Terrestrial acclimation and exercise lead to bone functional response in Polypterus senegalus pectoral fins.

The ability of bones to sense and respond to mechanical loading is a central feature of vertebrate skeletons. However, the functional demands imposed on terrestrial and aquatic animals differ vastly. The pectoral girdle of the basal actinopterygian fish Polypterus senegalus was previously shown to exhibit plasticity following terrestrial acclimation, but the pectoral fin itself has yet to be examined. We investigated skeletal plasticity in the pectoral fins of P. senegalus after exposure to terrestrial loading. Juvenile fish were divided into three groups: a control group was kept under aquatic conditions without intervention, an exercised group was also kept in water but received daily exercise on land, and a terrestrial group was kept in a chronic semi-terrestrial condition. After 5 weeks, the pectoral fins were cleared and stained with Alcian Blue and Alizarin Red to visualize cartilage and bone, allowing measurements of bone length, bone width, ossification and curvature to be taken for the endochondral radial bones. Polypterus senegalus fin bones responded most strongly to chronic loading in the terrestrial condition. Fish that were reared in a terrestrial environment had significantly longer bones compared with those of aquatic controls, wider propterygia and metapterygia, and more ossified metapterygia and medial radials, and they showed changes in propterygial curvature. Exercised fish also had longer and more ossified medial radials compared with those of controls. Polypterus senegalus fin bones exhibit plasticity in response to novel terrestrial loading. Such plasticity could be relevant for transitions between water and land on evolutionary scales, but key differences between fish and tetrapod bone make direct comparisons challenging.

Artificial turf infill associated with systematic toxicity in an amniote vertebrate.

Artificial athletic turf containing crumb rubber (CR) from shredded tires is a growing environmental and public health concern. However, the associated health risk is unknown due to the lack of toxicity data for higher vertebrates. We evaluated the toxic effects of CR in a developing amniote vertebrate embryo. CR water leachate was administered to fertilized chicken eggs via different exposure routes, i.e., coating by dropping CR leachate on the eggshell; dipping the eggs into CR leachate; microinjecting CR leachate into the air cell or yolk. After 3 or 7 d of incubation, embryonic morphology, organ development, physiology, and molecular pathways were measured. The results showed that CR leachate injected into the yolk caused mild to severe developmental malformations, reduced growth, and specifically impaired the development of the brain and cardiovascular system, which were associated with gene dysregulation in aryl hydrocarbon receptor, stress-response, and thyroid hormone pathways. The observed systematic effects were probably due to a complex mixture of toxic chemicals leaching from CR, such as metals (e.g., Zn, Cr, Pb) and amines (e.g., benzothiazole). This study points to a need to closely examine the potential regulation of the use of CR on playgrounds and artificial fields.

Integration and modularity of teleostean pectoral fin shape and its role in the diversification of acanthomorph fishes.

Phenotypic integration and modularity describe the strength and pattern of interdependencies between traits. Integration and modularity have been proposed to influence the trajectory of evolution, either acting as constraints or facilitators. Here, we examine trends in the integration and modularity of pectoral fin morphology in teleost fishes using geometric morphometrics. We compare the fin shapes of the highly diverse radiation of acanthomorph fishes to lower teleosts. Integration and modularity are measured using two-block partial least squares analysis and the covariance ratio coefficient between the radial bones and lepidotrichia of the pectoral fins. We show that the fins of acanthomorph fishes are more tightly integrated but also more morphologically diverse and faster evolving compared to nonacanthomorph fishes. The main pattern of shape covariation in nonacanthomorphs is concordant with the main trajectory of evolution between nonacanthomorphs and acanthomorphs. Our findings support a facilitating role for integration during the acanthomorph diversification. Potential functional consequences and developmental mechanisms of fin integration are discussed.

Phenotypic plasticity of muscle fiber type in the pectoral fins of Polypterus senegalus reared in a terrestrial environment.

