Differential antennal proteome comparison of adult honeybee drone, worker and queen (Apis mellifera L.)

There is growing evidence in the literature suggesting that caste differentiation in the stingless bee, Melipona scutellaris, and other bees in the genus Melipona, is triggered by environmental signals, particularly a primer pheromone. With the proper amount of food and a chemical stimulus, 25% of females emerge as queens, in agreement with a long-standing “two loci/two alleles model” proposed in the 1950s. We surmised that these larvae must be equipped with an olfactory system for reception of these chemical signals. Here we describe for the first time the diversity of antennal sensilla in adults and the morphology of larvae of M. scutellaris. Having found evidence for putative olfactory sensilla in larvae, we next asked whether olfactory proteins were expressed in larvae. Since the molecular basis of M. scutellaris is still unknown, we cloned olfactory genes encoding chemosensory proteins (CSP) and odorant-binding proteins (OBPs) using M. scutellaris cDNA template and primers designed on the basis CSPs and OBPs previously reported from the European honeybee, Apis mellifera. We cloned two CSP and two OBP genes and then attempted to express the proteins encoded by these genes. With a recombinant OBP, MscuOBP8, and a combinatorial single-chain variable fragment antibody library, we generated anti-MscuOBP8 monoclonal antibody. By immunohistochemistry we demonstrated that the anti-MscuOBP8 binds specifically to the MscuOBP8. Next, we found evidence that MscuOBP8 is expressed in M. scutellaris larvae and it is located in the mandibular region, thus further supporting the hypothesis of olfactory function in immature stages. Lastly, molecular modeling suggests that MscuOBP8 may function as a carrier of primer pheromones or other ligands.

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“…Sensilla placodea is involved in the reception of pheromones and other semiochemicals in Apis and Bombus bees [39, 51, 52]. Contrary to A .…”

Section: Resultsmentioning

confidence: 99%

Characterization of antennal sensilla, larvae morphology and olfactory genes of Melipona scutellaris stingless bee

et al. 2017

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“…The two most significant differences between ITB and RJB antennae were found in components of the oxidative phosphorylation pathway and in chemoreception. The antennal chemoreception requires a sophisticated, compartmentalized organization (Leal, ) and is energetically demanding (Fang et al, ). Antennae of RJBs contained higher amounts of proteins that function in oxidative phosphorylation and hydrogen ion transmembrane transport, which presumably enable ATP production.…”

Section: Discussionmentioning

confidence: 99%

Behavioural, physiological and molecular changes in alloparental caregivers may be responsible for selection response for female reproductive investment in honey bees

Han

et al. 2019

Molecular Ecology

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Reproductive investment is a central life history variable that influences all aspects of life. Hormones coordinate reproduction in multicellular organisms, but the mechanisms controlling the collective reproductive investment of social insects are largely unexplored. One important aspect of honey bee (Apis mellifera) reproductive investment consists of raising female‐destined larvae into new queens by alloparental care of nurse bees in form of royal jelly provisioning. Artificial selection for commercial royal jelly production over 40 years has increased this reproductive investment by an order of magnitude. In a cross‐fostering experiment, we establish that this shift in social phenotype is caused by nurse bees. We find no evidence for changes in larval signalling. Instead, the antennae of the nurse bees of the selected stock are more responsive to brood pheromones than control bees. Correspondingly, the selected royal jelly bee nurses are more attracted to brood pheromones than unselected control nurses. Comparative proteomics of the antennae from the selected and unselected stocks indicate putative molecular mechanisms, primarily changes in chemosensation and energy metabolism. We report expression differences of several candidate genes that correlate with the differences in reproductive investment. The functional relevance of these genes is supported by demonstrating that the corresponding proteins can competitively bind one previously described and one newly discovered brood pheromone. Thus, we suggest several chemosensory genes, most prominently OBP16 and CSP4, as candidate mechanisms controlling queen rearing, a key reproductive investment, in honey bees. These findings reveal novel aspects of pheromonal communication in honey bees and explain how sensory changes affect communication and lead to a drastic shift in colony‐level resource allocation to sexual reproduction. Thus, pheromonal and hormonal communication may play similar roles for reproductive investment in superorganisms and multicellular organisms, respectively.

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“…FABP (clone 1BH4 in reference Evans and Wheeler (1999)) is notable for its potentially differential expression between castes, as it is expressed in worker but not queen larvae (Evans and Wheeler 1999). More recently, differences in the expression of FABPs have been found in the drone, worker, and queen antennal proteomes (Fang et al 2012).…”

Section: Lipid Transport and Metabolism (I Category)mentioning

confidence: 99%

Two-dimensional gel proteome analysis of honeybee, Apis mellifera, worker red-eye pupa hemolymph

et al. 2013

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Apis mellifera Linnaeus is a holometabolous insect that undergoes complete metamorphosis in its nonfeeding pupal stage before transitioning to the adult stage. Its pupal stages are classifiable by the unique color pigmentation of its compound eyes and thorax; notably, there is a red-eye stage involving an unpigmented body that has a relatively short duration and is easy to recognize. The aim of the current study was to create a proteomic reference map of the worker red-eye pupa hemolymph. Hemolymph was collected from dorsal vessels using glass capillary tubes and was examined using pI 3-10 two-dimensional gel electrophoresis (2DE; 10 and 14 %) and matrix-assisted laser desorption/ionization (MALDI) time-of-flight (TOF)/TOF protein identification. This experimental approach allowed us to identify 129 different proteins organized into orthologous groups. Overall, the predominant category was posttranslational modifications, protein turnover and chaperones (23.3 % of the identified proteins). In addition, we identified proteins in the non-orthologous groups of olfaction (2.3 % of the identified proteins) and storage hexamerins (3.1 % of the identified proteins). Quantitatively, the major protein isoforms that were accurately identified via 10 % 2DE were four forms of storage hexamerin: the 110, 70a, 70b, and 70c forms. The most abundant enzymes identified were short-chain dehydrogenases/reductases with pivotal developmental roles in ecdysteroidogenesis and a sigma class glutathione-S-transferase that most likely serves as a major protectant against the by-products of oxidative stress. Many of the identified proteins are known to be involved in the mechanisms of metamorphosis. All of the identified proteins are useful as markers for future comparative physiological and developmental studies.

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Differential antennal proteome comparison of adult honeybee drone, worker and queen (Apis mellifera L.)

Cited by 26 publications

References 60 publications

Characterization of antennal sensilla, larvae morphology and olfactory genes of Melipona scutellaris stingless bee

Characterization of antennal sensilla, larvae morphology and olfactory genes of Melipona scutellaris stingless bee

Behavioural, physiological and molecular changes in alloparental caregivers may be responsible for selection response for female reproductive investment in honey bees

Two-dimensional gel proteome analysis of honeybee, Apis mellifera, worker red-eye pupa hemolymph

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