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Molecular analysis of neural crest formation.

Neural crest cells arise within the ectoderm during neurulation and give rise to most of the peripheral nervous system. Following neural tube closure, they come to lie within the dorsal neural tube from which they emerge and subsequently migrate extensively to numerous and characteristic sites. There, they differentiate into neurons and glia of the peripheral nervous system, cartilage and bone of the face, melanocytes and various other cell types. Fate mapping experiments have demonstrated that the neural crest arises at the juncture between presumptive epidermis and neural plate. However, injection of lineage tracer into individual cells reveals that single neural fold cells are not committed to a neural crest fate; rather these cells can form all ectodermal derivatives (epidermis, neural tube, neural crest). Inductive interactions between the neural and non-neural ectoderm can generate neural crest cells, suggesting that signals travel through the epidermis to generate neural crest cells prior to neural tube closure. Induction of the neural crest appears to be a multiphasic process and involves a combination of an early Wnt signal together with later functions of BMP signaling pathways. We have used a variety of molecular screens to isolate molecular constituents involved in neural crest formation. We have identified a secreted factor, Noelin-1, which is expressed in the prospective avian neural plate and may play a role making the neural tube competent to form neural crest. Noelin-1 mRNA is expressed in a graded pattern in the closing neural tube, with highest expression in the neural folds and no detectable expression at the ventral midline. Its expression precedes that of Slug, a zinc finger transcription factor that represents the earliest known neural crest marker gene. Over-expression of Noelin-1 using recombinant retroviruses causes an excess of neural crest emigration and prolongs the time that the neural tube is competent to generate and regenerate neural crest cells. These results support an important role for Noelin-1 in rendering the neural tube competent to respond to inductive cues to generate neural crest.

Animals↗

Early induction of neural crest cells: lessons learned from frog, fish and chick.

The identification of genes in Xenopus, chick and zebrafish expressed early in prospective neural crest (NC) cells has challenged the previous view that the NC is induced during the closure of the neural tube. We compare here the early inductive molecular mechanisms in different organisms and, despite observed differences, propose a general common model for NC induction.

Animals↗

Opponent activities of Shh and BMP signaling during floor plate induction in vivo.

We performed in vivo experiments in chick embryos that examined whether application of an exogenous source of Shh protein mimics the ability of the notochord to induce ectopic floor plate cells in the neural tube. Shh cannot act alone to induce a floor plate. However, coapplication of Shh and chordin, a BMP antagonist normally coexpressed with Shh in the notochord, results in a marked switch from dorsal to ventral cell fate, including a dramatic and widespread induction of floor plate cells. These data provide in vivo evidence that notochord-derived BMP antagonists may normally generate a permissive environment for the Shh-mediated induction of floor plate. Further experiments performed to address the source of BMPs that are inhibited by the action of chordin suggest that they derive specifically from the surface ectoderm and dorsal-most neuroepithelium. These data indicate that, at neural groove stages, dorsally derived BMPs affect ventral-most regions of the neural plate, suggesting a novel long-range action of BMPs. Together, these studies suggest that the balance of dorsally derived signals and notochord-derived signals determines the extent of floor plate cell induction.

Animals↗

A simple "neural induction" model with two interacting cleavage-arrested ascidian blastomeres.

A single anterior-animal blastomere, which includes the presumptive neural region in the eight-cell embryo of the Halocynthia, a protochordate, when dissociated, cleavage-arrested with cytochalasin B, and cultured in isolation, differentiated exclusively to epidermal type judging from membrane excitability and immunoreactivity. However, when the same blastomere was cultured in contact with a single anterior-vegetal blastomere, which includes the presumptive notochordal region, it displayed Na spikes and showed no expression of the epidermal antigen, suggesting that "neural induction" resulted in a single cell during the interaction with a single neighboring cell. This simple two-cell system can be used for further studies on the induction mechanism.

Animals↗

Regulations in the induction of the organized neural system in amphibian embryos.

Some of the recent data on the induction of the neural system in amphibian embryos are reviewed, utilizing a model, according to which two basic events regulate in this system: (1) ectodermal dorsalization, which occurs all over the induced region of the ectoderm and is responsible for the neural and mesectodermal pathways and (2) caudalization, which occurs only on the posterior level of dorsalized ectoderm and is responsible for the posterior mode of induced differentiation, functioning as a gradient with the apex at the posterior end of the embryo. Dorsalization of ectoderm can be caused by treatment with Con A or TPA, both of which are potential mitogens. Not only after the treatment with TPA, but also during normal dorsalization, the activation of protein kinase C occurs in responding cells. The possibility is suggested that an early step of mitogenic transmembrane signal transduction induced by a growth factor regulates dorsalization in intact embryos. Ectodermal dorsalization is responsible for the appearance of neuronal and glial cell lineages, and independent of the ECM network formed on the internal surface of the responding ectoderm during gastrulation. In caudalization, a series of experiments suggests that the regulatory role is played by the transcript of the mesodermal posterior homeobox gene, Xhox 3. The expression of this gene in time and location closely coincides with the pattern of convergent extension, one type of morphogenetic movement, which is expressed in a posterior-anterior gradient. This directed cell motility is responsible for the formation of the body axis of vertebrates, and was shown to be involved in caudalization by earlier induction experiments in urodele embryos.(ABSTRACT TRUNCATED AT 250 WORDS)

Amphibians↗

Brown adipose tissue as a derivative of mesoderm grafted below the kidney capsule. A model for differentiation of isolated rat mesoderm.

