Ozone exposure disrupts insect sexual communication.

Insect sexual communication often relies upon sex pheromones. Most insect pheromones, however, contain carbon-carbon double bonds and potentially degrade by oxidation. Here, we show that frequently reported increased levels of Anthropocenic ozone can oxidize all described male-specific pheromones of Drosophila melanogaster, resulting in reduced amounts of pheromones such as cis-Vaccenyl Acetate and (Z)-7-Tricosene. At the same time female acceptance of ozone-exposed males is significantly delayed. Interestingly, groups of ozone-exposed males also exhibit significantly increased levels of male-male courtship behaviour. When repeating similar experiments with nine other drosophilid species, we observe pheromone degradation and/or disrupted sex recognition in eight of them. Our data suggest that Anthropocenic levels of ozone can extensively oxidize double bonds in a variety of insect pheromones, thereby leading to deviations in sexual recognition.

The role of intrasexual competition on the evolution of male-male courtship display: a systematic review.

Background: Evidence of male-male courtship display is widespread across the animal kingdom. Yet, its function and evolutionary origin remain unclear. Here, we hypothesise that male-male courtship display evolved in response to selection pressure exerted by intrasexual competition during male-female courtship interactions. Intrasexual competition can be caused by bystander male pressure through eavesdropping and exploiting on displayer male's courtship interactions with females. This bystander pressure can lead to an audience effect by the displayer, who will change their courtship behaviour in the presence of bystanders and display directly towards them, even in the absence of females, as an intimidation strategy. In species where this selection pressure has taken place, we predict that the male courtship display will have a dual function: attract females and deter competitors. Therefore, we expected to find more evidence of bystander-related behaviours in species for which male-male courtship display is linked to intrasexual competition compared to species for which other explanatory hypotheses are more plausible (e.g., mistaken identity or courtship practice). Methodology: We conducted two systematic reviews to test this hypothesis. First, we conducted a search for studies of species with courtship display between males and of the hypotheses provided to explain this behaviour. Our goal was to identify the species with male-male courtship display and evidence of intrasexual competition. Second, among the species with male-male courtship display, we searched for evidence of bystander-related behaviours, i.e., articles referring to eavesdropping, exploitation, and audience effect during male-female courtship interactions. Our goal was to test whether species with intrasexual competition are also more likely to show bystander-related behaviours. Results: Although most studies reporting male courtship display towards other males do not suggest any explanatory hypothesis for this behaviour, the intrasexual competition hypothesis was largely mentioned and supported by some studies reviewed. Additionally, there is more evidence of eavesdropping and of all three bystander-related behaviours combined in species for which the intrasexual competition hypothesis was suggested. Conclusions: Overall, our review supports the hypothesis that intrasexual competition can play a key role in male courtship display evolution, namely that male-male courtship display may have evolved as a secondary function of male-female courtship interactions via bystander male pressure. However, our review also shows that despite the increasing interest in same-sex sexual behaviours, and male-male courtship display in particular, most studies were found to be merely descriptive, and the hypotheses they suggested to explain courtship display between males mostly speculative. This highlights an important gap in the literature. To clarify both the evolution and the function of male-male courtship display, this behaviour needs to be empirically studied more often. Our review can help advancing this research area, as it makes the 20 species with male-male courtship display for which the intrasexual competition hypothesis was suggested excellent candidates for empirical research.

Light Environment Interacts with Visual Displays in a Species-Specific Manner in Multimodal-Signaling Wolf Spiders.

AbstractLight availability is highly variable, yet predictable, over various timescales and is expected to play an important role in the evolution of visual signals. Courtship displays of the wolf spider genus Schizocosa always involve the use of substrate-borne vibrations; however, there is substantial variation in the presence and complexity of visual displays among species. To gain insight into the role the light environment plays in the evolution of courtship displays, we tested the function of visual courtship signaling across distinct light environments in four species of Schizocosa that vary in their degree of ornamentation and dynamic visual signals. We ran mating and courtship trials at three light intensities (bright, dim, and dark) and tested the hypothesis that ornamentation interacts with light environment. We also examined each species' circadian activity patterns. The effects of the light environment on courtship and mating varied between species, as did circadian activity patterns. Our results suggest that femur pigmentation may have evolved for diurnal signaling, whereas tibial brushes may function to increase signal efficacy under dim light. Additionally, we found evidence for light-dependent changes in selection on male traits, illustrating that short-term changes in light intensity have the potential for strong effects on the dynamics of sexual selection.

