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Gene targets and developmental regulation of the honeybee Apis mellifera ftz-f1

ABSTRACT: Ftz-f1 (Fushi tarazu transcription factor 1) is an orphan member of the nuclear hormone receptor superfamily, with one of its transcriptional variant (βFtz-f) being homolog of the vertebrate SF1. Ftz-f1 gene is induced after a 20E pulse, and allows for the expression of down-stream genes in processes leading to major switching in development, like those of metamorphosis. In honeybees, the expression of some genes (e.g. vitellogenin, vg; pro-phenoloxidase, pro-Po; juvenile hormone esterase, jhe) during the last part of pharate-adult development is known to be under hormonal control [increasing juvenile hormone titers (JH) in the presence of declining levels of ecdysteroids], both, in queens and workers. However, despite the USP involvement in the gene expression cascade promoted by JH, it does not seem to be the unique mediator of its activity, and, thus, we are still ignorant of the molecular factors regulating the expression of these genes, at least, during the critical period of pharate-adult development. Here, we show that Amftz-f1 has caste-specific transcription profile in fat bodies during this developmental stage, peaking just at the moment of the resuming of JH titers increase in the presence of low levels of ecdysteroids. Knock-down experiments (by RNAi) showed that the expression of genes essential to adult development (e.g. vg, and cuticular protein genes) depends on ftz-f1. Our results suggest that Amftz-f1 can also be considered a competence factor to the expression of several genes during pharate-adult development, and, thus, as a key actor in one of the last developmental processes leading to caste differentiation in A. mellifera.

ORGANISM(S): Apis mellifera

PROVIDER: GSE90037 | GEO | 2018/12/13

REPOSITORIES: GEO

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Project description:In honey bees, Vitellogenin (Vg) is hypothesized to be a major factor affecting hormone signaling, food-related behavior, immunity, stress resistance and lifespan. Likewise microRNAs play important roles in posttranscriptional gene regulation and affect many biological processes thereby showing many parallels to Vg functions. The molecular basis of Vg and microRNA interactions is largely unknown. Here, we exploited the well-established RNA interference (RNAi) protocol for Vg knockdown to investigate its effects on microRNA population in honey bee foragerM-bM-^@M-^Ys brain and fat body tissue. To identify microRNAs that are differentially expressed between tissues in control and knockdown foragers, we used M-BM-5ParafloM-BM-. microfluidic oligonucleotide microRNA microarrays. Our results show 76 and 74 miRNAs were expressed in the brain of control and knockdown foragers whereas 66 and 69 miRNAs were expressed in the fat body of control and knockdown foragers respectively. Target prediction identified potential seed matches for differentially expressed subset of microRNAs affected by Vg knockdown. These candidate genes are involved in a broad range of biological processes including insulin signaling, juvenile hormone (JH) and ecdysteroid signaling previously shown to affect foraging behavior. Thus, here we demonstrate a causal link between Vg expression-variation and variation in the abundance of microRNAs in different tissues with possible consequences for regulation of foraging behavior. We knocked down Vitellogenin (Vg) gene expression (using RNAi) in adult workers to identify potential downstream consequences on the expression of microRNA population in the fat body compared to control group (dsRNA-GFP injected bees). Six biological samples of fat body-derived small RNA fraction were prepared for each treatment group (dsRNA-Vg and dsRNA-GFP). Each biological sample contained pooled RNA from 5 unique individuals. Each fat body pool contained a total of 2 M-BM-5g of small RNA fraction, to which each of the 5 individuals contributed equally (400 ng). Pools were named as M-bM-^@M-^\control forager fat bodyM-bM-^@M-^] (GFFb) and M-bM-^@M-^\knockdown forager fat bodyM-bM-^@M-^] (VFFb), followed by a number from 1 to 6.

Project description:In honey bees, Vitellogenin (Vg) is hypothesized to be a major factor affecting hormone signaling, food-related behavior, immunity, stress resistance and lifespan. Likewise microRNAs play important roles in posttranscriptional gene regulation and affect many biological processes thereby showing many parallels to Vg functions. The molecular basis of Vg and microRNA interactions is largely unknown. Here, we exploited the well-established RNA interference (RNAi) protocol for Vg knockdown to investigate its effects on microRNA population in honey bee foragerM-bM-^@M-^Ys brain and fat body tissue. To identify microRNAs that are differentially expressed between tissues in control and knockdown foragers, we used M-BM-5ParafloM-BM-. microfluidic oligonucleotide microRNA microarrays. Our results show 76 and 74 miRNAs were expressed in the brain of control and knockdown foragers whereas 66 and 69 miRNAs were expressed in the fat body of control and knockdown foragers respectively. Target prediction identified potential seed matches for differentially expressed subset of microRNAs affected by Vg knockdown. These candidate genes are involved in a broad range of biological processes including insulin signaling, juvenile hormone (JH) and ecdysteroid signaling previously shown to affect foraging behavior. Thus, here we demonstrate a causal link between Vg expression-variation and variation in the abundance of microRNAs in different tissues with possible consequences for regulation of foraging behavior. We knocked down Vitellogenin (Vg) gene expression (using RNAi) in adult workers to identify potential downstream consequences on the expression of microRNA population in the brain compared to control group (dsRNA-GFP injected bees). Six biological samples of brain-derived small RNA fraction were prepared for each treatment group (dsRNA-Vg and dsRNA-GFP). Each biological sample contained pooled RNA from 5 unique individuals. Each brain pool contained a total of 1 M-BM-5g of small RNA fraction, to which each of the 5 individuals contributed equally (200 ng). Pools were named as M-bM-^@M-^\control forager brainM-bM-^@M-^] (GFBr) and M-bM-^@M-^\knockdown forager brainM-bM-^@M-^] (VFBr), followed by a number from 1 to 6.

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Gene targets and developmental regulation of the honeybee Apis mellifera ftz-f1

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