Carpal bone kinematics and ligament lengthening studied for the full range of joint movement

Present data on carpal kinematics and carpal ligament behaviour are limited to flexion and deviation movements of the hand. These motions do not represent all the wrist-joint motions which are important for the activities of daily living. The goal of this project was to obtain insight into carpal kinematics and carpal ligament behaviour during motions of the hand covering the full range of motion of the wrist joint.

The carpals and the ligaments of four wrist-joint specimens were provided with radiopaque markers. These joints were subjected to Röntgenstereophotogrammetric experimentation in a large number of hand positions to determine carpal positions and ligament lengths. The movements of the carpal bones were described by means of finite helical axes (FHA).

It was found that the movements of the carpals in the distal row closely resemble those of the hand. Conversely, the motions of the carpals of the proximal row appeared not to be directly proportional to the hand motions and exhibited clear out-of-plane movements. Furthermore, it could be shown that movements of the hand into the ulnodorsal quadrant of the full range of hand motion corresponds to larger helical rotations and translations for most of the carpals than when the hand was moved into any other quadrant. The maximal ligament length changes determined did not exceed the length changes reported for pure flexion and pure deviation movements of the hand.     

Autopodial development in the sea turtles Chelonia mydas and Caretta caretta

The manus and pes were studied using whole-mount and histological preparations of ontogenetic series of Chelonia mydas and Caretta caretta. Patterns of connectivity and sequences of chondrification events are similar to those reported for other turtle species, with respect to both the primary axis and the digital arch. There is no evidence of anterior condensations in the region distal to the radius and the tibia, supporting the hypothesis that the radiale and tibiale are absent in turtles. The three middle metacarpals are the first elements to start ossification in the manus of C. mydas, while ossification has not started in the pes. In the hatchling of C. mydas, most carpals have started ossification, whereas tarsals are mostly still cartilaginous. In C. caretta, the first carpals to ossify are the ulnare and intermedium, followed by the pisiform. Among metatarsals, the fifth hooked metatarsal is the last one to start ossification. The fibulare and intermedium fuse early in chondrogenesis, later becoming the astragalocalcaneum. Ossification in the carpals of C. caretta starts while tarsals are still cartilaginous. The derived autopodial proportions in each autopodium of adults are laid out at the condensation stage, and features that were present in basal turtles are absent at all stages examined (developmental penetrance). In contrast to this, conservatism is expressed in the presence of similar patterns of connectivity during early chondrogenesis, and in the development of overall proportions of the manus versus pes. As in adult anatomy, the development of the autopodium of marine turtles is a mosaic of derived and plesiomorphic features.     

Anatomy of the squirrel wrist: bones, ligaments, and muscles

Anatomical differences among squirrels are usually most evident in the comparison of flying squirrels and nongliding squirrels. This is true of wrist anatomy, probably reflecting the specializations of flying squirrels for the extension of the wing tip and control of it during gliding. In the proximal row of carpals of most squirrels, the pisiform articulates only with the triquetrum, but in flying squirrels there is also a prominent articulation between the pisiform and the scapholunate, providing a more stable base for the styliform cartilage, which supports the wing tip. In the proximal wrist joint, between these carpals and the radius and ulna, differences in curvature of articular surfaces and in the location of ligaments also correlate with differences in degree and kind of movement occurring at this joint, principally reflecting the extreme dorsal flexion and radial deviation of the wrist in flying squirrels when gliding. The distal wrist joint, between the proximal and distal rows of carpals, also shows most variation among flying squirrels, principally in the articulations of the centrale with the other carpal bones, probably causing the distal row of carpal bones to function more like a single unit in some animals. There is little variation in wrist musculature, suggesting only minor evolutionary modification since the tribal radiation of squirrels, probably in the early Oligocene. Variation in the carpal bones, particularly the articulation of the pisiform with the triquetrum and the scapholunate, suggests a different suprageneric grouping of flying squirrels than previously proposed by McKenna (1962) and Mein (1970). J. Morphol. 246:85-102, 2000. Published 2000 Wiley-Liss, Inc.     

