Orco-dependent survival of odorant receptor neurons in ants.

Olfaction is essential for complex social behavior in insects. To discriminate complex social cues, ants evolved an expanded number of odorant receptor (Or) genes. Mutations in the obligate odorant co-receptor gene orco lead to the loss of ~80% of the antennal lobe glomeruli in the jumping ant Harpegnathos saltator. However, the cellular mechanism remains unclear. Here, we demonstrate massive apoptosis of odorant receptor neurons (ORNs) in the mid to late stages of pupal development, possibly due to ER stress in the absence of Orco. Further bulk and single-nucleus transcriptome analysis shows that, although most orco-expressing ORNs die in orco mutants, a small proportion of them survive: They express ionotropic receptor (Ir) genes that form IR complexes. In addition, we found that some Or genes are expressed in mechanosensory neurons and nonneuronal cells, possibly due to leaky regulation from nearby non-Or genes. Our findings provide a comprehensive overview of ORN development and Or expression in H. saltator.

Influence of Host Plants and Tending Ants on the Cuticular Hydrocarbon Profile of a Generalist Myrmecophilous Caterpillar.

In myrmecophilous organisms, which live in symbiosis with ants, cuticular hydrocarbons (CHCs) play a pivotal role in interspecific communication and defense against chemical-oriented predators. Although these interactions form complex information webs, little is known about the influence of biotic environmental factors on the CHC profiles of myrmecophiles. Here, we analyzed the effect of different host plants and tending ants on the larval CHC profile of Synargis calyce (Lepidoptera: Riodinidae), a polyphagous species with facultative myrmecophily. Groups of caterpillars were fed individually with three host plant species (without tending ants), and with two tending ant species. Through gas chromatography analysis, we compared the cuticular profiles of treatments and found a high similarity between plants and caterpillars (65-82%), but a low similarity between caterpillars and their tending ants (30 - 25%). Cluster analysis showed that caterpillars, ants, and plants form distinct groups, indicating that S. calyce caterpillars have their own chemical profile. These results are similar to those observed for Lycaenidae caterpillars indicating that there is functional convergence in the chemical strategies used by myrmecophilous caterpillar species with similar ecology. Also, the results suggest that the cuticular compounds of S. calyce are primarily influenced by their host plants rather than their tending ants. Thus, we propose that these caterpillars present a trade-off between camouflage and directly informing their presence to ants, maintaining their unique chemical profile, though slightly affected by biotic environmental factors.

Ant may well destroy a whole dam: glycans of colonic mucus barrier disintegrated by gut bacteria.

The colonic mucus layer plays a critical role in maintaining the integrity of the colonic mucosal barrier, serving as the primary defense against colonic microorganisms. Predominantly composed of mucin 2 (MUC2), a glycosylation-rich protein, the mucus layer forms a gel-like coating that covers the colonic epithelium surface. This layer provides a habitat for intestinal microorganisms, which can utilize mucin glycans present in the mucus layer as a sustainable source of nutrients. Additionally, metabolites produced by the microbiota during the metabolism of mucus glycans have a profound impact on host health. Under normal conditions, the production and consumption of mucus maintain a dynamic balance. However, several studies have demonstrated that certain factors, such as dietary fiber deficiency, can enhance the metabolism of mucus glycans by gut bacteria, thereby disturbing this balance and weakening the mucus barrier function of the mucus layer. To better understand the occurrence and development of colon-related diseases, it is crucial to investigate the complex metabolic patterns of mucus glycosylation by intestinal microorganisms. Our objective was to comprehensively review these patterns in order to clarify the effects of mucus layer glycan metabolism by intestinal microorganisms on the host.

Transcriptional and post-transcriptional control of odorant receptor choice in ants.

