Biology subjects

McAfee, A.

Publications and source records attributed to McAfee, A..

5 recordsLinked to original sources

bioRxiv2018

Proteomic analysis of chemosensory organs in the honey bee parasite Varroa destructor: a comprehensive examination of the potential carriers for semiochemicals

The mite Varroa destructor is the major parasite of the honey bee and is responsible for great economical losses. The biochemical tools used by Varroa to detect semiochemicals produced by the host are still largely unknown. We have performed proteomic analysis on chemosensory organs of this species in order to identify putative soluble carriers for pheromones and other olfactory cues emitted by the host. In particular, we have analysed forelegs, mouthparts (palps, chelicera and hypostome) and the second pair of legs (as control tissue) in reproductive and phoretic stages of the Varroa life cycle. We identified 958 Varroa proteins, most of them common to organs and stages. Sequence analysis shows that four proteins can be assigned to the odorant-binding protein (OBP)-like class, which bear some similarity to insect OBPs, but so far are only reported in some Chelicerata. In addition, we have detected the presence of two proteins belonging to the Niemann-Pick family, type C2 (NPC2), which have been suggested to act as semiochemical carriers. This work contributes to elucidating the chemical communication systems in Varroa with the aim of understanding how detection of semiochemicals has evolved in terrestrial non-hexapod Arthropoda. Data are available via ProteomeXchange with identifier PXD008679.

zoology↗

bioRxiv2017

Results of follow-up experiments to "Odorant cues linked to social immunity induce lateralized antennal stimulation in honey bees (Apis mellifera L.)"

In 2017, we published the paper \"Odorant cues linked to social immunity induce lateralized antennal stimulation in honey bees (Apis mellifera L.)\" in Scientific Reports. Since then, we have performed three follow-up experiments which have either negative or contradictory results. Previously, we used electrophysiology to show that hygienic bees displayed significantly higher sensitivity to {beta}-ocimene when stimulated via their left antennae compared to their right. We repeated this assay using worker honey bees from a single hygienic colony and found, to our surprise, that the right antennae elicited higher sensitivity. We also previously attempted to identify a molecular basis for lateralization by using mass spectrometry-based proteomics to compare left and right antennal proteomes. Of the 1,845 proteins, none were differentially expressed. Here, we repeated this experiment but employed orthogonal peptide fractionation to increase proteome coverage to 3,114 proteins; however, still none were differentially expressed. Finally, we attempted to manipulate gene expression of a key antennal odorant binding protein linked to hygienic behaviour (OBP18) using RNA interference via antenna microinjection. We were not able to achieve long-lasting OBP18 knock-down, but comparing the proteomes of untreated, mock dsRNA-treated and OBP18 dsRNA-treated worker antennae revealed numerous off-target effects of the act of injecting alone. By openly reporting this data, we hope to set an example for information transparency.

molecular biology↗

bioRxiv2017

Death pheromones triggering hygienic behavior in honey bees (Apis mellifera L.)

Eusocial insects live in teeming societies with thousands of their kin. In this crowded environment, workers combat disease by removing or burying their dead or diseased nestmates. For honey bees, we found that hygienic brood-removal behavior is triggered by two odorants - {beta}-ocimene and oleic acid - which are released from brood upon freeze-killing. {beta}-ocimene is a co-opted pheromone that normally signals larval food-begging, whereas oleic acid is a conserved necromone across arthropod taxa. Interestingly, the odorant blend can induce hygienic behavior more consistently than either odorant alone. We suggest that the volatile {beta}-ocimene flags hygienic workers attention, while oleic acid is the death cue, triggering removal. Bees with high hygienicity detect and remove brood with these odorants faster than bees with low hygienicity, and both molecules are strong ligands for hygienic behavior-associated odorant binding proteins (OBP16 and OBP18). Odorants that induce low levels of hygienic behavior, however, are weak ligands for these OBPs. We are therefore beginning to paint a picture of the molecular mechanism behind this complex behavior, using odorants associated with freeze-killed brood as a model.

animal behavior and cognition↗

bioRxiv2017

A Varroa Destructor Protein Atlas Reveals Molecular Underpinnings Of Developmental Transitions And Sexual Differentiation

Varroa destructor is the most economically damaging honey bee pest, weakening colonies by simultaneously parasitizing bees and transmitting harmful viruses. Despite these impacts on honey bee health, surprisingly little is known about its fundamental molecular biology. Here we present a Varroa protein atlas crossing all major developmental stages (egg, protonymph, deutonymph and adult) for both male and female mites as a web-based interactive tool (http://foster.nce.ubc.ca/varroa/index.html). By intensity-based label-free quantitation, 1,433 proteins were differentially expressed across developmental stages, including two distinct viral polyproteins. Enzymes for processing carbohydrates and amino acids were among many of these differences as well as proteins involved in cuticle formation. Lipid transport involving vitellogenin was the most significantly enriched biological process in the foundress (reproductive female) and young mites. In addition, we found that 101 proteins were sexually regulated and functional enrichment analysis suggests that chromatin remodeling may be a key feature of sex determination. In a proteogenomic effort, we identified 519 protein-coding regions (169 of which were differentially expressed) supported by 1,464 peptides which were previously unannotated. Since this is a recurring trend with annotating genomes of non-model species, we analyzed their amino acid and nucleotide composition as well as their orthology to other species to suggest reasons why they may have been missed initially. Overall, this work provides a first-of-its-kind interrogation of the patterns of gene expression that govern the Varroa life cycle and the tools we have developed will support further research on this threatening honey bee pest.

developmental biology↗

bioRxiv2016

Odorant cues linked to social immunity induce lateralized antenna stimulation in honey bees (Apis mellifera L.)

Hygienic behaviour (HB) is a social immunity trait in honey bees (Apis mellifera L.) whereby workers detect, uncap and remove unhealthy brood, improving disease resistance in the colony. This is clearly economically valuable; however, the molecular mechanism behind it is not well understood. The freeze-killed brood (FKB) assay is the conventional method of HB selection, so we compared odour profiles of FKB and live brood. Surprisingly, we found that significantly more brood pheromone ({beta}-ocimene) was released from FKB. {beta}-ocimene abundance also positively correlated with HB, suggesting there could be a brood effect contributing to overall hygiene. We found that β ocimene stimulated worker antennae in a dose-dependent manner, with the left antennae responding significantly stronger than right antennae in hygienic bees, but not in non-hygienic bees. This suggests that HB depends not only on odour detection, but also lateralization of sensitivity. We also compared odour profiles of Varroa-infested brood to healthy brood and found an overall interactive effect between developmental stage and infestation, but specific odours did not drive these differences. Overall, the data we present here is an important foundation on which to build our understanding the molecular mechanism behind this complex behaviour.

biochemistry↗