Septin Defects Favour Symmetric Inheritance of the Budding Yeast Deceptive Courtship Memory.

Mnemons are prion-like elements that encode cellular memories of past cellular adaptations and do not spread to progenies during cell divisions. During the deceptive courtship in budding yeast, the Whi3 mnemon (Whi3mnem) condenses into a super-assembly to encode a mating pheromone refractory state established in the mother cell. Whi3mnem is confined to the mother cell such that their daughter cells have the ability to respond to the mating pheromone. Confinement of Whi3mnem involves its association with the endoplasmic reticulum membranes and the compartmentalization of these membranes by the lateral membrane diffusion barrier at the bud neck, the limit between the mother cell and the bud. However, during the first cell division after the establishment of the pheromone refractory state, this adaptation is more likely to be inherited by the daughter cell than in subsequent cell divisions. Here, we show that the first cell division is associated with larger daughter cells and cytokinesis defects, traits that are not observed in subsequent cell divisions. The cytoskeletal septin protein shows aberrant localisation in these divisions and the septin-dependent endoplasmic reticulum membrane diffusion barrier is weakened. Overall, these data suggest that cytokinesis defects associated with prolonged cell division can alter the confinement and inheritance pattern of a cellular memory.

Reproductive advantage of the winners of male-male competition in Drosophila prolongata.

In the resource-defence mating system, where males compete for limited resources to acquire females, male traits associated with fighting ability are selected, leading to the evolution of sexual dimorphism. However, the evolution of sexual dimorphism is also driven by other mechanisms, such as female selection. Therefore, to elucidate the evolutionary mechanisms of male traits, it is necessary to clarify their contribution to fitness through male-male competition. In this regard, it is surprising that numerous studies on sexually dimorphic species have assumed the resource-defence mating system without directly examining the relationship between resource-defending behaviour and mating success. In a sexually dimorphic fruit fly, Drosophila prolongata, the presence of the resource-defence mating system has been suggested, but technical difficulties had prevented spatial quantification of the resource-defending behaviour. In this study, by using an automated behaviour analysis tool previously developed, we located the occurrence of male-male competition and courtship to investigate their relationship in D. prolongata, considering the position of food resources. We found that the male-male competition led to the exclusive occupation of resources, increasing the courtship opportunities of the resource holders. These results illustrate the importance of resource-defending for reproductive success in D. prolongata.

The Drosophila dopamine 2-like receptor D2R (Dop2R) is required in the blood brain barrier for male courtship.

The blood brain barrier (BBB) has the essential function to protect the brain from potentially hazardous molecules while also enabling controlled selective uptake. How these processes and signaling inside BBB cells control neuronal function is an intense area of interest. Signaling in the adult Drosophila BBB is required for normal male courtship behavior and relies on male-specific molecules in the BBB. Here we show that the dopamine receptor D2R is expressed in the BBB and is required in mature males for normal mating behavior. Conditional adult male knockdown of D2R in BBB cells causes courtship defects. The courtship defects observed in genetic D2R mutants can be rescued by expression of normal D2R specifically in the BBB of adult males. Drosophila BBB cells are glial cells. Our findings thus identify a specific glial function for the DR2 receptor and dopamine signaling in the regulation of a complex behavior.

Evolutionary conservation and diversification of auditory neural circuits that process courtship songs in Drosophila.

Acoustic communication signals diversify even on short evolutionary time scales. To understand how the auditory system underlying acoustic communication could evolve, we conducted a systematic comparison of the early stages of the auditory neural circuit involved in song information processing between closely-related fruit-fly species. Male Drosophila melanogaster and D. simulans produce different sound signals during mating rituals, known as courtship songs. Female flies from these species selectively increase their receptivity when they hear songs with conspecific temporal patterns. Here, we firstly confirmed interspecific differences in temporal pattern preferences; D. simulans preferred pulse songs with longer intervals than D. melanogaster. Primary and secondary song-relay neurons, JO neurons and AMMC-B1 neurons, shared similar morphology and neurotransmitters between species. The temporal pattern preferences of AMMC-B1 neurons were also relatively similar between species, with slight but significant differences in their band-pass properties. Although the shift direction of the response property matched that of the behavior, these differences are not large enough to explain behavioral differences in song preferences. This study enhances our understanding of the conservation and diversification of the architecture of the early-stage neural circuit which processes acoustic communication signals.

