The biological bases of wanting have been characterized in mammals, but whether an equivalent wanting system exists in insects remains unknown. In this study, we focused on honey bees, which perform intensive foraging activities to satisfy colony needs, and sought to determine whether foragers leave the hive driven by specific expectations about reward and whether they recollect these expectations during their waggle dances. We monitored foraging and dance behavior and simultaneously quantified and interfered with biogenic amine signaling in the bee brain. We show that a dopamine-dependent wanting system is activated transiently in the bee brain by increased appetite and individual recollection of profitable food sources, both en route to the goal and during waggle dances. Our results show that insects share with mammals common neural mechanisms for encoding wanting of stimuli with positive hedonic value.

Chlorpyrifos is a widely used organophosphorus insecticide. The acute oral 24 h median lethal dose (LD50) value of chlorpyrifos in Apis mellifera and in Apis cerana was estimated to assess differential acute chlorpyrifos toxicity in the two bee species. The LD50 values of chlorpyrifos in A. mellifera and in A. cerana are 103.4 ng/bee and 81.8 ng/bee, respectively, which suggests A. cerana bees are slightly more sensitive than A. mellifera bees to the toxicity of chlorpyrifos. Doses half the acute LD50 of chlorpyrifos were selected to study behavioral changes in the two bee species using proboscis extension response assay. A. mellifera foragers treated with chlorpyrifos showed significantly lower response to the 10% sucrose solution compared to control bees after 2, 24 and 48 h. Chlorpyrifos significantly impaired the olfactory learning abilities and 2 h memory retention of forager bees regardless of honey bee species, which may affect the foraging success of bees exposed to chlorpyrifos.

The ethological study of dance behaviour has yielded some findings since Karl Von Frisch discovered and interpreted the ‘dance language’ in the honey bee. However, the function and role of long non-coding RNAs on dance behaviour are hardly known until now. In this study, the differential expression patterns of lncRNAs in the brains of waggling dancers and non-dancing bees were analysed by RNA sequencing. Furthermore, lncRNA-mRNA association analysis was constructed to decipher the waggle dance. The results of RNA sequencing indicated that a total of 2877 lncRNAs and 9647 mRNAs were detected from honey bee brains. Further comparison analysis displayed that two lncRNAs, MSTRG.6803.3 and XR_003305156.1, may be involved in the waggle dance. The lncRNA-mRNA association analysis showed that target genes of differentially expressed lncRNAs in the brains between waggling dancers and non-dancing bees were mainly annotated in biological processes related to metabolic process, signalling and response to stimulus and in molecular function associated with signal transducer activity, molecular transducer activity and binding. Nitrogen metabolism was likely implicated in the modulation of the waggle dance. Our findings contribute to further understanding the occurrence and development of waggle dance.

MicroRNAs (miRNAs) play a vital role in the nerve regulation of honey bees (Apis mellifera). This study aims to investigate the differences in expression of miRNAs in a honey bee’s brain for olfactory learning tasks and to explore their potential role in a honey bee’s olfactory learning and memory. In this study, 12 day old honey bees with strong and weak olfactory performances were utilized to investigate the influence of miRNAs on olfactory learning behavior. The honey bee brains were dissected, and a small RNA-seq technique was used for high-throughput sequencing. The data analysis of the miRNA sequences revealed that 14 differentially expressed miRNAs (DEmiRNAs) between the two groups, strong (S) and weak (W), for olfactory performance in honey bees were identified, which included seven up-regulated and seven down-regulated. The qPCR verification results of the 14 miRNAs showed that four miRNAs (miR-184-3p, miR-276-3p, miR-87-3p, and miR-124-3p) were significantly associated with olfactory learning and memory. The target genes of these DEmiRNAs were subjected to the GO database annotation and KEGG pathway enrichment analyses. The functional annotation and pathway analysis showed that the neuroactive ligand-receptor interaction pathway, oxidative phosphorylation, biosynthesis of amino acids, pentose phosphate pathway, carbon metabolism, and terpenoid backbone biosynthesis may be a great important pathway related to olfactory learning and memory in honey bees. Our findings together further explained the relationship between olfactory performance and the brain function of honey bees at the molecular level and provides a basis for further study on miRNAs related to olfactory learning and memory in honey bees.

The grooming behavior of honeybees serves as a crucial auto-protective mechanism against Varroa mite infestations. Compared to Apis mellifera, Apis cerana demonstrates more effective grooming behavior in removing Varroa mites from the bodies of infested bees. However, the underlying mechanisms regulating grooming behavior remain elusive. In this study, we evaluated the efficacy of the auto-grooming behavior between A. cerana and A. mellifera and employed RNA-sequencing technology to identify differentially expressed genes (DEGs) in bee brains with varying degrees of grooming behavior intensity. We observed that A. cerana exhibited a higher frequency of mite removal between day 5 and day 15 compared to A. mellifera, with day-9 bees showing the highest frequency of mite removal in A. cerana. RNA-sequencing results revealed the differential expression of the HTR2A and SLC17A8 genes in A. cerana and the CCKAR and TpnC47D genes in A. mellifera. Subsequent homology analysis identified the HTR2A gene and SLC17A8 gene of A. cerana as homologous to the HTR2A gene and SLC17A7 gene of A. mellifera. These DEGs are annotated in the neuroactive ligand–receptor interaction pathway, the glutamatergic synaptic pathway, and the calcium signaling pathway. Moreover, CCKAR, TpnC47D, HTR2A, and SLC17A7 may be closely related to the auto-grooming behavior of A. mellifera, conferring resistance against Varroa infestation. Our results further explain the relationship between honeybee grooming behavior and brain function at the molecular level and provide a reference basis for further studies of the mechanism of honeybee grooming behavior.