Showing papers in "Journal of Morphology in 2004"

Stages of embryonic development in the Atlantic cod Gadus morhua.

Abstract: The early development of the Atlantic cod, Gadus morhua was studied from fertilization until first-feeding. Multiple families were reared at 7 degrees C and a developmental staging series was prepared using morphological landmarks visible with the light microscope. Stages were named rather than numbered to allow for future additions and broadly grouped into larger time intervals called periods. The most useful staging features were found to be initially cell number, and later in development, somite number. The mean cell cycle time for the first six cleavages was 135 min and the linear regression equation for development of somites(s) over time (t) was s = 0.29t - 18.14. The segmentation period began at 220 h postfertilization (hpf), and unlike some other teleosts, the addition of new somites continued throughout the majority of embryonic development, until just prior to hatching. Hatching occurred at 256 hpf, after which individuals remained motionless at the water's surface, undergoing negative phototaxis only after the first day posthatch. The first-feeding stage was reached at the end of the third day posthatch, subsequent to development of a functional jaw and hindgut. This staging series provides an essential baseline reference for future experiments involving developing cod embryos and for the aquaculture industry.

Biomechanical modeling and sensitivity analysis of bipedal running ability. II. Extinct taxa

Abstract: I used a simple mathematical model of the inverse dynamics of locomotion to estimate the minimum muscle masses required to maintain quasi-static equilibrium about the four main limb joints at mid-stance of fast running. Models of 10 extant taxa (a human, a kangaroo, two lizards, an alligator, and five birds) were analyzed in various bipedal poses to examine how anatomy, size, limb orientation, and other model parameters influence running ability. I examined how the muscle masses required for fast running compare to the muscle masses that are actually able to exert moments about the hip, knee, ankle, and toe joints, to see how support ability varies across the limb. I discuss the assumptions and limitations of the models, using sensitivity analysis to see how widely the results differed with feasible parameter input values. Even with a wide range of input values, the models validated the analysis procedure. Animals that are known to run bipedally were calculated as able to preserve quasi-static equilibrium about their hindlimb joints at mid-stance, whereas non-bipedal runners (iguanas and alligators) were recognized as having too little muscle mass to run quickly in bipedal poses. Thus, this modeling approach should be reliable for reconstructing running ability in extinct bipeds such as nonavian dinosaurs. The models also elucidated how key features are important for bipedal running capacity, such as limb orientation, muscle moment arms, muscle fascicle lengths, and body size. None of the animals modeled had extensor muscle masses acting about any one joint that were 7% or more of their body mass, which provides a reasonable limit for how much muscle mass is normally apportioned within a limb to act about a particular joint. The models consistently showed that a key biomechanical limit on running ability is the capacity of ankle extensors to generate sufficiently large joint moments. Additionally, the analysis reveals how large ratite birds remain excellent runners despite their larger size; they have apomorphically large extensor muscles with relatively high effective mechanical advantage. Finally, I reconstructed the evolution of running ability in the clade Reptilia, showing that the ancestors of extant birds likely were quite capable runners, even though they had already reduced key hip extensors such as M. caudofemoralis longus.

Life stages and reproductive components of the Marmorkrebs (marbled crayfish), the first parthenogenetic decapod crustacean.

Abstract: Recently, we briefly reported on the first case of parthenogenesis in the decapod Crustacea which was found in the Marmorkrebs or marbled crayfish, a cambarid species of unknown geographic origin and species identity. Curiously, this animal is known only from aquarium populations, where it explosively propagates. By means of light and electron microscopic techniques we have now investigated the reproductive components of this crayfish, using more than 100 specimens ranging from hatchling to repeatedly spawned adult. Additionally, we documented its principal life stages. Our results revealed that the external sexual characters and also the gonads of the marbled crayfish are purely female, making this fast-reproducing species a good model for investigating female reproductive features in crayfish. Testicular tissues, ovotestes, or male gonoducts, gonopores, or gonopods were never found, either in small juveniles or large adult specimens, confirming the parthenogenetic nature of this crayfish. Parthenogenesis may have arisen spontaneously or by interspecific hybridization since Wolbachia-like feminizing microorganisms were not found in the ovaries. The external sexual characters of the marbled crayfish are first recognized in Stage 4 juveniles and are structurally complete approximately 2 months after hatching in specimens of approximately 2 cm total length. In the same life stage the ovary is fully differentiated as well, although the oocytes are in previtellogenic and primary vitellogenic stages only. The architecture of the mature ovary and also the synchronous maturation of cohorts of primary vitellogenic oocytes by secondary vitellogenesis are in general agreement with data published on ovaries of bisexual crayfish. New results were obtained with respect to the muscular nature of the ovarian envelope and its extensive proliferation after the first spawning, the distribution of hemal sinuses in the ovarian envelope and in the interstitium around the oogenetic pouches, the high transport activity of the follicle cells, and the colonization of oogenetic pouches by previtellogenic oocytes that originate in the germaria. Investigation of the nuclei of oocytes in the germaria and oogenetic pouches revealed no signs of meiosis, as usually found in females of bisexual decapods, suggesting that parthenogenesis in the marbled crayfish might be an apomictic thelytoky. The detection of new rickettsial and coccidian infections in the ovary and further organs raises fears that the marbled crayfish might endanger native European species by transmission of pathogens once escaped into the wild.

Blubber development in bottlenose dolphins (Tursiops truncatus).

Abstract: Blubber, the lipid-rich hypodermis of cetaceans, functions in thermoregulation, buoyancy control, streamlining, metabolic energy storage, and locomotion. This study investigated the development of this specialized hypodermis in bottlenose dolphins (Tursiops truncatus) across an ontogenetic series, including fetuses, neonates, juveniles, subadults, and adults. Blubber samples were collected at the level of the mid-thorax, from robust specimens (n = 25) that stranded along the coasts of North Carolina and Virginia. Blubber was dissected from the carcass and its mass, and the depth and lipid content at the sample site, were measured. Samples were prepared using standard histological methods, viewed by light microscopy, and digital images of blubber captured. Images were analyzed through the depth of the blubber for morphological and structural features including adipocyte size, shape, and numbers, and extracellular, structural fiber densities. From fetus to adult, blubber mass and depth increased proportionally with body mass and length. Blubber lipid content increased dramatically with increasing fetal length. Adult and juvenile blubber had significantly higher blubber lipid content than fetuses, and this increase was reflected in mean adipocyte size, which increased significantly across all robust life history categories. In juvenile, subadult, and adult dolphins, this increase in cell size was not uniform across the depth of the blubber, with the largest increases observed in the middle and deep blubber regions. Through-depth counts of adipocytes were similar in all life history categories. These results suggest that blubber depth is increased during postnatal growth by increasing cell size rather than cell number. In emaciated adults (n = 2), lipid mobilization, as evidenced by a decrease in adipocyte size, was localized to the middle and deep blubber region. Thus, in terms of both lipid accumulation and depletion, the middle and deep blubber appear to be the most metabolically dynamic. The superficial blubber likely serves a structural role important in streamlining the animal. This study demonstrates that blubber is not a homogeneous tissue through its depth, and that it displays life history-dependent changes in its morphology and lipid content. J. Morphol. 259:7–20, 2004. © 2003 Wiley-Liss, Inc.