Female preferences between incipient species of Drosophila athabasca reveal divergent closed functions, epistasis, and segregation of veiled preference alleles.

Theoretical models typically assume that speciation is driven by divergence in female preference functions for secondary sexual traits. However, little is known about how these functions diverge between incipient species. This study used song playback experiments and mate choice tests to characterize female preference functions for divergent male courtship songs between two incipient species of Drosophila athabasca complex; D. athabasca (WN) and Drosophila mahican (EA). The study focused on two male song parameters: pulses per burst (PPB) and inter-pulse interval (IPI). Preference functions for PPB were open-ended and lacked species-specificity. In contrast, preference functions for IPI revealed a closed shape, with peak preferences highly divergent between species. Thus, females imposed divergent and ongoing sexual selection on male IPI. Further work demonstrated variation in peak preferences and choosiness for IPI across genetic lines, while F1 hybrid females varied extensively in their mating preferences across different heterospecific line crosses. Finally, F2 female genotypes with foreign single-copy chromosome substitutions showed little to no change in preferences until three or four chromosomes were combined: indicating strong dominance epistasis. These results suggest "veiled" or hidden segregating female preference alleles for male speciation phenotypes. This may explain the rapid speciation observed in this species complex.

Male scent but not courtship vocalizations induce estrus in wild female house mice.

Exposure to males or male urinary scent can induce and accelerate the rate of female estrous cycling in house mice ("Whitten effect"), and this response has been replicated many times since its discovery over 60 years ago. Here, we tested whether exposing female mice to recordings of male courtship ultrasonic vocalizations (USVs) induces estrous cycling, and whether exposure to both male scent and USVs has a stronger effect than to either of these stimuli alone. We conducted our study with 60 wild-derived female house mice (Mus musculus musculus). After singly housing females for 14 days, we monitored estrous stages via vaginal cytology for two weeks while isolated from males or male stimuli. We continued monitoring estrus for two more weeks during experimental exposure to one of four different types of stimuli: (1) clean bedding and background noise playback (negative control); (2) recordings of male USVs (16 min per day) and clean bedding (male USV treatment); (3) soiled male bedding and background noise playback (male odor treatment; positive control); or (4) male USVs and soiled male bedding (male odor and USV treatment). Females were then paired with males to test whether any of the four treatments influenced female reproduction (especially latency to birth). We confirmed that exposure to male odor increased female cycling, as expected, but exposure to recordings of male USVs had no effect on estrus. Females exposed to both USVs and odor went through more cycles compared to controls, but did not differ significantly from exposure to male odor (and background noise). After pairing females with a male, females showing male odor-induced cycling produced their first litter sooner than controls, whereas USVs did not have such an effect. This is the first study to our knowledge to show that male odor induces estrus in wild house mice and to show functional effects on reproduction. Our results do not support the hypothesis that male vocalizations induce female estrus, although we suggest other approaches that could be used to further test this hypothesis.

Mutation of the Drosophila melanogaster serotonin transporter dSERT impacts sleep, courtship, and feeding behaviors.

The Serotonin Transporter (SERT) regulates extracellular serotonin levels and is the target of most current drugs used to treat depression. The mechanisms by which inhibition of SERT activity influences behavior are poorly understood. To address this question in the model organism Drosophila melanogaster, we developed new loss of function mutations in Drosophila SERT (dSERT). Previous studies in both flies and mammals have implicated serotonin as an important neuromodulator of sleep, and our newly generated dSERT mutants show an increase in total sleep and altered sleep architecture that is mimicked by feeding the SSRI citalopram. Differences in daytime versus nighttime sleep architecture as well as genetic rescue experiments unexpectedly suggest that distinct serotonergic circuits may modulate daytime versus nighttime sleep. dSERT mutants also show defects in copulation and food intake, akin to the clinical side effects of SSRIs and consistent with the pleomorphic influence of serotonin on the behavior of D. melanogaster. Starvation did not overcome the sleep drive in the mutants and in male dSERT mutants, the drive to mate also failed to overcome sleep drive. dSERT may be used to further explore the mechanisms by which serotonin regulates sleep and its interplay with other complex behaviors.