During development, mesoderm differentiates into connective tissue, cartilage, bone, muscle and kidney. In experimental conditions the developmental spectrum of mesoderm grafted below the kidney capsule is reduced so that mostly brown adipose tissue (BAT) appears. Since BAT is a particular tissue with a specific developmental pattern, the structural and functional characteristics of experimentally developed BAT were analyzed in the present study. Mesoderm from nine-day-old rat embryos was grafted below the kidney capsule of adult rats and one month later the BAT-containing tumors were analyzed. The ultrastructural and morphometrical data of BAT-containing tumors were the same as in BAT developed in situ. Tissue-specific mRNA for uncoupling protein (UCP) was expressed in BAT-containing tumors, and immuno-electron microscopical analysis showed that mitochondria of these brown adipocytes contained UCP. Injections of noradrenaline and exposure of BAT-tumor-bearing rats to cold stress increased both the amount of UCP and the expression of UCP mRNA in tumors of BAT; i.e., experimentally developed BAT entirely resembled standard BAT. It is proposed that mesoderm isolated and displaced below the kidney capsule lacks the inductive stimuli of ectoderm and endoderm, and as a result mesoderm can not realize the natural pattern of differentiation. Here, in a new environment, mesoderm is exposed to new stimuli which induce differentiation of mesoderm into BAT, probably through neuro-vascular elements from the medial side of the kidney (BAT area). Thus, although mesoderm contains a wide differentiation capacity, it can differentiate into only one type of tissue, depending on the presence and range of inductive stimuli.

Adipose Tissue, Brown↗

K562 cells: a source for embryonic globin chains.

A combination of DEAE-cellulose chromatography and reversed-phase high-performance liquid chromatography (HPLC) has been used to devise a method for generating large quantities of embryonic as well as fetal globin chains. The identity of these globin chains was further confirmed by their tryptic peptide mapping. This technique could, therefore, provide a reliable source for these polypeptides for both analytical and immunological purposes. Moreover, the study of human hemoglobin switching, particularly embryonic to fetal, has been greatly hampered by the absence of a suitable model. K562 cells, due to their potential for differential induction of embryonic and fetal hemoglobin synthesis, can thus be used for this purpose and the various hemoglobins produced can then be effectively monitored using this method.

Chromatography, DEAE-Cellulose↗

The primitive streak.

The emphasis of this review is on the primitive streak of the chick embryo, collated with such information as is available on the mouse embryo. Little modern work has been published on any reptile primitive streak. The following topics are considered: evolutionary significance; formation of the primitive streak; ingression and de-epithelialisation; the basal lamina; migration from the primitive streak of the endoderm and mesoderm; the role of the extracellular matrix; changes in cell adhesiveness; regression of the primitive streak and its role in body patterning; the primitive streak and induction.

Animals↗

Ets-mediated brain induction in embryos of the ascidian Halocynthia roretzi.

The larval ascidian brain (sensory vesicle) is located on the dorsal side of the trunk region and forms part of the anterior central nervous system. Sensory organs such as the otolith, ocellus, and hydrostatic-pressure organ reside in the brain. The brain coordinates the core roles of the larval nervous system. The brain is derived from anterior animal a-line blastomeres. The default fate of these blastomeres is epidermis, and the inductive signals from anterior vegetal blastomeres convert the fate into brain. It remains unclear, however, when these inductive interactions take place. To determine when, we examined whether partial embryos derived from brain-lineage blastomeres isolated at various stages express neural and epidermal marker genes. Partial embryos derived from brain-lineage blastomeres isolated after the 32-cell stage expressed all the neural marker genes examined. The expression of the epidermal marker gene was first reduced in partial embryos when blastomeres were isolated at the 64-cell stage. Moreover, the process for brain specification seemed to continue after the 110-cell stage. We also investigated the function of HrEts, an ascidian homolog of Ets transcription factors, to elucidate the molecular mechanism of brain induction. HrEts functions were inhibited by the use of antisense morpholino oligonucleotides. Loss of Ets functions resulted in loss of the expression of some of the neural marker genes and the ectopic expression of the epidermal marker gene in brain precursor cells. These results suggest that HrEts is an essential transcription factor that mediates ascidian brain induction.

Animals↗

An Ets transcription factor, HrEts, is target of FGF signaling and involved in induction of notochord, mesenchyme, and brain in ascidian embryos.

