Multiple axes of visual system diversity in Ithomiini, an ecologically diverse tribe of mimetic butterflies.

The striking structural variation seen in arthropod visual systems can be explained by the overall quantity and spatio-temporal structure of light within habitats coupled with developmental and physiological constraints. However, little is currently known about how fine-scale variation in visual structures arises across shorter evolutionary and ecological scales. In this study, we characterise patterns of interspecific (between species), intraspecific (between sexes) and intraindividual (between eye regions) variation in the visual system of four ithomiine butterfly species. These species are part of a diverse 26-million-year-old Neotropical radiation where changes in mimetic colouration are associated with fine-scale shifts in ecology, such as microhabitat preference. Using a combination of selection analyses on visual opsin sequences, in vivo ophthalmoscopy, micro-computed tomography (micro-CT), immunohistochemistry, confocal microscopy and neural tracing, we quantify and describe physiological, anatomical and molecular traits involved in visual processing. Using these data, we provide evidence of substantial variation within the visual systems of Ithomiini, including: (i) relaxed selection on visual opsins, perhaps mediated by habitat preference, (ii) interspecific shifts in visual system physiology and anatomy, and (iii) extensive sexual dimorphism, including the complete absence of a butterfly-specific optic neuropil in the males of some species. We conclude that considerable visual system variation can exist within diverse insect radiations, hinting at the evolutionary lability of these systems to rapidly develop specialisations to distinct visual ecologies, with selection acting at the perceptual, processing and molecular level.

Scleraxis genes are required for normal musculoskeletal development and for rib growth and mineralization in zebrafish.

Tendons are an essential part of the musculoskeletal system, connecting muscle and skeletal elements to enable force generation. The transcription factor scleraxis marks vertebrate tendons from early specification. Scleraxis-null mice are viable and have a range of tendon and bone defects in the trunk and limbs but no described cranial phenotype. We report the expression of zebrafish scleraxis orthologs: scleraxis homolog (scx)-a and scxb in cranial and intramuscular tendons and in other skeletal elements. Single mutants for either scxa or scxb, generated by clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9), are viable and fertile as adult fish. Although scxb mutants show no obvious phenotype, scxa mutant embryos have defects in cranial tendon maturation and muscle misalignment. Mutation of both scleraxis genes results in more severe defects in cranial tendon differentiation, muscle and cartilage dysmorphogenesis and paralysis, and lethality by 2-5 wk, which indicates an essential function of scleraxis for craniofacial development. At juvenile and adult stages, ribs in scxa mutants fail to mineralize and/or are small and heavily fractured. Scxa mutants also have smaller muscle volume, abnormal swim movement, and defects in bone growth and composition. Scleraxis function is therefore essential for normal craniofacial form and function and vital for fish development.-Kague, E., Hughes, S. M., Lawrence, E. A., Cross, S., Martin-Silverstone, E., Hammond, C. L., Hinits, Y. Scleraxis genes are required for normal musculoskeletal development and for rib growth and mineralization in zebrafish.

Giantin-knockout models reveal a feedback loop between Golgi function and glycosyltransferase expression.

The Golgi is the cellular hub for complex glycosylation, controlling accurate processing of complex proteoglycans, receptors, ligands and glycolipids. Its structure and organisation are dependent on golgins, which tether cisternal membranes and incoming transport vesicles. Here, we show that knockout of the largest golgin, giantin, leads to substantial changes in gene expression but only limited effects on Golgi structure. Notably, 22 Golgi-resident glycosyltransferases, but not glycan-processing enzymes or the ER glycosylation machinery, are differentially expressed following giantin ablation. This includes near-complete loss of function of GALNT3 in both mammalian cell and zebrafish models. Giantin-knockout zebrafish exhibit hyperostosis and ectopic calcium deposits, recapitulating phenotypes of hyperphosphatemic familial tumoral calcinosis, a disease caused by mutations in GALNT3. These data reveal a new feature of Golgi homeostasis: the ability to regulate glycosyltransferase expression to generate a functional proteoglycome.

Powered flight in hatchling pterosaurs: evidence from wing form and bone strength.

Competing views exist on the behaviour and lifestyle of pterosaurs during the earliest phases of life. A 'flap-early' model proposes that hatchlings were capable of independent life and flapping flight, a 'fly-late' model posits that juveniles were not flight capable until 50% of adult size, and a 'glide-early' model requires that young juveniles were flight-capable but only able to glide. We test these models by quantifying the flight abilities of very young juvenile pterosaurs via analysis of wing bone strength, wing loading, wingspan and wing aspect ratios, primarily using data from embryonic and hatchling specimens of Pterodaustro guinazui and Sinopterus dongi. We argue that a young Sinopterus specimen has been mischaracterised as a distinct taxon. The humeri of pterosaur juveniles are similar in bending strength to those of adults and able to withstand launch and flight; wing size and wing aspect ratios of young juveniles are also in keeping with powered flight. We therefore reject the 'fly-late' and 'glide-early' models. We further show that young juveniles were excellent gliders, albeit not reliant on specialist gliding. The wing forms of very young juveniles differ significantly from larger individuals, meaning that variation in speed, manoeuvrability, take-off angle and so on was present across a species as it matured. Juveniles appear to have been adapted for flight in cluttered environments, in contrast to larger, older individuals. We propose on the basis of these conclusions that pterosaur species occupied distinct niches across ontogeny.

