Abstract
In adult Drosophila, most of the hemocytes are macrophage-like cells (so called plasmatocytes), which serve various functions in organ homeostasis and immune defense. Ontogeny and functions are largely conserved between vertebrate and invertebrate macrophages. Hence, Drosophila offers a powerful genetic toolbox to study macrophage function and genetically modulate these cells. Technological advances in high-throughput sequencing approaches allowed to give an in-depth characterization of vertebrate macrophage populations and their heterogenous composition within different organs as well as changes in disease. Embryonic and larval hemocytes in Drosophila have been recently analyzed in single-cell RNA-sequencing (scRNA-seq) approaches during infection and steady state. These analyses revealed anatomical and functional Drosophila hemocyte subtypes dedicated to specific tasks. Only recently, the Fly Cell Atlas provided a whole transcriptomic single-cell atlas via single-nuclei RNA-sequencing (snRNA-seq) of adult Drosophila including many different tissues and cell types where hemocytes were also included. Yet, a specific protocol to isolate nuclei from adult hemocytes for snRNA-seq and study these cells in different experimental conditions was not available. In this chapter, we give a detailed protocol to purify hemocyte nuclei from adult Drosophila, which can be used in subsequent analyses such as snRNA-seq.
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Change history
22 December 2023
A correction has been published.
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Acknowledgments
We would like to acknowledge the Lighthouse Core Facility, Medical faculty, University of Freiburg and its staff, especially J. Bodinek-Wersing and U. Jagadeshwaran, for their assistance with the sorting of nuclei. The Kierdorf lab is supported by project grants of the Fritz Thyssen Foundation and of the DFG. The Kierdorf lab is further supported by the DFG through project grants within SFB/TRR167 (Project ID 259373024), CRC1479 (Project ID 441891347) and within Germany’s Excellence Strategy (grant no. CIBSS—EXC-2189, Project ID 390939984). Research in the Paeschke laboratory is fund by an ERC Stg Grant (638988-G4DSB) and by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) – Project-ID 369799452 – TRR237” as well as by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) under Germany’s Excellence Strategy – EXC2151 – 390873048.
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Hersperger, F., Kastl, M., Paeschke, K., Kierdorf, K. (2024). Hemocyte Nuclei Isolation from Adult Drosophila melanogaster for snRNA-seq. In: Mass, E. (eds) Tissue-Resident Macrophages. Methods in Molecular Biology, vol 2713. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3437-0_4
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DOI: https://doi.org/10.1007/978-1-0716-3437-0_4
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