In ascidian embryos, a fibroblast growth factor (FGF) signal induces notochord, mesenchyme, and brain formation. Although a conserved Ras/MAPK pathway is known to be involved in this signaling, the target transcription factor of this signaling cascade has remained unknown. We have isolated HrEts, an ascidian homolog of vertebrate Ets1 and Ets2, to elucidate the transcription factor involved in the FGF signaling pathway in embryos of the ascidian Halocynthia roretzi. Maternal mRNA of HrEts was detected throughout the entire egg cytoplasm and early embryos. Its zygotic expression started in several tissues, including the notochord and neural plate. Overexpression of HrEts mRNA did not affect the general organization of the tadpoles, but resulted in formation of excess sensory pigment cells. In contrast, suppression of HrEts function by morpholino antisense oligonucleotide resulted in severe abnormalities, similar to those of embryos in which the FGF signaling pathway was inhibited. Notochord-specific Brachyury expression at cleavage stage and notochord differentiation at the tailbud stage were abrogated. Formation of mesenchyme cells was also suppressed, and the mesenchyme precursors assumed muscle fate. In addition, expression of Otx in brain-lineage blastomeres was specifically suppressed. These results suggest that an Ets transcription factor, HrEts, is involved in signal transduction of FGF commonly in notochord, mesenchyme, and brain induction in ascidian embryos.

Amino Acid Sequence↗

Induction and initial patterning of the nervous system - the chick embryo enters the scene.

Until recently, almost everything known about the molecular controls of early neural development came from studies in amphibians. It is now possible to misexpress factors in chick embryos at relatively late stages in development, allowing careful dissection of the timing of cell interactions. This is starting to contribute significantly to our understanding of neural induction and early patterning.

Animals↗

The floor plate: multiple cells, multiple signals.

One of the key organizers in the CNS is the floor plate - a group of cells that is responsible for instructing neural cells to acquire distinctive fates, and that has an important role in establishing the elaborate neuronal networks that underlie the function of the brain and spinal cord. In recent years, considerable controversy has arisen over the mechanism by which floor plate cells form. Here, we describe recent evidence that indicates that discrete populations of floor plate cells, with characteristic molecular properties, form in different regions of the neuraxis, and we discuss data that imply that the mode of floor plate induction varies along the anteroposterior axis.

Animals↗

Gli2 mediation of hedgehog signals in slow muscle induction in zebrafish.

Zebrafish skeletal muscles are composed of two major types of muscle fibers, broadly classified as fast or slow fibers. Recent studies have demonstrated that members of the Hedgehog (Hh) family induce the formation of slow muscle fibers. Hedgehog signals are secreted proteins that function through the transcription factor Glis. We report here the characterization of a zebrafish Gli2 expression in slow and fast muscle cells and the study of the roles of Hedgehogs and Gli2 in zebrafish muscle development using two mutant strains; sonic-you (syu) and you-too (yot), respective for sonic hedgehog (shh) and Gli2 mutation. We have demonstrated that Shh and Gli2 mutation causes similar defects in slow muscle formation. There is, however, a difference in the degree of defect between these two mutants. In yot mutant embryos, development of slow muscles was completely blocked, whereas in syu mutant embryos, a small number of slow muscle cells could still form, suggesting that other Hhs were also involved in slow muscle induction. Induction of slow muscles by other Hhs appeared to require Gli2, because ectopic expression of Echidna hedgehog (Ehh) and Tiggy-winkle hedgehog (Twhh) failed to induce slow muscles in yot mutant embryos. Together, these data suggest that further Hhs, other than Shh, are also involved in the induction and differentiation of slow muscle cells and that Gli2 is required by Shh, Twhh, and Ehh, thus playing a key role in the induction and differentiation of slow muscle cells.

Animals↗

Endoderm development in vertebrates: fate mapping, induction and regional specification.

The formation of the vertebrate body plan begins with the differentiation of cells into three germ layers: ectoderm, mesoderm and endoderm. Cells in the endoderm give rise to the epithelial lining of the digestive tract, associated glands and respiratory system. One of the fundamental problems in developmental biology is to elucidate how these three primary germ layers are established from the homologous population of cells in the early blastomere. To address this question, ectoderm and mesoderm development have been extensively analyzed, but study of endoderm development has only begun relatively recently. In this review, we focus on the 'where', 'when' and 'how' of endoderm development in four vertebrate model organisms: the zebrafish, Xenopus, chick and mouse. We discuss the classical fate mapping of the endoderm and the more recent progress in characterizing its induction, segregation and regional specification.

Animals↗

Ectoderm induces muscle-specific gene expression in Drosophila embryos.

We have inhibited normal cell-cell interactions between mesoderm and ectoderm in wild-type Drosophila embryos, and have assayed the consequences on muscle development. Although most cells in gastrulation-arrested embryos do not differentiate, they express latent germ layer-specific genes appropriate for their position. Mesoderm cells require proximity to ectoderm to express several muscle-specific genes. We show that ventral ectoderm induces mesoderm cells to express nautilus (a MyoD homologue) and to differentiate somatic myofibers, whereas dorsal ectoderm induces mesoderm cells to express visceral and cardiac muscle-specific genes. Our findings suggest that muscle determination in Drosophila is regulated by induction between germ layers during gastrulation.

Animals↗