Biology subjects

Kronauer, D. J. C.

Publications and source records attributed to Kronauer, D. J. C..

12 recordsLinked to original sources

bioRxiv2026

Spontaneous neural activity in the developing ant olfactory system

Spontaneous neural activity is a pillar of mammalian neurodevelopment. In mammals, developing olfactory sensory neurons (OSNs) show spontaneous activity patterns that depend on the olfactory receptor (OR) they express. This activity is necessary for proper sorting of OSN axons into olfactory bulb glomeruli. This contrasts with the simpler olfactory system of the fruit fly Drosophila melanogaster, where antennal lobe glomeruli form without functional ORs. Compared to Drosophila, ants have a vastly expanded olfactory system and, unlike Drosophila, require functional ORs for proper development. This raises the possibility that ants, like mammals, rely on spontaneous neural activity. To test whether developing ant OSNs are spontaneously active, we performed live calcium imaging in translucent pupae of the clonal raider ant Ooceraea biroi, using a transgenic line expressing the genetically encoded calcium indicator GCaMP6s in OSNs. Developing O. biroi OSNs exhibit spontaneous calcium transients from the earliest stages of glomerulus formation, and features of this activity change over the course of metamorphosis. During early stages, activity is faster, varies widely between cells, and is uncorrelated across the OSN population. By mid-metamorphosis, activity has slowed, with longer calcium events and fewer peaks per cell. As pupae approach eclosion, a new mode appears, with rhythmic, sawtooth-shaped calcium events that synchronously engage OSNs in both antennae. This stage-dependent spontaneous activity suggests that the expanded ant olfactory system may use a developmental logic more akin to mammals than to Drosophila.

neuroscience↗

bioRxiv2026

Rare sex punctuates strict asexual reproduction in the clonal raider ant, Ooceraea biroi

While asexual species can often outcompete their sexual counterparts over ecological timescales, their long-term evolutionary success is hindered by a diminished ability to purge deleterious mutations and to adapt to changing environments. However, some asexual species persist for millions of years, and a major question in evolutionary biology is how they do so. One solution is to occasionally reproduce sexually, as has been shown in a handful of primarily asexual species. Here, we investigate the possibility of rare sex in the clonal raider ant, Ooceraea biroi. We report the whole-genome sequence of a previously uncharacterized clonal line and, using population genetic and phylogenetic analyses, show that it originated through sexual reproduction between two extensively studied clonal lines. The mitochondrial genome of this clonal line differs from that of the maternal clonal line at only a single nucleotide, suggesting that the sexual reproduction event occurred within the past few hundred years. These results demonstrate that sex occurs sporadically in the clonal raider ant, allowing it to generate new genetic combinations and potentially to overcome some of the costs of asexuality.

evolutionary biology↗

bioRxiv2025

A reference brain for the clonal raider ant

Ants exhibit remarkable collective and social behaviors, such as alloparental care1, chemical communication2, homing3, and cooperative group hygiene4. The clonal raider ant Ooceraea biroi is especially well-suited for investigating the neuronal and genetic underpinnings of these behaviors5. Unlike most ant species, O. biroi lacks a queen caste. Instead, colonies consist entirely of regular workers and slightly larger intercaste workers6. All workers reproduce in synchrony via parthenogenesis, giving rise to age-matched cohorts of clonally identical offspring7,8. This unique life history enables precise experimental control over age, genotype, and colony composition. These features have also facilitated the introduction of genetically encoded calcium indicators into O. biroi, enabling in vivo two-photon imaging to investigate the neural basis of social behaviors9. Despite its promise as a neuroscience model, the structure of the clonal raider ant brain has not been systematically characterized, and a representative reference brain does not exist. To address this gap, we imaged the brains of 40 age-matched, genetically identical individuals with confocal microscopy and, using 3D groupwise registration, generated the first reference brain for the species. We introduce a registration pipeline to align brains to this reference, facilitating the comparison of anatomical features across labeling experiments with high spatial precision. Unexpectedly, despite homogeneity in genotype, age, and external morphology, we discovered extensive interindividual variability across our collection of brain samples. This raises the possibility that behavioral division of labor in O. biroi is linked to individual differences in brain structure. This work provides a powerful resource for the emerging clonal raider ant neuroscience community and reveals novel features of the species neurobiology that may influence social behaviors and colony function.

