Drosophila embryos as model systems for monitoring bacterial infection in real time.

Drosophila embryos are well studied developmental microcosms that have been used extensively as models for early development and more recently wound repair. Here we extend this work by looking at embryos as model systems for following bacterial infection in real time. We examine the behaviour of inj...

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Autores principales:	Isabella Vlisidou, Andrea J Dowling, Iwan R Evans, Nicholas Waterfield, Richard H ffrench-Constant, Will Wood
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2009
Materias:	Immunologic diseases. Allergy RC581-607 Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/3351161658954512bb23d2f79c5194cb
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spelling	oai:doaj.org-article:3351161658954512bb23d2f79c5194cb2021-11-25T05:47:46ZDrosophila embryos as model systems for monitoring bacterial infection in real time.1553-73661553-737410.1371/journal.ppat.1000518https://doaj.org/article/3351161658954512bb23d2f79c5194cb2009-07-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/19609447/pdf/?tool=EBIhttps://doaj.org/toc/1553-7366https://doaj.org/toc/1553-7374Drosophila embryos are well studied developmental microcosms that have been used extensively as models for early development and more recently wound repair. Here we extend this work by looking at embryos as model systems for following bacterial infection in real time. We examine the behaviour of injected pathogenic (Photorhabdus asymbiotica) and non-pathogenic (Escherichia coli) bacteria and their interaction with embryonic hemocytes using time-lapse confocal microscopy. We find that embryonic hemocytes both recognise and phagocytose injected wild type, non-pathogenic E. coli in a Dscam independent manner, proving that embryonic hemocytes are phagocytically competent. In contrast, injection of bacterial cells of the insect pathogen Photorhabdus leads to a rapid 'freezing' phenotype of the hemocytes associated with significant rearrangement of the actin cytoskeleton. This freezing phenotype can be phenocopied by either injection of the purified insecticidal toxin Makes Caterpillars Floppy 1 (Mcf1) or by recombinant E. coli expressing the mcf1 gene. Mcf1 mediated hemocyte freezing is shibire dependent, suggesting that endocytosis is required for Mcf1 toxicity and can be modulated by dominant negative or constitutively active Rac expression, suggesting early and unexpected effects of Mcf1 on the actin cytoskeleton. Together these data show how Drosophila embryos can be used to track bacterial infection in real time and how mutant analysis can be used to genetically dissect the effects of specific bacterial virulence factors.Isabella VlisidouAndrea J DowlingIwan R EvansNicholas WaterfieldRichard H ffrench-ConstantWill WoodPublic Library of Science (PLoS)articleImmunologic diseases. AllergyRC581-607Biology (General)QH301-705.5ENPLoS Pathogens, Vol 5, Iss 7, p e1000518 (2009)
institution	DOAJ
collection	DOAJ
language	EN
topic	Immunologic diseases. Allergy RC581-607 Biology (General) QH301-705.5
spellingShingle	Immunologic diseases. Allergy RC581-607 Biology (General) QH301-705.5 Isabella Vlisidou Andrea J Dowling Iwan R Evans Nicholas Waterfield Richard H ffrench-Constant Will Wood Drosophila embryos as model systems for monitoring bacterial infection in real time.
description	Drosophila embryos are well studied developmental microcosms that have been used extensively as models for early development and more recently wound repair. Here we extend this work by looking at embryos as model systems for following bacterial infection in real time. We examine the behaviour of injected pathogenic (Photorhabdus asymbiotica) and non-pathogenic (Escherichia coli) bacteria and their interaction with embryonic hemocytes using time-lapse confocal microscopy. We find that embryonic hemocytes both recognise and phagocytose injected wild type, non-pathogenic E. coli in a Dscam independent manner, proving that embryonic hemocytes are phagocytically competent. In contrast, injection of bacterial cells of the insect pathogen Photorhabdus leads to a rapid 'freezing' phenotype of the hemocytes associated with significant rearrangement of the actin cytoskeleton. This freezing phenotype can be phenocopied by either injection of the purified insecticidal toxin Makes Caterpillars Floppy 1 (Mcf1) or by recombinant E. coli expressing the mcf1 gene. Mcf1 mediated hemocyte freezing is shibire dependent, suggesting that endocytosis is required for Mcf1 toxicity and can be modulated by dominant negative or constitutively active Rac expression, suggesting early and unexpected effects of Mcf1 on the actin cytoskeleton. Together these data show how Drosophila embryos can be used to track bacterial infection in real time and how mutant analysis can be used to genetically dissect the effects of specific bacterial virulence factors.
format	article
author	Isabella Vlisidou Andrea J Dowling Iwan R Evans Nicholas Waterfield Richard H ffrench-Constant Will Wood
author_facet	Isabella Vlisidou Andrea J Dowling Iwan R Evans Nicholas Waterfield Richard H ffrench-Constant Will Wood
author_sort	Isabella Vlisidou
title	Drosophila embryos as model systems for monitoring bacterial infection in real time.
title_short	Drosophila embryos as model systems for monitoring bacterial infection in real time.
title_full	Drosophila embryos as model systems for monitoring bacterial infection in real time.
title_fullStr	Drosophila embryos as model systems for monitoring bacterial infection in real time.
title_full_unstemmed	Drosophila embryos as model systems for monitoring bacterial infection in real time.
title_sort	drosophila embryos as model systems for monitoring bacterial infection in real time.
publisher	Public Library of Science (PLoS)
publishDate	2009
url	https://doaj.org/article/3351161658954512bb23d2f79c5194cb
work_keys_str_mv	AT isabellavlisidou drosophilaembryosasmodelsystemsformonitoringbacterialinfectioninrealtime AT andreajdowling drosophilaembryosasmodelsystemsformonitoringbacterialinfectioninrealtime AT iwanrevans drosophilaembryosasmodelsystemsformonitoringbacterialinfectioninrealtime AT nicholaswaterfield drosophilaembryosasmodelsystemsformonitoringbacterialinfectioninrealtime AT richardhffrenchconstant drosophilaembryosasmodelsystemsformonitoringbacterialinfectioninrealtime AT willwood drosophilaembryosasmodelsystemsformonitoringbacterialinfectioninrealtime
_version_	1718414451357515776

Drosophila embryos as model systems for monitoring bacterial infection in real time.

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