The development of the skull of the Egyptian cobra Naja h. haje (Squamata: Serpentes: Elapidae)

Background: The study of craniofacial development is important in understanding the ontogenetic processes behind morphological diversity. A complete morphological description of the embryonic skull development of the Egyptian cobra, Naja h. haje, is lacking and there has been little comparative discussion of skull development either among elapid snakes or between them and other snakes. Methodology/Principal Findings: We present a description of skull development through a full sequence of developmental stages of the Egyptian cobra, and compare it to other snakes. Associated soft tissues of the head are noted where relevant. The first visible ossification centres are in the supratemporal, prearticular and surangular, with slight ossification visible in parts of the maxilla, prefrontal, and dentary. Epiotic centres of ossification are present in the supraoccipital, and the body of the supraoccipital forms from the tectum posterior not the tectum synoticum. The venom glands are visible as distinct bodies as early at stage 5 and enlarge later to extend from the otic capsule to the maxilla level with the anterior margin of the eye. The gland becomes more prominent shortly before hatching, concomitant with the development of the fangs. The tongue shows incipient forking at stage 5, and becomes fully bifid at stage 6. Conclusions/significance: We present the first detailed staging series of cranial development for the Egyptian cobra, Naja h. haje. This is one of the first studies since the classical works of G. de Beer and W. Parker that provides a detailed description of cranial development in an advanced snake species. It allows us to correct errors and misinterpretations in previous accounts which were based on a small sample of specimens of uncertain age. Our results highlight potentially significant variation in supraoccipital formation among squamates and the need for further research in this area

Bone growth following demineralized bone matrix implantation requires angiogenesis

Angiogenesis is required for endochondral ossification during development and fracture healing; however the exact mechanisms and temporal relationship between the two processes remains unclear. In this study, we utilize an in vivo model of endochondral ossification in mice by implanting demineralized bone matrix (DBM) proximal to the femur to induce ectopic bone formation. TNP-470, a drug known to be anti-angiogenic, was used to inhibit vascularization during the time course of de novo bone formation in order to define the role of angiogenesis during the chondrogenic phase of endochondral bone formation. Day 2, day 8, and day 16 post-surgery were selected time points to represent pre-chondrogenic, chondrogenic, and bone mineralization stages, respectively. Plain x-ray and micro-CT analysis showed that inhibition of angiogenesis led to decreased mineralized tissue formation. Inhibited angiogenesis was confirmed with qRT-PCR. Most striking, however, is that while stem cells are recruited and committed to the chondrogenic lineage, subsequent chondrogenesis failed to progress based on the failure of Sox5 and Sox6 expression, which directs chondrocyte commitment. This expands the role for angiogenesis to a much earlier stage than currently thought and places the necessity of angiogenesis very early in the endochondral ossification process

Comparative developmental osteology of the seahorse skeleton reveals heterochrony amongst Hippocampus sp. and progressive caudal fin loss

Background: Seahorses are well known for their highly derived head shape, prehensile tail and armoured body. They belong to the family of teleosts known as Syngnathidae, which also includes the pipefishes, pipehorses and seadragons. Very few studies have investigated the development of the skeleton of seahorses because larvae are extremely difficult to obtain in the wild and breeding in captivity is rarely successful. Here we compare the developmental osteology of Hippocampus reidi over an ontogenetic series spanning the first 93 days after release from the brood pouch to that of a smaller series of Hippocampus; namely H. subelongatus. Results: We compare the osteology in these two species over growth to the published description of the dwarf species, H. zosterae. We show that ossification onset in H. subelongatus is earlier than in H. reidi, despite similar sizes at parturition. Interestingly, the timing of development of the bony skeleton in H. zosterae is similar to that of the larger species, H. subelongatus. Furthermore, we show that the growth rate of all three species is similar up until about 30 days post pouch release. From this stage onwards in the life history, the size of the dwarf species H. zosterae remains relatively constant whilst the other two species continue growing with an accelerated growth phase. Conclusion: This data together with a phylogenetic assessment suggests that there has been a heterochronic shift (a delay) in the timing of ossification in H. reidi and accelerated bonedevelopment in H. zosterae. That is, H. zosterae is not a developmentally truncated dwarf species but rather a smaller version of its larger ancestor, "a proportioned dwarf" species. Furthermore, we show that caudal fin loss is incomplete in Hippocampus seahorses. This study shows that these three species of Hippocampus seahorses have evolved (either directly or indirectly) different osteogenic strategies over the last 20-30 million years of seahorse evolution