neuroscience↗

bioRxiv2025

Transcriptional Interference Gates Monogenic Odorant Receptor Expression in Ants

Communication is crucial to social life, and in ants, it is mediated primarily through olfaction. Ants have more odorant receptor (OR) genes than any other group of insects, generated through tandem duplications that produce large genomic arrays of related genes. However, how olfactory sensory neurons produce a single functional OR from these arrays remains unclear. In ants, only mRNA from one OR in an array is exported into the cytoplasm, while upstream genes are silent and transcripts from downstream genes remain nuclear. Here, we show that non-canonical readthrough transcription in the downstream direction generates non-translated transcripts. We also find that OR promoters are bidirectional, producing antisense long non-coding RNAs that appear to suppress the expression of upstream genes. Finally, we present evidence that this regulatory architecture is conserved across ants and bees, suggesting that this mechanism for functionally monogenic OR expression is widespread in insects with expanded OR repertoires.

neuroscience↗

bioRxiv2025

Heterozygosity at a conserved candidate sex determination locus is associated with female development in the clonal raider ant

Sex determination is a developmental switch that triggers sex-specific developmental programs. This switch is "flipped" by the expression of genes that promote male- or female-specific development. Many lineages have evolved sex chromosomes that act as primary signals for sex determination. However, haplodiploidy (males are haploid and females are diploid), which occurs in ca. 12% of animal species, is incompatible with sex chromosomes. Haplodiploid taxa must, therefore, rely on other strategies for sex determination. One mechanism, "complementary sex determination" (CSD), uses heterozygosity as a proxy for diploidy. In CSD, heterozygosity at a sex determination locus triggers female development, while hemizygosity or homozygosity permits male development. CSD loci have been mapped in honeybees and two ant species, but we know little about their evolutionary history. Here, we investigate sex determination in the clonal raider ant, Ooceraea biroi. We identified a 46kb candidate CSD locus at which all females are heterozygous, but most diploid males are homozygous for either allele. As expected for CSD loci, the candidate locus has more alleles than most other loci, resulting in a peak of nucleotide diversity. This peak negligibly affects the amino acid sequences of protein-coding genes, suggesting that heterozygosity of a non-coding genomic sequence triggers female development. This locus is distinct from the CSD locus in honeybees but homologous to a CSD locus mapped in two distantly related ant species, implying that this molecular mechanism has been conserved since a common ancestor that lived approximately 112 million years ago.

evolutionary biology↗

bioRxiv2024

Pheromone representation in the ant antennal lobe changes with age

While the neural basis of age-related decline has been extensively studied (1-3), less is known about changes in neural function during the pre-senescent stages of adulthood. Adult neural plasticity is likely a key factor in social insect age polyethism, where individuals perform different tasks as they age and divide labor in an age-dependent manner (4-9). Primarily, workers transition from nursing to foraging tasks (5, 10), become more aggressive, and more readily display alarm behavior (11-16) as they get older. While it is unknown how these behavioral dynamics are neurally regulated, they could partially be generated by altered salience of behaviorally relevant stimuli (4, 6, 7). Here, we investigated how odor coding in the antennal lobe (AL) changes with age in the context of alarm pheromone communication in the clonal raider ant (Ooceraea biroi) (17). Similar to other social insects (11, 12, 16), older ants responded more rapidly to alarm pheromones, the chemical signals for danger. Using whole-AL calcium imaging (18), we then mapped odor representations for five general odorants and two alarm pheromones in young and old ants. Alarm pheromones were represented sparsely at all ages. However, alarm pheromone responses within individual glomeruli changed with age, either increasing or decreasing. Only two glomeruli became sensitized to alarm pheromones with age, while at the same time becoming desensitized to general odorants. Our results suggest that the heightened response to alarm pheromones in older ants occurs via increased sensitivity in these two core glomeruli, illustrating the importance of sensory modulation in social insect division of labor and age-associated behavioral plasticity.