Volant Fossil Vertebrates: Potential for Bioinspired Flight Technology.

Animal flight is ecologically important and has a long evolutionary history. It has evolved independently in many distantly related clades of animals. Powered flight has evolved only three times in vertebrates, making it evolutionarily rare. Major recent fossil discoveries have provided key data on fossil flying vertebrates and critical insights regarding the evolution and different arrangements of animal flight surfaces. Combined with new methodologies, these discoveries have paved the way for potentially expanding biomimetic and biologically inspired designs to incorporate lessons from fossil taxa. Here, we review the latest knowledge and literature regarding flight performance in fossil vertebrates. We then synthesise key elements to provide an overview of those cases where fossil flyers might provide new insights for applied sciences.

Morphometric assessment of pterosaur jaw disparity.

Pterosaurs were a successful group of Mesozoic flying reptiles. They were the first vertebrate group to achieve powered flight and varied enormously in morphology and ecology, occupying a variety of niches and developing specialized feeding strategies. Ecomorphological principles suggest this variation should be reflected by great morphological diversity in the lower jaw, given that the mandible served as the primary apparatus for prey acquisition. Here we present the first study of mandibular shape disparity in pterosaurs and aim to characterize major aspects of variation. We use a combination of geometric morphometric approaches, incorporating both outline analysis using elliptical Fourier analysis and semi-landmark approaches. Our results show that morphological convergence is prevalent and many pterosaurs, belonging to diverse dietary groups and subclades, overlap in morphospace and possessed relatively simple 'rod-shaped' jaws. There is no clear trend of size distributions in pterosaur mandibular morphospace, and larger forms are widely distributed. Additionally, there is limited functional signal within pterosaur lower jaw morphospace. Instead, the development of a large anterior ventral crest represents the major component of disparity. This suggests that a socio-sexual trait was a key driver for innovation in pterosaur lower jaw shape.

A small azhdarchoid pterosaur from the latest Cretaceous, the age of flying giants.

Pterosaur fossils from the Campanian-Maastrichtian of North America have been reported from the continental interior, but few have been described from the west coast. The first pterosaur from the Campanian Northumberland Formation (Nanaimo Group) of Hornby Island, British Columbia, is represented here by a humerus, dorsal vertebrae (including three fused notarial vertebrae), and other fragments. The elements have features typical of Azhdarchoidea, an identification consistent with dominance of this group in the latest Cretaceous. The new material is significant for its size and ontogenetic stage: the humerus and vertebrae indicate a wingspan of ca 1.5 m, but histological sections and bone fusions indicate the individual was approaching maturity at time of death. Pterosaurs of this size are exceedingly rare in Upper Cretaceous strata, a phenomenon commonly attributed to smaller pterosaurs becoming extinct in the Late Cretaceous as part of a reduction in pterosaur diversity and disparity. The absence of small juveniles of large species-which must have existed-in the fossil record is evidence of a preservational bias against small pterosaurs in the Late Cretaceous, and caution should be applied to any interpretation of latest Cretaceous pterosaur diversity and success.

Exploring the Relationship between Skeletal Mass and Total Body Mass in Birds.

Total body mass (TBM) is known to be related to a number of different osteological features in vertebrates, including limb element measurements and total skeletal mass. The relationship between skeletal mass and TBM in birds has been suggested as a way of estimating the latter in cases where only the skeleton is known (e.g., fossils). This relationship has thus also been applied to other extinct vertebrates, including the non-avian pterosaurs, while other studies have used additional skeletal correlates found in modern birds to estimate TBM. However, most previous studies have used TBM compiled from the literature rather than from direct measurements, producing values from population averages rather than from individuals. Here, we report a new dataset of 487 extant birds encompassing 79 species that have skeletal mass and TBM recorded at the time of collection or preparation. We combine both historical and new data for analyses with phylogenetic control and find a similar and well-correlated relationship between skeletal mass and TBM. Thus, we confirm that TBM and skeletal mass are accurate proxies for estimating one another. We also look at other factors that may have an effect on avian body mass, including sex, ontogenetic stage, and flight mode. While data are well-correlated in all cases, phylogeny is a major control on TBM in birds strongly suggesting that this relationship is not appropriate for estimating the total mass of taxa outside of crown birds, Neornithes (e.g., non-avian dinosaurs, pterosaurs). Data also reveal large variability in both bird skeletal and TBM within single species; caution should thus be applied when using published mass to test direct correlations with skeletal mass and bone lengths.