Wing Digits Research Articles

Selection on Phalanx Development in the Evolution of the Bird Wing.

The frameshift hypothesis is a widely accepted model of bird wing evolution. This hypothesis postulates a shift in positional values, or molecular-developmental identity, that caused a change in digit phenotype. The hypothesis synthesized developmental and paleontological data on wing digit homology. The “most anterior digit” (MAD) hypothesis presents an alternative view based on changes in transcriptional regulation in the limb. The molecular evidence for both hypotheses is that the MAD expresses Hoxd13 but not Hoxd11 and Hoxd12. This digit I “signature” is thought to characterize all amniotes. Here, we studied Hoxd expression patterns in a phylogenetic sample of 18 amniotes. Instead of a conserved molecular signature in digit I, we find wide variation of Hoxd11, Hoxd12, and Hoxd13 expression in digit I. Patterns of apoptosis, and Sox9 expression, a marker of the phalanx-forming region, suggest that phalanges were lost from wing digit IV because of early arrest of the phalanx-forming region followed by cell death. Finally, we show that multiple amniote lineages lost phalanges with no frameshift. Our findings suggest that the bird wing evolved by targeted loss of phalanges under selection. Consistent with our view, some recent phylogenies based on dinosaur fossils eliminate the need to postulate a frameshift in the first place. We suggest that the phenotype of the Archaeopteryx lithographica wing is also consistent with phalanx loss. More broadly, our results support a gradualist model of evolution based on tinkering with developmental gene expression.

An autoregulatory cell cycle timer integrates growth and specification in chick wing digit development.

A fundamental question is how proliferation and growth are timed during embryogenesis. Although it has been suggested that the cell cycle could be a timer, the underlying mechanisms remain elusive. Here we describe a cell cycle timer that operates in Sonic hedgehog (Shh)-expressing polarising region cells of the chick wing bud. Our data are consistent with Shh signalling stimulating polarising region cell proliferation via Cyclin D2, and then inhibiting proliferation via a Bmp2-p27kip1 pathway. When Shh signalling is blocked, polarising region cells over-proliferate and form an additional digit, which can be prevented by applying Bmp2 or by inhibiting D cyclin activity. In addition, Bmp2 also restores posterior digit identity in the absence of Shh signalling, thus indicating that it specifies antero-posterior (thumb to little finger) positional values. Our results reveal how an autoregulatory cell cycle timer integrates growth and specification and are widely applicable to many tissues.

Identification of a novel Afipia species isolated from an Indian flying fox.

An old world fruit bat Pteropus giganteus, held in captivity and suffering from necrosis of its wing digits, failed to respond to antibiotic therapy and succumbed to the infection. Samples submitted to the National Centre for Foreign Animal Disease were tested for viral infection. Vero E6 cells exhibited minor but unique cytopathic effects on second blind passage, and full CPE by passage four. Utilizing an unbiased random amplification technique from cell culture supernatant, we identified a bacterium belonging to the Bradyrhizobiaceae. Purification of cell culture supernatant on TY media revealed a slow growing bacterial isolate. In this study using electron microscopy, 16S rRNA gene analysis and whole genome sequencing, we identify a novel bacterial species associated with the site of infection belonging to the genus Afipia. This genus of bacteria is very diverse, with only a limited number of species characterized. Afipia felis, previously described as the etiological agent to cause cat scratch disease, and Afipia septicemium, most recently shown to cause disease in humans, highlight the potential for members of this genus to form a branch of opportunistic pathogens within the Bradyrhizobiaceae. Increased utilization of next generation sequencing and genomics will aid in classifying additional members of this intriguing bacterial genera.