Carpal ontogeny in Monodelphis domestica and Caluromys philander (Marsupialia)

Carpal bones have experienced numerous changes during marsupial evolution, even though their diversity and development remain poorly studied. The aim of this work was to document adult form and the pattern of mesenchymal tissue condensation and formation of chondrification and ossification centers in the hand of two marsupials. Two fundamental questions were asked: whether the loss of embryonic precursors was associated with the loss of adult elements, or whether there were developmental signs of ancestral mammalian elements that have been fused or lost in marsupial taxa. We were also interested to find out whether there is sexual dimorphismus in the carpals, as has been reported for some didelphids. Histological sections, cleared and stained specimens and macerated skeletons representing an ontogenetic series of Monodelphis domestica were used to document carpal development. Comparisons were made with perinatal stages of Caluromys philander and with adult specimens of other marsupials. A prenatal M. domestica in the 13th day after conception has a cell condensation that because of its position is homologized with a centrale, which is at birth already lost or fused. Neonatal M. domestica and C. philander have the number and arrangement of their adult carpal anatomy. Trapezium and trapezoid start ossification later than most other carpals, while pisiform and prepollex are the last to do so. Adult males of M. domestica have relatively larger and more robust pisiforms, compared to other carpals, than females. This sexual dimorphism develops relatively late as it was not recorded in male specimens around 160 days old. An extra sesamoid bone located just distal to the radius and proximo-palmar to the scaphoid was recorded in specimens of C. philander, C. derbianus and Didelphis virginiana.     

Limb development in the gekkonid lizard gonatodes albogularis: A reconsideration of homology in the lizard carpus and tarsus

Despite the attention squamate lizards have received in the study of digit and limb loss, little is known about limb morphogenesis in pentadactyl lizards. Recent developmental studies have provided a basis for understanding lizard autopodial element homology based on developmental and comparative anatomy. In addition, the composition and identity of some carpal and tarsal elements of lizard limbs, and reptiles in general, have been the theme of discussions about their homology compared to non‐squamate Lepidosauromorpha and basal Amniota. The study of additional embryonic material from different lizard families may improve our understanding of squamate limb evolution. Here, we analyze limb morphogenesis in the gekkonid lizard Gonatodes albogularis describing patterns of chondrogenesis and ossification from early stages of embryonic development to hatchlings. Our results are in general agreement with previous developmental studies, but we also show that limb development in squamates probably involves more chondrogenic elements for carpal and tarsal morphogenesis, as previously recognized on the grounds of comparative anatomy. We provide evidence for the transitory presence of distal carpale 1 and intermedium in the carpus and tibiale, intermedium, distal centralia, and distal tarsale 2 in the tarsus. Hence, we demonstrate that some elements that were believed to be lost in squamate evolution are conserved as transitory elements during limb development. However, these elements do not represent just phylogenetic burden but may be important for the morphogenesis of the lizard autopodium. J. Morphol., 2010. © 2010 Wiley‐Liss, Inc.     

Growth and shaping of metacarpal and carpal cartilage anlagen: application of morphometry to the development of short and long bone. A study of human hand anlagen in the fetal period

