Integrated K+ channel and K+Cl- cotransporter functions are required for the coordination of size and proportion during development.

The coordination of growth during development establishes proportionality within and among the different anatomic structures of organisms. Innate memory of this proportionality is preserved, as shown in the ability of regenerating structures to return to their original size. Although the regulation of this coordination is incompletely understood, mutant analyses of zebrafish with long-finned phenotypes have uncovered important roles for bioelectric signaling in modulating growth and size of the fins and barbs. To date, long-finned mutants identified are caused by hypermorphic mutations, leaving unresolved whether such signaling is required for normal development. We isolated a new zebrafish mutant, schleier, with proportional overgrowth phenotypes caused by a missense mutation and loss of function in the K+-Cl- cotransporter Kcc4a. Creation of dominant negative Kcc4a in wild-type fish leads to loss of growth restriction in fins and barbs, supporting a requirement for Kcc4a in regulation of proportion. Epistasis experiments suggest that Kcc4a and the two-pore potassium channel Kcnk5b both contribute to a common bioelectrical signaling response in the fin. These data suggest that an integrated bioelectric signaling pathway is required for the coordination of size and proportion during development.

Vision fine-tunes preparation for landing in the cane toad, Rhinella marina.

In toad hopping, the hindlimbs generate the propulsive force for take-off while the forelimbs resist the impact forces associated with landing. Preparing to perform a safe landing, in which impact forces are managed appropriately, likely involves the integration of multiple types of sensory feedback. In toads, vestibular and/or proprioceptive feedback is critical for coordinated landing; however, the role of vision remains unclear. To clarify this, we compare pre-landing forelimb muscle activation patterns before and after removing vision. Specifically, we recorded EMG activity from two antagonistic forelimb muscles, the anconeus and coracoradialis, which demonstrate distance-dependent onset timing and recruitment intensity, respectively. Toads were first recorded hopping normally and then again after their optic nerves were severed to remove visual feedback. When blind, toads exhibited hop kinematics and pre-landing muscle activity similar to when sighted. However, distance-dependent relationships for muscle activity patterns were more variable, if present at all. This study demonstrates that blind toads are still able to perform coordinated landings, reinforcing the importance of proprioceptive and/or vestibular feedback during hopping. But the increased variability in distance-dependent activity patterns indicates that vision is more responsible for fine-tuning the motor control strategy for landing.

Mineral homeostasis and regulation of mineralization processes in the skeletons of sharks, rays and relatives (Elasmobranchii).

Sharks, rays and other elasmobranch fishes are characterized by a skeletal type that is unique among living vertebrates, comprised predominantly of an unmineralized cartilage, covered by a thin outer layer of sub-millimeter, mineralized tiles called tesserae. The mineralized portion of the skeleton appears to grow only by apposition, adding material at the edges of each tessera; maintenance of non-mineralized joints between tesserae is therefore vital, with precise control of mineral deposition and inhibition at the many thousands of growth fronts in the skeleton. Yet, we have only scattered evidence as to how the elasmobranchs mineralize and grow their skeletons. In this review, we take an "environment to skeleton" approach, drawing together research from a vast range of perspectives to track calcium and phosphate from the typical elasmobranch habitats into and through the body, to their deposition at tesseral growth fronts. In the process, we discuss the available evidence for skeletal resorption capability, mineral homeostasis hormones, and nucleation inhibition mechanisms. We also outline relevant theories in crystal nucleation and typical errors in measurements of serum calcium and phosphate in the study of vertebrate biology. We assemble research that suggests consensus in some concepts in elasmobranch skeletal development, but also highlight the very large gaps in our knowledge, particularly in regards to endocrine functional networks and biomineralization mechanisms. In this way, we lay out frameworks for future directions in the study of elasmobranch skeletal biology with stronger and more comparative links to research in other disciplines and into other taxa.

Pre-landing wrist muscle activity in hopping toads.

Coordinated landing requires preparation. Muscles in the limbs important for decelerating the body should be activated prior to impact so that joints may be stiffened and limbs stabilized during landing. Moreover, because landings vary in impact force and timing, muscle recruitment patterns should be modulated accordingly. In toads, which land using their forelimbs, previous work has demonstrated such modulation in muscles acting at the elbow, but not at the shoulder. In this study, we used electromyography and high-speed video to test the hypothesis that antagonistic muscles acting at the wrists of toads are activated in advance of impact, and that these activation patterns are tuned to the timing and force of impact. We recorded from two wrist extensors: extensor carpi ulnaris (ECU) and extensor digitorum communis longus (EDCL), and two wrist flexors: flexor carpi ulnaris (FCU) and palmaris longus (PL). Each muscle was recorded in 4-5 animals (≥15 hops per animal). In all muscles, activation intensity was consistently greatest shortly before impact, suggesting the importance of these muscles during landing. Pre-landing recruitment intensity regularly increased with aerial phase duration (i.e. hop distance) in all muscles except PL. In addition, onset timing in both wrist flexors was also modulated with hop distance, with later onset times being associated with longer hops. Thus, activation patterns in major flexors and extensors of the wrist are tuned to hop distance with respect to recruitment intensity, onset timing or both.