Insects and mammals have independently evolved odorant receptor genes that are arranged in large genomic tandem arrays. In mammals, each olfactory sensory neuron chooses to express a single receptor in a stochastic process that includes substantial chromatin rearrangements. Here, we show that ants, which have the largest odorant receptor repertoires among insects, employ a different mechanism to regulate gene expression from tandem arrays. Using single-nucleus RNA sequencing, we found that ant olfactory sensory neurons choose different transcription start sites along an array but then produce mRNA from many downstream genes. This can result in transcripts from dozens of receptors being present in a single nucleus. Such rampant receptor co-expression at first seems difficult to reconcile with the narrow tuning of the ant olfactory system. However, RNA fluorescence in situ hybridization showed that only mRNA from the most upstream transcribed odorant receptor seems to reach the cytoplasm where it can be translated into protein, whereas mRNA from downstream receptors gets sequestered in the nucleus. This implies that, despite the extensive co-expression of odorant receptor genes, each olfactory sensory neuron ultimately only produces one or very few functional receptors. Evolution has thus found different molecular solutions in insects and mammals to the convergent challenge of selecting small subsets of receptors from large odorant receptor repertoires.

Targeted treatment of injured nestmates with antimicrobial compounds in an ant society.

Infected wounds pose a major mortality risk in animals. Injuries are common in the ant Megaponera analis, which raids pugnacious prey. Here we show that M. analis can determine when wounds are infected and treat them accordingly. By applying a variety of antimicrobial compounds and proteins secreted from the metapleural gland to infected wounds, workers reduce the mortality of infected individuals by 90%. Chemical analyses showed that wound infection is associated with specific changes in the cuticular hydrocarbon profile, thereby likely allowing nestmates to diagnose the infection state of injured individuals and apply the appropriate antimicrobial treatment. This study demonstrates that M. analis ant societies use antimicrobial compounds produced in the metapleural glands to treat infected wounds and reduce nestmate mortality.

Estrogen-related receptor functions via the 20-hydroxyecdysone and IIS/TOR signaling pathways to regulate the development and morphology changes of ant Polyrhachis vicina Roger (Hymenoptera, Formicidae).

Estrogen-related receptor (ERR) is a key regulator of insect growth, development, and metabolic processes in insects; however, the molecular mechanisms underlying its effects are not fully understood. We investigated roles of 20-hydroxyecdysone (20E) and insulin/insulin-like signaling/target of rapamycin (IIS/TOR) signaling pathways in the effects of PvERR on larval development, metamorphosis, and adult growth in ant Polyrhachis vicina Roger. PvFOXO expression levels depended on caste and developmental stage. PvERR RNAi significantly reduced the expression levels of IIS/TOR signaling pathway genes and 20E signaling pathway genes in fourth-instar larvae, pupae, females, and workers and significantly increased the expression levels of IIS/TOR signaling pathway genes PvFOXO and PvAkt in males. PvFOXO RNAi resulted in developmental defects and increased mortality. After PvFOXO RNAi, the expression of PvERR, 20E signaling pathway genes, and IIS/TOR signaling pathway genes decreased significantly in pupae, females, and workers and increased significantly in fourth-instar larvae. Exogenous 20E attenuated expression changes induced by PvFOXO RNAi in a sex- and stage-specific manner. These results indicate that ERR interacts with 20E and IIS/TOR signaling pathways to regulate caste determination, metamorphosis, and male fertility in P. vicina and that correlations between PvERR and PvFOXO are caste- and stage-specific.

Neuropeptide regulations on behavioral plasticity in social insects.

Social insects demonstrate remarkable behavioral flexibility in response to complex external and social environments. One of the most striking examples of this adaptability is the context-dependent division of labor among workers of bees and ants. Neuropeptides, the brain's most diverse group of messenger molecules, play an essential role in modulating this phenotypic plasticity related to labor division in social insects. Integrated omics research and mass spectrometry imaging technology have greatly accelerated the identification and spatiotemporal analysis of neuropeptides. Moreover, key roles of several neuropeptides in age- and caste-dependent behavioral plasticity have been uncovered. This review summarizes recent advances in the characterization, expression, distribution, and functions of neuropeptides in controlling behavioral plasticity in social insects, particularly bees and ants. The article concludes with a discussion of future directions and challenges in understanding the regulation of social behavior by neuropeptides.