Ecology and evolution of bird sounds.

Birds are among nature's most social animals. They are renowned for their group migrations, their cooperative foraging, their communal roosting, their synchronous breeding aggregations, their precise parent-offspring interactions, their coordinated group defenses and their intricate territorial and courtship rituals. In these and other contexts, and indeed in most moments of their lives, birds' capacities to navigate complex social demands and relationships can tip the balance between health or sickness, between reproductive success or failure, between life or death. It is thus no surprise that birds have evolved sophisticated systems for mediating social interactions. Foremost among these are communication signals, most evident in birds as songs, calls, color patterns and postural displays. These and other communication signals can transmit information about a signalers' status, motivations and possible future actions, for sensory and cognitive processing by signal receivers. In general, signals provide benefits for both signal senders and receivers. Consider the bright spring plumage of a male wood warbler who has established a breeding territory. The male's plumage pattern benefits him by signaling his 'ownership' of his territory, which can help him attract potential mates and preempt conflict with potential rivals. His plumage pattern also benefits his intended audiences: it informs females about the signaler's potential as a mate, and it informs other males about the degree of threat they might face were they to attempt a territory take-over. Communication signals are, in essence, a glue that maintains the cohesion of bird societies, and a currency that mediates sophisticated relationships within.

The masculinization steroid milieu caused by fluorene-9-bisphenol disrupts sex-typical courtship behavior in female zebrafish (Danio rerio).

In vertebrates, the behavior of congenital sex differences between males and females is highly dependent on steroid signals and hormonal milieu. The presence of endocrine disrupting chemicals (EDCs) in the environment generally plays a similar role to sex hormones, so its interference with aquatic organism population stability can not be ignored and is worth studying. Fluorene-9-bisphenol (BHPF) has been clarified as an endocrine disruptor on organisms by several studies but its mechanism in perturbation of courtship behavior of female zebrafish is not clear. Here, we proposed an automated multi-zebrafish tracking method quantifying the courtship process and reported that zebrafish females exposed to BHPF, are not receptive to males but rather court females, and lose normal ovarian function with an altered sex steroid milieu. Our results showed that BHPF damaged 17ß-estradiol synthesis by down-regulation of sox3 and cyp19a1a, linking apoptosis with ovary development and female fecundity. The down-regulated expression of estrogen signaling through an estrogen receptor, esr2b, caused the induction of masculinization of female courtship behavior and sexual preference in zebrafish females after BHPF treatment. This process might be mediated by inhibiting the transcription of a neuropeptide B (npb) in the brain. Our study reveals that the estrogen signaling pathway may play an important role in classical courtship behavior and sexual preference of zebrafish. This study provided evidence that anti-estrogenic chemical exposure caused adverse effects on the regulation of the brain-gonad-estrogen axis of aquatic organisms, which should be of concern and highlighted the importance of controlling environmental contamination.

Behavior and neural activation patterns of non-redundant visual and acoustic signaling during courtship in an African cichlid fish.

Animals evolve mechanisms to send and receive communication signals through multiple sensory channels during crucial behavioral contexts such as aggression and reproduction. This ensures the transmission of important context-dependent signals that supply either the same (redundant) or different (non-redundant) information to the receiver. Despite the importance of multimodal communication, there are relatively few species in which information on sender signals and receiver responses are known. Further, little is known about where context-dependent unimodal and multimodal information is processed in the brain to produce adaptive behaviors. We used the African cichlid, Astatotilapia burtoni, to investigate how unimodal and multimodal signals are processed within the female brain in a reproductive context. During courtship, dominant males produce low frequency sounds in conjunction with visual displays (quivers) directed towards receptive gravid females. We compared affiliation behaviors and neural activation patterns in gravid females exposed to visual, acoustic and visual-acoustic signals from courting dominant males. Females displayed reduced affiliation in auditory-only conditions, but similar affiliation during visual and visual-acoustic conditions, demonstrating that visual-acoustic signaling from males is non-redundant but vision dominates. Using the neural activation marker cfos, we identified differential activation in specific socially relevant brain nuclei between unimodal and multimodal conditions and distinct neural co-activation networks associated with each sensory context. Combined with our previous work on chemosensory signaling, we propose that A. burtoni represents a valuable vertebrate model for studying context-dependent behavioral and neural decision making associated with non-redundant multimodal communication.