neuroscience↗

bioRxiv2024

Unselfish meiotic drive maintains heterozygosity in a parthenogenetic ant

According to Mendels second law, chromosomes segregate randomly in meiosis. Non-random segregation is primarily known for cases of selfish meiotic drive in females, in which particular alleles bias their own transmission into the oocyte1,2. Here, we report a rare example of unselfish meiotic drive for crossover inheritance in the clonal raider ant, Ooceraea biroi. This species produces diploid offspring parthenogenetically via fusion of two haploid nuclei from the same meiosis3. This process should cause rapid genotypic degeneration due to loss of heterozygosity, which results if crossover recombination is followed by random (Mendelian) segregation of chromosomes4,5. However, by comparing whole genomes of mothers and daughters, we show that loss of heterozygosity is exceedingly rare, raising the possibility that crossovers are infrequent or absent in O. biroi meiosis. Using a combination of cytology and whole genome sequencing, we show that crossover recombination is, in fact, common, but that loss of heterozygosity is avoided because crossover products are faithfully co-inherited. This results from a programmed violation of Mendels law of segregation, such that crossover products segregate together rather than randomly. This discovery highlights an extreme example of cellular "memory" of crossovers, which could be a common yet cryptic feature of chromosomal segregation.

evolutionary biology↗

bioRxiv2022

Sparse and stereotyped encoding implicates a core glomerulus for ant alarm behavior

Ants communicate via large arrays of pheromones and possess expanded, highly complex olfactory systems, with antennal lobes in the brain comprising ~500 glomeruli. This expansion implies that odors could activate hundreds of glomeruli, which would pose challenges for higher order processing. To study this problem, we generated the first transgenic ants, expressing the genetically encoded calcium indicator GCaMP6s in olfactory sensory neurons. Using two-photon imaging, we mapped complete glomerular responses to four ant alarm pheromones. Alarm pheromones robustly activated [≤]6 glomeruli, and activity maps for the three pheromones inducing panic-alarm in our study species converged on a single glomerulus. These results demonstrate that, rather than using broadly tuned combinatorial encoding, ants employ precise, narrowly tuned, and stereotyped representation of alarm pheromone cues. The identification of a central sensory hub glomerulus for alarm behavior suggests that a simple neural architecture is sufficient to translate pheromone perception into behavioral outputs.

neuroscience↗

bioRxiv2022

The alarm pheromone and alarm response of the clonal raider ant

Ants communicate via an arsenal of different pheromones produced in a variety of exocrine glands. For example, ants release alarm pheromones in response to danger to alert their nestmates and to trigger behavioral alarm responses. Here we characterize the alarm pheromone and the alarm response of the clonal raider ant Ooceraea biroi, a species that is amenable to laboratory studies but for which no pheromones have been identified. During an alarm response, ants quickly become unsettled, leave their nest pile, and are sometimes initially attracted to the source of alarm, but ultimately move away from it. We find that the alarm pheromone is released from the head of the ant and identify the putative alarm pheromone as a blend of two compounds found in the head, 4-methyl-3-heptanone and 4-methyl-3-heptanol. These compounds are sufficient to induce alarm behavior alone and in combination. They elicit similar, though slightly different behavioral features of the alarm response, with 4-methyl-3-heptanone being immediately repulsive and 4-methyl-3-heptanol being initially attractive before causing ants to move away. The behavioral response to these compounds in combination is dose-dependent, with ants becoming unsettled and attracted to the source of alarm pheromone at low concentrations and repulsed at high concentrations. While 4-methyl-3-heptanone and 4-methyl-3-heptanol are known alarm pheromones in other more distantly related ant species, this is the first report of the chemical identity of a pheromone in O. biroi, and the first alarm pheromone identified in the genus Ooceraea. Identification of a pheromone that triggers a robust, consistent, and conserved behavior, like the alarm pheromone, provides an avenue to dissect the behavioral and neuronal mechanisms underpinning chemical communication.

