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Published July 11, 2024 | Version Published
Journal Article Open

Connectomic reconstruction of a female Drosophila ventral nerve cord

Creators

  • 1. ROR icon California Institute of Technology

Abstract

A deep understanding of how the brain controls behaviour requires mapping neural circuits down to the muscles that they control. Here, we apply automated tools to segment neurons and identify synapses in an electron microscopy dataset of an adult female Drosophila melanogaster ventral nerve cord (VNC)1, which functions like the vertebrate spinal cord to sense and control the body. We find that the fly VNC contains roughly 45 million synapses and 14,600 neuronal cell bodies. To interpret the output of the connectome, we mapped the muscle targets of leg and wing motor neurons using genetic driver lines2 and X-ray holographic nanotomography3. With this motor neuron atlas, we identified neural circuits that coordinate leg and wing movements during take-off. We provide the reconstruction of VNC circuits, the motor neuron atlas and tools for programmatic and interactive access as resources to support experimental and theoretical studies of how the nervous system controls behaviour.

Copyright and License

© The Author(s), under exclusive licence to Springer Nature Limited 2024.

Acknowledgement

The authors thank R. Wilson for financial support of S.G. during development of braincircuits.io (via U19NS104655 and R01NS129647 to R. Wilson); and R. Mann, H. Cheong, E. Ehrhardt, K. Eichler, G. Card and G. Jefferis for assistance with leg and wing MN identification. This work was supported by the beamtime LS3033 at ID16A beamline, ERC 852455 to A.P., a Searle Scholar Award, a Klingenstein-Simons Fellowship, a Pew Biomedical Scholar Award, a McKnight Scholar Award, a Sloan Research Fellowship, the New York Stem Cell Foundation, and a UW Innovation Award to J.C.T.; a Genise Goldenson Award to W.-C.A.L.; NIH U19NS104655 to J.C.T. and M.D.; NIH R01MH117808 to J.C.T., W.-C.A.L. and H.S.S. J.C.T. is a New York Stem Cell Foundation–Robertson Investigator.

Contributions

These authors contributed equally: Anthony Azevedo, Ellen Lesser, Jasper S. Phelps, Brandon Mark

These authors jointly supervised this work: Wei-Chung Allen Lee, John C. Tuthill

J.S.P., H.S.S., W.-C.A.L. and J.C.T. conceived the project. H.S.S., W.-C.A.L. and J.C.T. acquired funding. R.L., N.K., K.L., A.H., M.C., D.I., J.G., M.T. and C.S.J. performed the segmentation of electron microscopy data and supported the proofreading of the segmentation. S.D., F.C., C.S.-M. and D.B. deployed and supported the annotation software, CAVE. S.G. developed visualizations and analysis tools at braincircuits.io. A.K. manually traced leg MNs and annotated synapses in CATMAID. J.S.P. and W.-C.A.L. established guidelines for collaborative proofreading efforts. J.S.P. organized and oversaw the proofreading community. S.K. deployed code to identify cell nuclei and performed quality checks. A.A., E.L., J.S.P., B.M., L.E., A.M., A.K., C.J.D., S.A., S.-Y.J.L., B.P., A.C. and K.S.-K. proofread neurons in FANC and edited the paper. J.S.P., A.P. and A.A. gathered and annotated the XNH volume of the fly leg. A.S. designed and performed light microscopic imaging. A.A. identified leg MNs in FANC. E.L. identified wing MNs in FANC. M.D. advised the project and edited the paper. A.A., E.L., J.S.P., B.M. and L.E. analysed data. A.A., E.L. and J.C.T. wrote the paper, with input from the other authors.

Data Availability

The electron microscopy dataset and reconstructions are freely available1. The segmentation of the FANC dataset is available by joining the FANC community. Instructions on joining the FANC community can be found at https://github.com/htem/FANC_auto_recon/wiki#collaborative-community (ref. 80).

Code Availability

Details on code for alignment, segmentation, automatic detection of synapses and cell nuclei, and annotation software (CAVE) can be found via the citations in Methods, ‘Electron microscopy dataset alignment, segmentation and annotation’. Code for interacting with FANC can be found at https://github.com/htem/FANC_auto_recon (ref. 80). Code for analysis and figures is available at https://github.com/EllenLesser/Azevedo_Lesser_Phelps_Mark_2023/ (ref. 70).

Conflict of Interest

H.S.S. declares financial interests in Zetta AI. The remaining authors declare no competing interests.

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Additional details

Identifiers

PMCID
PMC11348827

Related works

Describes
Journal Article: https://rdcu.be/e5wfS (ReadCube)

Funding

National Institutes of Health
U19NS104655
National Institutes of Health
R01NS129647
European Research Council
852455
Chicago Community Trust
Searle Scholar
Esther A. & Joseph Klingenstein Fund
Simons Foundation
Pew Charitable Trusts
McKnight Brain Research Foundation
Alfred P. Sloan Foundation
New York Stem Cell Foundation
University of Washington
Harvard University
Genise Goldenson Fund
National Institutes of Health
U19NS104655
National Institutes of Health
R01MH117808

Dates

Available
2024-06-26
Version of record

Caltech Custom Metadata

Caltech groups
Division of Biology and Biological Engineering (BBE) , GALCIT , Tianqiao and Chrissy Chen Institute for Neuroscience
Publication Status
Published