Circles in the sea: annual courtship "torus" behaviour of basking sharks Cetorhinus maximus identified in the eastern North Atlantic Ocean.

Groups of basking sharks engaged in circling behaviour are rarely observed, and their function remains enigmatic in the absence of detailed observations. Here, underwater and aerial video recordings of multiple circling groups of basking sharks during late summer (August and September 2016-2021) in the eastern North Atlantic Ocean showed groups numbering between 6 and 23 non-feeding individuals of both sexes. Sharks swam slowly in a rotating "torus" (diameter range: 17-39 m), with individuals layered vertically from the surface to a maximum depth of 16 m. Within a torus, sharks engaged in close-following, echelon, close-flank approach or parallel-swimming behaviours. Measured shark total body lengths were 5.4-9.5 m (mean LT : 7.3 m ± 0.9 s.d.; median: 7.2 m, n = 27), overlapping known lengths of sexually mature males and females. Males possessed large claspers with abrasions that were also observed on female pectoral fins. Female body colouration was paler than that of males, similar to colour changes observed during courtship and mating in other shark species. Individuals associated with most other members rapidly (within minutes), indicating toroidal behaviours facilitate multiple interactions. Sharks interacted through fin-fin and fin-body contacts, rolling to expose the ventral surfaces to following sharks, and breaching behaviour. Toruses formed in late summer when feeding aggregations in zooplankton-rich thermal fronts switched to non-feeding following and circling behaviours. Collectively, the observations explain a courtship function for toruses. This study highlights northeast Atlantic coastal waters as a critical habitat supporting courtship reproductive behaviour of endangered basking sharks, the first such habitat identified for this species globally.

New species of tree cricket Oecanthus Serville, 1831 (Orthoptera: Grylloidea) from Southern Brazilian Atlantic Forest, with bioacoustics.

Tree Crickets, Oecanthus Serville, 1831 have a worldwide distribution, and are known as Tree Crickets because they are common in the treetops, even though many species occur in different strata of forest, and also in open fields. In this work we describe a new species of Oecanthus from Southern Brazil, based on the most significant diagnostic structures in the ocanthine taxonomy, such as the phallic sclerites and metanotal gland morphology, as well as calling song. The new species herein described has a distinct faint red spot between the eyes, a metanotal gland with a triangular posterior median lobe with the apex curved up and forward, the median lophi of pseudepiphallus slightly elongate with an U-shaped invagination, and the ectophallic distal arc prolongation with the posterior apex U-bifurcated. The calling song stands out among the local stridulating crickets for alternate a regular and an irregular train of chirps. Illustrations of the external morphology, information about type specimens, material examined, measurements, and spectrogram of the calling and courtship songs is provided.

The potential for the evolution of thermally sensitive courtship behaviours in the treehopper, Enchenopa binotata.

The ability of animals to adapt to warming will depend on the evolutionary potential of thermally sensitive traits. The number of studies measuring the quantitative genetics of a wide variety of thermally sensitive traits has steadily increased; however, no study has yet investigated the quantitative genetics of thermal sensitivity for courtship traits. Since courtship often precedes mating, the ability of these traits to respond to warming may impact reproduction and therefore population persistence. Here, we use classic quantitative genetics breeding design to estimate heritability of various aspects of the thermal sensitivity of courtship behaviours in the treehopper Enchenopa binotata. We generated individual-level thermal courtship activity curves for males and females and measured levels of genetic variation in the thermal sensitivity of courtship activity. We found low heritability with 95% credible intervals that did not approach zero for most traits. Levels of genetic variation were highest in traits describing thermal tolerance. We also found some evidence for genetic correlations between traits within but not across sexes. Together, our results suggest that the range of temperatures over which these treehoppers actively court can evolve, although it remains unclear whether adaptation can happen quickly enough to match the speed of warming.

The doublesex gene regulates dimorphic sexual and aggressive behaviors in Drosophila.

