Unknown,Transcriptomics,Genomics,Proteomics

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Analysis of the honey bee brain transcriptome from nurses and forager bees

ABSTRACT: New insights into the transcriptional regulation of behavioral plasticity in honey bees gained by analyzing brain genes expression with the CAGEscan technique that involves identification of specific transcription factors, cis regulatory motifs and alternate transcriptional start sites Examination of 2 different types of Honey Bee Apis Mellifera samples (Nurse and Foragers)

ORGANISM(S): Apis mellifera

SUBMITTER: Vladimir Bajic

PROVIDER: E-GEOD-64315 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description: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).

Project description:Honey bees move through a series of in-hive tasks (M-bM-^@M-^\nursingM-bM-^@M-^]) to outside tasks (M-bM-^@M-^\foragingM-bM-^@M-^]) that coincident with an intense increase in metabolic activity. Social context can cause worker bees to speed up, or slow down this process and foragers may revert back to their earlier in hive tasks accompanied by reversion to earlier physiological states. To determine if the transcriptional profile of forager bees can revert, or if the effects of flight on gene expression are irreversible, we used whole-genome microarrays. Brain tissue and flight muscle exhibited independent patterns of expression during behavioral transitions, with patterns of expression in the brain reflecting both age and behavior, while flight muscle exhibited primarily age-related patterns of expression. Our data suggest that the transition from little to no flight (nurse) to intense flight (forager), rather than the amount of flight has a major effect on gene expression. Following behavioral reversion there was a partial reversion in gene expression but some aspects of forager expression patterns, such as those for genes involved in immune function, remained. These data suggest an epigenetic control and energy balance role in honey bee functional senescence. Brains and thoraces from the same individuals of all behavioral groups were compared on a total of 132 arrays. The samples were hybridized against each other using a loop design. The groups tested are outlined as follows: Typical aged nurse 'YN' (8-10 days old; <1 day flight experience), Precocious forager 'PF' (8 to 10 days old; 2 to 3 days flight experience), Overaged nurse 'ON' (19 to 22 days old; < 1 day flight experience), Forager - low flight 'TFL' (19 to 22 days old; 2 to 3 days flight experience), Forager - high flight 'TFH' ( 19 to 22 days old; 7 to 9 days flight experience), Forager - old 'OF' (25 to 26 days old; 10 to 12 days flight experience), Reverted nurse 'RN' (25 to 26 days old; 7 to 9 days flight experience). The comparisons are outlined as follows: YN:ON (6 arrays), ON:RN (6 arrays), RN:TFH (6 arrays), TFH:TFL (6 arrays), TFL:PF (6 arrays), PF:YN (6 arrays), ON:TFL (12 arrays), YN:OF (6 arrays), OF:RN (6 arrays), RN:YN (6 arrays). Each comparison was done for individual brains and thoraces. Total: 132 arrays

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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Analysis of the honey bee brain transcriptome from nurses and forager bees

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