Unknown,Transcriptomics,Genomics,Proteomics

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RNA-seq of antennal tissue of male and female honey bees (Apis mellifera) at 2 different time points of the day

ABSTRACT: In this RNA-seq study, we compared the antennal transcriptomes of sexually mature drones (males) and time-trained foragers (females) of Apis mellifera collected at different times of day and different activity states. The goals of our project was to provide a more comprehensive description of gene expression differences between drone and forager antennae. Most studies on insect antennal transcriptome still focus on identifying and reporting genes involved in odorant binding and detection. In contrast, we also aimed at identifying so far unrecognised molecules, not directly involved in odorant detection, but likely playing an important role in peripheral olfactory processing. We also wanted to explore whether daily changes in gene expression and correlations between gene expression levels and behavioral activity might be a fruitful approach to identify additional genes involved in odorant reception. Apis mellifera drones perform mating flight in the afternoon around 14:00 hour (h) in Bangalore, India. During their daily mating flight activity, drones were caught at the hive entrance and color marked on the thorax. On the next day color-marked drones were collected at two different time points: 9:00 (inactive) and 14:00 h (active). Honey bee foragers can be trained to visit a food source at a specific time of the day. In this study, an A. mellifera colony was transferred in an enclosed outdoor flight cage to train the foragers to visit a feeder at a specific time of the day. A sucrose reward (1M sucrose solution) was presented either from 8:00 to 10:00 h (morning training) or from 13:00 to 15:00 h (afternoon training) for 10 consecutive days. On the 8th, 9th and 10th day of training, foragers visiting the feeder were marked on their thorax with different colors, one type of color each day, to identify the frequently visiting foragers. On the 11th day, the feeder was not presented and the foragers that had all 3 color marks were collected at 9:00 and 14:00 h. All the samples were immediately flash frozen in liquid nitrogen. Collected samples were transferred from liquid nitrogen onto dry ice and the entire antennae (i.e. scape, pedicel and flagellum) were cut off. We pooled 10 antennae from 5 bees per sample and extracted total RNA using Trizol method. Antennal transcriptomes of drones (n=3 per time point), morning-trained foragers (n=2 per time point) and afternoon-trained foragers (n=2 per time point) were sequenced at 2 different time points (9:00 h and 14:00 h).

INSTRUMENT(S): Illumina HiSeq 2500

ORGANISM(S): Apis mellifera

SUBMITTER: Rikesh Jain

PROVIDER: E-MTAB-8489 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Olfaction and chemical communication are fundamental for coordination in insect societies, especially in species forming large colonies. Reproductive and task partitioning suggest that colony members belonging to different castes or subcastes or performing different tasks during their life (polyethism) may produce different semiochemicals and be differently sensitive to the variety of pheromones and/or environmental odours. The main peripheral olfactory organs are the antennal chemosensilla, where the early olfactory processes take place. At this stage, members of two different families of soluble chemosensory proteins (Odorant Binding Proteins -OBPs-, Chemosensory Proteins -CSPs-) show a remarkable affinity for different odorants and act as carriers while a further family, the Niemann Pick type C2 proteins (NPC2) may have a similar function, although this has not been fully demonstrated. Sensillar lymph also contains Odorant degrading enzymes (ODEs) which are involved in inactivation through degradation of the chemical signals, once the message is conveyed. Despite their importance in chemical communication, poor is known about how proteins involved in peripheral olfaction and, more generally antennal proteins, differ in honeybees of different caste, task and age. Here we investigate for the first time, using a shotgun proteomic approach, the antennal profile of honeybee of different castes (queens and workers) and of workers performing different tasks (nursers, guards and foragers) by controlling for the potential confounding effect of age. Regarding olfactory proteins, major differences were observed between queens and workers, some of which were found to be more abundant in queens (OBP3, OBP18, NPC2-1) and others to be more abundant in workers (OBP15, OBP21, CSP1, CSP3); while between workers performing different tasks, OBP14 was more abundant in nurses with respect to guards and foragers, and two ODEs were more abundant in guards with respect to workers of 2nd week. Apart from proteins involved in olfaction, we have found that the antennal proteomes are mainly characterized by castes and tasks, while age has a limited effect on antennal protein profile.

Project description:While Apis cerana cerana, like Apis mellifera, undergoes a behavioral transition from in-hive nursing to outdoor foraging duties, nothing is known about the genes underlying this social signal-triggered aged-related transition in this species. Here, we simultaneously sequenced the head transcriptomes of the 7-day-old normal nurses (N7BY), 18- and 22-day-old normal foragers (N18CJ and N22CJ), 7-day-old precocious foragers (Tq7CJ) and 22-day-old overaged or reverted nurses (Tq22BY) of A. cerana cerana by RNA-seq and made a 3-tier comparison (from pairwise to group-wise and between-group) to unravel the genes associated with this transition. Six pairwise comparisons revealed 165-492 differentially expressed genes between nurses vs. foragers. Subsequent 3 group-wise and 1 between-group comparisons narrowed the transition-associated genes down to 18 nurse- and 41 forager-unique genes and 29 (14 and 15 genes upregulated in nurses and foragers, respectively) differentially expressed genes between the 3 types of foragers and 2 types of nurses. The uniquely expressed genes are usually low-abundance long noncoding RNAs, transcription factors, transcription coactivators, RNA-binding proteins, kinases or phosphatases involved in signaling transduction and/or gene expression regulation, whereas the differentially expressed genes are often high-abundance downstream genes that directly perform the tasks of nurses or foragers, such as major royal jelly proteins for nurses and the genes involved in sugar/protein digestion, lipids/fatty acids metabolism, plant allelochemicals detoxification and defense against pathogens and predators for foragers. Mapping of the clean reads to the published A. mellifera genome uncovered that the 3 types of foragers had a greater percentage of reads from annotated exons and intergenic regions, whereas the 2 types of nurses had a greater percentage of reads from introns. Taken together, these results suggest that the reciprocal nurse-forager behavioral transition of the A. cerana cerana is regulated by a social signal-triggered intron-exon/intergenic epigenetic shift and the resulted transcriptional shift of the nurse- and forager-associated genes.

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RNA-seq of antennal tissue of male and female honey bees (Apis mellifera) at 2 different time points of the day

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