Pattern and rate of ovary differentiation with reference to somatic development in anuran amphibians

Abstract: The rate of somatic development of anuran amphibians is only roughly correlated with the rate of gonad differentiation and varies among species. The somatic stage of a tadpole often does not reflect its age, which seems to be crucial for gonad differentiation rate. We compared the morphology and differentiation of developing ovaries at the light and electron microscopy level, with reference to somatic growth and age of a female. Our observations were performed on 12 species of six families (Rana lessonae, R. ridibunda, R. temporaria, R. arvalis, R. pipiens, R. catesbeiana, Bombina bombina, Hyla arborea, Bufo bufo. B. viridis, Xenopus laevis, Pelobates fuscus) and compared with the results obtained by other authors. This allowed us to describe the unified pattern of anuran female gonad differentiation. Ovary differentiation was divided into 10 stages: I-III, undifferentiated gonad; IV, sexual differentiation; V, first nests of meiocytes; VI, first diplotene oocytes; VII-IX, increasing number of diplotene oocytes and decreasing number of oogonia and nests; X, fully developed ovary composed of diplotene oocytes with rudimental patches of oogonia. We distinguished three types of ovary differentiation rate: basic (most species), retarded (genus Bufo), and accelerated (green frogs of the subgenus Pelophylax genus Rana).

Pulmonary pneumaticity in the postcranial skeleton of extant aves: a case study examining Anseriformes.

Abstract: Anseriform birds were surveyed to examine how the degree of postcranial pneumaticity varies in a behaviorally and size-diverse clade of living birds. This study attempts to extricate the relative effects of phylog- eny, body size, and behavioral specializations (e.g., diving, soaring) that have been postulated to influence the extent of postcranial skeletal pneumaticity. One hundred anseri- form species were examined as the focal study group. Methods included latex injection of the pulmonary appa- ratus followed by gross dissection or direct examination of osteological specimens. The Pneumaticity Index (PI) is introduced as a means of quantifying and comparing post- cranial pneumaticity in a number of species simulta- neously. Phylogenetically independent contrasts (PICs) were used to examine the relationship between body size and the degree of postcranial pneumaticity throughout the clade. There is a high degree of similarity (i.e., clade- specificity) within most anseriform subgroups. As a whole, Anseriformes demonstrate no significant relationship be- tween relative pneumaticity and body size, as indicated by regression analysis of body mass on PI. It is apparent, however, that many clades of diving ducks do exhibit lower PIs than their nondiving relatives. By exclusion of diving taxa from analyses, a significant positive slope is observed and the hypothesis of relatively higher pneuma- ticity in larger-bodied birds is only weakly supported. However, low correlations indicate that factors other than body size account for much of the variation observed in relative pneumaticity. Pneumaticity profiles were mapped onto existing phylogenetic hypotheses. A reduction in the degree of postcranial pneumaticity occurred indepen- dently in at least three anseriform subclades specialized for diving. Finally, enigmatic pneumatic features located in distal forelimb elements of screamers (Anhimidae) re- sult from invasion of bone by a network of subcutaneous air sac diverticula spreading distally along the wings. J. Morphol. 261:141-161, 2004. © 2004 Wiley-Liss, Inc.

Relationships of loading history and structural and material characteristics of bone: development of the mule deer calcaneus.

Abstract: If a bone's morphologic organization exhibits the accumulated effects of its strain history, then the relative contributions of a given strain stimulus to a bone's development may be inferred from a bone's hierarchical organization. The artiodactyl calcaneus is a short cantilever, loaded habitually in bending, with prevalent compression in the cranial (Cr) cortex, tension in the caudal (Cd) cortex, and shear in the medial and lateral cortices (i.e., neutral axis). Artiodactyl calcanei demonstrate unusually heterogeneous structural and material organization between these cortices. This study examines potential relationships between developmental morphologic variations and the functional strain distribution of the deer calcaneus. One calcaneus was obtained from each of 36 (fetus to adult) wild deer. Predominant collagen fiber orientation (CFO), microstructural characteristics, mineral content (% ash), and geometric parameters were determined from transversely cut segments. Radiographs were examined for arched trabeculae, which may reflect tension/compression stress trajectories. Results showed that cross-sectional shape changes with age from quasi-circular to quasi-elliptical, with the long axis in the cranial–caudal direction of habitual bending. Cranial (“compression”) cortical thickness increased at a greater rate than the Cd (“tension”) cortex. Fetal bones exhibited arched trabeculae. Percent ash was not uniform (Cr > Cd), and this disparity increased with age (absolute differences: 2.5% fetuses, 4.3% adults). Subadult bones showed progressively more secondary osteons and osteocyte lacunae in the Cr cortex, but the Cd cortex tended to have more active remodeling in the subadult and adult bones. Nonuniform Cr:Cd CFO patterns first consistently appear in the subadults, and are correlated with secondary bone formation and habitual strain mode. Medial and lateral cortices in these groups exhibited elongated secondary osteons. These variations may represent “strain-mode-specific” (i.e., tension, compression, shear) adaptations. The heterogeneous organization may also be influenced by variations in longitudinal strain magnitude (highest in the Cr cortex) and principal strain direction—oblique in medial-lateral cortices (where shear strains also predominate). Other factors such as local reductions in longitudinal strain may influence the increased remodeling activity of the Cd cortex. Some structural variations, such as arched trabeculae, that are established early in ontogeny may be strongly influenced by genetic- or epigenetic-derived processes. Material variations, such as secondary osteon population densities and CFO, which appear later, may be products of extragenetic factors, including microdamage. J. Morphol. 259:281–307, 2004. Published 2004 Wiley-Liss, Inc.

