Oral Bacterial Infection and Shedding in Drosophila melanogaster

Autor: Siva-Jothy, Jonathon A, Prakash, Arun, Vasanthakrishnan, Radhakrishnan B, Monteith, Katy M, Vale, Pedro F
Přispěvatelé: University of Edinburgh, Institut de Génétique et Développement de Rennes (IGDR), Université de Rennes (UR)-Centre National de la Recherche Scientifique (CNRS)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), 095831, Wellcome Trust, United Kingdom, Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )-Centre National de la Recherche Scientifique (CNRS)-Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)
Jazyk: angličtina
Rok vydání: 2018
Předmět:
Zdroj: Journal of visualized experiments : JoVE
Journal of visualized experiments : JoVE, 2018, 135, ⟨10.3791/57676⟩
Journal of visualized experiments : JoVE, JoVE, 2018, ⟨10.3791/57676⟩
Siva-Jothy, J, Prakash, A, Vasanthakrishnan, R B, Monteith, K M & Vale, P 2018, ' Oral Bacterial Infection and Shedding in Drosophila Melanogaster ', Journal of Visualized Experiments (JoVE), vol. 135, e57676 . https://doi.org/10.3791/57676
Journal of Visualized Experiments : JoVE
ISSN: 1940-087X
DOI: 10.3791/57676⟩
Popis: The fruit fly Drosophila melanogaster is one of the best developed model systems of infection and innate immunity. While most work has focused on systemic infections, there has been a recent increase of interest in the mechanisms of gut immunocompetence to pathogens, which require methods to orally infect flies. Here we present a protocol to orally expose individual flies to an opportunistic bacterial pathogen (Pseudomonas aeruginosa) and a natural bacterial pathogen of D. melanogaster (Pseudomonas entomophila). The goal of this protocol is to provide a robust method to expose male and female flies to these pathogens. We provide representative results showing survival phenotypes, microbe loads, and bacterial shedding, which is relevant for the study of heterogeneity in pathogen transmission. Finally, we confirm that Dcy mutants (lacking the protective peritrophic matrix in the gut epithelium) and Relish mutants (lacking a functional immune deficiency (IMD) pathway), show increased susceptibility to bacterial oral infection. This protocol, therefore, describes a robust method to infect flies using the oral route of infection, which can be extended to the study of a variety genetic and environmental sources of variation in gut infection outcomes and bacterial transmission.
Databáze: OpenAIRE
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