Anatomy, morphology and evolution of the patella in squamate lizards and tuatara (Sphenodon punctatus)

The patella (kneecap) is the largest and best-known of the sesamoid bones, postulated to confer biomechanical advantages including increasing joint leverage and reinforcing the tendon against compression. It has evolved several times independently in amniotes, but despite apparently widespread occurrence in lizards, the patella remains poorly characterised in this group and is, as yet, completely undescribed in their nearest extant relative Sphenodon (Rhynchocephalia). Through radiography, osteological and fossil studies we examined patellar presence in diverse lizard and lepidosauromorph taxa, and using computed tomography, dissection and histology we investigated in greater depth the anatomy and morphology of the patella in 16 lizard species and 19 Sphenodon specimens. We have found the first unambiguous evidence of a mineralised patella in Sphenodon, which appears similar to the patella of lizards and shares several gross and microscopic anatomical features. Although there may be a common mature morphology, the squamate patella exhibits a great deal of variability in development (whether from a cartilage anlage or not, and in the number of mineralised centres) and composition (bone, mineralised cartilage or fibrotendinous tissue). Unlike in mammals and birds, the patella in certain lizards and Sphenodon appears to be a polymorphic trait. We have also explored the evolution of the patella through ancestral state reconstruction, finding that the patella is ancestral for lizards and possibly Lepidosauria as a whole. Clear evidence of the patella in rhynchocephalian or stem lepidosaurian fossil taxa would clarify the evolutionary origin(s) of the patella, but due to the small size of this bone and the opportunity for degradation or loss we could not definitively conclude presence or absence in the fossils examined. The pattern of evolution in lepidosaurs is unclear but our data suggest that the emergence of this sesamoid may be related to the evolution of secondary ossification centres and/or changes in knee joint conformation, where enhancement of extensor muscle leverage would be more beneficial.Sophie Regnault, Marc E. H. Jones, Andrew A. Pitsillides, John R. Hutchinso

Teratogenic test of Pandanus conoideus var. yellow fruit extract to development of rat embryo (Rattus norvegicus)

Muna L, Astirin OP, Sugiyarto. 2010. Uji teratogenik ekstrak Pandanus conoideus varietas buah kuning terhadap perkembangan embrio tikus putih (Rattus norvegicus). Nusantara Bioscience 2: 126-134. Penelitian ini betujuan untuk mengkaji pengaruh pemberian ekstrak Pandanus conoideus Lam. var. buah kuning terhadap persentase fetus hidup, kematian intrauterus, berat dan panjang fetus, keadaan morfologi fetus, serta struktur skeleton fetus tikus putih. Dalam penelitian ini digunakan 25 tikus bunting yang dibagi menjadi lima kelompok secara acak, sehingga masing-masing kelompok terdiri dari lima ekor tikus. Setiap kelompok diberi dosis yang berbeda. P1 (kontrol) diberi 1 mL minyak wijen, P2, P3, P4 dan P5 diberi ekstrak masing-masing: 0,02 mL, 0,04 mL, 0,08 mL dan 0,16 mL. Ekstrak tersebut diberikan secara oral pada kebuntingan hari ke 5 sampai hari ke 17 (fase organogenesis). Pengamatan dilakukan pada hari ke 18 dengan cara bedah sesar untuk mengambil fetus dari uterus. Morfologi fetus diamati setelah fetus dikeluarkan dari uterus, sedangkan untuk pengamatan struktur skeleton dibuat preparat wholemount dengan pewarnaan ganda Alcian blue dan Allizarin Red-S. Hasil percobaan dianalisis dengan ANAVA satu jalur. Hasil penelitian menunjukkan bahwa pemberian ekstrak tidak berpengaruh terhadap persentase fetus hidup, kematian intrauterus, serta berat dan panjang fetus (P≥0,05). Pemberian ekstrak pada induk mengakibatkan kecacatan skeleton (lordosis) fetus pada dosis 0,16 mL dan menghambat osifikasi fetus. Kata kunci: Pandanus conoideus var. buah kuning, teratogenik, tikus putih

