Kevin M. Middleton

Differential response to a selective cannabinoid receptor antagonist (SR141716: rimonabant) in female mice from lines selectively bred for high voluntary wheel-running behaviour.

Exercise is a naturally rewarding behaviour in human beings and can be associated with feelings of euphoria and analgesia. The endocannabinoid system may play a role in the perception of neurobiological rewards during and after prolonged exercise. Mice from lines that have been selectively bred for high voluntary wheel running (high runner or HR lines) may have evolved neurobiological mechanisms that increase the incentive salience of endurance-type exercise. Here, we test the hypothesis that endocannabinoid signalling has been altered in the four replicate HR lines as compared with four nonselected control lines. After 18 days of acclimation to cages with attached wheels, we injected mice with rimonabant (SR141716), a selective cannabinoid CB1 receptor antagonist. During the time of normal peak running, each mouse received, in a randomized order, intraperitonial injection of rimonabant (0.1 or 3.0 mg/kg) or vehicle, over 9 days. Drug response was quantified as wheel revolutions, time and speed 10–70 min postinjection. Rimonabant decreased running in all mice; however, female HR mice differentially decreased running speed and distance (but not time) as compared with control females. We conclude that altered endocannabinoid signalling plays a role in the high wheel running of female HR mice.

The effect of body size on the wing movements of pteropodid bats, with insights into thrust and lift production

SUMMARY In this study we compared the wing kinematics of 27 bats representing six pteropodid species ranging more than 40 times in body mass (Mb=0.0278–1.152 kg), to determine whether wing posture and overall wing kinematics scaled as predicted according to theory. The smallest species flew in a wind tunnel and the other five species in a flight corridor. Seventeen kinematic markers on the midline and left side of the body were tracked in three dimensions. We used phylogenetically informed reduced major axis regression to test for allometry. We found that maximum wingspan (bmax) and maximum wing area (Smax) scaled with more positive allometry, and wing loading (Qs) with more negative allometry (bmax∝Mb0.423; Smax∝Mb0.768; Qs∝Mb0.233) than has been reported in previous studies that were based on measurements from specimens stretched out flat on a horizontal surface. Our results suggest that larger bats open their wings more fully than small bats do in flight, and that for bats, body measurements alone cannot be used to predict the conformation of the wings in flight. Several kinematic variables, including downstroke ratio, wing stroke amplitude, stroke plane angle, wing camber and Strouhal number, did not change significantly with body size, demonstrating that many aspects of wing kinematics are similar across this range of body sizes. Whereas aerodynamic theory suggests that preferred flight speed should increase with mass, we did not observe an increase in preferred flight speed with mass. Instead, larger bats had higher lift coefficients (CL) than did small bats (CL∝Mb0.170). Also, the slope of the wingbeat period (T) to body mass regression was significantly more shallow than expected under isometry (T∝Mb0.180), and angle of attack (α) increased significantly with body mass [α∝log(Mb)7.738]. None of the bats in our study flew at constant speed, so we used multiple regression to isolate the changes in wing kinematics that correlated with changes in flight speed, horizontal acceleration and vertical acceleration. We uncovered several significant trends that were consistent among species. Our results demonstrate that for medium- to large-sized bats, the ways that bats modulate their wing kinematics to produce thrust and lift over the course of a wingbeat cycle are independent of body size.

Functional significance of genetic variation underlying limb bone diaphyseal structure.

Limb bone diaphyseal structure is frequently used to infer hominin activity levels from skeletal remains, an approach based on the well-documented ability of bone to adjust to its loading environment during life. However, diaphyseal structure is also determined in part by genetic factors. This study investigates the possibility that genetic variation underlying diaphyseal structure is influenced by the activity levels of ancestral populations and might also have functional significance in an evolutionary context. We adopted an experimental evolution approach and tested for differences in femoral diaphyseal structure in 1-week-old mice from a line that had been artificially selected (45 generations) for high voluntary wheel running and non-selected controls. As adults, selected mice are significantly more active on wheels and in home cages, and have thicker diaphyses. Structural differences at 1 week can be assumed to primarily reflect the effects of selective breeding rather than direct mechanical stimuli, given that the onset of locomotion in mice is shortly after Day 7. We hypothesized that if genetically determined diaphyseal structure reflects the activity patterns of members of a lineage, then selected animals will have relatively larger diaphyseal dimensions at 1 week compared to controls. The results provide strong support for this hypothesis and suggest that limb bone cross sections may not always only reflect the activity levels of particular fossil individuals, but also convey an evolutionary signal providing information about hominin activity in the past.