Comparative analysis of masseter fiber architecture in tree‐gouging (Callithrix jacchus) and nongouging (Saguinus oedipus) callitrichids

Abstract: Common marmosets (Callithrix jacchus) and cotton-top tamarins (Saguinus oedipus) (Callitrichidae, Primates) share a broadly similar diet of fruits, insects, and tree exudates. Common marmosets, however, differ from tamarins by actively gouging trees with their anterior teeth to elicit tree exudate flow. During tree gouging, marmosets produce relatively large jaw gapes, but do not necessarily produce relatively large bite forces at the anterior teeth. We compared the fiber architecture of the masseter muscle in tree-gouging Callithrix jacchus (n = 10) to nongouging Saguinus oedipus (n = 8) to determine whether the marmoset masseter facilitates producing these large gapes during tree gouging. We predict that the marmoset masseter has relatively longer fibers and, hence, greater potential muscle excursion (i.e., a greater range of motion through increased muscle stretch). Conversely, because of the expected trade-off between excursion and force production in muscle architecture, we predict that the cotton-top tamarin masseter has more pinnate fibers and increased physiological cross-sectional area (PCSA) as compared to common marmosets. Likewise, the S. oedipus masseter is predicted to have a greater proportion of tendon relative to muscle fiber as compared to the common marmoset masseter. Common marmosets have absolutely and relatively longer masseter fibers than cotton-top tamarins. Given that fiber length is directly proportional to muscle excursion and by extension contraction velocity, this result suggests that marmosets have masseters designed for relatively greater stretching and, hence, larger gapes. Conversely, the cotton-top tamarin masseter has a greater angle of pinnation (but not significantly so), larger PCSA, and higher proportion of tendon. The significantly larger PCSA in the tamarin masseter suggests that their masseter has relatively greater force production capabilities as compared to marmosets. Collectively, these results suggest that the fiber architecture of the common marmoset masseter is part of a suite of features of the masticatory apparatus that facilitates the production of relatively large gapes during tree gouging. J. Morphol. 261:276–285, 2004. © 2004 Wiley-Liss, Inc.

Structure and function of the horn shark (Heterodontus francisci) cranium through ontogeny: Development of a hard prey specialist

Abstract: The horn sharks (Heterodontidae: Chon- drichthyes) represent one of four independent evolutions of durophagy in the cartilaginous fishes. We used high- resolution computed tomography (CT scanning) to visual- ize and quantify the mineralized tissue of an ontogenetic series of horn sharks. CT scanning of neonatal through adult California horn sharks (Heterodontus francisci) con- firmed that this technique is effective for examining min- eralized tissue in even small (10 mm) specimens. The jaw joint is among the first areas to become mineralized and is the most heavily mineralized area in the cranium of a neonatal horn shark. The hyoid is also well mineralized, although the poorly mineralized molariform teeth indicate that the neonatal animal may be a suction feeder on softer prey. The symphysis of the jaws never mineralizes, in sharp contrast to the condition in the hard prey-crushing stingrays. Digitally reslicing the CT scans along the jaws allowed measurement of the second moment of area (Ina). Assuming that the jaws are made of the same material at all ages, Ina is an indicator of the flexural stiffness of the jaws. In all sizes of shark the lower jaws were stiffer than the upper and the stiffness increased in the area of the molariform teeth. The central region of the jaws, where the rami meet, support cuspidate grasping teeth and has the lowest Ina. The spotted eagle ray (Aetobatus narinari), a hard prey-crushing stingray, shows a different pattern of flexural stiffness, with the peak at the central part of the jaws where the prey is reduced between flattened tooth plates. Although the eagle ray jaws have a higher Ina than the horn shark, they are also far more heavily min- eralized. When the relative amounts of mineralization are taken into account, horn sharks do better with what min- eral they have than does the eagle ray. With a tight jaw joint and loose mandibular symphysis, as well as nearly opposite patterns of stiffness in the jaws, it is clear that two of the clades of hard prey specialists use very different methods for cracking the hard prey problem. J. Morphol.

Phylogenetic comparison of serotonin-immunoreactive neurons in representatives of the Chilopoda, Diplopoda, and Chelicerata: implications for arthropod relationships.

Abstract: The phylogenetic relationships within the Arthropoda have been discussed controversially for more than a century. Comparative studies on structure and development of the nervous system have contributed important arguments to this discussion. Arthropods have individually identifiable neurons that can be used as characters in phylogenetic studies. In the present report, the arrangement of serotonin-immunoreactive neurons in the ventral nerve cord was examined in seven representatives of the Chelicerata, Chilopoda, and Diplopoda. The goal of this analysis was to determine whether number, arrangement, and axonal morphology of the serotonergic neurons in these groups are similar to the pattern found in representatives of the Hexapoda and Crustacea, as explored in a previous study. The results indicate that the pattern in the seven species examined here does not correspond to that present in the Hexapoda and Crustacea. In particular, the pattern in Chilopoda and Diplopoda is clearly different from that of the Hexapoda. The hexapodan pattern most closely resembles that of the Crustacea. These findings are discussed with regard to recent reports on the mechanisms of neurogenesis in these taxa. Furthermore, the proposed ground patterns of the various groups are reconstructed and the characters are plotted on two competing hypotheses of arthropod phylogeny, the traditional Tracheata hypothesis and an alternative hypothesis derived from molecular and recent morphological data, the Tetraconata concept. The data discussed in this article moderately support the Tetraconata hypothesis.

Structure of long bones in mammals.

Abstract: Techniques for staining (silver, osmium, metal sulfides, ink) and microphotography (epi-illumination) of polished bone surfaces have been developed to visualize the three-dimensional structure of the shafts of mammalian long bones. Bone is a two-compartment system with capillaries and some kinds of connective tissue in one compartment separated from fibers of bone collagen, often forming lamellae, in the other. Laminar bone consists of stacks of lamellae separated by vascular spaces containing capillary network sheets. It is deposited at the periosteal and endosteal surfaces. Osteonic bone, well described in the literature, consists of cylinders of lamellae with central vascular spaces. The primary structure of the shafts of mammalian long bones is laminar and laminae often remain as the main component. Secondary osteons are a replacement within laminae. As laminar bones mature, some of the irregular longitudinal capillary spaces in the network sheets enlarge and become less crooked to form secondary osteons. Parts of the random networks become ordered longitudinal ones, resulting in collapse of those network spaces not converted to osteons. The residual capillaries become bloodless, making the surviving network spaces difficult to resolve. This may account for them being overlooked in descriptions of bone structure. For example, laminar bone occurs with osteonic bone in the human femur, although it is rarely figured. Nearly mature bones switch the kind of primary bone deposited at the peripheral (periosteal) surface from laminar to primary osteonic. J. Morphol. 262:546–565, 2004. © 2004 Wiley-Liss, Inc.

Brood care in freshwater crayfish and relationship with the offspring's sensory deficiencies.

