The Journal of Experimental Biology

In behavioral biology, many models have been proposed to explain how communication systems evolve. Within neuroethology, the principle of sender-receiver matching has spurred much research in auditory communication, but fewer studies on mechanosensory communication. We investigated sender-receiving matching in mechanosensory communication, focusing on the "hula", a courtship behavior that produces an undulating movement of the tail, in axolotls (Ambystoma mexicanum), an aquatic salamander. We characterized typical courtship behaviors, then quantified tail-motion parameters (speed, sweep angle, and elevation angle) from males as they performed the hula. We then constructed a "Robotail", a robotic device that mimics the physical and motion properties of the male tail during courtship. Interestingly, females initially responded to the Robotail as if it were a prey item, an effect that was mitigated by the addition of male whole-body odorants. We examined female behavioral responses to changes in individual Robotail movement parameters and found that speed and sweep angle were important to locomotion. Females transitioned between locomotor states more often when exposed to combinations of wide sweep angles and fast speeds from the Robotail, which males perform moderately or rarely, perhaps reflecting a preference for vigorous movements. We then assessed neural responses to stimuli generated by the Robotail by recording from the anterodorsal lateral line nerve (ADLLn), which innervates the mechanosensory neuromasts on the snout. The female ADLLn responded most vigorously when stimulated with moderate sweep angles and speeds, parameters often used by courting males. Thus, our behavioral results support a receiver bias model but our neurophysiological results support a sender-receiver matching model within mechanosensory communication during courtship in axolotls. Our results also provide novel evidence that mechanosensory cues generated during hula behavior in salamanders play a role in courtship.

The ability of animals to adjust their heading, i.e. to turn, is essential for all walking animals. While several studies have addressed how leg movement or inter-leg coordination may change during turning, relatively little is known about how turning-related changes scale with turn magnitude. Here, we used spontaneous and visually induced turns of unrestrained walking stick insects to test (i) how high-level parameters of unrestrained turning scale with low-level parameters of leg movement, and (ii) the effect of visual guidance on turning parameters. To this end, we used a step change in stationary landmark position in an open-field arena to constrain timing and magnitude of target-directed turns. These visually guided turns were compared with spontaneous turns in an all-white condition. We show that visually induced turns were walked at a larger forward velocity and had fewer short steps than spontaneous turns. The scaling of turning responses was dominated by an increase in turning duration (factor 1.87) rather than turning speed (factor 1.32). Increased rotational velocity correlated with reduced forward velocity, though with flexible timing of both effects. These changes were accompanied by larger shifts in step direction, as well as an increased asymmetry of step types between inner and outer legs, suggesting a mix of distinct turning strategies, that depend on overall turn angle. Future models on six-legged locomotion should thus consider the incorporation of more than one mechanism to govern turning.

Vestibular neurons are core elements of the pathways involved in vestibulo-motor functions, such as vestibulo-spinal and vestibulo-ocular reflexes. To meet behavioral needs, electrophysiological neuronal properties are adequately adapted to the sensory-motor computation sustaining these distinct vestibular reflexes. During frog metamorphosis, there is a complete reorganization of the posturo-locomotor system while the oculomotor system remains minimally changed, probably associated to so far unknown changes in vestibular neuronal properties. We used this unique model to investigate the central developmental mechanisms underlying such a reconfiguration of vestibular-associated behaviors. Central vestibular neurons exhibit two types of electrophysiological phenotypes: tonic neurons with a continuous discharge and phasic neurons with a transitory discharge mainly due to the activation of Kv1.1 channel. Electrophysiological recordings and Kv1.1 immunolabeling of vestibulospinal (VS) and vestibulo-ocular (VO) neurons at both larval and juvenile stages revealed that the majority of VS neurons exhibited a tonic discharge in larvae but a phasic discharge in juvenile, while VO neurons remained mainly tonic throughout development. Changes in phasic and tonic neurons proportions in VS population are partly explained by neurogenesis. But we provide evidences that an electrophysiological phenotype switch is a concomitant developmental mechanism participating in the maturation of these central vestibular neurons. All together our results showed that the maturation process in central vestibular neuronal groups is highly related to the metamorphosis-induced remodeling of vestibulo-motor functions they are involved in, with the ultimate purpose of ensuring an adequate adaptation of neuronal elements properties to the developmental changes of behavioral constrains.

