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Apis mellifera (honey bee) antennae proteomics investigating lateralization and dsRNA microinjection

ABSTRACT: Previously, we attempted to identify the molecular basis for lateralization in honey bee workers by comparing protein expression between left and right antennae of hygienic bees using mass spectrometry-based quantitative proteomics (PXD005242). We identified 1,845 unique proteins but found no significant differences; therefore, we suggested this may be because of insufficient depth of coverage. To improve proteome coverage, we fractionated the peptides from left and right antennae from 4 highly hygienic colonies and repeated the comparison. Here, we identified 3,114 unique proteins (a 69% improvement), which is among the highest proteome coverage achieved in honey bees. However, we still did not identify significant differences. We also compared the proteomes of antennae that were injected with mock dsRNA, dsRNA targeting OBP18, and uninjected. We found a strong effect of microinjecting alone, but no knockdown of our target.

INSTRUMENT(S): micrOTOF

ORGANISM(S): Apis Mellifera (ncbitaxon:7460)

SUBMITTER: Leonard Foster

PROVIDER: MSV000081790 | MassIVE | Thu Dec 07 14:23:00 GMT 2017

REPOSITORIES: MassIVE

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Project description:BACKGROUND: Social insects, such as honey bees, use molecular, physiological and behavioral responses to combat pathogens and parasites. The honey bee genome contains all of the canonical insect immune response pathways, and several studies have demonstrated that pathogens can activate expression of immune effectors. Honey bees also use behavioral responses, termed social immunity, to collectively defend their hives from pathogens and parasites. These responses include hygienic behavior (where workers remove diseased brood) and allo-grooming (where workers remove ectoparasites from nestmates). We have previously demonstrated that immunostimulation causes changes in the cuticular hydrocarbon profiles of workers, which results in altered worker-worker social interactions. Thus, cuticular hydrocarbons may enable workers to identify sick nestmates, and adjust their behavior in response. Here, we test the specificity of behavioral, chemical and genomic responses to immunostimulation by challenging workers with a panel of different immune stimulants (saline, Sephadex beads and Gram-negative bacteria E. coli). RESULTS: While only bacteria-injected bees elicited altered behavioral responses from healthy nestmates compared to controls, all treatments resulted in significant changes in cuticular hydrocarbon profiles. Immunostimulation caused significant changes in expression of hundreds of genes, the majority of which have not been identified as members of the canonical immune response pathways. Furthermore, several new candidate genes that may play a role in cuticular hydrocarbon biosynthesis were identified. Finally, we identified common genes regulated by pathogen challenge in honey bees and other insects, suggesting that immune responses are conserved at the molecular level. CONCLUSIONS: These studies suggest that honey bee genomic responses to immunostimulation are substantially broader than expected, and may mediate the behavioral changes associated with social immunity by orchestrating changes in chemical signaling.

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Apis mellifera (honey bee) antennae proteomics investigating lateralization and dsRNA microinjection

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