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Tools allowing independent visualization and genetic manipulation of Drosophila melanogaster macrophages and surrounding tissues

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Gyoergy, Attila, Roblek, Marko, Ratheesh, Aparna, Valoskova, Katarina, Belyaeva, Vera, Wachner, Stephanie, Matsubayashi, Yutaka, Sánchez-Sánchez, Besaiz J., Stramer, Brian and Siekhaus, Daria E. (2018) Tools allowing independent visualization and genetic manipulation of Drosophila melanogaster macrophages and surrounding tissues. G3: Genes, Genomes, Genetics, 8 (3). g3.300452.2017. doi:10.1534/g3.117.300452 ISSN 2160-1836.

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Official URL: http://dx.doi.org/10.1534/g3.117.300452

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Abstract

Drosophila melanogaster plasmatocytes, the phagocytic cells among hemocytes, are essential for immune responses, but also play key roles from early development to death through their interactions with other cell types. They regulate homeostasis and signaling during development, stem cell proliferation, metabolism, cancer, wound responses, and aging, displaying intriguing molecular and functional conservation with vertebrate macrophages. Given the relative ease of genetics in Drosophila compared to vertebrates, tools permitting visualization and genetic manipulation of plasmatocytes and surrounding tissues independently at all stages would greatly aid a fuller understanding of these processes, but are lacking. Here, we describe a comprehensive set of transgenic lines that allow this. These include extremely brightly fluorescing mCherry-based lines that allow GAL4-independent visualization of plasmatocyte nuclei, the cytoplasm, or the actin cytoskeleton from embryonic stage 8 through adulthood in both live and fixed samples even as heterozygotes, greatly facilitating screening. These lines allow live visualization and tracking of embryonic plasmatocytes, as well as larval plasmatocytes residing at the body wall or flowing with the surrounding hemolymph. With confocal imaging, interactions of plasmatocytes and inner tissues can be seen in live or fixed embryos, larvae, and adults. They permit efficient GAL4-independent Fluorescence-Activated Cell Sorting (FACS) analysis/sorting of plasmatocytes throughout life. To facilitate genetic studies of reciprocal signaling, we have also made a plasmatocyte-expressing QF2 line that, in combination with extant GAL4 drivers, allows independent genetic manipulation of both plasmatocytes and surrounding tissues, and GAL80 lines that block GAL4 drivers from affecting plasmatocytes, all of which function from the early embryo to the adult.

Item Type: Journal Article
Subjects: Q Science > QL Zoology
Q Science > QP Physiology
Q Science > QR Microbiology
Divisions: Faculty of Science, Engineering and Medicine > Medicine > Warwick Medical School > Biomedical Sciences > Cell & Developmental Biology
Faculty of Science, Engineering and Medicine > Medicine > Warwick Medical School > Biomedical Sciences
Faculty of Science, Engineering and Medicine > Medicine > Warwick Medical School
Library of Congress Subject Headings (LCSH): Macrophages, Drosophila melanogaster -- Genetics, Blood cells, Small interfering RNA
Journal or Publication Title: G3: Genes, Genomes, Genetics
Publisher: Genetics Society of America
ISSN: 2160-1836
Official Date: 1 March 2018
Dates:
DateEvent
1 March 2018Published
31 December 2017Accepted
Volume: 8
Number: 3
Article Number: g3.300452.2017
DOI: 10.1534/g3.117.300452
Status: Peer Reviewed
Publication Status: Published
Access rights to Published version: Open Access (Creative Commons)
Date of first compliant deposit: 8 October 2018
Date of first compliant Open Access: 9 October 2018
RIOXX Funder/Project Grant:
Project/Grant IDRIOXX Funder NameFunder ID
2P40 OD-010949-10A1National Institutes of Healthhttp://dx.doi.org/10.13039/100000002
P40 OD-018537National Institutes of Healthhttp://dx.doi.org/10.13039/100000002
DASI_FWF01_P29638SAustrian Science Fundhttp://dx.doi.org/10.13039/501100002428
GA-2012-32950H2020 Marie Skłodowska-Curie Actionshttp://dx.doi.org/10.13039/100010665
LSC16_021NÖ Forschungs- und Bildungsges.m.b.H. (NFB)http://viaf.org/viaf/184958581
UNSPECIFIEDÖsterreichischen Akademie der WissenschaftenUNSPECIFIED
334077/IRTIMH2020 Marie Skłodowska-Curie Actionshttp://dx.doi.org/10.13039/100010665

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