Understanding how animals produce a versatile locomotor repertoire requires unraveling the interplay between higher centers, decentralized locomotor circuits, and sensory feedback. However, the principles governing their integration remain elusive. We investigated amphibious centipedes through stepwise neural lesions and neuromechanical modeling. Behavioral experiments revealed that while decentralized circuits autonomously generate coordination, the brain and subesophageal ganglion provide situational flexibility, such as modulating trunk undulation and initiating leg folding. Integrating these findings, our model demonstrated how higher centers selectively inhibit or release lower circuit dynamics. Simulations verified that varying only a few descending control parameters reproduces transitions between slow walking, fast walking, and swimming. This work may capture the essence of the locomotor circuitry that harnesses decentralized self-organization to coordinate the bodys large degrees of freedom.

Predictions of skeletal muscle energy consumption under a diverse range of muscle contractile conditions are critical for improving our understanding of locomotion. Existing mathematical models, while capturing the mechanical dependence of energy consuming processes, neglect the time-dependent behaviour and recovery costs associated with regenerating ATP. This time-dependence is important for predicting the energetic response of muscles during repetitive or cyclical tasks like locomotion, where muscle undergoes many contraction cycles. This study presents a novel model to predict energetic rates based on physiological processes: Ca2+ transport costs, cross-bridge cycling costs, and ATP regeneration. Previous mathematical models include the dependence on Ca2+ transport and cross-bridge cycling, but neglect the time-dependent response and the subsequent recovery of ATP following the contraction. Model parameters were obtained from existing data on isolated muscle preparations, and predicted energetic rates were validated on separate datasets across a range of contractile conditions including dynamic, sub-maximal, and twitch contractions. The time-dependent model was able to capture the influence of contraction frequency on peak energetic rates and the time-course of energetic recovery observed experimentally. The model captures key physiological processes while maintaining a minimal number of free parameters and low computational cost. This enables generalisability across muscles and species, and implementation into larger scale musculoskeletal models.

Take-off is a fast and energy-efficient strategy for bipedal animals, such as birds, to achieve rapid movement; however, how muscle physiology scales to govern this universal behavior remains unresolved. Research in other species physiologies is not readily applicable. As a result, important questions, whether theropod dinosaurs such as Tyrannosaurus rex were capable of jumping, remain unanswered. In this article, we coupled Lagrangian dynamics with Hills muscle equations and developed new experimental methods to quantify joint rotational stiffness and damping, thereby enabling a systematic description of lower-limb mechanics. The approach establishes a novel kinetic framework that links muscle contractile properties to lower-limb performance without invoking control optimization. Animal observations and tabletop mechanisms validate the framework. The mechanics model reveals that the take-off time of about 0.1 s across body masses of 0.003 to 90 kg is achievable, as heavier birds generate proportionally higher reaction forces. Additionally, Tyrannosaurus rex should be capable of jumping, based on the available physiology data. Beyond evolutionary insights, our framework provides a new methodology for analyzing the mechanical properties of biological joints and informing the design of scalable bio-inspired robots.

Although muscle spindle sensory signals have been extensively studied, little is known about how and why muscle spindle firing is modulated by the central nervous system during movement. Specialized motor neurons to the muscle spindle, i.e. gamma motor neurons, can profoundly alter spindle firing during behavior, but technological limitations hinder our ability to record gamma motor and muscle spindle sensory signals during most behaviors. We used a biophysical model of a muscle spindle within a muscle-tendon unit to simulate how gamma drive may modulate muscle spindle Ia firing during locomotion. Based on a few available recordings from decerebrate animals, we demonstrate that our model, tuned to passive stretch conditions, can reproduce profound changes in muscle spindle firing in response to identical joint motions in locomotor vs. relaxed stretch conditions. Our model can discover phasic patterns of two types of gamma motor neuron drive based on recorded muscle spindle Ia firing and joint motion. By simulating perturbations, we conclude that: 1) sinusoidal activation of static gamma motor neurons during locomotion, encoding intended movement, modulates muscle spindle signals such that they act as sensorimotor feedback signals based on errors from the intended muscle fascicle length; 2) phasic on/off activation of dynamic gamma motor neurons during locomotion acts as an event detector, heightening muscle spindle Ia responses to discrete perturbations. As such, their muscle-within-muscle structure allows the muscle spindle to act as a highly tunable physical internal model of muscle state to guide movement. Our model supports proposed but as-yet-untested theories of muscle spindle function and offers a framework for extending the testing of muscle spindle function to active, behavioral conditions.

