drosophila embryos as model systems for monitoring bacterial infection in real time果蝇胚胎作为模型系统,实时监测细菌感染.pdfVIP

Drosophila Embryos as Model Systems for Monitoring Bacterial Infection in Real Time 1. 2. 1 1 Isabella Vlisidou , Andrea J. Dowling , Iwan R. Evans , Nicholas Waterfield , Richard H. 2 1 ffrench-Constant , Will Wood * 1 Department of Biology and Biochemistry, University of Bath, Bath, United Kingdom, 2 School of Biological Sciences, University of Exeter in Cornwall, Penryn, United Kingdom Abstract 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 constit