Correspondence to Robert Huber or William W Ja.

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Competing interests

The authors declare no competing financial interests.

Integrated supplementary information

Supplementary Figure 1 Comparison of data between Expresso and ARC.

(a) Total ingestion volume and calorie intake for 33 minutes on the indicated diet following 24 hours of starvation, measured in the Expresso. n = 20 male flies. Data and figure were adapted from Yapici et al. 2016. (b) Replication of Expresso experiment using the ARC. n = 15 Canton-S males. Box and whiskers represent mean with 95% confidence intervals; blue circles represent mean ± s.e.m. a adapted with permission from ref. 22, Elsevier.

Supplementary Figure 2 ARC data output structure.

Mock data portraying 7 reads from the ARC. Each row represents a single read and each column is organized by the repeating structure shown. Labels (red) do not appear in the output file, whereas data (black) will appear in a tab-delimited format. The final column (Stimulus Delivered) will only appear in the data file if the optional arousal threshold module is run.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1 and 2. (PDF 332 kb)

Supplementary Data

Compressed archive of ARC software. Archive contains seven .stl files for 3D printing of ARC components, jArduino firmware, the JavaGrinders framework and the ARC modules (ARCController and ARCControllerMultiCam), the Noah analysis program (Python-based), and a sample ARC data set of 30 Canton-S males over 2 d. (ZIP 28593 kb)

Video guide to using the ARC.

Overview of the procedure. (MP4 31023 kb)

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Murphy, K., Park, J., Huber, R. et al. Simultaneous measurement of sleep and feeding in individual Drosophila. Nat Protoc 12, 2355–2359 (2017). https://doi.org/10.1038/nprot.2017.096

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