Mealybug salivary microbes inhibit induced plant defenses.

BACKGROUND: Phenacoccus solenopsis is a polyphagous invasive mealybug that caused serious damage to crops worldwide. Phloem-sucking hemipterans are known to carry symbiotic microbes in their saliva. However, the role of salivary bacteria of P. solenopsis in modulating plant defenses remains limited. Exploring the impact of salivary bacteria on plant defense responses will contribute to the development of new targets for efficient control of invasive mealybugs. RESULTS: Salivary bacteria of the invasive mealybug P. solenopsis can suppress herbivore-induced plant defenses and thus enhance mealybug fitness. Mealybugs treated with an antibiotic showed decreased weight gain, fecundity and survival. Untreated mealybugs suppressed jasmonic acid (JA)-regulated defenses but activated salicylic acid (SA)-regulated defenses in cotton plants. In contrast, antibiotic-treated mealybugs triggered JA-responsive gene expression and JA accumulation, and showed shortened phloem ingestion. Reinoculating antibiotic-treated mealybugs with Enterobacteriaceae or Stenotrophomonas cultivated from mealybug saliva promoted phloem ingestion and fecundity, and restored the ability of mealybugs to suppress plant defenses. Fluorescence in situ hybridization visualization revealed that Enterobacteriaceae and Stenotrophomonas colonize salivary glands and are secreted into the mesophyll cells and phloem vessels. Exogenous application of the bacterial isolates to plant leaves inhibited JA-responsive gene expression and activated SA-responsive gene expression. CONCLUSION: Our findings imply that symbiotic bacteria in the saliva of the mealybug play an important role in manipulating herbivore-induced plant defenses, enabling this important pest to evade induced plant defenses and promoting its performance and destructive effects on crops. © 2023 Society of Chemical Industry.

Protective role of trehalose in the Namib desert ant, Ocymyrmex robustior.

Over recent decades, increasing attention has been paid to how low-molecular-weight molecules affect thermal tolerance in animals. Although the disaccharide sugar trehalose is known to serve as a thermal protectant in unicellular organisms, nothing is known about its potential role in insects. In this study, we investigated the effect of trehalose on heat tolerance in the Namib desert ant, Ocymyrmex robustior, one of the most thermotolerant animals found in terrestrial ecosystems. First, we tested whether a trehalose-supplemented diet increased worker survival following exposure to heat stress. Second, we assessed the degree of protein damage by comparing protein aggregation levels for trehalose-supplemented workers and control workers. Third, we compared the expression levels of three genes involved in trehalose metabolism. We found that trehalose supplementation significantly enhanced worker heat tolerance, increased metabolic levels of trehalose and reduced protein aggregation under conditions of heat stress. Expression levels of the three genes varied in a manner that was consistent with the maintenance of trehalose in the hemolymph and tissues under conditions of heat stress. Altogether, these results suggest that increased trehalose concentration may help protect Namib desert ant individuals against heat stress. More generally, they highlight the role played by sugar metabolites in boosting tolerance in extremophiles.

Identification, cloning, and characterization of a novel chitinase from leaf-cutting ant Atta sexdens: An enzyme with antifungal and insecticidal activity.

Chitinases are enzymes that degrade chitin, a polysaccharide found in the exoskeleton of insects, fungi, yeast, and internal structures of other vertebrates. Although chitinases isolated from bacteria, fungi and plants have been reported to have antifungal or insecticide activities, chitinases from insects with these activities have been seldomly reported. In this study, a leaf-cutting ant Atta sexdens DNA fragment containing 1623 base pairs was amplified and cloned into a vector to express the protein (AsChtII-C4B1) in Pichia pastoris. AsChtII-C4B1, which contains one catalytic domain and one carbohydrate-binding module (CBM), was secreted to the extracellular medium and purified by ammonium sulfate precipitation followed by nickel column chromatography. AsChtII-C4B1 showed maximum activity at pH 5.0 and 55 °C when tested against colloidal chitin substrate and maintained >60% of its maximal activity in different temperatures during 48 h. AsChtII-C4B1 decreased the survival of Spodoptera frugiperda larvae fed with an artificial diet that contained AsChtII-C4B1. Our results have indicated that AsChtII-C4B1 has a higher effect on larva-pupa than larva-larva molts. AsChtII-C4B1 activity targets more specifically the growth of filamentous fungus than yeast. This work describes, for the first time, the obtaining a recombinant chitinase from ants and the characterization of its insecticidal and antifungal activities.