Abstract: Prolonged brood care is one of the evolution- ary clues for the successful colonization of freshwater hab- itats by freshwater crayfish (Astacida). By means of mac- rophotography, light microscopy, and scanning electron microscopy we investigated all phases of brood care in freshwater crayfish, with particular emphasis on the mor- phological structures involved. We selected the recently discovered parthenogenetic marbled crayfish (species identity not yet known) as a model organism due to its fast reproduction and high resistance to handling stress. In order to examine if there is a causal relationship between brood care and the developmental status of the offspring's sensory apparatus, we additionally investigated major sense organs of juvenile Stages 1-5 in comparison with those of the adults. Brood care in the marbled crayfish is characterized by initial and final "active" phases domi- nated by specific maternal or juvenile behavior and a medial "passive" phase based more on the action of tem- porarily developed structures rather than on behavior. The most remarkable feature of this period, which in- cludes permanent carrying of the eggs and the first two juvenile stages under the mother's abdomen, is safeguard- ing of hatching by a telson thread that keeps the helpless newborn hatchlings linked to the egg cases on the mater- nal pleopods and thus prevents them from being lost. Further important transient structures are the recurved hooks on the first pereiopods of Stage 1 and 2 juveniles that are used to firmly attach these nonfeeding stages to the mother's abdomen. In hatchlings all sense organs nec- essary for an independent life, such as eyes, olfactory aesthetascs, gustatory fringed setae, hydrodynamic recep- tor hairs, and statocysts are not developed or are under- developed, making brood care indispensable. Most of these sense organs appear in Stage 2 juveniles, but only from Stage 3, the first freelancing and feeding stage, are all sense organs well developed and operating, thus reducing brood care in this final period to temporary provisioning of shelter. Brooding of the eggs and postembryonic brood care are to some extent also found in other freshwater Decapoda like freshwater crabs and aeglid anomurans, but safeguarding of hatching is confined to the Astacida only. This sophisticated mode of passive brood care is unique in the animal kingdom and is apparently related to the sensory deficiencies of the first juvenile stage. J. Mor- phol. 262:566 -582, 2004. © 2004 Wiley-Liss, Inc.

Mechanical functions of setae from the mouth apparatus of seven species of decapod crustaceans.

Abstract: The mouthpart setae of seven species of decapods were examined with macro-video recordings and scanning electron microscopy. The general mechanical (nonsensory) functions of the different mouthparts are described and an account of their setation is given. This offers the possibility to determine the mechanical functions of the different types of setae. Pappose setae do not participate in food handling but in general make setal barriers. Plumose setae likewise do not contact food objects but assist in current generation. Papposerrate setae are rare but they were seen to assist in pushing food particles into the mouth. Serrulate setae are very common and mainly participate in gentle food handling and grooming. Serrate setae are used for more rough food manipulation and grooming. The roughest shredding, tearing, and manipulation of prey items are handled by the cuspidate setae. Simple setae seem to be divided into two populations with very different functions. On the maxillipeds of Panulirus argus they are used for shredding, tearing, and holding the food objects, but on the basis of maxilla 2 of three other species they appear to have very little mechanical influence and only when handling small prey items. The functional scheme seems to be consistent within the Decapoda.

The spermatozoon of Eurasian murine rodents: Its morphological diversity and evolution.

Abstract: The murine rodents are the most speciose subfamily of mammals. Here the morphology of the spermatozoon, as determined by scanning and transmission electron microscopy of representative species from four Eurasian clades, is described. Much interspecific variability in all components of the spermatozoon was found to occur, although most species have a bilaterally flattened sperm head with a single apical hook of variable length and orientation. Ultrastructural observations indicate that this apical hook invariably contains a nuclear projection as well as a large extension of the subacrosomal cytoskeleton, as a perforatorium rostrally, and a complex asymmetrical acrosomal extension. These spermatozoa also have relatively long tails that are attached to the lower concave surface of the sperm head. Uniquely, in species in the Apodemus clade, the apical hook is orientated caudally. In a few species a highly derived sperm head morphotype that does not contain an apical hook is present. These sperm heads vary in morphology from being globular in two species of Bandicota, to bilaterally flattened and paddle-shaped in Tokudaia and Micromys. In spermatozoa of the latter two genera the subacrosomal cytoskeleton, which is less extensive than in species with a hooked sperm head, forms an apical extension, but that is not the case in Bandicota. In all species where the sperm head lacks an apical hook the acrosome is more symmetrical. The sperm tail is much shorter in these species, with attachment to the head occurring on the ventral surface in Tokudaia and basal in Micromys and the two species of Bandicota. As the sperm head morphotype with a complex apical hook is present in all the major clades of murine rodents, it is likely to be a plesiomorphic character within each of these clades, with the nonhooked sperm heads, which vary greatly in structure between species of the different lineages, probably being independently derived. The ultrastructural organization of the sperm head of Bandicota, but not those of Micromys or Tokudaia, suggest divergence in some of the morphological events associated with sperm-egg interaction at the time of fertilization.

Pectoral fin and girdle development in the basal actinopterygians Polyodon spathula and Acipenser transmontanus.

Abstract: The pectoral fins of Acipenseriformes possess endoskeletons with elements homologous to both the fin radials of teleosts and the limb bones of tetrapods. Here we present a study of pectoral fin development in the North American paddlefish, Polyodon spathula, and the white sturgeon, Acipenser transmontanus, which reveals that aspects of both teleost and tetrapod endoskeletal patterning mechanisms are present in Acipenseriformes. Those elements considered homologous to teleost radials, the propterygium and the mesopterygial radials, form via subdivision of an initially chondrogenic plate of mesenchymal cells called the endoskeletal disc. In Acipenseriformes, elements homologous to the sarcopterygian metapterygium develop separately from the endoskeletal disc as an outgrowth of the endoskeletal shoulder girdle that extends into the posterior margin of the finbud. As in tetrapods, the elongating metapterygium and the metapterygial radials form in a proximal to distal order as discrete condensations from initially nonchondrogenic mesenchyme. Patterns of variation seen in the Acipenseriform fin also correlate with putative homology: all variants from the "normal" fin bauplan involved the metapterygium and the metapterygial radials alone. The primary factor distinguishing Polyodon and Acipenser fin development from each other is the composition of the endoskeletal extracellular matrix. Proteoglycans (visualized with Alcian Blue) and Type II collagen (visualized by immunohistochemistry) are secreted in different places within the mesenchymal anlage of the fin elements and girdle and at different developmental times. Acipenseriform pectoral fins differ from the fins of teleosts in the relative contribution of the endoskeleton and dermal rays. The fins of Polyodon and Acipenser possess elaborate endoskeletons overlapped along their distal margins by dermal lepidotrichia. In contrast, teleost fins generally possess relatively small endoskeletal radials that articulate with the dermal fin skeleton terminally, with little or no proximodistal overlap.

Embryological aspects of inducible morphological defenses in Daphnia.

