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Published October 22, 2018 | public
Journal Article Open

Imaging neural activity in the ventral nerve cord of behaving adult Drosophila


To understand neural circuits that control limbs, one must measure their activity during behavior. Until now this goal has been challenging, because limb premotor and motor circuits have been largely inaccessible for large-scale recordings in intact, moving animals—a constraint that is true for both vertebrate and invertebrate models. Here, we introduce a method for 2-photon functional imaging from the ventral nerve cord (VNC) of behaving adult Drosophila melanogaster. We use this method to reveal patterns of activity across nerve cord populations during grooming and walking and to uncover the functional encoding of moonwalker ascending neurons (MANs), moonwalker descending neurons (MDNs), and a previously uncharacterized class of locomotion-associated A1 descending neurons. Finally, we develop a genetic reagent to destroy the indirect flight muscles and to facilitate experimental access to the VNC. Taken together, these approaches enable the direct investigation of circuits associated with complex limb movements.

Additional Information

© 2018 The Author(s). This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. Received 13 March 2018; Accepted 02 October 2018; Published 22 October 2018. Data availability: The source data used for the analyses in this study are available on the Harvard Dataverse on a 'public repository'. Analysis code and sample datasets can be found 'here'. We thank B.J. Dickson (Janelia Research Campus, VA) for MDN-1-Gal4 and MAN-Gal4 fly strains. We thank G. Rubin (Janelia Research Campus, VA) for DNa01-Gal4, DNb06-Gal4, DNg13-Gal4, and DNg16-Gal4 fly strains. A.C. acknowledges support from the National Institutes of Health (R01HL124091). M.H.D. acknowledges support from the National Institute of Neurological Disorders and Stroke of the National Institutes of Health (U01NS090514). P.R. acknowledges support from the Swiss National Science Foundation (31003A_175667). Author Contributions: C.L.C. generated strains; performed experiments; analyzed data, L.H. performed experiments; analyzed data, M.C.V. generated strains; performed experiments; analyzed data, D.F. designed and implemented the motion-compensation algorithm F.A. analyzed data, M.U. designed the motion-compensation algorithm, A.C. designed and supervised the project, M.H.D. designed and supervised the project, P.R. conceived of, designed, and supervised the project; performed experiments; analyzed data, All authors contributed to writing the paper. The authors declare no competing interests.

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