Birdsong mediates territory acquisition and mate choice. In agonistic interactions, local songs generally elicit stronger responses than songs from more distant populations. However, the molecular mechanisms associated with differential responses to local vs. foreign songs are poorly understood. We addressed this knowledge gap by combining behavioral assays in the field with blood transcriptomic analysis, using a within-subjects design to ask whether male song sparrows (Melospiza melodia) show differential gene expression when exposed to playback of local and foreign songs. Transcriptomic profiles reflected the difference in behavioral response to local vs. foreign songs, with individuals exposed to local songs showing greater expression of genes associated with song perception and production, anti-inflammatory responses and energy metabolism. Our study suggests that changes in expression of key molecular pathways correlate with behavioral responses to geographic song variation, providing insight into the potential mechanisms regulating signal recognition and response to social challenges. HighlightsO_LIGene expression in sparrow blood was measured after simulated territorial intrusion. C_LIO_LIStronger response to local songs was associated with differential gene expression. C_LIO_LISong-associated genes (FOXP2, NRXN1) had higher expression when birds heard local songs. C_LIO_LIGene expression in the blood contains potential biomarkers of song recognition. C_LI

Parental care can have pervasive effects on offsprings neurodevelopment. Parent-offspring interactions are often modulated by the neuropeptide oxytocin, which is responsible for the development of social bonds. The development of the oxytocinergic system is dependent on the quality of parental care during the post-natal phase. However, it is yet unknown how post-natal direct interactions can influence the development of the oxytocinergic pathway. Here we tested how an obligate parental care behaviour, tadpole transport in poison frogs, influences the development of the oxytocinergic pathway. To this end, we quantified whole brain expression of oxytocin receptor and oxytocin precursor throughout three developmental stages of A. femoralis tadpoles, before, during and after tadpole transport. Our results show an overall downregulation during tadpole transport, which indicates that during transport tadpoles enter a dormant state to slow down development until they are placed in water. Interestingly, the expression of oxytocin precursor did not vary between the three developmental stages. This might indicate that oxytocin is being recruited during transport, but does not lead to neurodevelopmental changes. In sum, here we present the first evidence of a dormant state during tadpole transport which might be an adaptive response to the terrestrial reproduction in poison frogs.

Body size and limb posture vary widely across mammals and are expected to shape locomotor stability, yet direct comparative evidence remains limited. Here, we tested whether smaller, crouched mammals exhibit greater lateral dynamic stability than larger, more upright species by comparing treadmill walking in mice and cats at dynamically similar speeds. Using kinematic analyses and size normalized measures of stability, we show that mice are substantially more laterally stable than cats. This increased stability is associated with relatively wider step widths and more crouched limb posture, indicating that support geometry and posture play dominant roles in stabilizing locomotion. Despite these differences, both species regulate lateral balance on a step-by-step basis, as revealed by relationships between center of mass motion and subsequent adjustments of the border of support. Our findings demonstrate that locomotor stability does not scale simply with body size but depends critically on posture dependent strategies that differ across species. These results identify lateral stability as a key factor of locomotor adaptation and suggest that crouched postures in small mammals may reduce reliance on active neural control while enhancing robustness in complex environments. SUMMARY STATEMENTLateral dynamic stability during quadrupedal locomotion depends primarily on limb posture and support geometry rather than body size. Smaller mammals achieve greater stability through crouched postures and wider step widths, whereas larger mammals operate closer to stability limits and rely more heavily on active control.

