Unknown,Transcriptomics,Genomics,Proteomics

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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

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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.

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

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