Dorsal induction from dorsal vegetal cells in Xenopus occurs after mid-blastula transition.
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We performed some experiments to investigate the temporal and spatial details of the dorsal induction exerted by dorsal vegetal cells in Xenopus embryo. Two dorsal vegetal cells (D1 cells) were transplanted into the ventral vegetal region of a recipient at the 32-cell stage. At various times after transplantation, the ventral animal-equatorial part was explanted and cultured. The explants isolated 5.5 h after transplantation (time 5.5) elongated and formed somites. In RT-PCR analysis, the expression of dorsal gene, chordin was activated in the explants isolated after time 4.0 (about the 4000-cell stage which corresponds to the mid blastula transition (MBT)) at control stage 10. In another series of experiments, ventral animal-equatorial and dorsal vegetal parts were isolated from the 4000-cell stage embryos and they were combined for 2.0-2.5 h. These ventral animal-equatorial explants elongated and formed somites. The chordin expression was also observed in the explants. But the 32- and 256-cell stage dorsal vegetal cells failed to exert the dorsalizing activity within the 2.0-2.5 h of the conjugation. These results suggest that 2 h contact after MBT is necessary and sufficient for the dorsal induction from the dorsal vegetal cells and it occurs as a result of the zygotic gene expression. Consistent with this idea, the zygotic dorsal genes, siamois and chordin were expressed on the upper regions of the transplanted D1 descendants at stage 10. Furthermore, this region began to gastrulate when the D1 cell was transplanted with upside-down orientation. Our data indicate that the upper region of the D1 descendants by itself act as the Spemann organizer rather than the Nieuwkoop center. (+info)
Neuroectodermal specification and regionalization of the Spemann organizer in Xenopus.
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During gastrulation in Xenopus convergence and extension movements, mediated by mediolateral intercalations, are the driving force for early neural plate morphogenesis. Here we show that the winged helix transcriptional regulator, Xfd-12' is dynamically expressed in medial neural plate precursors that undergo convergence and extension movements. These medial neuraxial progenitors are specified in and beyond the Spemann organizer prior to specification of the basal anlage of the neural plate. The initiation of Xfd-12' expression coincides with the induction of mesendoderm by Nodal-related growth factors at the late blastula stage. Comparative expression analysis suggests that cellular rearrangements at the pre-gastrulation stage account for regionalization of the Spemann organizer into head and trunk organizer compartments, the latter in which medial neural plate progenitors reside. While the maintenance of Xfd-12' expression in the dorsal non-involuting marginal zone requires FGF signalling, its subsequent positioning along the medial aspect of the neuraxis depends on signalling by Wnt and Nodal-related family members. Based on these findings we propose that XFD-12' is a trunk organizer component that might control convergence and extension movements of medial neural plate precursors during gastrulation. (+info)
Uncx4.1 is required for the formation of the pedicles and proximal ribs and acts upstream of Pax9.
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The expression of the homeobox gene Uncx4.1 in the somite is restricted to the caudal half of the newly formed somite and sclerotome. Here we show that mice with a targeted mutation of the Uncx4.1 gene exhibit defects in the axial skeleton and ribs. In the absence of Uncx4.1, pedicles of the neural arches and proximal ribs are not formed. In addition, dorsal root ganglia are disorganized. Histological and marker analysis revealed that Uncx4.1 is not necessary for somite segmentation. It is required to maintain the condensation of the caudal half-sclerotome, from which the missing skeletal elements are derived. The loss of proximal ribs in Pax1/Pax9 double mutants and the data presented here argue for a role of Uncx4.1 upstream of Pax9 in the caudolateral sclerotome. Our results further indicate that Uncx4.1 may be involved in the differential cell adhesion properties of the somite. (+info)
The paired homeobox gene Uncx4.1 specifies pedicles, transverse processes and proximal ribs of the vertebral column.
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The axial skeleton develops from the sclerotome, a mesenchymal cell mass derived from the ventral halves of the somites, segmentally repeated units located on either side of the neural tube. Cells from the medial part of the sclerotome form the axial perichondral tube, which gives rise to vertebral bodies and intervertebral discs; the lateral regions of the sclerotome will form the vertebral arches and ribs. Mesenchymal sclerotome cells condense and differentiate into chondrocytes to form a cartilaginous pre-skeleton that is later replaced by bone tissue. Uncx4.1 is a paired type homeodomain transcription factor expressed in a dynamic pattern in the somite and sclerotome. Here we show that mice homozygous for a targeted mutation of the Uncx4.1 gene die perinatally and exhibit severe malformations of the axial skeleton. Pedicles, transverse processes and proximal ribs, elements derived from the lateral sclerotome, are lacking along the entire length of the vertebral column. The mesenchymal anlagen for these elements are formed initially, but condensation and chondrogenesis do not occur. Hence, Uncx4.1 is required for the maintenance and differentiation of particular elements of the axial skeleton. (+info)
The Oak Ridge Polycystic Kidney (orpk) disease gene is required for left-right axis determination.
