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Activation of an innate immune response in large numbers of permeabilized Drosophila embryos
Stockholm University, Faculty of Science, Department of Molecular Biology and Functional Genomics.
Stockholm University, Faculty of Science, Department of Molecular Biology and Functional Genomics.
2011 (English)In: Developmental and Comparative Immunology, ISSN 0145-305X, E-ISSN 1879-0089, Vol. 35, no 3, 263-266 p.Article in journal (Refereed) Published
Abstract [en]

Innate immunity in Drosophila involves the inducible expression and synthesis of antimicrobial peptides. We have previously shown that not only Drosophila larvae and adults, but also embryos, are capable of mounting an immune response after injection of bacterial substances. To simplify genetic dissection of the signaling pathways involved in immune-gene regulation we developed a procedure for permeabilization of large number of embryos and subsequent infiltration with bacterial fragments. This approach, which promoted expression of CecropinA1- and Diptericin-driven β-gal expression in the epidermis of more than 90% of the treated embryos, will enable analysis of mutants that are embryonic lethal. Thus, genes that are involved in essential pleiotrophic functions, in addition to being candidates in immune-regulation will be amenable for analysis of their involvement in the fly's immune defense.

Place, publisher, year, edition, pages
2011. Vol. 35, no 3, 263-266 p.
Keyword [en]
Drosophila; Innate immunity, Antimicrobial peptide, NF-kappaB, Genetic screens, Insects
National Category
Biological Sciences
URN: urn:nbn:se:su:diva-64253DOI: 10.1016/j.dci.2010.11.002ISI: 000289871800005PubMedID: 21075135OAI: diva2:456474
Available from: 2011-11-14 Created: 2011-11-14 Last updated: 2012-01-19Bibliographically approved
In thesis
1. Signaling pathways in Drosophila immunity
Open this publication in new window or tab >>Signaling pathways in Drosophila immunity
2011 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Drosophila relies on innate immunity to protect itself from its hostile environment throughout its life cycle. Despite the remarkable progress in understanding many aspects of Drosophila immunity, there are still big gaps in our knowledge. The general aim of this thesis was to gain a better understanding about the regulatory mechanisms controlling gene expression in Drosophila, with a focus on immunity. 

To enable isolation of Drosophila genes involved in immunity, we developed a method that allows visualization of immune gene expression in large number of embryos.  Reporter gene expression in wild type and mutant embryos was used to validate this approach, which should be a valuable complement to existing genetic and RNAi screens. 

Cactus, the Drosophila IκB protein, is known as a cytoplasmic inhibitor of Dif and Dorsal. We discovered that Cactus is also present in the cell nucleus. In response to Toll pathways signaling, cytoplasmic Cactus degrades rapidly in a proteasome-dependent manner, while a nuclear form of Cactus is stable and persists throughout signaling. This suggests that Cactus also has a function in the nucleus.

A genome-wide RNAi-based screen was performed in cultured S2 cells. Several novel components of NF-κB pathways were isolated as putative regulators of Drosophila immunity. One of them, the G protein-coupled receptor kinase-2 (Gprk2), was shown to be required for Drosomycin expression and for resistance to infection. Gprk2 interacts with Cactus, but is not required for Cactus degradation upon Toll pathway activation.  

The dpld/wech gene was previously found to affect periferal nervous system development. Here, we show that wech belongs to the LIN-41 subclade of the TRIM protein superfamily, and contains target sites for microRNAs. Genetic and cell transfection assays confirmed that wech expression is regulated by the microRNA let-7. This seems to be a conserved regulatory mechanism throughout the LIN-41 subclade. 

Place, publisher, year, edition, pages
Stockholm: Department of Molecular Biology and Functional Genomics, Stockholm University, 2011. 67 p.
Drosophila immunity, signaling pathways, Cactus, NFκB
National Category
Natural Sciences
Research subject
Molecular Biology
urn:nbn:se:su:diva-64256 (URN)978-91-7447-398-8 (ISBN)
Public defence
2011-12-15, Nordenskiöldsalen, Geovetenskapens hus, Svante Arrhenius väg 12, Stockholm, 10:00 (English)

At the time of the doctoral defense, the following paper was unpublished and had a status as follows: Paper 2: Manuscript.

Available from: 2011-11-23 Created: 2011-11-14 Last updated: 2013-12-06Bibliographically approved

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Esfahani, Shiva SeyedoleslamiEngström, Ylva
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