Erythropoietin elevates but not voluntary wheel running in mice

SUMMARY Voluntary activity is a complex trait, comprising both behavioral (motivation, reward) and anatomical/physiological (ability) elements. In the present study, oxygen transport was investigated as a possible limitation to further increases in running by four replicate lines of mice that have been selectively bred for high voluntary wheel running and have reached an apparent selection limit. To increase oxygen transport capacity, erythrocyte density was elevated by the administration of an erythropoietin (EPO) analogue. Mice were given two EPO injections, two days apart, at one of two dose levels (100 or 300 μg kg–1). Hemoglobin concentration ([Hb]), maximal aerobic capacity during forced treadmill exercise () and voluntary wheel running were measured. [Hb] did not differ between high runner (HR) and non-selected control (C) lines without EPO treatment. Both doses of EPO significantly (P<0.0001) increased [Hb] as compared with sham-injected animals, with no difference in [Hb] between the 100 μg kg–1 and 300 μg kg–1 dose levels (overall mean of 4.5 g dl–1 increase). EPO treatment significantly increased by ∼5% in both the HR and C lines, with no dose × line type interaction. However, wheel running (revolutions per day) did not increase with EPO treatment in either the HR or C lines, and in fact significantly decreased at the higher dose in both line types. These results suggest that neither [Hb] per se nor is limiting voluntary wheel running in the HR lines. Moreover, we hypothesize that the decrease in wheel running at the higher dose of EPO may reflect direct action on the reward pathway of the brain.

Membrane muscle function in the compliant wings of bats

Unlike flapping birds and insects, bats possess membrane wings that are more similar to many gliding mammals. The vast majority of the wing is composed of a thin compliant skin membrane stretched between the limbs, hand, and body. Membrane wings are of particular interest because they may offer many advantages to micro air vehicles. One critical feature of membrane wings is that they camber passively in response to aerodynamic load, potentially allowing for simplified wing control. However, for maximum membrane wing performance, tuning of the membrane structure to aerodynamic conditions is necessary. Bats possess an array of muscles, the plagiopatagiales proprii, embedded within the wing membrane that could serve to tune membrane stiffness, or may have alternative functions. We recorded the electromyogram from the plagiopatagiales proprii muscles of Artibeus jamaicensis, the Jamaican fruit bat, in flight at two different speeds and found that these muscles were active during downstroke. For both low- and high-speed flight, muscle activity increased between late upstroke and early downstroke and decreased at late downstroke. Thus, the array of plagiopatagiales may provide a mechanism for bats to increase wing stiffness and thereby reduce passive membrane deformation. These muscles also activate in synchrony, presumably as a means to maximize force generation, because each muscle is small and, by estimation, weak. Small differences in activation timing were observed when comparing low- and high-speed flight, which may indicate that bats modulate membrane stiffness differently depending on flight speed.

Wingbeat kinematics and motor control of yaw turns in Anna's hummingbirds (Calypte anna)

SUMMARY The biomechanical and neuromuscular mechanisms used by different animals to generate turns in flight are highly variable. Body size and body plan exert some influence, e.g. birds typically roll their body to orient forces generated by the wings whereas insects are capable of turning via left–right wingbeat asymmetries. Turns are also relatively brief and have low repeatability, with almost every wingbeat serving a different function throughout the change in heading. Here we present an analysis of Annas hummingbirds (Calypte anna) as they fed continuously from an artificial feeder revolving around the outside of the animal. This setup allowed for examination of sustained changes in yaw without requiring any corresponding changes in pitch, roll or body position. Hummingbirds sustained yaw turns by expanding the wing stroke amplitude of the outer wing during the downstroke and by altering the deviation of the wingtip path during both downstroke and upstroke. The latter led to a shift in the inner–outer stroke plane angle during the upstroke and shifts in the elevation of the stroke plane and in the deviation of the wingtip path during both strokes. These features are generally more similar to how insects, as opposed to birds, turn. However, time series analysis also revealed considerable stroke-to-stroke variation. Changes in the stroke amplitude and the wingtip velocity were highly cross-correlated, as were changes in the stroke deviation and the elevation of the stroke plane. As was the case for wingbeat kinematics, electromyogram recordings from pectoral and wing muscles were highly variable, but no correlations were found between these two features of motor control. The high variability of both kinematic and muscle activation features indicates a high level of wingbeat-to-wingbeat adjustments during sustained yaw. The activation timing of the muscles was more repeatable than the activation intensity, which suggests that the former may be constrained by harmonic motion and that the latter may play a large role in kinematic adjustments. Comparing the revolution frequency of the feeder with measurements of free flight yaws reveals that feeder tracking, even at one revolution every 2 s, is well below the maximum yaw capacity of the hummingbirds.

Hummingbirds control turning velocity using body orientation and turning radius using asymmetrical wingbeat kinematics

Turning in flight requires reorientation of force, which birds, bats and insects accomplish either by shifting body position and total force in concert or by using left–right asymmetries in wingbeat kinematics. Although both mechanisms have been observed in multiple species, it is currently unknown how each is used to control changes in trajectory. We addressed this problem by measuring body and wingbeat kinematics as hummingbirds tracked a revolving feeder, and estimating aerodynamic forces using a quasi-steady model. During arcing turns, hummingbirds symmetrically banked the stroke plane of both wings, and the body, into turns, supporting a body-dependent mechanism. However, several wingbeat asymmetries were present during turning, including a higher and flatter outer wingtip path and a lower more deviated inner wingtip path. A quasi-steady analysis of arcing turns performed with different trajectories revealed that changes in radius were associated with asymmetrical kinematics and forces, and changes in velocity were associated with symmetrical kinematics and forces. Collectively, our results indicate that both body-dependent and -independent force orientation mechanisms are available to hummingbirds, and that these kinematic strategies are used to meet the separate aerodynamic challenges posed by changes in velocity and turning radius.

