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DsGFP as exogenous control on RNAi assays and its effects on gene expression in Apis mellifera

ABSTRACT: Double stranded RNAs are used to induce gene silencing in functional studies. In Apis mellifera green fluorescent protein (GFP) dsRNA (dsGFP) has been used as an exogenous control as its sequence has no homology in honeybee genome. However, some undesirable effects are observed after dsGFP treatment. A microarray approach comparing gene expression differences between untreated and dsGFP treated groups, containing honeybees workers in two different developmental stages, pre-pupae and light-brown eyed pupae, were used to test the reliability of dsGFP as a control for RNAi experiments. According to these microarrays results dsGFP can be used as a control in RNAi assays as long as the affected genes are taken into account in the analysis.

ORGANISM(S): Apis mellifera

PROVIDER: GSE43193 | GEO | 2013/01/10

SECONDARY ACCESSION(S): PRJNA184873

REPOSITORIES: GEO

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dsGFP as exogenous control on RNAi assays and its effects on gene expression in Apis mellifera

Project description:Double stranded RNAs are used to induce gene silencing in functional studies. In Apis mellifera green fluorescent protein (GFP) dsRNA (dsGFP) has been used as an exogenous control as its sequence has no homology in honeybee genome. However, some undesirable effects are observed after dsGFP treatment. A microarray approach comparing gene expression differences between untreated and dsGFP treated groups, containing honeybees workers in two different developmental stages, pre-pupae and light-brown eyed pupae, were used to test the reliability of dsGFP as a control for RNAi experiments. According to these microarrays results dsGFP can be used as a control in RNAi assays as long as the affected genes are taken into account in the analysis. Analysis used loop-design to compare dsGFP treated and noon-treated workers in pre-pupae and light-brown-eyed pupae stages.

2013-01-10 | E-GEOD-43193 | biostudies-arrayexpress

Comparison of Apis mellifera workers and queens brain development at fourth instar larvae

Project description:In Apis mellifera, the female eggs can develop into workers or queen depending on the diet offered during early development. The outputs of the developed honeybee females are two morphs with particular morphological traits and related physiology. The differential feeding regime experienced by the queen and the worker larvae of the honeybee Apis mellifera shapes a complex endocrine response cascade that ultimately sets up differences in brain morphologies. Herein we report on aspects of brain morphogenesis during larval development and the brain gene expression signature of fourth instar larvae (L4) of both castes, a developmental stage characterized by the greatest differences in juvenile hormone (JH) titers between castes Using results from the hybridization of whole genome-based oligonucleotide arrays with RNA samples from brain of fourth instar larvae honeybees of both castes we present a list of differentially expressed genes.

2012-12-31 | GSE39239 | GEO

Comparison of Apis mellifera workers and queens brain development at fourth instar larvae

2012-12-31 | E-GEOD-39239 | biostudies-arrayexpress

Comparison of Apis mellifera workers and queens hind leg development at prepupal stages

Project description:In Apis mellifera, the female eggs can develop into workers or queen depending on the diet offered during early development. The outputs of the developed honeybee females are two morphs with particular morphological traits and related physiology. Among the specialized structures in workers the hind tibia forms the corbicula or the pollen basket, a smooth region surrounded by a row of a long scopal hairs, used for carrying pollen and other materials to the nest. This morphological trait and the respective behaviour are absent in queens. Herein we show details of the initial steps of hind legs morphogenesis in honeybee castes. Using results from the hybridization of whole genome-based oligonucleotide arrays with RNA samples from hind leg imaginal discs of pre-pupa honeybees of both castes we present a list of differentially expressed genes.

2012-02-15 | GSE34293 | GEO

Comparison of Apis mellifera workers and queens hind leg development at prepupal stages

Project description:In Apis mellifera, the female eggs can develop into workers or queen depending on the diet offered during early development. The outputs of the developed honeybee females are two morphs with particular morphological traits and related physiology. Among the specialized structures in workers the hind tibia forms the corbicula or the pollen basket, a smooth region surrounded by a row of a long scopal hairs, used for carrying pollen and other materials to the nest. This morphological trait and the respective behaviour are absent in queens. Herein we show details of the initial steps of hind legs morphogenesis in honeybee castes. Using results from the hybridization of whole genome-based oligonucleotide arrays with RNA samples from hind leg imaginal discs of pre-pupa honeybees of both castes we present a list of differentially expressed genes. Analysis used one dye-swap combination to compare workers and queens hind leg development at prepupal stages when juvenile hormone titers is much higher in queens

2012-02-15 | E-GEOD-34293 | biostudies-arrayexpress

Diversity of populations of Deformed wing virus in the honeybee (Apis mellifera) pupae overtly infected by injection of DWV virus preparations from overtly and covertly infected honeybees to haemolymph.

