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Effects of dsRNA-GFP as exogenous control on honey bee RNAi assays: gene expression changes in workers

ABSTRACT: A green fluorescence protein (GFP)-derived dsRNA (dsRNA-GFP) has been used as an exogenous control for Apis mellifera RNAi assays by multiple research groups. Its sequence does not share any significant homology with any known honey bee genes. Although dsRNA-GFP is not expected to trigger an RNAi response in treated bees, undesirable effects on gene expression, pupal pigmentation or developmental timing have been routinely observed. To better understand the multiple molecular and phenotypic effects of dsRNA-GFP in honey bees and to evaluate its use as a control for RNAi studies, we examined the impact of dsRNA-GFP on global gene expression patterns in developing workers. We found that dsRNA-GFP causes large-scale changes in gene expression associated with multiple biological processes. Furthermore, dsRNA-GFP exposure tended to preferentially decrease, rather than increase, expression of genes compared to controls.

ORGANISM(S): Apis mellifera

PROVIDER: GSE41004 | GEO | 2013/01/10

SECONDARY ACCESSION(S): PRJNA184875

REPOSITORIES: GEO

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Effects of dsRNA-GFP as exogenous control on honey bee RNAi assays: gene expression changes in workers

Project description:A green fluorescence protein (GFP)-derived dsRNA (dsRNA-GFP) has been used as an exogenous control for Apis mellifera RNAi assays by multiple research groups. Its sequence does not share any significant homology with any known honey bee genes. Although dsRNA-GFP is not expected to trigger an RNAi response in treated bees, undesirable effects on gene expression, pupal pigmentation or developmental timing have been routinely observed. To better understand the multiple molecular and phenotypic effects of dsRNA-GFP in honey bees and to evaluate its use as a control for RNAi studies, we examined the impact of dsRNA-GFP on global gene expression patterns in developing workers. We found that dsRNA-GFP causes large-scale changes in gene expression associated with multiple biological processes. Furthermore, dsRNA-GFP exposure tended to preferentially decrease, rather than increase, expression of genes compared to controls. Gene expression differences were analyzed using a dye-swap design. Second instar worker larvae received 0.5 M-NM-<g of dsRNA-GFP. Non-treated and dsRNA-GFP treated 7 day-old workers (W7d) were sampled.

2013-01-10 | E-GEOD-41004 | biostudies-arrayexpress

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Transcription profiling of genes related to ovary activation in worker honey bee brains

Project description:This experiment examines differences in gene expression between wildtype and an experimental strain (anarchistic) of worker honey bees (Apis mellifera). Mature wildtype and anarchistic workers tend to have non-active and active ovaries, respectively. Thus young workers from these strains are expected to show differential expression at loci involved in the regulation of worker reproduction, which occurs via arrhenotokous parthenogenesis.

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Trancriptome analysis of honey bees (Apis mellifera) infected with Nosema ceranae

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Project description:Extensive annual losses of honey bees (Apis mellifera L.) represent a global problem for agriculture and biodiversity. The parasitic mite Varroa destructor in association with viral co-infections plays a key role in this phenomenon; however, the precise mechanisms are still unclear. We employed a unique combination of transcriptomic, proteomic, metabolomic, and functional analyses to elucidate the effects of Varroa parasitisation. We focused on complex differences between parasitised and unparasitised ten-days old honey bee workers collected from identical colonies before overwintering. Honey bees exposed to mite parasitation during their development revealed alterations in transcriptome and proteome related to immunity, oxidative stress, olfactory recognition, metabolism of sphingolipids and RNA regulatory mechanisms. Specifically, immune reactions and sphingolipids metabolism were strongly up-regulated in parasitised honey bees; whereas olfactory recognition and oxidative stress pathways were down-regulated compared to unparasitised bees. Additionally, the metabolomic analysis confirmed the depletion of nutrients, decreased energy stores and generally disrupted metabolism of parasitised workers, as previously reported. By virtue of comprehensive omics-based analysis, we define the key changes in the honey bee facing Varroa parasitism and suggest possible mechanisms underlying its detrimental effects. This study provides a theoretical basis for future efforts in efficient control strategies against Varroa mites.

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