Skeletal morphology of the middle Eocene swift Scaniacypselus and the evolutionary history of true swifts (Apodidae)

New specimens of Scaniacypselus szarskii, a stem group representative of the Apodidae (true swifts) from the middle Eocene Messel oilshale in Germany, are described. These fossils show that Scaniacypselus differs from crown group Apodidae in a number of distinct and previously unrecognized osteological features. Notably, the sternum of Scaniacypselus is shorter than that of extant true swifts, the ulna proportionally longer, the carpometacarpus shorter, and the internal index process on the proximal phalanx of the major wing digit less developed. In details of its humerus morphology, Scaniacypselus is distinguished from all extant apodiform birds. Scaniacypselus further has a much shorter tarsometatarsus than most crown group Apodidae. It is hypothesized that short legs are plesiomorphic for Apodidae as a total group (stem group and crown group representatives), but that crown group Apodidae primitively have an elongated tarsometatarsus as an adaptation for clinging to vertical surfaces. The less specialized wing and pectoral girdle morphology suggests that Scaniacypselus was probably not as aerial as extant Apodidae. The differences in foot morphology indicate that it was more arboreal than its living relatives and had different breeding and roosting habits. Crown group Apodiformes probably diverged well after the middle Eocene, and the derived nesting behavior may have contributed to their evolutionary success.

Homologies and homeotic transformation of the theropod 'semilunate' carpal.

The homology of the ‘semilunate' carpal, an important structure linking non-avian and avian dinosaurs, has been controversial. Here we describe the morphology of some theropod wrists, demonstrating that the ‘semilunate' carpal is not formed by the same carpal elements in all theropods possessing this feature and that the involvement of the lateralmost distal carpal in forming the ‘semilunate' carpal of birds is an inheritance from their non-avian theropod ancestors. Optimization of relevant morphological features indicates that these features evolved in an incremental way and the ‘semilunate' structure underwent a lateral shift in position during theropod evolution, possibly as a result of selection for foldable wings in birds and their close theropod relatives. We propose that homeotic transformation was involved in the evolution of the ‘semilunate' carpal. In combination with developmental data on avian wing digits, this suggests that homeosis played a significant role in theropod hand evolution in general.

New developmental evidence supports a homeotic frameshift of digit identity in the evolution of the bird wing.

BackgroundThe homology of the digits in the bird wing is a high-profile controversy in developmental and evolutionary biology. The embryonic position of the digits cartilages with respect to the primary axis (ulnare and ulna) corresponds to 2, 3, 4, but comparative-evolutionary morphology supports 1, 2, 3. A homeotic frameshift of digit identity in evolution could explain how cells in embryonic positions 2, 3, 4 began developing morphologies 1, 2, 3. Another alternative is that no re-patterning of cell fates occurred, and the primary axis shifted its position by some other mechanism. In the wing, only the anterior digit lacks expression of HoxD10 and HoxD12, resembling digit 1 of other limbs, as predicted by 1, 2, 3. However, upon loss of digit 1 in evolution, the most anterior digit 2 could have lost their expression, deceitfully resembling a digit 1. To test this notion, we observed HoxD10 and HoxD12 in a limb where digit 2 is the most anterior digit: The rabbit foot. We also explored whether early inhibition of Shh signalling in the embryonic wing bud induces an experimental homeotic frameshift, or an experimental axis shift. We tested these hypotheses using DiI injections to study the fate of cells in these experimental wings.ResultsWe found strong transcription of HoxD10 and HoxD12 was present in the most anterior digit 2 of the rabbit foot. Thus, we found no evidence to question the use of HoxD expression as support for 1, 2, 3. When Shh signalling in early wing buds is inhibited, our fate maps demonstrate that an experimental homeotic frameshift is induced.ConclusionAlong with comparative morphology, HoxD expression provides strong support for 1, 2, 3 identity of wing digits. As an explanation for the offset 2, 3, 4 embryological position, the homeotic frameshift hypothesis is consistent with known mechanisms of limb development, and further proven to be experimentally possible. In contrast, the underlying mechanisms and experimental plausibility of an axis shift remain unclear.

