A BMP-mediated transcriptional cascade involving Cash1 and Tlx-3 specifies first-order relay sensory neurons in the developing hindbrain.

The divergent homeobox-containing transcription factor, Tlx-3 (also known as Hox11L2/Rnx), is required for proper formation of first-order relay sensory neurons in the developing vertebrate brainstem. To date, however, the inductive signals and transcriptional regulatory cascade underlying their development are poorly understood. We previously isolated the chick Tlx-3 homologue and showed it is expressed early (i.

Specification of dorsoventral polarity in the embryonic chick mesencephalon and its presumptive role in midbrain morphogenesis.

The chick midbrain is subdivided into functionally distinct ventral and dorsal domains, tegmentum and optic tectum. In the mature tectum, neurons are organized in layers, while they form discrete nuclei in the tegmentum. Dorsoventral (DV) specification of the early midbrain should thus play a crucial role for the organization of the neuronal circuitry in optic tectum and tegmentum.

A conserved role for retinoid signaling in vertebrate pancreas development.

Retinoic acid (RA) signaling plays critical roles in the regionalization of the central nervous system and mesoderm of all vertebrates that have been examined. However, to date, a role for RA in pancreas and liver development has only been demonstrated for the teleost zebrafish. Here, we demonstrate that RA signaling is required for development of the pancreas but not the liver in the amphibian Xenopus laevis and the avian quail.

The role of Engrailed in establishing the dorsoventral axis of the chick limb.

Expression and mutation analyses in mice suggest that the homeobox-containing gene Engrailed (En) plays a role in dorsoventral patterning of the limb. During the initial stages of limb bud outgrowth, En-1 mRNA and protein are uniformly distributed throughout the ventral limb bud ectoderm. Limbs of En-1(-/-) mice display a double dorsal phenotype suggesting that normal expression of En-1 in the ventral ectoderm is required to establish and/or maintain ventral limb characteristics.

A role for FGF-8 in the initiation and maintenance of vertebrate limb bud outgrowth.

Background: The outgrowth of the vertebrate limb bud is the result of a reciprocal interaction between the mesenchyme and a specialized region of the ectoderm, the apical ectodermal ridge (AER), which overlies it. Signals emanating from the AER act to maintain the underlying mesenchyme, called the progress zone, in a highly proliferative and undifferentiated state. Removal of the AER results in the cessation of limb bud growth, thus causing limb truncation.

Independent assignment of antero-posterior and dorso-ventral positional values in the developing chick hindbrain.

Background: Cell patterning in the developing central nervous system seem to involve a coordinate system of positional information, in which specific fates are assigned to multipotent precursor cells by positional signals acting on the antero-posterior and dorso-ventral axes of the neural tube. Before neurons differentiate in the hindbrain, it becomes subdivided antero-posteriorly into a series of developmental compartments, the rhombomeres. When the rhombomeres are delineated from each other by interfaces at which cell mixing is transiently restricted, they are determined for expression of specific selector Hox genes that may encode aspects of their individual identity.

Quail neural crest cells cannot read positional values in the dorsal trunk feathers of the chicken embryo.

If quail neural crest cells are grafted to the chick, they migrate into the feathers of the host and produce melanin pigment. In one study, the dorsal trunk feathers of the chimaera were found to have quail-like pigment patterns. This was interpreted in terms of a positional information model.

Integration of new embryonic nephrons into the kidney.

The current report summarizes our experiments exploring the feasibility of creating a chimeric kidney, that is, an organ constituted by cells derived from more than one fertilized ovum. The overall strategy has been to obtain donor renal tissue from avian and murine embryos and to implant this into the host avian mesonephric mesoderm or into the cortex of murine neonatal kidney. In both models, donor cells were distinguished from the host by the presence of characteristic nuclear or cytoplasmic markers.

The spatial and temporal distribution of polarizing activity in the flank of the pre-limb-bud stages in the chick embryo.

The presence of polarizing activity in the limb buds of developing avian embryos determines the pattern of the anteroposterior axis of the limbs in the adult. Maps of the spatial distribution and the strength of the signal within limb buds of different stages are well documented. Polarizing activity can also be found in Hensen's node in the early embryo.

Pigment patterns in neural crest chimeras constructed from quail and guinea fowl embryos.

The pattern of pigmentation in bird embryos is determined by the spatial organization of melanocyte differentiation. Some of the results from recent, neural crest transplantation experiments support a model based on a prepattern in the feathers; others could be interpreted in terms of a nonspecific pattern resulting from a failure of the crest cells to read the positional values in another species. To distinguish between these possibilities, the crucial test is to construct chimeras from two species with different pigment patterns.