Muscle fiber types in the pectoral fins of fishes have rarely been examined, despite their morphological and functional diversity. Here, we describe the distribution of fast and slow muscle fibers in the pectoral fins of Polypterus senegalus, an amphibious, basal actinopterygian. Each of the four muscle groups examined using mATPase staining showed distinct fiber-type regionalization. Comparison between fish raised in aquatic and terrestrial environments revealed terrestrially reared fish possess 28% more fast muscle compared with aquatically reared fish. The pattern of proximal-distal variation in the abductors differed, with a relative decrease in fast muscle fibers near the pectoral girdle in aquatic fish compared with an increase in terrestrial fish. Terrestrially reared fish also possess a greater proportion of very small diameter fibers, suggesting that they undergo more growth via hyperplasia. These observations may be a further example of adaptive plasticity in Polypterus, allowing for greater bursts of power during terrestrial locomotion.

Locomotor flexibility of Polypterus senegalus across various aquatic and terrestrial substrates.

Amphibious fishes show wide variation in form and function. Examination of terrestrial locomotion in fishes has largely focused on highly specialized taxa. From an evolutionary perspective we are interested in how relatively unspecialized fishes locomote when exposed to different terrestrial environments. In this study, we explore the locomotory repertoire of the basal actinopterygian Polypterus senegalus. We describe its terrestrial locomotory strategies on different surfaces and compare them with steady aquatic locomotion. Our study is the first to describe axial and appendicular-based terrestrial locomotion across a range of substrates in this species and shows two distinct terrestrial gaits in P. senegalus, each with substantial variation. One uses body undulation against complex surfaces and the other incorporates alternating, supportive pectoral fins to achieve a tetrapod-like gait. Gait use is correlated with substrate. P. senegalus uses body and pectoral fins in diverse ways, showing opportunistic and flexible strategies of navigating novel environments.

Polypterus and the evolution of fish pectoral musculature.

Polypterus, a member of the most primitive living group of ray-finned fishes, has demonstrated the ability to perform fin-assisted terrestrial locomotion, a behavior that indicates a complex pectoral musculoskeletal system. Review of the literature reveals that many aspects of the pectoral muscular anatomy of Polypterus are still unclear, with a number of conflicting descriptions. We provide a new interpretation of the pectoral musculature using soft tissue-enhanced microCT scanning and gross anatomical dissection. The results demonstrate a complex musculature, with six independent muscles crossing the glenoid-fin joint. Comparisons with other bony-fish (Osteichthyes), including both ray-finned (Actinopterygii) and lobed-fin (Sarcopterygii) fish, indicate the presence of novel muscles within Polypterus: coracometapterygialis I+II and the zonopropterygialis medialis. Examination of these muscular additions in the context of osteichthyan phylogeny indicates that this represents a previously unrecognized event in the evolution of pectoral musculature in Osteichthyes. Despite its phylogenetic position as a basal actinopterygian, the musculature of Polypterus has more similarities both anatomically and functionally with that of sarcopterygians. This anatomy, along with other features of Polypterus anatomy such as lobed fins, ventral paired lungs, and a large spiracle, may make it a good model for inferences of stem tetrapod locomotion.

Developmental plasticity and the origin of tetrapods.

The origin of tetrapods from their fish antecedents, approximately 400 million years ago, was coupled with the origin of terrestrial locomotion and the evolution of supporting limbs. Polypterus is a member of the basal-most group of ray-finned fish (actinopterygians) and has many plesiomorphic morphologies that are comparable to elpistostegid fishes, which are stem tetrapods. Polypterus therefore serves as an extant analogue of stem tetrapods, allowing us to examine how developmental plasticity affects the 'terrestrialization' of fish. We measured the developmental plasticity of anatomical and biomechanical responses in Polypterus reared on land. Here we show the remarkable correspondence between the environmentally induced phenotypes of terrestrialized Polypterus and the ancient anatomical changes in stem tetrapods, and we provide insight into stem tetrapod behavioural evolution. Our results raise the possibility that environmentally induced developmental plasticity facilitated the origin of the terrestrial traits that led to tetrapods.