animal behavior and cognition↗

bioRxiv2022

Microbial symbionts are shared between ants and their associated beetles

The transmission of microbial symbionts across individuals and generations can be critical for animal development and survival. Likewise, the transmission of microbes across closely interacting species could also affect host biology. Army ants (Formicidae: Dorylinae) and their hundreds of closely associated insect species (myrmecophiles) can provide a unique insight into interspecific symbiont sharing. Here, we compared the microbiota of workers and larvae of the army ant Eciton burchellii with those of 13 myrmecophile beetle species using 16S rRNA amplicon sequencing. We found that the previously characterized symbionts of army ant workers (Unclassified Firmicutes and Unclassified Entomoplasmatales) were largely absent from ant larvae and from myrmecophiles, whose microbial communities were usually dominated by Rickettsia, Wolbachia, Rickettsiella, and/or Weissella. Strikingly, different species of myrmecophiles and ant larvae often shared identical 16S rRNA genotypes of common bacteria. In particular, army ant larvae, some workers, and several myrmecophile species often hosted identical Weissella (Lactobacillales), based on 16S rRNA and also protein-coding gene sequences. Also, we found high relatedness between some newly characterized Weissella and animal-associated strains from aquatic and marine habitats. Looking more broadly, we found Weissella OTUs in 11.6% of samples from nearly all habitats and environments characterized by the Earth Microbiome Project. Together, our data show that unrelated but closely interacting species can share much of their microbiota. The high relatedness of strains found across such disparate hosts as ants, beetles, trout, and whales suggests that some versatile microbes move between hosts and habitats despite few opportunities for direct interaction.

ecology↗

bioRxiv2020

Colony expansions underlie the evolution of army ant mass raiding

Collective behavior emerges from local interactions between group members, and natural selection can fine-tune these interactions to achieve different collective outcomes. However, at least in principle, collective behavior can also evolve via changes in group-level parameters. Here, we show that army ant mass raiding, an iconic collective behavior in which many thousands of ants spontaneously leave the nest to go hunting, has evolved from group raiding, in which a scout directs a much smaller group of ants to a specific target. We describe the structure of group raids in the clonal raider ant, a close relative of army ants. We find that the coarse structure of group raids and mass raids is highly conserved, and that army ants and their relatives likely follow similar behavioral rules, despite the fact that their raids differ strikingly in overall appearance. By experimentally increasing colony size in the clonal raider ant, we show that mass raiding gradually emerges from group raiding without altering individual behavioral rules. This suggests a simple mechanism for the evolution of army ant mass raids, and more generally that scaling effects may provide an alternative mechanism for evolutionary transitions in complex collective behavior.

evolutionary biology↗

bioRxiv2016

The adaptive significance of phasic colony cycles in army ants

Army ants are top arthropod predators in tropical forests around the world. The colonies of many army ant species undergo stereotypical behavioral and reproductive cycles, alternating between brood care and reproductive phases. In the brood care phase, colonies contain a cohort of larvae that are synchronized in their development and have to be fed. In the reproductive phase larvae are absent and oviposition takes place. Despite these colony cycles being a striking feature of army ant biology, their adaptive significance is unclear. Here we use a modelling approach to show that cyclic reproduction is favored under conditions where per capita foraging costs decrease with the number of larvae in a colony (\"High Cost of Entry\" scenario), while continuous reproduction is favored under conditions where per capita foraging costs increase with the number of larvae (\"Resource Exhaustion\" scenario). We argue that the former scenario specifically applies to army ants, because large raiding parties are required to overpower prey colonies. However, once raiding is successful it provides abundant food for a large cohort of larvae. The latter scenario, on the other hand, will apply to non-army ants, because in those species local resource depletion will force workers to forage over larger distances to feed large larval cohorts. Our model provides the first quantitative framework for understanding the adaptive value of phasic colony cycles in ants.

evolutionary biology↗