Application of synthetic photostable retinoids induces novel limb and facial phenotypes during chick embryogenesisin vivo

We have recently developed a range of synthetic retinoid analogues which include the compounds EC23 and EC19. They are stable on exposure to light and are predicted to be resistant to the normal metabolic processes involved in the inactivation of retinoids in vivo. Based on the position of the terminal carboxylic acid groups in the compounds we suggest that EC23 is a structural analogue of all-trans retinoic acid (ATRA), and EC19 is an analogue of 13-cis retinoic acid. Their effects on the differentiation of pluripotent stem cells has been previously described in vitro and are consistent with this hypothesis. We present herein the first description of the effects of these molecules in vivo. Retinoids were applied to the anterior limb buds of chicken embryos in ovo via ion-exchange beads. We found that retinoid EC23 produces effects on the wing digits similar to ATRA, but does so at two orders of magnitude lower concentration. When larger quantities of EC23 are applied, a novel phenotype is obtained involving production of multiple digit 1s on the anterior limb. This corresponds to differential effects of ATRA and EC23 on sonic hedgehog (shh) expression in the developing limb bud. With EC23 application we also find digit 1 phenotypes similar to thumb duplications described in the clinical literature. EC23 and ATRA are shown to have effects on the entire proximal–distal axis of the limb, including hitherto undescribed effects on the scapula. This includes suppression of expression of the scapula marker Pax1. EC23 also produces effects similar to those of ATRA on the developing face, producing reductions of the upper beak at concentrations two orders of magnitude lower than ATRA. In contrast, EC19, which is structurally very similar to EC23, has novel, less severe effects on the face and rarely alters limb development. EC19 and ATRA are effective at similar concentrations. These results further demonstrate the ability of retinoids to influence embryonic development. Moreover, EC23 represents a useful new tool to investigate developmental processes and probe the mechanisms underlying congenital abnormalities in vertebrates including man.

Cenozoic phoenicopteriform birds from central Asia

A new phoenicopteriform bird, Palaelodus kurochkini sp. nov., is described from the terminal middle Miocene of Mongolia (the Sharga locality) on the basis of a partial tibiotarsus and tentatively associated fragmentary tarsometatarsus, carpometacarpus and proximal phalanx of the major wing digit. This is the first record of the extinct phoenicopteriform family Palaelodidae in Asia and shows that the genus Palaelodus is one of few taxa in the Sharga fauna to have a wide distribution range. Two other fossils of Phoenicopteriform birds from the Cenozoic of Central Asia are also restudied in this work; Agnopterus turgaiensis from the early Oligocene of Kazakhstan is provisionally placed within the extant family Phoenicopteridae, and Cygnopterus lambrechti, also from the Lower Oligocene of Kazakhstan, is identified as belonging to an indeterminate phoenicopteriform family.

Mathematical modelling of digit specification by a sonic hedgehog gradient

The three chick wing digits represent a classical example of a pattern specified by a morphogen gradient. Here we have investigated whether a mathematical model of a Shh gradient can describe the specification of the identities of the three chick wing digits and if it can be applied to limbs with more digits. We have produced a mathematical model for specification of chick wing digit identities by a Shh gradient that can be extended to the four digits of the chick leg with Shh-producing cells forming a digit. This model cannot be extended to specify the five digits of the mouse limb. Our data suggest that the parameters of a classical-type morphogen gradient are sufficient to specify the identities of three different digits. However, to specify more digit identities, this core mechanism has to be coupled to alternative processes, one being that in the chick leg and mouse limb, Shh-producing cells give rise to digits; another that in the mouse limb, the cellular response to the Shh gradient adapts over time so that digit specification does not depend simply on Shh concentration.

Digit loss in archosaur evolution and the interplay between selection and constraints

