Article | Published:

Optogenetic control of Drosophila using a red-shifted channelrhodopsin reveals experience-dependent influences on courtship

Nature Methods volume 11, pages 325332 (2014) | Download Citation

Abstract

Optogenetics allows the manipulation of neural activity in freely moving animals with millisecond precision, but its application in Drosophila melanogaster has been limited. Here we show that a recently described red activatable channelrhodopsin (ReaChR) permits control of complex behavior in freely moving adult flies, at wavelengths that are not thought to interfere with normal visual function. This tool affords the opportunity to control neural activity over a broad dynamic range of stimulation intensities. Using time-resolved activation, we show that the neural control of male courtship song can be separated into (i) probabilistic, persistent and (ii) deterministic, command-like components. The former, but not the latter, neurons are subject to functional modulation by social experience, which supports the idea that they constitute a locus of state-dependent influence. This separation is not evident using thermogenetic tools, a result underscoring the importance of temporally precise control of neuronal activation in the functional dissection of neural circuits in Drosophila.

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Acknowledgements

We thank K. Deisseroth (Stanford University) and B. Pfeiffer (Janelia Farm Research Campus) for plasmids. Fly stocks were generously provided by the Bloomington Stock Center, A. Fiala (Georg-August-Universität Göttingen), G.M. Rubin, L.L. Looger, B.J. Dickson (Janelia Farm Research Campus) and P.A. Garrity (Brandeis University). We also thank members of the Anderson lab for helpful discussion and sharing of flies. H.K.I. was supported by the Nakajima Foundation. J.Y.L. was funded by Foundation of Research, Science and Technology New Zealand. The project was supported by grants from the US National Institutes of Health to R.Y.T. (NS027177) and to D.J.A. (R01DA031389-03). D.J.A. and R.Y.T. are supported by the Howard Hughes Medical Institute.

Author information

Author notes

    • Hidehiko K Inagaki
    •  & Yonil Jung

    These authors contributed equally to this work.

Affiliations

  1. Howard Hughes Medical Institute, Pasadena, California, USA.

    • Hidehiko K Inagaki
    • , Yonil Jung
    • , Eric D Hoopfer
    • , Allan M Wong
    • , Roger Y Tsien
    •  & David J Anderson
  2. Division of Biology, California Institute of Technology, Pasadena, California, USA.

    • Hidehiko K Inagaki
    • , Yonil Jung
    • , Eric D Hoopfer
    • , Allan M Wong
    • , Neeli Mishra
    •  & David J Anderson
  3. Department of Pharmacology, University of California, San Diego, La Jolla, California, USA.

    • John Y Lin
    •  & Roger Y Tsien

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Contributions

H.K.I. and D.J.A. designed the experiments. H.K.I., Y.J. and N.M. performed behavioral experiments. H.K.I. and A.M.W. created the transgenic flies. H.K.I. performed physiological experiments. E.D.H. provided P1-GAL4. J.Y.L. and R.Y.T. provided the ReaChR reagent and advice on its biophysical properties. H.K.I. and Y.J. performed the data analysis. H.K.I. and D.J.A. prepared the figures and wrote the paper.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to David J Anderson.

Supplementary information

PDF files

  1. 1.

    Supplementary Text and Figures

    Supplementary Figures 1–3, Supplementary Tables 1–4 and Supplementary Note

Text files

  1. 1.

    Supplementary Software

    Software to control LED and camera for behavioral experiments

Videos

  1. 1.

    Activation of Gr5a neurons with ReaChR

    This movie shows representative PER behavior triggered by activation of Gr5a neurons with ReaChR. The first half of the movie shows activation with pulsing light-stimuli (100 msec pulse width, 1 Hz, 627 nm), and the last half of the movie shows activation with a continuous light-stimulus (627 nm). The white light appears on the right side of the movie is the light from the indicator IR LED (850 nm), which the flies cannot see.

  2. 2.

    Activation of HB9 GAL4 neurons with ReaChR

    This movie shows representative side-walking (first half of the movie) and paralysis (last half of the movie) triggered by activation of HB9-GAL4 neurons with ReaChR (continuous, 530 nm).

  3. 3.

    Activation of Crz GAL4 neurons with ReaChR

    This movie shows representative ejaculation behavior triggered by activation of Crz-GAL4 neurons with ReaChR (continuous, 530 nm). Note that Crz-GAL4; UAS-ReaChR flies bend their abdomen and extrude their genitals from the abdomen. At the end of movie (around 14 sec) the fly ejaculates and stops bending it abdomen. Note that although the control fly on the right (empty-GAL4; UAS-ReaChR) shows abdominal bending several times, it is less frequent and not accompanied by genital extrusion or ejaculation.

  4. 4.

    Activation of Fru GAL4 neurons with ReaChR

    This movie shows representative wing extension (first half of the movie) and paralysis (last half of the movie) triggered by activation of Fru-GAL4 neurons with ReaChR (continuous, 530 nm).

  5. 5.

    Activation of P1 and pIP10 neurons with ReaChR

    This movie shows representative wing extension behavior triggered by activation of P1 neurons (first half of the movie) and pIP10 neurons (last half of the movie) with ReaChR (continuous, 530 nm).

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DOI

https://doi.org/10.1038/nmeth.2765

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