A histological and morphometric analysis of human metacarpal and carpal anlagen between the 16th and 22nd embryonic weeks was carried out with the aim of studying the establishment of the respective anlage architecture. No differences in the pattern of growth were documented between the peripheral and central zones of the metacarpal epiphyses and those of the carpals. The regulation of longitudinal growth in long bone anlagen occurred in the transition zone between the epiphysis and the diaphysis (homologous to the metaphyseal growth plate cartilage in more advanced developmental stage of the bone). Comparative zonal analysis was conducted to assess the chondrocyte density, the mean chondrocyte lacunar area, the paired chondrocyte polarity in the orthogonal longitudinal and transverse planes, and the lacunar shape transformation in the metacarpal. In transition from epiphysis to diaphysis chondrocyte density decreased and mean lacunar area increased. No significant differences in the chondrocyte maturation cycle were observed between proximal/distal metacarpal epiphyses and the carpal anlagen. The number of paired chondrocyte oriented along the growth vector was significantly higher in both proximal/distal transition zones between epiphysis and diaphysis. Human metacarpals shared with experimental models (like mice and nonmammal tetrapods) an early common chondrocyte maturation cycle but with a different timing due to the slower embryonic and fetal developmental rate of human anlagen.     

Genotyping of Ochrobactrum spp. by AFLP analysis

AFLP was used to analyze the genetic diversity among Ochrobactrum strains. AFLP patterns showed a great genomic variability that separated the samples into three distinct clusters. Ochrobactrum intermedium was found to be closely related to Brucella abortus S99.     

The mole's thumb -- evolution of the hand skeleton in talpids (Mammalia)

Osteological specimens representing 15 out of the 16 currently recognized talpid genera were examined and scored for seven discrete morphological characters of the hand. The phylogenetic distribution of these characters was studied in the context of alternative hypotheses of talpid relationships, using three species of shrews and a hedgehog as outgroups. All talpids show a similar number and arrangement of carpal bones. The most obvious differences concern the presence of additional sesamoid bones, the relative size of the os falciforme when present, and the degree of fusion of the scaphoid and lunate in the proximal carpal row. Marked differences in the relative length and proportions of the metacarpals also exist. The development of the carpals in Talpa europaea was studied through examination of histological sections of the hand of an embryo and a neonate. Whereas carpal anatomy in the neonate mirrors the arrangement and proportions of the adult, in the embryo the scaphoid and lunate are still separate, there are no signs of the os falciforme, and the size proportions of metacarpals to carpals are obviously different to those of the adult. A prehallux or tibial sesamoid, serial homologue to the os falciforme or prepollex (a radial sesamoid), does not have an obvious functional role, and its presence might be the result of a common epigenetic control in the hand and the foot resulting in a non-adaptive structure in the latter.     

Appendicular skeleton in Bachia bicolor (Squamata: Gymnophthalmidae): osteology,limb reduction and postnatal skeletal ontogeny

The osteology of the appendicular skeleton and its postnatal development are described in Bachia bicolor, a serpentiform lizard with reduced limbs. The pectoral girdle is well developed and the forelimb consists of a humerus, ulna, radius, five carpal elements (ulnare, radiale, distal carpals 4–3, centrale), four metacarpals (II, III, IV, V) and phalanges (phalangeal formula X‐2‐2‐2‐2). In the hindlimb, the femur is small and slender, and articulates distally with a series of ossified amorphous and extremely reduced elements that correspond to a fibula, tibia and proximal and distal tarsals 4 and 3. The pelvic girdle consists of ischium, pubis and ilium, but its two halves are widely separated; the ilium is the least reduced element. We describe the ossification and development during postnatal skeletal ontogeny, especially of epiphyseal secondary centres, ossifications of carpal elements, apophyseal ossifications and sesamoids. Compared to other squamates, B. bicolor shows an overall reduction in limb size, an absence of skeletal elements, a fusion of carpal elements, an early differentiation of apophyseal centres, and a low number of sesamoids and apophyseal centres. These observations suggest that the reductions are produced by heterochronic changes during postnatal development and probably during embryonic development; therefore the appendicular skeleton exhibits a pattern of paedomorphic features.     