Liars, medicine, and compassion.

This paper defends an account of compassion and argues for the centrality of compassion to the proper practice of medicine. The argument proceeds by showing that failures of compassion can lead to poor medical treatment and disastrous outcomes. Several case studies are discussed, exemplifying the difference between compassionate and noncompassionate responses to patients seeking help. Arguments are offered in support of approaching reports of persistent pain with a trusting attitude, rather than distrust or skepticism. The article concludes by suggesting educational improvements to encourage compassion.

Biomechanics and control of landing in toads.

Anything that jumps must land, but unlike during jumping when muscles produce energy to accelerate the body into the air, controlled landing requires muscles to dissipate energy and decelerate the body. Among anurans, toads (genus Bufo) exhibit highly coordinated landing behaviors, using their forelimbs to stabilize the body after touch-down as they lower their hindlimbs to the ground. Moreover, toads land frequently, as they cover distances by stringing together long series of relatively short hops. We have been using toads as a model to understand the biomechanics and motor control strategies of coordinated landing. Our results show that toads prepare for landing differently depending on how far they hop. For example, the forelimbs are extended farther prior to impact after long hops than after short ones. Such kinematic alterations are mirrored by predictable modulation of the recruitment intensity of forelimb muscles before impact, such that longer hops lead to higher levels of pre-landing recruitment of muscles. These differences in kinematics and muscular activity help to control the most flexed configuration of the elbow that is achieved after impact, which in turn constrains the extent to which muscles involved in dissipating energy are stretched. Indeed, a combination of in vivo and in vitro experiments has shown that the elbow-extending anconeus muscle, which is stretched during landing as the elbow flexes, rarely reaches lengths longer than those on the plateau of the muscle's length-tension curve (where damage becomes more likely). We have also been studying how movements of the hindlimbs after take-off help to stabilize animals during landing. In particular, the immediate and rapid flexion of a toad's knees after take-off leads to a repositioning of the animal's center of mass (COM) that better aligns it with ground-reaction forces (GRFs) at impact and reduces torques that would destabilize the animal. Finally, recent work on sensory feedback involved in preparation for landing demonstrates that vision is not required for coordinated landing. Toads can effectively utilize proprioceptive and/or vestibular information during take-off to help inform themselves about landing conditions, but may also use other sensory modalities after take-off to modulate landing behavior.

Pelvic girdle shape predicts locomotion and phylogeny in batoids.

In terrestrial vertebrates, the pelvic girdle can reliably predict locomotor mode. Because of the diminished gravitational effects on positively buoyant bony fish, the same relationship does not appear to exist. However, within the negatively buoyant elasmobranch fishes, benthic batoids employ pelvic fin bottom-walking and punting as primary or supplementary forms of locomotion. Therefore, in this study, we employed geometric and linear morphometrics to investigate if their pelvic girdles exhibit shape characteristics similar to those of sprawling terrestrial vertebrates. We tested for correlates of pelvic girdle shape with 1) Order, 2) Family, 3) Swim Mode, and/or 4) Punt Mode. Landmarks and semilandmarks were placed along outlines of dorsal views of 61 batoid pelvic girdles (3/3 orders, 10/13 families, 35/72 genera). The first three relative warps explained 88.45% of the variation among individuals (P < 0.01%). Only Order and Punt Mode contained groups that were all significantly different from each other (P < 0.01%). Discriminant function analyses indicated that the majority of variation within each category was due to differences in extension of lateral and prepelvic processes and puboischiac bar angle. Over 60% of the original specimens and 55% of the cross-validated specimens were correctly classified. The neutral angle of the propterygium, which articulates with the pelvic girdle, was significantly different among punt modes, whereas only pectoral fin oscillators had differently shaped pelvic girdles when compared with batoids that perform other swimming modes (P < 0.01). Pelvic girdles of batoids vary greatly, and therefore, likely function in ways not previously described in teleost fishes. This study illustrates that pelvic girdle shape is a good predictor of punt mode, some forms of swimming mode, and a species' Order. Such correlation between locomotor style and pelvic girdle shape provides evidence for the convergent evolution of morphological features that support both sprawled-gait terrestrial walking and aquatic bottom-walking.