Most animal species display dimorphic sexual behaviors and male-biased aggressiveness. Current models have focused on the male-specific product from the fruitless (fruM) gene, which controls male courtship and male-specific aggression patterns in fruit flies, and describe a male-specific mechanism underlying sexually dimorphic behaviors. Here we show that the doublesex (dsx) gene, which expresses male-specific DsxM and female-specific DsxF transcription factors, functions in the nervous system to control both male and female sexual and aggressive behaviors. We find that Dsx is not only required in central brain neurons for male and female sexual behaviors, but also functions in approximately eight pairs of male-specific neurons to promote male aggressiveness and approximately two pairs of female-specific neurons to inhibit female aggressiveness. DsxF knockdown females fight more frequently, even with males. Our findings reveal crucial roles of dsx, which is broadly conserved from worms to humans, in a small number of neurons in both sexes to establish dimorphic sexual and aggressive behaviors.

Nutritional condition affects tergal gland secretion and courtship success of male cockroaches.

An integral part of the courtship sequence of the German cockroach (Blattella germanica) involves the male raising his wings to expose tergal glands on his dorsum. When a female cockroach feeds on the secretion of these glands, she is optimally positioned for mating. Core chemical components have been identified, but the effect of male diet on the quality of the tergal gland secretion remains unexplored. After validating the pivotal role of tergal feeding in mating, we starved or fed reproductively mature males for one week. We then paired each male with a sexually receptive female and observed their interactions through an infrared-sensitive camera. While starvation had no effect on male courtship behavior, it did influence the duration of female tergal feeding and mating outcomes. Females fed longer on the gland secretion of fed males, and fed males experienced greater mating success than starved males (73.9% vs. 48.3%, respectively). These results suggest that the quality of the tergal gland secretions, and by association mating success, are dependent on the nutritional condition of the male.

Acoustic and postural displays in a miniature and transparent teleost fish, Danionella dracula.

Acoustic behavior is widespread across vertebrates, including fishes. We report robust acoustic displays during aggressive interactions for a laboratory colony of Danionella dracula, a miniature and transparent species of teleost fish closely related to zebrafish (Danio rerio), which are hypothesized to be sonic based on the presence of a hypertrophied muscle associated with the male swim bladder. Males produce bursts of pulsatile sounds and a distinct postural display - extension of a hypertrophied lower jaw, a morphological trait not present in other Danionella species - during aggressive but not courtship interactions. Females show no evidence of sound production or jaw extension in such contexts. Novel pairs of size-matched or -mismatched males were combined in resident-intruder assays where sound production and jaw extension could be linked to individuals. In both dyad contexts, resident males produced significantly more sound pulses than intruders. During heightened sonic activity, the majority of the highest sound producers also showed increased jaw extension. Residents extended their jaw more than intruders in size-matched but not -mismatched contexts. Larger males in size-mismatched dyads produced more sounds and jaw extensions compared with their smaller counterparts, and sounds and jaw extensions increased with increasing absolute body size. These studies establish D. dracula as a sonic species that modulates putatively acoustic and postural displays during aggressive interactions based on residency and body size, providing a foundation for further investigating the role of multimodal displays in a new model clade for neurogenomic and neuroimaging studies of aggression, courtship and other social interactions.

Endocrine modulation of primary chemosensory neurons regulates Drosophila courtship behavior.

The decision to engage in courtship depends on external cues from potential mates and internal cues related to maturation, health, and experience. Hormones allow for coordinated conveyance of such information to peripheral tissues. Here, we show Ecdysis-Triggering Hormone (ETH) is critical for courtship inhibition after completion of copulation in Drosophila melanogaster. ETH deficiency relieves post-copulation courtship inhibition (PCCI) and increases male-male courtship. ETH appears to modulate perception and attractiveness of potential mates by direct action on primary chemosensory neurons. Knockdown of ETH receptor (ETHR) expression in GR32A-expressing neurons leads to reduced ligand sensitivity and elevated male-male courtship. We find OR67D also is critical for normal levels of PCCI after mating. ETHR knockdown in OR67D-expressing neurons or GR32A-expressing neurons relieves PCCI. Finally, ETHR silencing in the corpus allatum (CA), the sole source of juvenile hormone, also relieves PCCI; treatment with the juvenile hormone analog methoprene partially restores normal post-mating behavior. We find that ETH, a stress-sensitive reproductive hormone, appears to coordinate multiple sensory modalities to guide Drosophila male courtship behaviors, especially after mating.