Evolution involves interplay between natural selection and developmental constraints. This is seen, for example, when digits are lost from the limbs during evolution. Extant archosaurs (crocodiles and birds) show several instances of digit loss under different selective regimes, and show limbs with one, two, three, four or the ancestral number of five digits. The 'lost' digits sometimes persist for millions of years as developmental vestiges. Here we examine digit loss in the Nile crocodile and five birds, using markers of three successive stages of digit development. In two independent lineages under different selection, wing digit I and all its markers disappear. In contrast, hindlimb digit V persists in all species sampled, both as cartilage, and as Sox9- expressing precartilage domains, 250 million years after the adult digit disappeared. There is therefore a mismatch between evolution of the embryonic and adult phenotypes. All limbs, regardless of digit number, showed similar expression of sonic hedgehog (Shh). Even in the one-fingered emu wing, expression of posterior genes Hoxd11 and Hoxd12 was conserved, whereas expression of anterior genes Gli3 and Alx4 was not. We suggest that the persistence of digit V in the embryo may reflect constraints, particularly the conserved posterior gene networks associated with the zone of polarizing activity (ZPA). The more rapid and complete disappearance of digit I may reflect its ZPA-independent specification, and hence, weaker developmental constraints. Interacting with these constraints are selection pressures for limb functions such as flying and perching. This model may help to explain the diverse patterns of digit loss in tetrapods. Our study may also help to understand how selection on adults leads to changes in development.

Correction: Bambi and Sp8 Expression Mark Digit Tips and Their Absence Shows That Chick Wing Digits 2 and 3 Are Truncated

Correction: Bambi and Sp8 Expression Mark Digit Tips and Their Absence Shows That Chick Wing Digits 2 and 3 Are Truncated

Bambi and Sp8 Expression Mark Digit Tips and Their Absence Shows That Chick Wing Digits 2 and 3 Are Truncated

An often overlooked aspect of digit development is the special nature of the terminal phalanx, a specialized structure with characteristics distinct from other phalanges, for example the presence of ectodermal derivatives such as nails and claws. Here, we describe the unique ossification pattern of distal phalanges and characteristic gene expression in the digit tips of chick and duck embryos. Our results show that the distal phalanx of chick wing digit 1 is a genuine tip with a characteristic ossification pattern and expression of Bambi and Sp8; however, the terminal phalanx of digits 2* and 3 is not a genuine tip, and these are therefore truncated digits. Bambi and Sp8 expression in the chick wing provides a direct molecular assessment of digit identity changes after experimental manipulations of digit primordia. In contrast, digits 1 and 2 of the duck wing both possess true tips. Although chick wing-tip development was not rescued by application of Fgf8, this treatment induced the development of extra phalanges. Grafting experiments show that competence for tip formation, including nails, is latent in the interdigital tissue. Our results deepen understanding of the mechanisms of digit tip formation, highlighting its developmental autonomy and modular nature, with implications for digit reduction or loss during evolution. * Numbering of wing digits is 1, 2, 3 from anterior to posterior.

On wing disparity and morphological variation of the Santana Group pterosaurs

Pterosaurs were widely spread throughout the Mesozoic Era, populating the whole world. Among this great diversity, two groups are commonly found in Brazil: the Anhangueridae and Tapejaridae. These can be mainly identified by cranial synapomorphies. However, because of the fragility of the pterosaur skeleton and rarity of the fossilisation process, the fossils found are usually incomplete, which hampers a proper taxonomic identification of the specimens. The specific proportions of these two groups of pterosaurs were obtained from bibliographic data and measurements of specimens. Eight Anhangueridae-like and seven Tapejaridae were used: Anhanguera piscator, Anhanguera santanae, Anhanguera spielbergi, Araripesaurus castilhoi, Barbosania gracilisrostris and three Anhangueridae sp. indet.; Sinopterus dongi, Tapejara wellnhoferi and five Tapejaridae sp. indet. We find that proportions of the humerus, wing metacarpal, first phalanx of the wing digit, femur and tibia are sufficient to identify partial remains of Araripe pterosaurs. A principal component analysis shows that each clade has different, non-overlapping scores in the studied ratios and these can be used with precision. Specific bone ratios for fast identification of anhanguerids and tapejarids are given, opening a broader way to diagnostic fragmentary bones.

Comment on "Embryological evidence identifies wing digits in birds as digits 1, 2, and 3."

AbstractTamura et al. (Science, 11 Febuary 2011, p. 753) claim that the three avian wing digits should be identified as digits I-III based on new embryological data and suggest that these identifications help to remove the conflict between the paleontological data and developmental data concerning the homology of the avian digits. However, their results are not novel, and the authors fail to address the critical problems relating to this interesting issue.