Project description:Experiment was designed (i) to analyse the strain composition of Deformed wing virus (DWV) populations in covertly and overtly infected honeybees (Apis mellifera) from Varroa-free and Varroa-infested colonies, and (ii) to determine abundance of the DWV strains following direct injection of the DWV preparations from covertly and overtly infected bees to the bee pupae haemolymph in the absence of Varroa destructor mites. Experiment included isolation of DWV preparations from the following bees: covertly-infected bees from Varroa-free colony, covertly infected bees exposed orally to the Varroa-selected DWV strains, and the overtly infected Varroa-exposed bees. Honeybee pupae were experimentally injected with those DWV preparations and sampled 4 days post injection following development of overt DWV infection. A series of the DWV cDNA fragment covering complete DWV genomic RNA sequences were amplified by RT-PCR using RNA extracted from virus preparations and the injected pupae. The cDNA preparations were sequenced using next generation(Illumina HighSeq 2000) paired-end sequencing to obtain data on the DWV strain composition.

2013-09-01 | E-MTAB-1675 | biostudies-arrayexpress

Honey bee: Apis mellifera syriaca

Project description:Apis mellifera syriaca is the native honeybee subspecies of Jordan and much of the Middle East. It expresses behavioral adaptations to a regional climate with very high temperatures, nectar dearth in summer, attacks of the Oriental wasp Vespa orientalis and in most cases it is resistant to varroa mites. The Thorax control sample of A. m. syriaca in this experiment was originally collected and stored since 2001 from Wadi Ben Hammad a remote valley in the southern region of Jordan. Using morphometric and Mitochondrial DNA markers it was proved that bees from this area had show higher similarity than other samples collected from the Middle East as represented by reference samples collected in 1952 by Brother Adam. The samples L1-L5 are collected from the National Center for Agricultural Research and Extension breading apiary which was originally established for the conservation of Apis mellifera syriaca. Goal was to use the genetic information in the breeding for varroa resistant bees and to determine the successfulness of this conservation program. Project funded by USAID-MERC grant number: TA-MOU-09-M29-075.

2016-02-01 | GSE43554 | GEO

Transcription profiling by array of brains from Apis mellifera mellifera and Apis mellifera ligustica honey bees of different ages

Project description:Expression profiling of honey bees brains as a function of genotype ( Apis mellifera mellifera/Apis mellifera ligustica) and age

2010-12-31 | E-TABM-952 | biostudies-arrayexpress

Apis mellifera mellifera

Project description:RNAi DWV Apis mellifera

| PRJNA336281 | ENA

Transcriptome analysis of Apis mellifera antennae reveals molecular divergence underlying the division of labor in worker bees

Project description:Olfaction system plays a fundamental role in mediating insect behavior. Besides, the division of queen, worker and drone, honeybee also exhibit an age-dependent division of labor. Worker bees perform discrete sets of behaviors throughout their lifespan. These behavioral states rely on the sense of the environments and chemical communications via their olfactory system - antennae. However, the olfactory adaption mechanism of workers in these processes of behavioral development is still unclear. In this study, we conducted a comprehensive and quantitative analysis of gene expression in Apis mellifera antenna of newly emerged workers, nurses, foragers, and defenders using RNA-seq. We found that antennae tissues continue to develop after transformation from pupae to adult. Additionally, we identified both developmental and labor-division specific expressed genes. We validated the unexpected discovery of major royal jelly protein genes, which are highly and specifically expressed in nurse honeybee workers. We further identified and validated that significant alternative splicing events are also involved in the development and division of labor. These findings provided a comprehensive transcriptome profile and new perspective into the molecular mechanism underlying honeybee division of labor.

2024-10-24 | GSE221246 | GEO

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