Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Caste-specific expression patterns of olfactory related genes in the leaf-cutting ant, Atta vollenweideri

ABSTRACT: To allow estimation of the complexity and gene expression differences of the antennal transcriptome between sexes as well as between castes, microarrays were designed based on an assembly of A. vollenweideri antennal sequence data from all sexes and castes. The microarrays were hybridized with samples generated from the respecitve antennal tissues, with four independent sample pools per sex and caste. Custom 2x105 k Agilent microarrays (Agilent Technologies, Palo Alto, CA) were designed based on the antennal transcriptome data of A.vollenweideri. For all contigs with an identified ORF by automatic annotation, two 60mer oligo probes were designed, with two additional 60mer oligo probes for opposite reading direction for those with unknown ORF, thus resulting in 4 oligo probes per contig. Additionally, we added three 60mer oligo probes for each identified and revised OR coding gene, resulting in a total of 5 oligo probes per OR coding contig. For design and arrangement of the Agilent microarrays, the eArray software (Agilent Technologies, Palo Alto, CA) was used. For sex- and caste-specific antennal microarray hybridizations, RNA from antennae of 50 (Males) to 300 (Tiny Workers) individuals was pooled per preparation, and four pools (replicates) each were dissected per sex and caste, respectively. Double purified total RNA was added to Agilent Technologies spike-in RNA and labeld using QuickAmp Amplification kit (Agilent Technologies) and the Kreatech ULS Fluorescent Labeling Kit with cyanine 3-CTP dye following the manufacturerÂ´s instructions. Amplified cRNA samples were used for microarray hybridization, scanned with the Agilent Microarray Scanner and data was extracted from TIFF images with Agilent Feature Extraction software version 9.1. Raw data output files were analyzed using the GeneSpring GX11 and GeneSifter microarray analysis softwares.

ORGANISM(S): Atta vollenweideri

SUBMITTER: Heiko Vogel

PROVIDER: E-GEOD-48824 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

ACCESS DATA

Similar Datasets

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: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).

Dataset Information

Caste-specific expression patterns of olfactory related genes in the leaf-cutting ant, Atta vollenweideri

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure