Project description:The innate immune response of insects relies on several humoral and cellular mechanisms that require the activation of circulating proteases in the hemolymph to be functional. Here, we analyzed the gelatinase and caseinase activities of Drosophila larval hemolymph under normal and pathogenic conditions (bacterial lipopolysaccharides or endoparasitoid Leptopilina boulardi) using in gel zymography. Gelatinase activity was more intense than caseinase activity and qualitative and quantitative variations were observed between D. melanogaster strains and Drosophila species. Mass spectrometry identified a large number of serine proteases in gel bands equivalent to the major gelatinase and caseinase bands and of these, the most abundant and redundant were Tequila and members of the Jonah and Trypsin protease families. However, hemolymph from Tequila null mutant larvae showed no obvious changes in zymographic bands. Nor did we observe any significant changes in hemolymph gelatinases activity 24 h after injection of bacterial lipopolysaccharides or after oviposition by endoparasitoid wasps. These data confirmed that many serine proteases are present in Drosophila larval hemolymph but those with gelatinase and caseinase activity may not change drastically during the immune response.
Project description:The unsurpassed simplicity of the fruitfly’s airway epithelium, that is made of homogenous epithelial cells only, favours its use as a model to study general features and response characteristics of airway epithelia in general. All epithelial cells are able to launch an immune response as characterized by the expression of antimicrobial peptide genes. Infection induces a complex change in the expression profile of these epithelial cells. Outstanding are a priming of the immune system and the launch of a survival program, presumably to counteract infection induced apoptotic signals, which comprises the concurrent expression of known longevity genes such as dFoxo, and dThor. In regions of the airway epithelium with strong immune reactions, a complex remodelling of the airways can be observed, which is characterized by metaplasia and presumably also by hyperplasia of the affected epithelial cells. At the transcriptional level, this reorganization of the airway epithelium is mirrored by a recapitulation of genetic programs that are characteristic for early phases of airway development. Taken together, the response characteristics of the fly’s airway epithelium towards infections discloses features that are known from inflammatory diseases of the human lung, thus opening the opportunity to study fundamental aspects of these diseases in the fly. Keywords: Ectopic activation of IMD, larval airways, third instar larvae, Gal4/UAS-system, two-colour microarrays
Project description:The process of metamorphosis relies on choreography of temporal and spatial cues that mediate, in which inappropriate activation of many of these genes results in growth defects, developmental delay and death. Using RNA interference, we have unraveled an essential role for the insect specific dopamine 1(D1)-like receptor Dop1R2 at the late larval/prepupal stage. We used microarrays to evaluate differentially expressed genes during the early/pale pupal stage and identified up-regulation of families of functionally related genes, including ecdysone induced genes, stress and defense/immune response genes, and those that underlie tissue morphogenesis.
Project description:The process of metamorphosis relies on choreography of temporal and spatial cues that mediate, in which inappropriate activation of many of these genes results in growth defects, developmental delay and death. Using RNA interference, we have unraveled an essential role for the insect specific dopamine 1(D1)-like receptor Dop1R2 at the late larval/prepupal stage. We used microarrays to evaluate differentially expressed genes during the early/pale pupal stage and identified up-regulation of families of functionally related genes, including ecdysone induced genes, stress and defense/immune response genes, and those that underlie tissue morphogenesis. Drosophila Dop1R2 RNAi or w1118 control early/pale pupae were collected from crosses maintained at 29C. These tissues were processed for hybridization on Affymetrix microarrays (DrosGenome1). Results were used to assess differential expression of genes in Dop1R2 RNAi verses control flies.