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Transcuticular optical imaging of stimulus-evoked neural activities in the Drosophila peripheral nervous system

Nature Protocols volume 5, pages 1229–1235 (2010)Cite this article

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Abstract

The nervous system of Drosophila is widely used to study neuronal signal processing because the activities of neurons can be controlled and monitored by cell type–specific expression of genetically encoded actuator and sensor proteins. Measuring neural activities in adult flies, however, usually requires surgical approaches to penetrate the firm and pigmented cuticular exoskeleton. Interfering with this exoskeleton is critical in the case of the peripheral nervous system (PNS), as sensory neurons are often located directly beneath the cuticle and are associated with specialized stimulus-receiving and -conducting cuticular structures. In this article, we describe how the activities of these neurons can be probed nondestructively through the cuticle if a genetically encoded fluorescent protein sensor with strong baseline fluorescence is used. The method is exemplified for mechanosensory neurons in the adult antenna but can also be applied to many other PNS neurons, as is shown for the femoral chordotonal organ located in the fly's leg.

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Figure 1: Transcuticular visualization of neural activity in the Drosophila antenna.
Figure 2: Time course of calcium-dependent fluorescence changes.
Figure 3: Mechanically evoked calcium signals in femoral chordotonal organ (FCO) neurons.

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Acknowledgements

We thank J.T. Albert, E. Buchner, M. Dübbert, K. Ito, K. Öchsner and T. Völler for fly strains and technical support. This work was supported by the Alexander von Humboldt Foundation and the Japan Society for the Promotion of Science (to A.K.); the DFG Research Centre for the Molecular Physiology of the Brain, the Volkswagen-Foundation and the BMBF Bernstein Network for Computational Neuroscience (to M.C.G.); and the DFG (SFB 554 to A.F.).

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Author notes

  1. Azusa Kamikouchi and Robert Wiek: These authors contributed equally to this work.

Authors and Affiliations

  1. School of Life Sciences, Tokyo University of Pharmacy and Life Sciences, Hachioji, Tokyo, Japan

    Azusa Kamikouchi

  2. Department of Cellular Neurobiology, Georg-August-University of Göttingen, c/o Max Planck Institute for Experimental Medicine, Göttingen, Germany

    Robert Wiek, Thomas Effertz & Martin C Göpfert

  3. Department of Molecular Neurobiology of Behaviour, Georg-August-University of Göttingen, c/o European Neuroscience Institute, Göttingen, Germany

    André Fiala

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  1. Azusa Kamikouchi
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  3. Thomas Effertz
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  4. Martin C Göpfert
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Contributions

A.K. and M.C.G. designed the experiments; A.K., T.E., R.W. and A.F. conducted the experiments. A.K., M.C.G and A.F. wrote the article. M.C.G and A.F. supervised the work. All authors discussed the concepts and results, and commented on the article.

Corresponding authors

Correspondence to Martin C Göpfert or André Fiala.

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Kamikouchi, A., Wiek, R., Effertz, T. et al. Transcuticular optical imaging of stimulus-evoked neural activities in the Drosophila peripheral nervous system. Nat Protoc 5, 1229–1235 (2010). https://doi.org/10.1038/nprot.2010.85

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