Unraveling the secretion mechanism of the curious nectaries in Gentianaceae.

Unusual nectaries were anatomically described as being usual traits for Gentianaceae. They are small, avascularized, and formed by 3 to 5 rosette cells with labyrinthine walls around one central cell. Such as nectaries have been reported for stems, petals, and sepals of different species of the family, however, there is no information on the mechanisms involved with the synthesis and release of secretion. Thus, this work aimed to unravel the mechanism of secretion and exudation of nectar for these curious nectaries using Calolisianthus speciosus as a model. Samples were processed according to standard methods for light and electron microscopy. Leaf and sepal nectaries were described, as were those of the apex of petals where ants were observed patrolling a darkened region. The enzymatic method was used for the detection of sugars, proteins, and amino acids in leaf and sepal exudates. The nectaries of petals of C. speciosus are similar to those of its leaves, sepals, and stem, although their activities are asynchronous. Polysaccharides were detected on the labyrinthine walls of rosette cells and protein in the opposite region of the cytoplasm. Labyrinthine walls increase the contact surface between rosette cells and the central cell, allowing for the transfer of secretion. After accumulation, the secretion is released to the subcuticular space of the central cell through disruption of the cuticle. The secretion and exudation of nectar were elucidated and involve distinct organelles.

Insulin signaling in the long-lived reproductive caste of ants.

In most organisms, reproduction is correlated with shorter life span. However, the reproductive queen in eusocial insects exhibits a much longer life span than that of workers. In Harpegnathos ants, when the queen dies, workers can undergo an adult caste switch to reproductive pseudo-queens (gamergates), exhibiting a five-times prolonged life span. To explore the relation between reproduction and longevity, we compared gene expression during caste switching. Insulin expression is increased in the gamergate brain that correlates with increased lipid synthesis and production of vitellogenin in the fat body, both transported to the egg. This results from activation of the mitogen-activated protein kinase (MAPK) branch of the insulin signaling pathway. By contrast, the production in the gamergate developing ovary of anti-insulin Imp-L2 leads to decreased signaling of the AKT/forkhead box O (FOXO) branch in the fat body, which is consistent with their extended longevity.

Social environment affects sensory gene expression in ant larvae.

Social insects depend on communication to regulate social behaviour. This also applies to their larvae, which are commonly exposed to social interactions and can react to social stimulation. However, how social insect larvae sense their environment is not known. Using RNAseq, we characterized expression of sensory-related genes in larvae of the ant Formica fusca, upon exposure to two social environments: isolation without contact to other individuals, and stimulation via the presence of other developing individuals. Expression of key sensory-related genes was higher following social stimulation, and larvae expressed many of the same sensory-related genes as adult ants and larvae of other insects, including genes belonging to the major insect chemosensory gene families. Our study provides first insights into the molecular changes associated with social information perception in social insect larvae.

Eusociality is linked to caste-specific differences in metabolism, immune system, and somatic maintenance-related processes in an ant species.