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Analysis of several mutations in the mouse is providing useful insights into the nature of the genes required for the establishment of the left-right axis during early development. Here we describe a new targeted allele of the mouse Tg737 gene, Tg737(Delta)2-3(beta)Gal), which causes defects in left-right asymmetry and other abnormalities during embryogenesis. The Tg737 gene was originally identified based on its association with the mouse Oak Ridge Polycystic Kidney (orpk) insertional mutation, which causes polycystic kidney disease and other defects. Complementation tests between the original orpk mutation and the new targeted knock-out mutation demonstrate that Tg737(Delta)2-3(beta)Gal) behaves as an allele of Tg737. The differences in the phenotype between the two mutations suggest that the orpk mutation is a hypomorphic allele of the Tg737 gene. Unlike the orpk allele, where all homozygotes survive to birth, embryos homozygous for the Tg737(Delta)2-3(beta)Gal) mutation arrest in development at mid-gestation and exhibit neural tube defects, enlargement of the pericardial sac and, most notably, left-right asymmetry defects. At mid-gestation the direction of heart looping is randomized, and at earlier stages in development lefty-2 and nodal, which are normally expressed asymmetrically, exhibit symmetrical expression in the mutant embryos. Additionally, we determined that the ventral node cells in mutant embryos fail to express the central cilium, which is a characteristic and potentially functional feature of these cells. The expression of both Shh and Hnf3(beta) is downregulated in the midline at E8.0, indicating that there are significant alterations in midline development in the Tg737(Delta)2-3(beta)Gal) homozygous embryos. We propose that the failure of ventral node cells to fully mature alters their ability to undergo differentiation as they migrate out of the node to contribute to the developing midline structures. Analysis of this new knockout allele allows us to define a critical role for the Tg737 gene during early embryogenesis. We have named the product of the Tg737 gene Polaris, which is based on the various polarity related defects associated with the different alleles of the Tg737 gene. (+info)
Traumatic spondylolisthesis of the axis: treatment rationale based on the stability of the different fracture types.
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Thirty-nine consecutive patients, 22 male and 17 female with an average age of 37.6 years, with traumatic spondylolisthesis of the axis were reviewed. The cause of injury in 75% of the patients was a road traffic accident. The fractures were classified according to Effendi et al., the type II fractures were further divided into three subgroups: flexion, extension and listhesis injuries. There were 10 type I (25.7%) and 29 type II fractures (74.4%); of these, 12 (30.8%) were classified as flexion-type, 2 (5.1%) as extension-type and 15 (38.5%) as listhesis-type. We did not identify any case of type III injury. Overall, 43.5% of the patients had sustained a significant head or chest trauma, with the highest incidence for type II listhesis injuries. Significant neurological deficits occurred in four patients (10.3%); in all four,the fracture was classified as a type II listhesis. All ten type I injuries were successfully treated with a cervical orthosis. Ten of the 12 type II flexion injuries demonstrated significant angulation. Two were treated with internal stabilisation, in seven with a halo device and one with a minerva plaster of Paris (PoP). Healing was uneventful in all ten patients. For the remaining two stable type II flexion injuries, application of a hard collar was adequate, as was the case for the two stable type II extension injuries. Six of the 15 type II spondylolisthesis injuries underwent primary internal stabilisation, and healing was uneventful in all cases. In four (44.4%) of the nine injuries that were primarily treated with a halo device/minerva PoP, secondary operative stabilisation had to be performed. The classification of Effendi et al. provides a complete description of the different fractures. However, further distinction of the type II injuries regarding their stability is mandatory. Type II spondylolisthesis injuries are unstable, with a high number of associated injuries, a great potential for neurological compromise and significant complications associated with non-operative treatment. The majority of type II extension and type II flexion injuries can be successfully treated with nonrigid external immobilisation. (+info)
Quantitative anatomy of the lateral masses of the atlas and axis vertebrae.
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The study was carried out to determine the safe site of entry and the appropriate trajectory of the screw implantation in the lateral masses of atlas (Cl) and axis (C2) during their fixation using the plate and screw technique. Fifty dried specimens of atlas and axis vertebrae were studied. Various dimensions of the lateral masses were quantitatively measured, laying stress on their relationship with the vertebral artery foramen. As the vertebral artery foramen was present entirely in the transverse process in all specimens, screw implantation in the facet of atlas was relatively safe. Best direction of screw implantation in the facet of atlas was observed to be 15 degrees medial to sagittal plane and 15 degrees superior to axial plane. It should be implanted from the middle of the posterior surface of facet. Vertebral artery foramen formed a deep groove in the undersurface of a majority of superior facets of axis. In 15% facets, vertebral artery foramen occupied the entire undersurface of the superior facet. Safe angle for screw implantation in the facet of axis through its pedicle was seen to be 40 degrees medial to sagittal plane and 20 degrees superior to axial plane. Safe site of screw entry in the axis was superior and medial third of the posterior surface of the pedicle. Quality of cancellous bone in the lateral masses in the proposed trajectory of screw in Cl and C2 was good, providing an excellent purchase of the screw. (+info)
Axis-inducing activities and cell fates of the zebrafish organizer.
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We have investigated axis-inducing activities and cellular fates of the zebrafish organizer using a new method of transplantation that allows the transfer of both deep and superficial organizer tissues. Previous studies have demonstrated that the zebrafish embryonic shield possesses classically defined dorsal organizer activity. When we remove the morphologically defined embryonic shield, embryos recover and are completely normal by 24 hours post-fertilization. We find that removal of the morphological shield does not remove all goosecoid- and floating head-expressing cells, suggesting that the morphological shield does not comprise the entire organizer region. Complete removal of the embryonic shield and adjacent marginal tissue, however, leads to a loss of both prechordal plate and notochord. In addition, these embryos are cyclopean, show a significant loss of floor plate and primary motorneurons and display disrupted somite patterning. Motivated by apparent discrepancies in the literature we sought to test the axis-inducing activity of the embryonic shield. A previous study suggested that the shield is capable of only partial axis induction, specifically being unable to induce the most anterior neural tissues. Contrary to this study, we find shields can induce complete secondary axes when transplanted into host ventral germ-ring. In induced secondary axes donor tissue contributes to notochord, prechordal plate and floor plate. When explanted shields are divided into deep and superficial fragments and separately transplanted we find that deep tissue is able to induce the formation of ectopic axes with heads but lacking posterior tissues. We conclude that the deep tissue included in our transplants is important for proper head formation. (+info)