Abstract: Many cases of predator-induced morphological plasticity in daphnids are well studied examples of inducible defenses. However, little is known about the early development of these sometimes conspicuous traits. We compared for the first time in five different Daphnia species the embryonic development of predator-induced and noninduced animals using scanning electron microscopy (SEM). We observed significant morphological changes in the last embryonic stage in helmet formation in Daphnia cucullata and in neck-pedestal development in Daphnia pulex. In contrast, no morphological changes could be found during embryogenesis between induced and noninduced Daphnia lumholtzi, D. longicephala, and D. ambigua. Strategies for initiating the defensive traits differ among Daphnia species because of trade-offs between environmental requirements and developmental constraints. Some general features of Daphnia embryonic development are described using SEM. All Daphnia embryos have to shed at least three different membranes before leaving the brood pouch of the mother. After the embryos shed the third membrane, chemosensillae that are likely able to detect predator-released chemicals are exposed to the olfactory environment.

Three-dimensional model of the feline hindlimb.

Abstract: The production of coordinated movement requires that the nervous system control muscle forces to generate joint torques required for control of the limb endpoint in Euclidean space. The nervous system must be capable of transformation between Euclidean space and joint space and between joint space and muscle space to determine what set of muscle activations is required to generate a desired motion (Lacquaniti and Maioli, 1994). Knowledge of these transformations is required to understand the basis of locomotion (Prochazka, 1996; Rossignol, 1996; Zernicke and Smith, 1996; Duysens et al., 2000; Stein et al., 2000) and other forms of voluntary movement (Mussa-Ivaldi et al., 1985; Flash, 1987; Feldman and Levin, 1995; Ghez et al., 1995; Gordon et al., 1995; Gottlieb et al., 1997; Kargo and Giszter, 2000; Ting et al., 2000). These transformations are based on the architecture and mechanics of the musculoskeletal system. Details of this information are available for humans (Delp et al., 2001), non-human primates (Cheng and Scott, 2000) and other animals, (Kargo et al., 2002; Kargo and Rome, 2002). The cat has been an important experimental subject for studies of normal and abnormal motor control (Macpherson, 1988; Pratt and Loeb, 1991; Prochazka, 1996; Rossignol, 1996; Macpherson and Fung, 1999), but detailed architectural information is not available in comprehensive form. The goal of this paper is to report our studies of the musculoskeletal anatomy of the feline hindlimb and a corresponding mathematical model to predict the transformations described above. Perhaps the first issue that arises in the construction of such a model is interspecimen variability. To generalize from one or a few measurements to a stereotypical or canonical anatomy, the variation between specimens must be small. If interspecimen variation is large, then the description of a sample is less likely to be representative of any individual. Interspecimen variability may also offer a means to understand the observed inter-individual variation in motor control patterns (McCollum et al., 1995). Another concern is the choice of kinematic constraints to be imposed on the model. These have previously ranged from simple hinges in anatomical planes (Mussa-Ivaldi et al., 1985) to screw-displacement axes (Woltring et al., 1985). The former is computationally straightforward, but practically limited. The latter is most useful in describing a specific observed motion for inverse dynamics, and either may introduce non-physiological constraints on the system. A compromise between these extremes has been suggested (Hollister et al., 1992), in which a joint is approximated as a combination of hinges representing physiological planes defined by joint morphology, rather than anatomical planes defined by body posture. This approach allows the computational simplicity of single degree-of-freedom joints, and extends the range over which that approximation closely represents the actual motion. The purpose of the work presented in this paper was to construct a three dimensional model of the feline hindlimb. Specific attention is focused on interspecimen variation in muscle paths and the use of joint axes.

Taste bud form and distribution on lips and in the oropharyngeal cavity of cardinal fish species (Apogonidae, Teleostei), with remarks on their dentition.

Abstract: The oral dentition and type and number of taste buds (TB) on the lips and in the oropharyngeal cavity were compared by means of SEM in 11 species of cardinal fishes (Apogonidae) belonging to five genera. The occurrence of a dense cover of skin papillae on the lips of some species (e.g., Apogon frenatus), as well as differences in structure of vomer, tongue, and palatinum, expose additional morphological characters important for clarification of the taxonomy of this group of fishes. Differences are also revealed in the type of dentition, such as on the vomer and epi-hypopharyngeal bones. Strong and dense dentition of the anterior part of the oral cavity and a high number of TB on this site in species feeding on larger prey (e.g., Cheilodipterus spp) is compared to the relatively feeble jaw armor and richness of TB on the more pharyngeal site in species feeding on smaller prey (e.g., Apogon angustatus, A. frenatus). In addition to the three types of TB (Types I–III) previously described from various teleost fish, a fourth type (Type IV), comprising very small buds, was found in some cardinal fish (Apogon angustatus, A. frenatus). The various TB are distributed from the lips to the pharyngeal bones, on the breathing valves, tongue, palatinum, and pharyngeal bones; their number and type on the various sites differ in the different species. In all species studied the Types I and II TB, elevated above the surrounding epithelium, dominated the lips and anterior part of mouth, while Types III and IV, which end apically at the level with the epithelium, dominated the more posterior pharyngeal region. The highest number of TB, around 24,600, were found in Fowleria variegata, a typical nocturnal species, and the lowest in the diurnal and crepuscular Apogon cyanosoma (1,660) and Cheilodipterus quinquestriatus (2,400). Differences are also revealed in the type of dentition, such as on the vomer and epi-hypopharyngeal bones. The number of TB increased with growth of the fishes. The differences in the total number of TB and their distribution in the oropharyngeal cavity in the various species indicates possible different mechanisms of foraging and food-recognition. J. Morphol. 259:316–327, 2004. © 2004 Wiley-Liss, Inc.

Anatomy and functional morphology of the feeding apparatus of the lesser electric ray, Narcine brasiliensis (Elasmobranchii: Batoidea).

Abstract: Protrusion of the jaws during feeding is common in Batoidea (rays, skates, sawfishes, and guitarfishes), members of which possess a highly modified jaw suspension. The lesser electric ray, Narcine brasiliensis, preys primarily on polychaete annelids using a peculiar and highly derived mechanism for jaw protraction. The ray captures its prey by protruding its jaws beneath the substrate and generating subambient buccal pressure to suck worms into its mouth. Initiation of this protrusion is similar to that proposed for other batoids, in that the swing of the distal ends of the hyomandibulae is transmitted to Meckel's cartilage. A "scissor-jack" model of jaw protrusion is proposed for Narcine, in which the coupling of the upper and lower jaws, and extremely flexible symphyses, allow medial compression of the entire jaw complex. This results in a shortening of the distance between the right and left sides of the jaw arch and ventral extension of the jaws. Motion of the skeletal elements involved in this extreme jaw protrusion is convergent with that described for the wobbegong shark, Orectolobus maculatus. Narcine also exhibits asymmetrical protrusion of the jaws from the midline during processing, accomplished by unequal depression of the hyomandibulae. Lower jaw versatility is a functional motif in the batoid feeding mechanism. The pronounced jaw kinesis of N. brasiliensis is partly a function of common batoid characteristics: euhyostylic jaw suspension (decoupling the jaws from the hyoid arch) and complex and subdivided cranial musculature, affording fine motor control. However, this mechanism would not be possible without the loss of the basihyal in narcinid electric rays. The highly protrusible jaw of N. brasiliensis is a versatile and maneuverable feeding apparatus well-suited for the animal's benthic feeding lifestyle.