AO_SCPLOWBSTRACTC_SCPLOWMuscle efficiency decreases with increasing size, largely due to a relative decrease in its mechanical output. Muscle mechanical output depends on its activation, strain, and strain rate and thus varies between different muscles within a limb during locomotion. Distinct muscle coordination patterns are required for efficient cycling, and so we would expect that the coordination patterns for efficient cycling or indeed locomotion would change across animal sizes. We tested whether muscle coordination would change with muscle size using data derived from human cycling: this paradigm allowed for controlled changes in both crank torque and cadence, allowing the multifactorial problem of muscle power output to be decomposed. We used kinematic and pedal data from 12 cyclists undergoing steady pedalling at cadences from 80 to 140 r.p.m. and generated musculoskeletal simulations of their movements. We introduced novel multisegment muscle models in the simulation that incorporated the internal muscle mass and thus accounted for the scaling effects of muscle tissue inertia. We solved the simulations for the muscle activity that was required to minimise the metabolic cost during cycling for each condition. The masses of the muscle models were scaled across five orders of magnitude. The predicted muscle activations were classified by Principal Component analysis to identify whether the coordination of muscle activity was modulated across models with different sized muscles. Analysis of variance revealed significant changes in coordination at the large-scale factors. This study shows how the coordination of muscle activity during locomotion will likely change across a range of body sizes due to the non-linear effects of the inertial mass within the muscle tissues.

Many taxa have evolved heteromorphic sex chromosomes like the XY system found in mammals. In additional to the sex determination gene which directs development of the gonad into an ovary or testis, sex chromosomes can have drastically different gene content, leading to substantial genetic differences between genetic males and females beyond their gonad identity. Studying the effects of these genetic differences is challenging, as the sex chromosomes and sex determination gene are inherited together, so the effects of genetic differences between the X and Y cannot be easily isolated from the hormonal differences produced by the ovary and testis. The threespine stickleback fish has a heteromorphic XY sex chromosome system and a wide range of well documented sex differences in morphology and behaviors, including complex mating behaviors and male-only parental care. Through genetic manipulation of amhy, the newly identified sex determination gene in threespine stickleback, we are able to generate gonadal males and females with either the XX or XY sex chromosome complement and analyze the separate effects of gonadal sex and sex chromosome complement on sexually dimorphic gene expression. We find that sex chromosomes have a larger effect on gene expression than gonadal sex in somatic tissues, while gonadal sex has a larger effect on expression in the gonads. We also find that the X and Y chromosomes are enriched for genes which show differential expression between females and males. Our findings demonstrate the significant biological impact of sex chromosomes outside of primary sex determination and showcase the utility of the threespine stickleback for studying the genetic basis of sex differences.

Human walking is often considered an inverted pendulum during single support, suggesting conservative dynamics. Gait consists of discrete steps connected by mechanically costly transitions. We examine how step length, walking speed, and work capacity jointly constrain walking mechanics. Using a powered simple walking model, minimum speed required to complete a step of given length is derived based on gravitational work; below this threshold, forward progression becomes mechanically infeasible, and the next heel-strike occurs early, producing shorter steps. Comparisons with empirical step length-speed relationships show that humans walk at higher speeds and require greater push-off work, indicating energy dissipation. We extend pendular dynamics by incorporating hip torque, a linearized axial force model, and muscle intervention. This framework reproduces key GRF features, including the M-shaped profile, without prescribing force trajectories a priori. Fitted parameters suggest reduced average loading (CBaseline < 1), active mid-stance unloading (Am < 0), and narrowly timed muscle action (small{sigma} m). Parameter studies show that increasing step length or speed increases transition work and peak forces, while hip torque timing indicates mechanical cost is minimized when energy modulation occurs after mid-stance. These findings indicate that preferred walking speed emerges from feasibility and work-capacity constraints, not energetic optimality alone.