Double anterior chick limb buds and models for cartilage rudiment specification.

Most models for the specification of the skeletal elements in the developing limb bud are based on a chemical specification well before overt cartilage differentiation. By contrast, a physico-mechanical model proposes that the process of condensation--an early feature of cartilage differentiation--is itself the basis for patterning the elements. The models thus make quite different predictions as to when the rudiments are specified.

In situ hybridization reveals differential spatial distribution of mRNAs for type I and type II collagen in the chick limb bud.

During limb development, type I collagen disappears from the region where cartilage develops and synthesis of type II collagen, which is characteristic of cartilage, begins. In situ hybridization using antisense RNA probes was used to investigate the spatial localization of type I and type II collagen mRNAs. The distribution of the mRNA for type II collagen corresponded well with the pattern of type II collagen synthesis, suggesting control at the level of transcription and mRNA accumulation.

Positional signalling and the development of the humerus in the chick limb bud.

The positional signal model for specification of the cartilaginous elements in limb development has been tested by examining the effect on the humerus of grafting a polarizing region to different positions along the anteroposterior axis of the limb bud at stage 16. The humerus between the host and grafted polarizing region was largely normal though there were variations in width, particularly the distal epiphysis. The humerus often showed mirror-image symmetry along the anteroposterior axis.

The differentiation of normal and muscle-free distal chick limb bud mesenchyme in micromass culture.

Distal chick wing bud mesenchyme from stages 19 to 27 embryos has been grown in micromass culture. The behavior of cultures comprising mesenchyme located within 350 microns of the apical ectodermal ridge (distal zone mesenchyme) was compared to that of cultures of the immediately proximal mesenchyme (subdistal zone cultures). In cultures of the distal mesenchyme from stages 21-24 limbs, all of the cells stained immunocytochemically for type II collagen within 3 days, indicating ubiquitous chondrogenic differentiation.

Positional signalling by Hensen's node when grafted to the chick limb bud.

Hensen's node from stage 4 to stage 10 shows polarizing activity when grafted to the anterior margin of the chick limb bud. It can specify additional digits though its action is somewhat attenuated when compared with the effect of a grafted polarizing region. At stage 10 the activity disappears from the node and is found both posterior to the node and in the future wing region of the flank.

Growth and morphogenesis of the fibula in the chick embryo.

The development of the avian fibula was studied both histologically and experimentally. It was found that from the onset of chondrogenesis, the fibula possessed a smaller diameter than the neighbouring tibia. The truncated growth of the fibula was a result of the loss of its distal epiphysis between stages 27-31.

Positional signalling along the anteroposterior axis of the chick wing. The effect of multiple polarizing region grafts.

We have proposed that positional information along the anteroposterior axis is specified by a signal from the polarizing region and that position may be specified by the concentration of a diffusible morphogen. While this model can account for a variety of results it is now clear that a model based on intercalation by growth of positional values can do the same. The distinction between the two models lies in whether a grafted polarizing region can alter existing positional values and in the distance over which it exerts its influence.

Effects of biochemical inhibitors on positional signalling in the chick limb bud.

In 3- to 4-day embryonic chick limb buds, a region at the posterior margin of the limb, the zone of polarizing activity, appears to be responsible for signalling positional information along the anteroposterior axis. Our experiments were designed to test which biosynthetic processes are required for polarizing activity. We have treated polarizing regions with biochemical inhibitors, and then assayed their abilities to induce limb reduplications when grafted into anterior sites on host limb buds and also measured their capacities for protein, RNA, and DNA synthesis.

Muscle-forming potential of the non-somitic cells of the early avian limb bud.

It has recently been shown that the musculature of the chick wing arises by migration of cells from the somites, and that on morphological grounds this process begins at about stage 14. We have carried out grafts of wing anlagen separate from the somites from quail donors to the extra-embryonic coelom of chicks, and find that anlagen from as early as stage 10 (11 pairs of somites) can give rise to muscle. We discuss the possible reasons for this finding, and conclude that in the absence of the cells normally giving rise to the musculature, mesodermal cells themselves can give rise to muscle.

Somite formation in the early chick embryo following grafts of Hensen's node.

Quail grafts of Hensen's node were examined for their potential to induce somites in chick blastoderms. The origin of the structures induced depended on the distance of the graft from the host's midline. Nodes placed at the margin of the area pellucida resulted in structures differentiated from the cells of the graft, whereas medially the graft organized host cells to form rows of somites.

Polarity reversal in hydra by oligomycin.

Intact hydra treated for 24 h with oligomycin gradually lose their head structures and the distal ends form feet. Grafting experiments show that the distal ends of treated animals induce proximal structures.