Predicting the bending properties of long bones: Insights from an experimental mouse model

OBJECTIVES Analyses of bone cross-sectional geometry are frequently used by anthropologists and paleontologists to infer the loading histories of past populations. To address some underlying assumptions, we investigated the relative roles of genetics and exercise on bone cross-sectional geometry and bending mechanics in three mouse strains: high bone density (C3H/He), low bone density (C57BL/6), and a high-runner strain homozygous for the Myh4Minimsc allele (MM). METHODS AND MATERIALS Weanlings of each strain were divided into exercise (wheel) or control (sedentary) treatment groups for a 7-week experimental period. Morphometrics of the femoral mid-diaphysis and mechanical testing were used to assess both theoretical and ex vivo bending mechanics. RESULTS Across all measured morphological and bending traits, we found relatively small effects of exercise treatment compared to larger and more frequent interstrain differences. In the exercised group, total distance run over the experimental period was not a predictor of any morphological or bending traits. Cross-sectional geometry did not accurately predict bone response to loading. DISCUSSION Results from this experimental model do not support hypothesized associations among extreme exercise, cross-sectional geometry, and bending mechanics. Our results suggest that analysis of cross-sectional geometry alone is insufficient to predict loading response, and questions the common assumption that cross-sectional geometry differences are indicative of differential loading history.

Visual Sensory Signals Dominate Tactile Cues during Docked Feeding in Hummingbirds

Animals living in and interacting with natural environments must monitor and respond to changing conditions and unpredictable situations. Using information from multiple sensory systems allows them to modify their behavior in response to their dynamic environment but also creates the challenge of integrating different, and potentially contradictory, sources of information for behavior control. Understanding how multiple information streams are integrated to produce flexible and reliable behavior is key to understanding how behavior is controlled in natural settings. Natural settings are rarely still, which challenges animals that require precise body position control, like hummingbirds, which hover while feeding from flowers. Tactile feedback, available only once the hummingbird is docked at the flower, could provide additional information to help maintain its position at the flower. To investigate the role of tactile information for hovering control during feeding, we first asked whether hummingbirds physically interact with a feeder once docked. We quantified physical interactions between docked hummingbirds and a feeder placed in front of a stationary background pattern. Force sensors on the feeder measured a complex time course of loading that reflects the wingbeat frequency and bill movement of feeding hummingbirds, and suggests that they sometimes push against the feeder with their bill. Next, we asked whether the measured tactile interactions were used by feeding hummingbirds to maintain position relative to the feeder. We created two experimental scenarios—one in which the feeder was stationary and the visual background moved and the other where the feeder moved laterally in front of a white background. When the visual background pattern moved, docked hummingbirds pushed significantly harder in the direction of horizontal visual motion. When the feeder moved, and the background was stationary, hummingbirds generated aerodynamic force in the opposite direction of the feeder motion. These results suggest that docked hummingbirds are using visual information about the environment to maintain body position and orientation, and not actively tracking the motion of the feeder. The absence of flower tracking behavior in hummingbirds contrasts with the behavior of hawkmoths, and provides evidence that they rely primarily on the visual background rather than flower-based cues while feeding.

Hip joint articular soft tissues of non-dinosaurian Dinosauromorpha and early Dinosauria: evolutionary and biomechanical implications for Saurischia

ABSTRACT Dinosauromorphs evolved a wide diversity of hind limb skeletal morphologies, suggesting highly divergent articular soft tissue anatomies. However, poor preservation of articular soft tissues in fossils has hampered any follow-on functional inferences. We reconstruct the hip joint soft tissue anatomy of non-dinosaurian dinosauromorphs and early dinosaurs using osteological correlates derived from extant sauropsids and infer trends in character transitions along the theropod and sauropodomorph lineagues. Femora and pelves of 107 dinosauromorphs and outgroup taxa were digitized using 3D imaging techniques. Key transitions were estimated using maximum likelihood ancestral state reconstruction. The hips of dinosauromorphs possessed wide a disparity of soft tissue morphologies beyond the types and combinations exhibited by extant archosaurs. Early evolution of the dinosauriform hip joint was characterized by the retention of a prominent femoral hyaline cartilage cone in post-neonatal individuals, with the cartilage cone independently reduced within theropods and sauropodomorphs. The femur of Dinosauriformes possessed a fibrocartilage sleeve on the metaphysis, which surrounded a hyaline core. The acetabulum of Dinosauriformes possessed distinct labrum and antitrochanter structures. In sauropodomorphs, hip congruence was maintained by thick hyaline cartilage on the femoral head, whereas theropods relied on acetabular tissues such as ligaments and articular pads. In particular, the craniolaterally ossified hip capsule of non- Avetheropoda neotheropods permitted mostly parasagittal femoral movements. These data indicate that the dinosauromorph hip underwent mosaic evolution within the saurischian lineage and that sauropodomorphs and theropods underwent both convergence and divergence in articular soft tissues, correlated with transitions in body size, locomotor posture, and joint loading.