Comparative anatomy of the superfamily Myliobatoidea (Chondrichthyes) with some comments on phylogeny.

Abstract: The anatomy of species belonging to the superfamily Myliobatoidea was examined with the aim of better determining their phylogenetic relationships. A wide variation among genera was observed in skeletal anatomy, despite the fact that they all share a common morphological pattern. However, variation among species of the same genus was low, excepting Mobula. Dorsal musculature showed a substantial consistency, except for the epiaxialis muscle, which was larger in rhinopterids and mobulids. Variation in the ventral muscles was low among species of the same genus, but considerable among different genera. Mobulids have a reduction in ventral muscles, while rhinopterids and myliobatoids show an increase in muscular mass. A consensus tree shows a basal split into two groups. The first includes the family Gymnuridae with the genera Gymnura and Aetoplatea; this group is supported by seven synapomorphies, including: 27(1) ceratobranchialis fused proximally, 36(1) anterior lateral processes present in the synarcual, 52(0) quadratomandibularis internal muscle present. The second group is composed of the family Myliobatidae (Myliobatis, Aetomylaeus, Aetobatus, Rhinoptera, Mobula, and Manta), this group is supported by 11 synapomorphies, including: 5(1) first postorbital process fused with the second, 21(1) fused mandibular symphysis, 24(1) first hypobranchial cartilage absent, 48(2) epiaxialis muscle inserted in the cranial orbital region, 73(1) pectoral fins joined behind the orbital region. This study concluded that myliobatoids (Myliobatis, Aetomylaeus, and Aetobatus) integrate a monophyletic group which, unlike other phylogenies previously obtained, is the sister group of rhinopterids (Rhinoptera). Mobulids (Mobula and Manta) are the sister group of myliobatoids-rhinopterids. J. Morphol. 262:517–535, 2004. © 2004 Wiley-Liss, Inc.

Morphological observation of antler regeneration in red deer (Cervus elaphus).

Abstract: Deer antler offers a unique opportunity to explore how nature solves the problem of mammalian appendage regeneration. Annual antler renewal is an example of epimorphic regeneration, which is known to take place through initial blastema formation. Detailed examination of the early process of antler regeneration, however, has thus far been lacking. Therefore, we con- ducted morphological observations on antler regenera- tion from naturally cast and artificially created pedicle/ antler stumps. On the naturally cast pedicle stumps, early antler regeneration underwent four distinguish- able stages (with the Chinese equivalent names): cast- ing of previous hard antlers (oil lamp bowl), early wound healing (tiger eye), late wound healing and early regeneration (millstone), and formation of main beam and brown tine (small saddle). Overall, no cone-shaped regenerate, a common feature to blastema-based regen- eration, was observed. Taken together with the exami- nation on the sagittal plane of each regenerating stage sample, we found that there are considerable overlaps between late-stage wound healing and the establish- ment of posterior and anterior growth centers. Obser- vation of antler regeneration from the artificially cre- ated stumps showed that the regeneration potential of antler remnants was significantly reduced compared with that of pedicle tissue. Interestingly, the distal por- tion of a pedicle stump had greater regeneration poten- tial than the proximal region, although this differential potential may not be constitutive, but rather caused by whether or not pedicle antlerogenic tissue becomes closely associated with the enveloping skin at the cut plane. Antler formation could take place from the distal peripheral tissues of an antler/pedicle stump, without the obvious participation of the entire central bony por- tion. Overall, our morphological results do not support the notion that antler regeneration takes place through the initial formation of a blastema; rather, it may be a stem cell-based process. J. Morphol. 262:731-740, 2004. © 2004 Wiley-Liss, Inc.

Structural fiber reinforcement of keel blubber in harbor porpoise (Phocoena phocoena).

Abstract: This study investigated the functional morphology of the blubber that forms the caudal keels of the harbor porpoise (Phocoena phocoena). Blubber is a pliant biocomposite formed by adipocytes and structural fibers composed of collagen and elastic fibers. Caudal keels are dorsally and ventrally placed triangular wedges of blubber that define the hydrodynamic profile of the porpoise tailstock. Mechanical tests on carcasses demonstrate that when keels are bent, they strain nonuniformly along their lengths, with highest strains just caudal to the dorsal fin and lowest at the insertion of the flukes. Therefore, caudal keels undergo nonuniform longitudinal deformation while maintaining a stable, triangular cross-sectional shape. Polarizing and transmitted light microscopy techniques were used to investigate blubber's 3D fiber architecture along the length of the dorsal keel. The triangular cross-sectional shape of the keel appears to be maintained by structural fibers oriented to act as tensile stays. The construction of the blubber composite is regionally specific :structural fiber densities and diameters are higher in the relatively stiff caudal region of the keel than in the more deformable cranial keel region. The orientations of structural fibers also change along the length of the keel. Cranially, no fibers are oriented along the long axis, whereas a novel population of longitudinally oriented fibers reinforces the keel at the insertion of the flukes. Thus, differences in the distribution and orientation of structural fibers contribute to the regionally specific mechanical properties of the dorsal keel.

Ontogeny of tracheal system structure: a light and electron-microscopy study of the metathoracic femur of the American locust, Schistocerca americana

Abstract: Does oxygen delivery become more challenging for insects as they increase in size? To partially test this hypothesis, we used quantitative light and electron microscopy to estimate the oxygen delivery capacity for two steps of tracheal oxygen delivery within the metathoracic femur (jumping leg) for 2nd instar (about 47 mg) and adult (about 1.7 g) locusts, Schistocerca americana. The fractional cross-sectional areas of the major tracheae running longitudinally along the leg were similar in adults and 2nd instars; however, since the legs of adults are longer, the mass-specific diffusive conductances of these tracheae were 4-fold greater in 2nd instars. Diffusive gas exchange longitudinally along the leg is easily possible for 2nd instars but not adults, who have many air sacs within the femur. Mitochondrial content fell proximally to distally within the femur in 2nd instars but not adults, supporting the hypothesis that diffusion was more important for the former. Lateral diffusing capacities of the tracheal walls were 12-fold greater in adults than 2nd instars. This was primarily due to differences in the smallest tracheal class (tracheoles), which had thinner epidermal and cuticular layers, greater surface to volume ratios, and greater mass-specific surface areas in adults. Adults also had greater mitochondrial contents, larger cell sizes and more intracellular tracheae. Thus, larger insects do not necessarily face greater problems with oxygen delivery; adult grasshoppers have superior oxygen delivery systems and greater mass-specific aerobic capacities in their legs than smaller/younger insects.