The shift to terrestrial environments in ancestrally aquatic animals is often associated with key physiological and physical changes, including shifts in respiratory physiology and in some cases, even the evolution of completely novel respiratory structures. Examining how respiration operates across a gradient of submersion states in ancestrally aquatic terrestrial animals may shed light on how complex biological traits shift under different selective regimes. In this work, we begin exploring respiration in terrestrially-adapted land crabs that still use their gills to respire while underwater. We tested the relationship between aquatic respiratory rates, body size, and sex in red devil vampire crabs (Geosesarma hagen) at two ecologically-relevant temperatures. We found small females respire more than small males at 28{degrees}C, while large females respire more than large males at 21{degrees}C. Additionally, body size is a significant factor affecting respiratory rates of both sexes at 21{degrees}C and warmer temperatures significantly increase respiration in small crabs of both sexes. Interactions between these factors also led to emerging trends that can be explained by both physiological rules, such as reproductive investment and surface-to-volume ratios and heat transfer. We also report a temperature coefficient (Q10) of 1.52 for this species, showing an expected 52% change in respiratory and metabolic rate for every 10{degrees}C increase. This work also demonstrates the importance of understanding how and to what extent biological variables like sex and body size interact with abiotic environmental factors when measuring physiological traits in ectothermic invertebrate animals.

In sex-role reversed species, females are socially polyandrous and compete for multiple mates, whereas males conduct the majority of parental care. To understand the extent to which physiological differences between females and males are shaped by sex roles, we examined sex differences in gene expression in sex-role reversed northern jacanas (Jacana spinosa). Given that females compete for mating opportunities, and males cycle between courtship and parental care, we predicted that transcriptomic profiles would be more similar between females and courting males, in contrast to female and parenting males. Leveraging a high quality de novo genome assembly, we conducted RNA-seq on two brain regions associated with the regulation of social behavior: the preoptic area of the hypothalamus and the nucleus taeniae. The majority of genes differentially expressed between the sexes were male-biased. Of these male-biased genes, the majority were located on the Z-chromosome. Contrary to our prediction, the greatest difference in autosomal gene expression was between females and courting males, in the preoptic area of the hypothalamus. Several differentially expressed genes related to elements of hormone signaling that are likely to be behaviorally salient, including higher expression of androgen receptor in females relative to parenting males, and higher expression of prolactin receptor in males, regardless of breeding stage. Some sex-associated gene networks were also associated with competitive traits, whereas others were associated with aggressive behaviors, regardless of sex. Few genes were differentially expressed between courting and parenting males, yet some nonetheless had connections to behavioral endocrinology, including prolactin, thyroid and insulin-like growth factor pathways. Our investigation of sex differences in gene expression can help to reveal the molecular mechanisms underlying female competition and male parental care in socially polyandrous species. We conclude that social polyandry is not a simple reversal in the direction of sex-biased gene expression in the brain, but rather a result of complex genetic and hormonal interactions that warrants further study.

Biological phase changes provoked by stress, such as vitrification or gel-sol transitions, enable many organisms, including extremotolerant tardigrades, to enter quiescent states and survive extreme environmental conditions. Protein-driven phase transitions are hypothesized to produce large-scale changes in intracellular viscosity, allowing tardigrades to survive extreme stresses such as desiccation. We report that the tardigrade Hypsibius exemplaris undergoes both large-scale and local increases in intracellular viscosity following exposure to anoxic and hyperosmotic stress. Such dramatic shifts in cellular viscosity would be expected to enhance cellular resilience to physical force. Indeed, we found that tardigrades can survive, behave normally, and reproduce after exposure to the highest simulated hypergravity (HG) achievable in an ultracentrifuge (one million times Earths gravity). In contrast, Caenorhabditis elegans, a similarly sized animal, does not survive these extreme forces owing to loss of cellular integrity. Remarkably, tardigrades frozen during exposure to extreme hypergravitational force show minimal disruption of fine cellular ultrastructure and little evidence of stratification of cellular components whose density varies by nearly a factor of two. Further, exposure to anoxia, hyperosmotic stress, and HG all result in a large increase in reactive oxygen species (ROS), which is required for survival under these extreme environments. Inhibition of NADPH oxidase (NOX) suppresses survival both to HG and hyperosmotic stress. Our findings suggest that intracellular viscosity changes in response to multiple extreme stresses may underlie the resilience of these animals to extraordinary physical stress, and that survival in or recovery from these states relies on ROS signaling via NADPH oxidase. Significance StatementTardigrades are renowned for surviving conditions that are lethal to nearly all other life forms. We reveal two mechanisms that support this resilience: intracellular viscosity changes and NADPH oxidase-mediated ROS signaling. Through direct assessment of the effects of altered cellular material properties, found that tardigrades are resilient to forces up to one million times Earths gravity, establishing them as the most hypergravity-resistant animal currently known.