The social organization of many primate, bird and rodent species and the role of individuals within that organization are associated with specific individual physiological traits. However, this association is perhaps most pronounced in eusocial insects (e.g., termites, ants). In such species, genetically close individuals show significant differences in behavior, physiology, and life expectancy. Studies addressing the metabolic changes according to the social role are still lacking. We aimed at understanding how sociality could influence essential molecular processes in a eusocial insect, the black garden ant (Lasius niger) where queens can live up to ten times longer than workers. Using mass spectrometry-based analysis, we explored the whole metabolome of queens, nest-workers and foraging workers. A former proteomics study done in the same species allowed us to compare the findings of both approaches. Confirming the former results at the proteome level, we showed that queens had fewer metabolites related to immunity. Contrary to our predictions, we did not find any metabolite linked to reproduction in queens. Among the workers, foragers had a metabolic signature reflecting a more stressful environment and a more highly stimulated immune system. We also found that nest-workers had more digestion-related metabolites. Hence, we showed that specific metabolic signatures match specific social roles. Besides, we identified metabolites differently expressed among behavioral castes and involved in nutrient sensing and longevity pathways (e.g., sirtuins, FOXO). The links between such molecular pathways and aging being found in an increasing number of taxa, our results confirm and strengthen their potential universality.

Biomarkers in a socially exchanged /fluid reflect colony maturity, behavior, and distributed metabolism.

In cooperative systems exhibiting division of labor, such as microbial communities, multicellular organisms, and social insect colonies, individual units share costs and benefits through both task specialization and exchanged materials. Socially exchanged fluids, like seminal fluid and milk, allow individuals to molecularly influence conspecifics. Many social insects have a social circulatory system, where food and endogenously produced molecules are transferred mouth-to-mouth (stomodeal trophallaxis), connecting all the individuals in the society. To understand how these endogenous molecules relate to colony life, we used quantitative proteomics to investigate the trophallactic fluid within colonies of the carpenter ant Camponotus floridanus. We show that different stages of the colony life cycle circulate different types of proteins: young colonies prioritize direct carbohydrate processing; mature colonies prioritize accumulation and transmission of stored resources. Further, colonies circulate proteins implicated in oxidative stress, ageing, and social insect caste determination, potentially acting as superorganismal hormones. Brood-caring individuals that are also closer to the queen in the social network (nurses) showed higher abundance of oxidative stress-related proteins. Thus, trophallaxis behavior could provide a mechanism for distributed metabolism in social insect societies. The ability to thoroughly analyze the materials exchanged between cooperative units makes social insect colonies useful models to understand the evolution and consequences of metabolic division of labor at other scales.

Division of labor is essential for cooperation, because groups can achieve more when individuals specialize in different tasks. This happens across the natural world, from different cells in organisms performing specific roles, to the individuals in an ant colony carrying out diverse duties. In both of these systems, individuals work together to ensure the survival of the collective unit ­ the body or the colony ­ instead of competing against each other. One of the main ways division of labor is evident within these two systems is regarding reproduction. Both in the body and in an ant colony, only one or a few individual units can reproduce, while the rest provide support. In the case of ant colonies, only queens and males reproduce, while the young workers nurse the brood and older workers forage for food. This intense cooperation requires close communication between individual units ­ in the case of some species of ants, by sharing fluids mouth-to-mouth. These fluids contain food but also many molecules produced by the ants themselves, including proteins. Given that both individuals and the colony as a whole change as they age ­ with workers acquiring new roles, and new queens and males only reared once the colony is mature ­ it is likely that the proteins transmitted in the fluid also change. To better understand whether the lifecycles of individuals and the age of the colony affect the fluids shared by carpenter ants Camponotus floridanus, Hakala et al. examined the ant-produced proteins in these fluids. This revealed differences in the proteins shared by young and mature colonies, and young nurse ants and older forager ants. In young colonies, the fluids contained proteins involved in fast sugar processing; while in mature colonies, the fluids contained more proteins to store nutrients, which help insect larvae grow into larger individuals, like queens. Young worker ants, who spend their time nursing the brood, produced more anti-aging proteins. This may be because these ants are in close contact with the queen, who lives much longer than the rest of the ants in the colony. Taken together, these observations suggest that ants divide the labor of metabolism, as well as work and reproduction. Dividing the labor of metabolism among individuals is one more similarity between ants and the cells of a multicellular organism, like a fly or a human. Division of labor allows the sharing of burden, with some individuals lightening the load of others. Understanding how ants achieve this by sharing fluids could shed new light on this complex exchange at other scales or in other organisms. By matching proteins to life stages, researchers have a starting point to examine individual molecules in more detail.

