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Specification of the Drosophila Orcokinin A neurons by combinatorial coding

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Abstract

The central nervous system contains a daunting number of different cell types. Understanding how each cell acquires its fate remains a major challenge for neurobiology. The developing embryonic ventral nerve cord (VNC) of Drosophila melanogaster has been a powerful model system for unraveling the basic principles of cell fate specification. This pertains specifically to neuropeptide neurons, which typically are stereotypically generated in discrete subsets, allowing for unambiguous single-cell resolution in different genetic contexts. Here, we study the specification of the OrcoA-LA neurons, characterized by the expression of the neuropeptide Orcokinin A and located laterally in the A1-A5 abdominal segments of the VNC. We identified the progenitor neuroblast (NB; NB5-3) and the temporal window (castor/grainyhead) that generate the OrcoA-LA neurons. We also describe the role of the Ubx, abd-A, and Abd-B Hox genes in the segment-specific generation of these neurons. Additionally, our results indicate that the OrcoA-LA neurons are “Notch Off” cells, and neither programmed cell death nor the BMP pathway appears to be involved in their specification. Finally, we performed a targeted genetic screen of 485 genes known to be expressed in the CNS and identified nab, vg, and tsh as crucial determinists for OrcoA-LA neurons. This work provides a new neuropeptidergic model that will allow for addressing new questions related to neuronal specification mechanisms in the future.

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Data Availability

The data that support the findings of this study are available from the corresponding author upon reasonable request.

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Acknowledgements

We are grateful to the Developmental Studies Hybridoma Bank at the University of Iowa and The Bloomington Stock Center for sharing antibodies, fly lines, and DNAs.

Funding

This work was supported by a grant from the MINECO (BFU2016-78327-P) to J.B-S and The University of Queensland, Australia, to ST.

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Authors and Affiliations

  1. Departamento de Biología, Facultad de Ciencias, Universidad Autónoma de Madrid Cantoblanco, E 28049, Madrid, Spain

    Irene Rubio-Ferrera, Luis Clarembaux-Badell, Pablo Baladrón-de-Juan, Miguel Ángel Berrocal-Rubio, Ignacio Monedero Cobeta & Jonathan Benito-Sipos

  2. Departamento de Fisiología, Facultad de Medicina, Universidad Autónoma de Madrid Cantoblanco, E 28049, Madrid, Spain

    Ignacio Monedero Cobeta

  3. Department of Anatomy and Physiology, The University of Melbourne, Melbourne, VIC, Australia

    Miguel Ángel Berrocal-Rubio

  4. School of Biomedical Sciences, The University of Queensland, Brisbane, QLD, 4072, Australia

    Stefan Thor

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  1. Irene Rubio-Ferrera
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Contributions

IFRC, PBD, LCB, and MBR performed experiments. IFRC, PBD, and LCB performed the statistical analysis. IMC conducted the analysis of the RNA-seq data. IFRC, PBD, LCB, IMC, ST, and JBS compiled the figures. ST, IMC, and JBS planned and supervised the project. IMC, ST, and JBS wrote the manuscript.

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Correspondence to Ignacio Monedero Cobeta or Jonathan Benito-Sipos.

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Rubio-Ferrera, I., Clarembaux-Badell, L., Baladrón-de-Juan, P. et al. Specification of the Drosophila Orcokinin A neurons by combinatorial coding. Cell Tissue Res 391, 269–286 (2023). https://doi.org/10.1007/s00441-022-03721-x

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