New Spontaneous Model of Fibrodysplasia Ossificans Progressiva

We report the first known example of spontaneous, naturally occurring fibrodysplasia ossificans progressiva (FOP) in a mammal. The Southeast Asian mouse deer of the genus _Tragulus_ (Artiodactyla: Tragulidae) have an osseous sheath covering the lower back and upper thigh region consistent with the clinical definition of FOP. This heterotophic bone deposition is sex related apparently with a genetic basis - it only occurs in males and is lacking in females; it is present in all adults males, including both wild obtained and zoo bred animals. _Tragulus_ may offer the opportunity to examine many of the disease's most significant attributes experimentally

Teratogenic effects of gabapentin on the skeletal system of Balb/C mice fetuses

Objectives: To evaluate the effects of gabapentin )GBP( administration on mice fetuses. Methods: This study was carried out in Birjand University of Medical Sciences during 2008. Thirty Balb/c pregnant mice were divided randomly into 3 groups: 2 experimental groups that received 25 mg/kg )I( and 50 mg/kg )II( of GBP intraperitoneally for the first 15 days of pregnancy, and a control group that received normal saline. External observations of day 18 fetuses and skeleton double staining were performed. Results: Both experimental groups showed similar disorders that can be categorized as the following: 1( decrease of fetal body weight and increase of fetal resorption, 2( macroscopic malformations, and 3( skeletal malformations. Fetal body weights were significantly lower, and fetus resorptions were significantly higher in both treated groups compared to the control group. Macroscopic malformations included exencephaly, limbs defects, brachygnathia, vertebral column deformity, and fetuses with severe retarded growth. Skeletal malformations included delayed ossification, scoliosis, calvaria deformity, and mandibular hypoplasia. Conclusion: This study revealed that GBP can induce previously unreported severe malformations if it is used continuously during the implantation, neurulation, and organogenesis stages of pregnancy. Therefore, it is suggested that great caution should be exercised in using GBP during the early stages of pregnancy until further studies are performed to better understand these effects

Ontogeny of the osteocranium in the African catfish, Clarias gariepinus burchell (1822) (Siluriformes: Clariidae): Ossification sequence as a response to functional demands

A

Histological evidence for a supraspinous ligament in sauropod dinosaurs

Supraspinous ossified rods have been reported in the sacra of some derived sauropod dinosaurs. Although different hypotheses have been proposed to explain the origin ofthis structure, histological evidence has never been provided to support or reject any of them. In order to establish its origin, we analyse and characterize the microstructure of thesupraspinous rod of two sauropod dinosaurs from the Upper Cretaceous of Argentina. The supraspinous ossified rod is almost entirely formed by dense Haversian bone. Remains ofprimary bone consist entirely of an avascular tissue composed of two types of fibre-like structures, which are coarse and longitudinally (parallel to the main axis of the element) oriented. These structures are differentiated on the basis of their optical properties under polarized light. Very thin fibrous strands are also observed in some regions. These small fibres are all oriented parallel to one another but perpendicular to the element main axis. Histological features of the primary bone tissue indicate that the sacral supraspinous rod corresponds to an ossified supraspinous ligament. The formation of this structure appears to have been a non-pathological metaplastic ossification, possibly induced by the continuous tensile forces applied to the element.Fil: Cerda, Ignacio Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte. Instituto de Investigación en Paleobiología y Geología; Argentina. Universidad Nacional de Río Negro; ArgentinaFil: Casal, Gabriel. Universidad Nacional de la Patagonia; ArgentinaFil: Martínez, Rubén Darío. Universidad Nacional de la Patagonia ; ArgentinaFil: Ibiricu, Lucio Manuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Nacional Patagónico; Argentin