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Long-term In Vivo Tracking of Inflammatory Cell Dynamics Within Drosophila Pupae

Weavers, H; Franz, A; Wood, W; Martin, P; (2018) Long-term In Vivo Tracking of Inflammatory Cell Dynamics Within Drosophila Pupae. Journal of Visualized Experiments , 136 , Article e57871. 10.3791/57871. Green open access

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Abstract

During the rapid inflammatory response to tissue damage, cells of the innate immune system are quickly recruited to the injury site. Once at the wound, innate immune cells perform a number of essential functions, such as fighting infection, clearing necrotic debris, and stimulating matrix deposition. In order to fully understand the diverse signaling events that regulate this immune response, it is crucial to observe the complex behaviors of (and interactions that occur between) multiple cell lineages in vivo, and in real-time, with the high spatio-temporal resolution. The optical translucency and the genetic tractability of Drosophila embryos have established Drosophila as an invaluable model to live-image and dissect fundamental aspects of inflammatory cell behavior, including mechanisms of developmental dispersal, clearance of apoptotic corpses and/or microbial pathogens, and recruitment to wounds. However, more recent work has now demonstrated that employing a much later stage in the Drosophila lifecycle – the Drosophila pupa – offers a number of distinct advantages, including improved RNAi efficiency, longer imaging periods, and significantly greater immune cell numbers. Here we describe a protocol for imaging wound repair and the associated inflammatory response at the high spatio-temporal resolution in live Drosophila pupae. To follow the dynamics of both re-epithelialization and inflammation, we use a number of specific in vivo fluorescent markers for both the epithelium and innate immune cells. We also demonstrate the effectiveness of photo-convertible fluorophores, such as Kaede, for following the specific immune cell subsets, to track their behavior as they migrate to, and resolve from, the injury site.

Type: Article
Title: Long-term In Vivo Tracking of Inflammatory Cell Dynamics Within Drosophila Pupae
Location: United States
Open access status: An open access version is available from UCL Discovery
DOI: 10.3791/57871
Publisher version: http://doi.org/10.3791/57871
Language: English
Additional information: © 2018 Creative Commons Attribution 3.0 License (https://creativecommons.org/licenses/by/3.0/).
Keywords: Animals, Drosophila, Inflammation, Microscopy, Confocal, Pupa
UCL classification: UCL
UCL > Provost and Vice Provost Offices
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences > Cell and Developmental Biology
URI: https://discovery.ucl.ac.uk/id/eprint/10073341
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