Carbon dioxide levels in initial nests of the leaf-cutting ant Atta sexdens (Hymenoptera: Formicidae).

Claustral foundation of nests by Atta sexdens Forel (Hymenoptera: Formicidae) involves great effort by its queens, solely responsible for the cultivation of the fungus and care for her offspring at this stage. The minimum workers, after 4 months, open access to the external environment to foraging plants to cultivate the symbiotic fungus, which decomposes the plant fragments and produces gongilidea nodules as food for the individuals in the colony. Colony gas exchange and decomposition of organic matter in underground ant nests generate carbon dioxide (CO2) emitted into the atmosphere. We described the carbon dioxide concentration in colonies in the field. The objective was to evaluate the carbon dioxide concentration in initial A. sexdens colonies, in the field, and their development. The CO2 level was also measured in 4-month-old colonies in the field, using an open respirometric system fitted with an atmospheric air inlet. The CO2 level of the respirometric container was read by introducing a tube into the nest inlet hole and the air sucked by a peristaltic pump into the CO2 meter box. The CO2 concentration in the initial colony was also measured after 4 months of age, when the offspring production (number of eggs, larvae, pupae and adult workers) stabilized. Ten perforations (15 cm deep) was carried out in the adjacent soil, without a nest of ants nearby, to determine the concentration of CO2. The composition of the nests in the field was evaluated after excavating them using a gardening shovel and they were stored in 250 ml pots with 1 cm of moistened plaster at the bottom. The CO2 concentration was higher in field nest than in adjacent soil. The concentration of carbon dioxide in A. sexdens nests in the field is higher than in those in the soil, due to the production of CO2 by the fungus garden and colony.

Color lightness of velvet ants (Hymenoptera: Mutillidae) follows an environmental gradient.

Color traits are highly influenced by environmental conditions along the distributional range of many species. Studies on the variation of animal coloration across different geographic gradients are, therefore, fundamental for a better understanding of the ecological and evolutionary processes that shape color variation. Here, we address whether color lightness in velvet ants (Hymenoptera: Mutillidae) responds to latitudinal gradients and bioclimatic variations, testing three ecogeographic rules: The Thermal melanism hypothesis; the Photoprotection hypothesis; and Gloger's rule. We test these hypotheses across the New World. We used photographs of 482 specimens (n = 142 species) of female mutillid wasps and extracted data on color lightness (V). We analyzed whether variation in color is determined by bioclimatic factors, using Phylogenetic Generalized Least Square analysis. Our explanatory variables were temperature, ultraviolet radiation, humidity, and forest indicators. Our results were consistent with the Photoprotection hypothesis and Gloger's rule. Species with darker coloration occupied habitats with more vegetation, higher humidity, and UV-B radiation. However, our results refute one of the initial hypotheses suggesting that mutillids do not respond to the predictions of the Thermal melanism hypothesis. The results presented here provide the first evidence that abiotic components of the environment can act as ecological filters and as selective forces driving the body coloration of velvet ants. Finally, we suggest that studies using animals with melanin-based colors as a model for mimetic and aposematic coloration hypotheses consider that this coloration may also be under the influence of climatic factors and not only predators.

Comparison of gene expression in the red imported fire ant (Solenopsis invicta) under different temperature conditions.

