Summary Ants exhibit cooperative behaviors and advanced forms of sociality that depend on pheromone-mediated communication. Odorant receptor neurons (ORNs) express specific odorant receptors (ORs) encoded by a dramatically expanded gene family in ants. In most eusocial insects, only the queen can transmit genetic information, restricting genetic studies. In contrast, workers in Harpegnathos saltator ants can be converted into gamergates (pseudoqueens) that can found entire colonies. This feature facilitated CRISPR-Cas9 generation of germline mutations in orco, the gene that encodes the obligate co-receptor of all ORs. orco mutations should significantly impact olfaction. We demonstrate striking functions of Orco in odorant perception, reproductive physiology and social behavior plasticity. Surprisingly, unlike in other insects, loss of OR functionality also dramatically impairs development of the antennal lobe where ORNs project. Therefore, the development of genetics in Harpegnathos establishes this ant species as a model organism to study the complexity of eusociality.

Coronaviruses (CoVs) assemble at endoplasmic reticulum Golgi intermediate compartment (ERGIC) membranes and egress from cells in cargo vesicles. Only a few molecules of the envelope (E) protein are assembled into virions. The role of E in morphogenesis is not fully understood. The cellular localization and dynamics of mouse hepatitis CoV A59 (MHV) E protein were investigated to further understanding of its role during infection. E protein localized in the ERGIC and Golgi with the amino and carboxy termini in the lumen and cytoplasm, respectively. E protein does not traffic to the cell surface. MHV was genetically engineered with a tetracysteine tag at the carboxy end of E. Fluorescence recovery after photobleaching (FRAP) showed that E is mobile in ERGIC/Golgi membranes. Correlative light electron microscopy (CLEM) confirmed the presence of E in Golgi cisternae. The results provide strong support that E proteins carry out their function(s) at the site of budding/assembly.

In many ant species, sibling larvae follow alternative ontogenetic trajectories that generate striking variation in morphology and behavior among adults. These organism-level outcomes are often determined by environmental rather than genetic factors. Therefore, epigenetic mechanisms may mediate the expression of adult polyphenisms. We produced the first genome-wide maps of chromatin structure in a eusocial insect and found that gene-proximal changes in histone modifications, notably H3K27 acetylation, discriminate two female worker and male castes in Camponotus floridanus ants and partially explain differential gene expression between castes. Genes showing coordinated changes in H3K27ac and RNA implicate muscle development, neuronal regulation, and sensory responses in modulating caste identity. Binding sites of the acetyltransferase CBP harbor the greatest caste variation in H3K27ac, are enriched with motifs for conserved transcription factors, and show evolutionary expansion near developmental and neuronal genes. These results suggest that environmental effects on caste identity may be mediated by differential recruitment of CBP to chromatin. We propose that epigenetic mechanisms that modify chromatin structure may help orchestrate the generation and maintenance of polyphenic caste morphology and social behavior in ants.

Dominance rank in animal societies is correlated with changes in both reproductive physiology and behavior. In some social insects, dominance status is used to determine a reproductive division of labor, where a few colony members reproduce while most remain functionally sterile. Changes in reproduction and behavior in this context must be coordinated through crosstalk between the brain and the reproductive system. We investigated a role for biogenic amines in forming this connection in the ant Harpegnathos saltator. In this species, workers engage in an elaborate dominance tournament to establish a group of reproductive workers termed gamergates. We analyzed biogenic amine content in the brains of gamergates, insideworkers and foragers under stable colony conditions and found that gamergates had the highest levels of dopamine. Dopamine levels were also positively correlated with increased ovarian activity among gamergates. Next, we experimentally induced workers to compete in a reproductive tournament to determine how dopamine may be involved in the establishment of a new hierarchy. Dopamine levels rose in aggressive workers at the start of a tournament, while workers that were policed by their nestmates (a behavior that inhibits ovarian activity) showed a rapid decline in dopamine. In addition to dopamine, levels of serotonin and tyramine differed among castes, and these changes could contribute to differences in caste-specific behavioral patterns observed among non-reproductive workers. Overall, these results provide support that biogenic amines link changes in behavior and dominance with reproductive activity in H. saltator as well as drive differences in worker task performance.

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