Brain and sense organ anatomy and histology in hemoglobinless Antarctic icefishes (Perciformes: Notothenioidei: Channichthyidae)

Abstract: The Channichthyidae, one of five Antarctic notothenioid families, includes 16 species and 11 genera. Most live at depths of 200-800 m and are a major component of fish biomass in many shelf areas. Channichthyids are unique among adult fishes in possessing pale white blood containing a few vestigal erythrocytes and no hemoglobin. Here we describe the brains of seven species and special sense organs of eight species of channichthyids. We emphasize Chionodraco hamatus and C. myersi, compare these species to other channichthyids, and relate our findings to what is known about brains and sense organs of red-blooded notothenioids living sympatrically on the Antarctic shelf. Brains of channichthyids generally resemble those of their bathydraconid sister group. Among channichthyids the telencephalon is slightly regressed, resulting in a stalked appearance, but the tectum, corpus cerebellum, and mechanoreceptive areas are well developed. Interspecific variation is present but slight. The most interesting features of channichthyid brains are not in the nervous tissue but in support structures: the vasculature and the subependymal expansions show considerable elaboration. Channichthyids have large accessory nasal sacs and olfactory lamellae are more numerous than in other notothenioids. The eyes are relatively large and laterally oriented with similar duplex (cone and rod) retinae in all eight species. Twin cones are the qualitatively dominant photoreceptor in histological sections and, unlike bathydraconids, there are no species with rod-dominated retinae. Eyes possess the most extensive system of hyaloid arteries known in teleosts. Unlike the radial pattern seen in red-blooded notothenioids and most other teleosts, channichthyid hyaloid arteries arise from four or five main branches and form a closely spaced anastomosing series of parallel channels. Cephalic lateral line canals are membranous and some exhibit extensions (canaliculi), but canals are more ossified than those of deeper-living bathydraconids. We conclude that, with respect to the anatomy and histology of the neural structures, the brain and sensory systems show little that is remarkable compared to other fishes, and exhibit little diversification within the family. Thus, the unusual habitat and a potentially deleterious mutation resulting in a hemoglobinless phenotype are reflected primarily in expansion of the vasculature in the brain and eye partially compensating for the absence of respiratory pigments. Neural morphology gives the impression that channichthyids are a homogeneous and little diversified group.

Renal organs of cephalopods: a habitat for dicyemids and chromidinids.

Abstract: The renal organs of 32 species of cephalopods (renal appendage of all cephalopods, and renal and pancreatic appendages in decapods) were examined for parasite fauna and for histological comparison. Two phylogenetically distant organisms, dicyemid mesozoans and chromidinid ciliates, were found in 20 cephalopod species. Most benthic cephalopods (octopus and cuttlefish) were infected with dicyemids. Two pelagic cephalopod species, Sepioteuthis lessoniana and Todarodes pacificus, also harbored dicyemids. Chromidinid ciliates were found only in decapods (squid and cuttlefish). One dicyemid species was found in branchial heart appendages of Rossia pacifica. Dicyemids and chromidinids occasionally occurred simultaneously in Euprymna morsei, Sepia kobiensis, S. peterseni, and T. pacificus. The small-sized cephalopod species, Idiosepius paradoxus and Octopus parvus, harbored no parasites. Comparative histology revealed that the external surface of renal organs varies morphologically in various cephalopod species. The small-sized cephalopod species have a simple external surface. In contrast, the medium- to large-sized cephalopod species have a complex external surface. In the medium- to large-sized cephalopod species, their juveniles have a simple external surface of the renal organs. The external surface subsequently becomes complicated as they grow. Dicyemids and chromidinids attach their heads to epithelia or insert their heads into folds of renal appendages, pancreatic appendages, and branchial heart appendages. The rugged and convoluted external surface provides a foothold for dicyemids and chromidinids with a conical head. They apparently do not harm these tissues of their host cephalopods.

Evolutionary transformation from muscular to hydraulic movements in spider (Arachnida, Araneae) genitalia: a study based on histological serial sections.

Abstract: The male genitalia of 107 spider species representing 73 families were serially sectioned and studied with an emphasis on muscles moving the genital bulb. As a rule, most non-Entelegynae have two bulbal muscles, most Entelegyne have none, but many exceptions occur. Variation also occurs with regard to origin and attachment of bulbal muscles. There appears to be a trend towards a shift of the origin from proximal (Liphistius, Atypus) to more distal palpal segments (Haplogynae). In most Entelegynae the muscular movement is replaced by hydraulic movement caused by expanding membranes (hematodochae). Hematodochae probably permit increased bulbal rotation and movements of higher complexity. New evidence is presented arguing against Palpimanidae being representatives of Entelegynae. Bulbal glands other than those discharging into the sperm duct (previously known in Amaurobiidae and Dictynidae only) are described in several entelegyne families.

From head to tail: The myoseptal system in basal actinopterygians

Abstract: Experimental studies indicated that myomeres play several functional roles during swimming. Some of the functions in question are thought to change rostrocaudally, e.g., anterior myomeres are thought to generate forces, whereas posterior myomeres are thought to transmit forces. In order to determine whether these putative functions are reflected in myoseptal morphology we carried out an analysis of the myoseptal system that includes epaxial and hypaxial myosepta of all body regions for the first time. We combined clearing and staining, microdissections, polarized light microscopy, SEM technique, and length measurements of myoseptal parts to reveal the spatial arrangement, collagen fiber architecture, and rostrocaudal gradients of myosepta. We included representatives of the four basal actinopterygian clades to evaluate our findings in an evolutionary and in a functional context. Our comparison revealed a set of actinopterygian groundplan features. This includes a set of specifically arranged myoseptal tendons (epineural, epipleural, lateral, and myorhabdoid tendons) in all epaxial and postanal hypaxial myosepta. Only preanal hypaxial myosepta lack tendons and exclusively consist of mediolateral fibers. Laterally, myosepta generally align with the helically wound fibers of the dermis in order not to limit the body's maximum curvature. Medially, the relationship of myosepta to vertebrae clearly differs from a 1:1 relationship : A myoseptum attaches to the anterior margin of a vertebra, turns caudally, and traverses at least three vertebrae in an almost horizontal orientation in all body regions. By this arrangement, horizontal multiple layers of myosepta are formed along the trunk dorsal and ventral to the horizontal septum. Due to their reinforcement by epineural or epipleural tendons, these multiple layers are hypothesized to resist the radial expansion of underlying muscle fibers and thus contribute to modulation of body stiffness. Rostrocaudally, a dorsoventral symmetry of epaxial and hypaxial myosepta in terms of spatial arrangement and collagen fiber architecture is gradually developed towards the postanal region. Furthermore, the rostrocaudal extension of myosepta measured between anterior and posterior cones gradually increases. This myoseptal region is reinforced by longitudinal fibers of lateral tendons. Furthermore, the percentage of connective tissue in a cross section increases. These morphological data indicate that posterior myosepta are equipped for multisegmental force transmission towards the caudal fin. Anteriormost myosepta have reinforced and elongated dorsal posterior cones. They are adequately designed to transmit epaxial muscular forces to the neurocranium in order to cause its elevation during suction feeding. J. Morphol. 259:155–171, 2004. © 2003 Wiley-Liss, Inc.