The red imported fire ant (RIFA), Solenopsis invicta Buren is native to South America and is known as a global problematic invasive species. This study focused on the molecular response of RIFA by comparing gene expression profiles after exposing ants to low (10 °C) and high (40 °C) temperature stress and comparing them to untreated controls (30 °C). A total of 99,085 unigenes (the clustered non-redundant transcripts that are filtered from the longest assembled contigs) were obtained, of which 19,154 were annotated with gene descriptions, gene ontology terms, and metabolic pathways. 86 gene ontology (GO) functional sub-groups and 23 EggNOG terms resulted. Differentially expressed genes (DEGs) with log2FC ≥ 10 were screened and were compared at different temperatures. We found 203, 48, and 66 specific DEGs co-regulated at 10, 20, and 40 °C. Comparing transcriptome profiles for differential gene expression resulted in various DE genes, including cytochrome P450, NADH dehydrogenase subunit 1, cuticle protein and heat shock protein (HSP), which have previously been reported to be involved in cold and high temperature resistance. GO analysis revealed that antioxidant activity is up-regulated under high temperature stress. We verified the RNA-seq data by qPCR on 20 up- and down-regulated DEGs. These findings provide a basis for future understanding of the adaptation mechanisms of RIFA and the molecular mechanisms underlying the response to low and high temperatures.

Mandibular muscle troponin of the Florida carpenter ant Camponotus floridanus: extending our insights into invertebrate Ca2+ regulation.

Ants use their mandibles for a variety of functions and behaviors. We investigated mandibular muscle structure and function from major workers of the Florida carpenter ant Camponotus floridanus: force-pCa relation and velocity of unloaded shortening of single, permeabilized fibres, primary sequences of troponin subunits (TnC, TnI and TnT) from a mandibular muscle cDNA library, and muscle fibre ultrastructure. From the mechanical measurements, we found Ca2+-sensitivity of isometric force was markedly shifted rightward compared with vertebrate striated muscle. From the troponin sequence results, we identified features that could explain the rightward shift of Ca2+-activation: the N-helix of TnC is effectively absent and three of the four EF-hands of TnC (sites I, II and III) do not adhere to canonical sequence rules for divalent cation binding; two alternatively spliced isoforms of TnI were identified with the alternatively spliced exon occurring in the region of the IT-arm α-helical coiled-coil, and the N-terminal extension of TnI may be involved in modulation of regulation, as in mammalian cardiac muscle; and TnT has a Glu-rich C-terminus. In addition, a structural homology model was built of C. floridanus troponin on the thin filament. From analysis of electron micrographs, we found thick filaments are almost as long as the 6.8 µm sarcomeres, have diameter of ~ 16 nm, and typical center-to-center spacing of ~ 46 nm. These results have implications for the mechanisms by which mandibular muscle fibres perform such a variety of functions, and how the structure of the troponin complex aids in these tasks.

Gene Coexpression Network Reveals Highly Conserved, Well-Regulated Anti-Ageing Mechanisms in Old Ant Queens.

Evolutionary theories of ageing predict a reduction in selection efficiency with age, a so-called "selection shadow," due to extrinsic mortality decreasing effective population size with age. Classic symptoms of ageing include a deterioration in transcriptional regulation and protein homeostasis. Understanding how ant queens defy the trade-off between fecundity and lifespan remains a major challenge for the evolutionary theory of ageing. It has often been discussed that the low extrinsic mortality of ant queens, that are generally well protected within the nest by workers and soldiers, should reduce the selection shadow acting on old queens. We tested this by comparing strength of selection acting on genes upregulated in young and old queens of the ant, Cardiocondyla obscurior. In support of a reduced selection shadow, we find old-biased genes to be under strong purifying selection. We also analyzed a gene coexpression network (GCN) with the aim to detect signs of ageing in the form of deteriorating regulation and proteostasis. We find no evidence for ageing. In fact, we detect higher connectivity in old queens indicating increased transcriptional regulation with age. Within the GCN, we discover five highly correlated modules that are upregulated with age. These old-biased modules regulate several antiageing mechanisms such as maintenance of proteostasis, transcriptional regulation, and stress response. We observe stronger purifying selection on central hub genes of these old-biased modules compared with young-biased modules. These results indicate a lack of transcriptional ageing in old C. obscurior queens, possibly facilitated by strong selection at old age and well-regulated antiageing mechanisms.