Biometric study of ruminant carpal bones and implications for phylogenetic relationships

The mammalian Carpus is a complex of several small bones with multiple interactions during walking. Therefore, it is highly probable that different mammalian families developed distinctive constellations in their Carpi, which could be useful for biometric identification of phylogenetic groupings. The carpal bones of nineteen extant ruminant species (nine bovid, nine cervid, and one moschid) have been investigated to search for biometric traits which are diagnostic for the three families. Additionally, we searched for diverging functional adaptations in the carpal constellations. Therefore, measurements have been taken from the five main carpals, which are carrying the body weight. As a sesamoid bone, Os carpi accessorium was excluded. After transformation of the data into their natural logarithms, multivariate methods of factor analyses and discriminant analyses were performed for each bone. Bivariate plots of the factor scores allowed a clear separation of bovids and cervids. The only one species of the Moschidae (Moschus moschiferus) lies closer to the cervids than to the bovids. The grouping is due to phylogenetic relationships and not due to functional differences in the groups or differing habitat preferences. Generally, the carpals of cervids are more slender and higher in contrast to the bulky and flat carpals in bovids. This approach could be used to assign isolated carpal bones found in fossil sites to their ruminant family.     

Phytogeny, Variation, and Morphological Integration

Studies of intra and interspecific variability of urodele mesopodiahave revealed biases in the production of novel phenotypes;observed patterns of variability represent a small subset ofthe possible arrangements. The major trend in the phylogeneticdiversification of the salamander limb is the reduction or lossof carpal or tarsal elements, and these patterns are often highlyhomoplastic. Mesopodial arrangements that are homoplastic arecommonly encountered in studies of intrapopulational variation.Patterns of variation, when integrated with developmental biology,inform about the relationships between morphological integrationand homology. The extensive conservation of primitive developmentaland genetic interactions suggest that the individuation of structuresmaynot be complete and that homologies, themselves, may be partial.     

Loss of carpal elements in crocodilian limb evolution: morphogenetic model corroborated by palaeobiological data

The comparison of bone homology between the manus of an Early Cretaceous fossil crocodile and that of the extant species Alligator mississippiensis supports explicidy, for the first time, the hypothesis of carpal loss in crocodilian limb evolution. This hypothesis, based on a developmental model of the organization of the tetrapod limb, is in accordance with the fossil evidence, and may supersede traditional Haeckelian views based on recapitulatory paradigms. The homologous relationships of carpal elements reveal the existence of two carpal patterns- one plesiomorphic and one apomorphic-in the crocodilian lineage. Phylogenetic change is explained causally by alterations of the osteogenesis of the distal carpals 2 and 3, which remain unossified in extant crocodile adults. This implies that crocodilian limb evolution is constrained by a process of paedomorphosis. This modification of the architecture of the crocodilian hand is a terminal event of its evolutionary history, affecting only eusuchian crocodiles. The results of this study contest the traditional view that the skeletal pattern of the crocodilian limb has been conserved unchanged since the Triassic.     

New Developmental Evidence Clarifies the Evolution of Wrist Bones in the Dinosaur–Bird Transition

From early dinosaurs with as many as nine wrist bones, modern birds evolved to develop only four ossifications. Their identity is uncertain, with different labels used in palaeontology and developmental biology. We examined embryos of several species and studied chicken embryos in detail through a new technique allowing whole-mount immunofluorescence of the embryonic cartilaginous skeleton. Beyond previous controversy, we establish that the proximal–anterior ossification develops from a composite radiale+intermedium cartilage, consistent with fusion of radiale and intermedium observed in some theropod dinosaurs. Despite previous claims that the development of the distal–anterior ossification does not support the dinosaur–bird link, we found its embryonic precursor shows two distinct regions of both collagen type II and collagen type IX expression, resembling the composite semilunate bone of bird-like dinosaurs (distal carpal 1+distal carpal 2). The distal–posterior ossification develops from a cartilage referred to as “element x,” but its position corresponds to distal carpal 3. The proximal–posterior ossification is perhaps most controversial: It is labelled as the ulnare in palaeontology, but we confirm the embryonic ulnare is lost during development. Re-examination of the fossil evidence reveals the ulnare was actually absent in bird-like dinosaurs. We confirm the proximal–posterior bone is a pisiform in terms of embryonic position and its development as a sesamoid associated to a tendon. However, the pisiform is absent in bird-like dinosaurs, which are known from several articulated specimens. The combined data provide compelling evidence of a remarkable evolutionary reversal: A large, ossified pisiform re-evolved in the lineage leading to birds, after a period in which it was either absent, nonossified, or very small, consistently escaping fossil preservation. The bird wrist provides a modern example of how developmental and paleontological data illuminate each other. Based on all available data, we introduce a new nomenclature for bird wrist ossifications.     