Comment on “Embryological evidence identifies wing digits in birds as digits 1, 2, and 3.”

Comment on “Embryological evidence identifies wing digits in birds as digits 1, 2, and 3.”

Transcriptomic analysis of avian digits reveals conserved and derived digit identities in birds

Morphological characters are the result of developmental gene expression. The identity of a character is ultimately grounded in the gene regulatory network directing development and thus whole-genome gene expression data can provide evidence about character identity. This approach has been successfully used to assess cell-type identity. Here we use transcriptomic data to address a long-standing uncertainty in evolutionary biology, the identity of avian wing digits. Embryological evidence clearly identifies the three wing digits as developing from digit positions 2, 3 and 4 (ref. 6), whereas palaeontological data suggest that they are digits I, II and III. We compare the transcriptomes of the wing and foot digits and find a strong signal that unites the first wing digit with the first foot digit, even though the first wing digit develops from embryological position 2. Interestingly, our transcriptomic data of the posterior digits show a higher degree of differentiation among forelimb digits compared with hindlimb digits. These data show that in the stem lineage of birds the first digit underwent a translocation from digit position 1 to position 2, and further indicate that the posterior wing digits have unique identities contrary to any model of avian digit identity proposed so far.

Identity of the avian wing digits: Problems resolved and unsolved

Controversy over bird wing digit identity has been a touchstone for various ideas in the phylogeny of birds, homology, and developmental evolution. This review summarizes the past 10 years of progress toward understanding avian digit identity. We conclude that the sum of evidence supports the Frame Shift Hypothesis, indicating that the avian wing digits have changed anatomical location. Briefly, the derivation of birds from theropod dinosaurs and the positional identities of the avian wing digits as 2, 3, and 4¹ are no longer in question. Additionally, increasing evidence indicates that the developmental programs for identity of the wing digits are of digits I, II, and III. Therefore, the attention moves from whether the digit identity frame shift occurred, to what the mechanisms of the frame shift were, and when in evolution it happened. There is considerable uncertainty about these issues and we identify exciting new research directions to resolve them.

Embryological Evidence Identifies Wing Digits in Birds as Digits 1, 2, and 3

The identities of the digits of the avian forelimb are disputed. Whereas paleontological findings support the position that the digits correspond to digits one, two, and three, embryological evidence points to digit two, three, and four identities. By using transplantation and cell-labeling experiments, we found that the posteriormost digit in the wing does not correspond to digit four in the hindlimb; its progenitor segregates early from the zone of polarizing activity, placing it in the domain of digit three specification. We suggest that an avian-specific shift uncouples the digit anlagen from the molecular mechanisms that pattern them, resulting in the imposition of digit one, two, and three identities on the second, third, and fourth anlagens.

Digit Identity in Bird Wing

For many years, there has been considerable controversy regarding bird wing digit identity. Embryological evidence has suggested that these digits represent digits 2, 3, and 4 in the pentadactyl ground state, whereas the fossil record supports a pattern of posterior digit reduction in the theropod

Reappraisal of Eocypselus—a stem group apodiform from the early Eocene of Northern Europe

Eocypselus vincenti is a small swift-like bird from the early Eocene of England and Denmark, whose phylogenetic affinities are controversial. Here, a new skeleton from the Danish Fur Formation is reported, in which for the first time the skull and pelvis are preserved. Further described are previously overlooked features of the other Fur Formation specimens, which bear on the systematic position of the species. It is detailed that E. vincenti falls outside crown group Apodiformes and represents the earliest diverging apodiform bird, with its osteology providing a link between that of apodiform birds and their morphologically disparate sister taxon, the Aegothelidae (owlet-nightjars). The poorly developed processus internus indicis on the proximal phalanx of the major wing digit indicates that Eocypselus had less elongated outer primaries than extant Apodiformes. Together with differences in the pectoral girdle, such as the lack of a coracoid/furcula articulation, this suggests that the fossil taxon was less well adapted than its extant relatives to feeding on the wing.