retro-Tango enables versatile retrograde circuit tracing in Drosophila

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Abstract

Transsynaptic tracing methods are crucial tools in studying neural circuits. Although a couple of anterograde tracing methods and a targeted retrograde tool have been developed in Drosophila melanogaster, there is still need for an unbiased, user-friendly, and flexible retrograde tracing system. Here we describe retro-Tango, a method for transsynaptic, retrograde circuit tracing and manipulation in Drosophila. In this genetically encoded system, a ligand-receptor interaction at the synapse triggers an intracellular signaling cascade that results in reporter gene expression in presynaptic neurons. Importantly, panneuronal expression of the elements of the cascade renders this method versatile, enabling its use not only to test hypotheses but also to generate them. We validate retro-Tango in various circuits and benchmark it by comparing our findings with the electron microscopy reconstruction of the Drosophila hemibrain. Our experiments establish retro-Tango as a key method for circuit tracing in neuroscience research.

Data availability

The R code used for analysis is available at: https://github.com/anthonycrown/retrotango

Article and author information

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Altar Sorkaç

Department of Neuroscience, Brown University, Providence, United States

Competing interests
The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0002-0739-6314
Rareș A Moșneanu

Department of Neuroscience, Brown University, Providence, United States

Competing interests
The authors declare that no competing interests exist.
Anthony M Crown

Department of Neuroscience, Brown University, Providence, United States

Competing interests
The authors declare that no competing interests exist.
Doruk Savaş

Department of Neuroscience, Brown University, Providence, United States

Competing interests
The authors declare that no competing interests exist.
Angel M Okoro

Department of Neuroscience, Brown University, Providence, United States

Competing interests
The authors declare that no competing interests exist.
Ezgi Memiş

Department of Neuroscience, Brown University, Providence, United States

Competing interests
The authors declare that no competing interests exist.
Mustafa Talay

Department of Neuroscience, Brown University, Boston, United States

Competing interests
The authors declare that no competing interests exist.
Gilad Barnea

Department of Neuroscience, Brown University, Providence, United States

For correspondence
gilad_barnea@brown.edu

Competing interests
The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0001-6842-3454

Funding

National Institute of Mental Health (RF1MH123213)

Gilad Barnea

National Institute on Deafness and Other Communication Disorders (F31DC019540)

Anthony M Crown

Brown University Carney Institute for Brain Science (Suna Kirac Fund for Brain Science)

Doruk Savaş
Ezgi Memiş

Brown University Carney Institute for Brain Science (Graduate award in brain science)

Doruk Savaş

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Copyright

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.