Immunohistochemical localization of LIM mineralization protein 1 during mouse molar development

LIM mineralization protein 1 (LMP-1) is an essential positive regulator of osteoblast differentiation, maturation and bone formation. Our previous investigations on the distribution of LMP-1 in mature human teeth indicated that LMP-1 might play a role in the odontoblast differentiation and dentin matrix mineralization. The aim of the present study was to use immunohistochemistry to determine the expression of LMP-1 during tooth development in mouse molars. In embryonic and postnatal Kunming mice, LMP-1 protein was expressed during molar development, but the expression levels and patterns differed at various developmental stages. At embryonic day 13.5 (E13.5), LMP-1 was found in the enamel organ. At E14.5, LMP-1 was detected in the entire enamel organ and in the underlying mesenchyme. At E16.5, LMP-1 was observed in the inner and outer enamel epithelium and the stratum intermedium. The expression also converged at the cusps in the dental papilla. At E18.5 and postnatal day 2.5 (P2.5), LMP-1 was restricted to the stratum intermedium, in differentiating dental papilla cells at cusps, while it disappeared in terminal differentiated ameloblasts and odontoblasts. At P13.5, no positive staining was detected in the odontoblasts or in the dental pulp cells. Therefore, LMP-1 showed spatiotemporal expression patterns during molar development and might participate in molar crown morphogenesis and odontoblast differentiation at late molar development.     

A comparison between methods of calculating skeletal age (Greulich-Pyle)

The skeletal age of each individual bone of the hand-wrist was assessed in serial radiographs of 169 Melbourne children. Seven different methods were used to obtain area skeletal ages from these bone skeletal ages. Methods employing arithmetic means of all the bone skeletal ages, excluding only extreme values, excluding those derived from the carpals or from other selected bones yielded similar means during the age range studied in each sex. There were large, but not significant, differences between the means derived from averages of the skeletal ages of the most and least mature bones and those derived from all the bone skeletal ages. A system of weighting and exclusion of bone skeletal ages led to the recording of area skeletal ages that differed from the others and fluctuated markedly for individuals until ossification had occurred in every carpal bone. The findings suggest that the exclusion of the skeletal ages of the carpal bones or the use of selected skeletal ages could lead to quicker assessments without real changes in means and variability.     

Expression and localization of Nell-1 during murine molar development

Nel-like molecule-1 (Nell-1) is a recently discovered secreted protein that plays an important role in osteoblast differentiation, bone formation, and bone regeneration. However, its expression and distribution during tooth development are largely unknown. The aim of this study was to investigate the expression patterns of Nell-1 during murine molar development by immunohistochemistry. Nell-1 protein was expressed during molar development in embryonic and postnatal Kunming mice, but its expression levels and patterns at various developmental stages differed. At embryonic day 13.5 (E13.5) and E14.5, Nell-1 was found in both the entire enamel organ and the underlying mesenchyme. At E16.5, it was detected in the inner and outer enamel epithelia, stratum intermedium, secondary enamel knot, and dental papilla. At E18.5, Nell-1 was expressed in the differentiating ameloblasts, differentiating odontoblasts, and stratum intermedium. Positive staining was also found in the outer enamel epithelium. At postnatal day 2.5 (P2.5), P5, and P7, Nell-1 appeared in the secretory and mature ameloblasts and odontoblasts (odontoblastic bodies and processes) as well as immature enamel. Hertwig’s epithelial root sheath also stained positively at P7. At P13.5, positive staining was restricted to the reduced dental epithelium and odontoblasts, whereas Nell-1 disappeared in the mature enamel. During tooth eruption, Nell-1 was observed only in the odontoblastic bodies, odontoblastic processes, and endothelial cells of blood vessels. The spatiotemporal expression patterns of Nell-1 during murine tooth development suggest that it might play an important role in ameloblast and odontoblast differentiation, secretion and mineralization of the extracellular enamel matrix, molar crown morphogenesis, as well as root formation.     