Variation in mitochondrial aerobic metabolism has been suggested to underlie variation in interindividual performance. Mitochondrial efficiency quantifies, directly or indirectly, the amount of adenosine triphosphate (ATP) produced relative to O2 consumption. High mitochondrial efficiency is theoretically beneficial by providing more ATP per amount of resource consumed, but may come at the cost of increased reactive oxygen species (ROS) production damaging tissues through oxidative stress. Mitochondrial efficiency is a plastic trait but how it changes through postnatal development remains unknown. We hypothesized that strong selective pressure could lead to an increased mitochondrial efficiency to support fast growth but incur an oxidative cost. We tested this hypothesis by quantifying mitochondrial aerobic metabolism, efficiency and ROS production through postnatal growth in Japanese quail (Coturnix japonica), in two highly aerobic tissues: skeletal and cardiac muscles. Mitochondrial efficiency was indeed higher during peak growth in both tissues, but this was surprisingly associated with markedly lower ROS production. This high efficiency was likely achieved via both a lower proton leak and a higher contribution of complex I to respiration. These results show that enhancing mitochondrial efficiency may be important to support growth, but suggest the presence of unexpected ROS mitigation processes during early-life growth.

Muscle mass significantly influences skeletal muscle behaviour, potentially explaining why traditional massless Hill-type models struggle to predict the forces generated by larger muscles during dynamic, submaximal contractions. However, the applicability of mass-enhanced Hill-type models in human locomotion remains unexplored. Here, we compared the predicted force from a 1D mass-enhanced Hill-type muscle model with a traditional 1D massless Hill-type muscle model across a range of experimentally measured human movements. Kinematic and electromyographic data were collected from twenty participants performing locomotor tasks and supplemented with existing cycling data. Muscle size was geometrically scaled by factors from 0.1 to 10, which causes lengths to be scaled proportionally, cross-sectional area and peak isometric force F0 with the square, and mass with the cube of the factor. Muscle tissue mass (inertia) and cadence increased the differences between mass-enhanced and massless predictions of force and power. At high cadence and the largest scale, the normalized root mean square difference between force traces reached 7% of F0, (averaged across muscles). However, differences between models were minimal (<1%) at human-sized scale 1. Real muscle additionally deforms in 3D, we still do not know the extent to which this extra dimensionality affects muscle forces for these human movements.

Despite its ubiquity in natural flows, the effects of turbulence on fish locomotion and behavior remain poorly understood. The prevailing hypothesis is that these effects depend on the spatial and temporal scales of the turbulence relative to the fishs size and swimming speed. But in conventional facilities, turbulence usually increases with mean flow, which forces higher swimming speeds and can leave these relative scales unchanged. We therefore present a novel experimental facility that leverages a jet array to decouple the turbulence from the mean flow and systematically control its scales. This approach allows the ratio of turbulent to fish inertial scales to be varied over an order of magnitude, providing a controlled framework for quantifying fish-turbulence interactions. The facility also supports experiments probing strategies fish may use to cope with turbulence, including collective behaviors. Insights from this work have broader implications for ecological studies and engineering applications, including the design of effective fishways and bio-inspired underwater vehicles.

Animals fight for resources to obtain fitness benefits; most contests are intrasexual, and males tend to fight more than females. Although the genetic basis of male aggression is well studied, we know little about the genetic variation of female aggression. Female aggression varies with reproductive status and is potentially influenced not only by her genotype, but also by the genotype of her mate. Here we measured both male and female aggression in a set of Drosophila melanogaster inbred lines by competing each line against a standard competitor. Aggression varied among lines for both sexes, but male and female aggression were not correlated. Female aggression for many lines increased with mating, as expected, but not all lines changed aggression. However, when females were mated to males of different lines, male genotype did not affect the post-mating change in aggression, suggesting that ejaculate-mediated effects do not vary across these lines. The aggression level of the standard opponent was positively correlated with that of focal individuals indicating that individuals modulate their behavior according to the genotype of their opponent.