Development of the nasal chemosensory organs in two terrestrial anurans: The directly developing frog, Eleutherodactylus coqui (Anura: Leptodactylidae), and the metamorphosing toad, Bufo americanus (Anura: Bufonidae)

Abstract: Nearly all vertebrates possess an olfactory organ but the vomeronasal organ is a synapomorphy for tetrapods. Nevertheless, it has been lost in several groups of tetrapods, including aquatic and marine animals. The present study examines the development of the olfactory and vomeronasal organs in two terrestrial anurans that exhibit different developmental modes. This study compares the development of the olfactory and vomeronasal organs in metamorphic anurans that exhibit an aquatic larva (Bufo americanus) and directly developing anurans that have eliminated the tadpole (Eleutherodactylus coqui). The olfactory epithelium in larval B. americanus is divided into dorsal and ventral branches in the rostral and mid-nasal regions. The larval olfactory pattern in E. coqui has been eliminated. Ontogeny of the olfactory system in E. coqui embryos starts to vary substantially from the larval pattern around the time of operculum development, the temporal period when the larval stage is hypothesized to have been eliminated. The nasal anatomy of the two frogs does not appear morphologically similar until the late stages of embryogenesis in E. coqui and the terminal portion of metamorphosis in B. americanus. Both species and their respective developing offspring, aquatic tadpoles and terrestrial egg/embryos, possess a vomeronasal organ. The vomeronasal organ develops at mid-embryogenesis in E. coqui and during the middle of the larval period in B. americanus, which is relatively late for neobatrachians. Development of the vomeronasal organ in both frogs is linked to the developmental pattern of the olfactory system. This study supports the hypothesis that the most recent common ancestor of tetrapods possessed a vomeronasal organ and was aquatic, and that the vomeronasal organ was retained in the Amphibia, but lost in some other groups of tetrapods, including aquatic and marine animals. J Morphol. 261:225–248, 2004. © 2004 Wiley-Liss, Inc.

Ovarian histology of the placentotrophic Mabuya mabouya (Squamata, Scincidae).

Abstract: Ovarian structure and folliculogenesis of females at different reproductive stages are described for the viviparous placentotrophic lizard Mabuya mabouya. The small ovaries have a thin wall formed by the ovarian epithelium and a thin tunica albuginea. One to two germinal beds that contain numerous oogonia and developing primordial follicles are derived from the ovarian epithelium and are next to the ovarian hilum. The ovarian cortex contains follicles at different stages of development, corpora lutea, and atretic follicles. The yolk nucleus and Balbiani complex were not evident in the ooplasm of previtellogenic follicles. The follicular epithelium of these follicles is polymorphic, as in other species of Squamata, but the larger cells are spherical and monolayered rather than pyriform. The zona radiata of the preovulatory follicles is less developed than in lecithotrophic species. These features suggest a decrease in metabolic and absorptive processes during follicular growth. In preovulatory follicles (1.5–1.8 mm diameter), primordial yolk vacuoles and small cortical granules are deposited in the ooplasm instead of fatty yolk platelets, so that only one stage of vitellogenesis is observed. Polyovular atretic follicles occur in some females. Follicular atresia is minimal for preovulatory follicles, but is more frequent in follicles with polymorphic epithelia. In the corpus luteum, the luteal tissue is formed from granulosa cells and luteolysis occurs during the late gastrula – late neurula embryonic stages. Thus, the maintenance of gestation from the pharyngula to preparturition stages seems to be related to secretion of extraluteal progesterone, possibly of placental origin. These observed ovarian features are related to the high degree of placental complexity of this species and show that the evolution of advanced placentotrophy in species of this lineage has been accompanied by concomitant changes in ovarian function. J. Morphol. 259:90–105, 2004. © 2003 Wiley-Liss, Inc.

Patterns of spatial and temporal visceral arch muscle development in the Mexican axolotl (Ambystoma mexicanum).

Abstract: The development of the vertebrate head is a complex process involving interactions between a multitude of cell types and tissues. This thesis describes the development of the cranial neural crest and of the visceral arch muscles in the head of two species. One, the Mexican axolotl (Ambystoma mexicanum), is a basal tetrapod, whereas the other, the Australian lungfish (Neoceratodus forsteri), belongs to the Dipnoi, the extant sister group of the Tetrapoda. The migration of neural crest cells, which form most of the bones and connective tissues in the head, and the morphogenesis of the jaw, was determined in the Mexican axolotl. It was shown that both the upper and lower jaws form from ventral condensations of neural crest cells in the mandibular arch. The dorsal condensation, earlier considered to give rise to the upper jaw, was shown to form the trabecula cranii.The normal spatio-temporal development of visceral arch muscles was investigated in both the Mexican axolotl and the Australian lungfish. In axolotl, the muscles tended to start forming almost simultaneously in all visceral arches at their future origins and extend towards their future insertions at the onset of muscle fibre formation. In lungfish, fibres formed simultaneously throughout most of each muscle anlage in the first and second visceral arch, but were delayed in the branchial arches. The anlagen were first observed at their future insertion, from which they developed towards future origins. To test the ability of neural crest cells to pattern the visceral arch muscles, migrating crest cells were extirpated from axolotl embryos, which resulted in a wide range of muscle malformations. In most cases, the muscles appeared in the right position but were small and extended in abnormal directions. This shows that neural crest cells are responsible not for the position of the muscles but for their correct anatomical pattern. Fate mapping showed that connective tissue surrounding myofibers is, at least partly, neural crest derived.In conclusion, the work presented in this thesis shows that although early development may map out the patterns of later development, the differences between axolotl and lungfish head development are not seen until during morphogenesis. Further investigation of morphogenesis is needed to explain the great variation of head morphology seen in vertebrates today.