The evolutionary history of Drosophila buzzatii. XXXV. Inversion polymorphism and nucleotide variability in different regions of the second chromosome

Inversions are portions of a chromosome where the gene order is reversed relative to a standard reference orientation. Because of reduced levels of recombination in heterokaryotypes, inversions have a potentially important effect on patterns of nucleotide variability in those genomic regions close to, or included in, the inverted fragments. Here we report sequence variation at three anonymous regions (STSs) located at different positions in relation to second-chromosome inversion breakpoints in 29 isochromosomal lines derived from an Argentinean population of Drosophila buzzatii. In agreement with previous findings in Drosophila, gene flux (crossing over and/or gene conversion) between arrangements seems to appreciably increase as we approach the middle sections of inversion 2j, and patterns of nucleotide variability within, as well as genetic differentiation between chromosome arrangements, are comparable to those observed at the molecular marker outside the inverted fragments. On the other hand, nucleotide diversity near the proximal breakpoint of inversion 2j is reduced when contrasted with that found at the other regions, particularly in the case of derived inverted chromosomes. Using the data from the breakpoint, we estimate that the inversion polymorphism is approximately 1.63 N generations old, where N is the effective population size. An excess of low-frequency segregating polymorphisms is detected; mostly in the ancestral 2st arrangement and probably indicating a population expansion that predates the coalescent time of inversion 2j. Heterogeneity in mutation rates between the markers linked to the inversions may be sufficient to explain the different levels of nucleotide diversity observed. When considered in the context of other studies on patterns of variation relative to physical distance to inversion breakpoints, our data appear to be consistent with the conclusion that inversions are unlikely to be "long-lived" balanced polymorphisms.     

Autopodial skeleton evolution in side-necked turtles (Pleurodira)

Carpal and tarsal anatomy was documented based on the observation of dry skeletons of adult specimens representing 25 species in 15 genera and on data taken from the literature. In addition, histological sections and cleared and double‐stained autopodia of recently hatched and juvenile specimens representing seven chelid and pelomedusoid species were studied. There is much more morphological diversity in the manus than in the pes. Variation in autopodial skeletons includes: the astragalus and calcaneum are either separated or fused; fusion of distal carpals 3–4−5 or just 4–5; number of centralia in the carpus; and presence/absence of a pisiform and of an accessory radial element. The widespread and probably basal phalangeal formula for Pleurodira is 2.3.3.3.3. Deviations are Pelomedusa subrufa, exhibiting a reduction to 2.2.2.2.2, Pelusios spp. with one phalanx less in digit I and for one species in digit V as well, and Acanthochelys pallidipectoris with an additional phalanx in the fourth finger. Six discrete characters itemizing some of the morphological variation observed were plotted on a composite pleurodire phylogeny, revealing not only homoplastic patterns but also the utility of some characters in supporting the monophyly of several clades. The pisiform is the last carpal element to ossify in Chelus fimbriatus. We hypothesize that the so‐called fifth hooked metatarsal represents the fusion of distal tarsal 5 with metatarsal V. The accessory radial element that was occasionally present in the turtles examined may represent an atavism of the otherwise lost radiale of turtles.