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Profiling tissue-specific modulation of gene expression in response to insulin signalling in Drosophila

ABSTRACT: We investigated tissue-specific regulation of gene expression in a long lived Drosophila IIS mutant (dilp2-3,5) compared to its control (wDah). As in the majority of Drosophila laboratory strains, the endosymbiont Wolbachia was present; in the absence of Wolbachia, life extension through dilp2-3,5 is abrogated (Grönke et al. 2010). To control for this, we additionally included strains of the same genotypes that lacked Wolbachia (dilp2-3,5T, wDahT). We quantified gene expression concurrently on the level of the proteome and the transcriptome, in four key tissues: brain, gut, fat body, and muscle. Proteome quantification was carried out on five biological replicates per experimental group. Corresponding transcriptome quantification was carried out on three biological replicates per experimental group, and published on GEO (accession number *GSE122190*)

INSTRUMENT(S): Q Exactive

ORGANISM(S): Drosophila Melanogaster (fruit Fly)

TISSUE(S): Brain, Gut, Muscle, Thorax, Fat Body

SUBMITTER: Robert Sehlke

LAB HEAD: Linda Partridge

PROVIDER: PXD011589 | Pride | 2021-04-21

REPOSITORIES: pride

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Tissue-specific modulation of gene expression in response to lowered insulin signalling in <i>Drosophila</i>.

Tain Luke Stephen LS Sehlke Robert R Meilenbrock Ralf Leslie RL Leech Thomas T Paulitz Jonathan J Chokkalingam Manopriya M Nagaraj Nagarjuna N Grönke Sebastian S Fröhlich Jenny J Atanassov Ilian I Mann Matthias M Beyer Andreas A Partridge Linda L

eLife 20210421

Reduced activity of the insulin/IGF signalling network increases health during ageing in multiple species. Diverse and tissue-specific mechanisms drive the health improvement. Here, we performed tissue-specific transcriptional and proteomic profiling of long-lived <i>Drosophila dilp2-3,5</i> mutants, and identified tissue-specific regulation of >3600 transcripts and >3700 proteins. Most expression changes were regulated post-transcriptionally in the fat body, and only in mutants infected with th ...[more]

PMID: 33879316

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Project description:The Drosophila-Wolbachia system is being used to study the molecular nature of the interactions between a host and a symbiont. This system offers a unique opportunity for such a study since the Drosophila genome sequence is available, several Wolbachi genomes will also be available soon and there are at least three known Wolbachia strains infecting Drosophila: a) mod+ strain that induces cytoplasmic incompatibility, b) mod- strain that cannot induce cytoplasmic incompatibility, and c) popcorn strain, a virulent strain which reduces in half the adult lifespan of Drosophila due to its massive proliferation in adult brain. The Drosophila-Wolbachia interaction manifests itself in 3 main ways; first, destruction of the CNS in infected adults, second, induction of some kind of modification or imprinting in the male germ-line resulting in an early failure in embryonic development, (cytoplasmic incompatability (CI)) and third, modification of the female germ-line resulting in resistance to modified sperm. We are interested in identifying Drosophila genes with changes in expression due to Wolbachia infection. We have generated a series of isogenic fly lines (those being used in the IGF P-element project) which we have infected with Wolbachia strains, infection is readily cured by growth on medium containing tetracycline. Thus, we have equivalent genetic background with and without the parasite. We have tested all of the transgenic lines for the level of CI and find strain-specific levels ranging from 0-50%. We also have a strain of D. simulans that shows over 95% CI. Plan: For our initial experiments we wish to make 4 comparisons, in all cases 2 day old males will be collected and for each comparison we will isolate 3 independent biological replicates: Melanogaster no CI [tet] x Melanogaster no CI [+wol] Melanogaster high CI [tet] x Melanogaster high CI [+wol] We will therefore identify genes with changed expression levels in the male upon Wolbachia infection by comparing the melanogaster strains with high or no CI in the presence of tetracycline and Wolbachia. We also hope to identify similar genes in simulans (where we expect the magnitude of the effect to be larger), differences between melanogaster and simulans are controlled for in the mel v sim comparison.

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