MBDs mediate methylation, deacetylation and transcriptional repression.
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Biomedical subjects
Publications and source records attributed to A Snape.
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Recent studies in the chick have indicated that rhombomeres (r) are segments that underlie the patterning of hindbrain nerves. These segments may also be important for the specification of branchial arch structures since alternating rhombomeres, r2, r4 and r6, each contribute crest to a specific arch. Krox-20 has been implicated in the segmental patterning of the hindbrain in the mouse by its expression prior to segment formation in alternating domains, which later correspond to r3 and r5. Here, we describe the sequence and developmental expression of the Xenopus Krox-20 gene, XKrox-20. Alternating domains of XKrox-20 expression appear in the early neurula, later correspond to r3 and r5, and persist until late tadpole stages. In contrast to this conserved spatial expression in rhombomeres, we find a pattern in the neural crest of Xenopus that appears different from that found in the mouse: expression occurs in crest that migrates from r5 into the third visceral arch. We speculate that this may reflect a distinct route of neural crest migration due to anatomical differences between these systems, rather than a difference in the site of origin of Krox-20-expressing crest.
The molecules involved in the commitment of Xenopus cells to particular germ layers are unknown. The question has been investigated for the cells of the blastula in in vivo cell transplantation assays and in vitro aggregation assays. Using the former technique, we have shown that vegetal cells become committed before gastrulation, even when placed in inappropriate sites. We could find no evidence of regional determination within the endoderm germ layer at the early gastrula stage. In aggregation assays, animal and vegetal cells sorted incompletely at the mid-blastula stage and more efficiently at the gastrula stage. Their behaviour may be mediated by differential expression of adhesion molecules, which so far remain unidentified.
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In order to compare their states of commitment with their normal developmental fate, single vegetal pole cells from early Xenopus embryos were labeled and transplanted into the blastocoels of host embryos. In a previous study we showed, using this single cell transplantation assay, that vegetal pole cells become committed to endoderm by the early gastrula stage. In this paper we examine some properties of the commitment process. First, we show that it is gradual. When vegetal blastomeres are taken from progressively older embryos an increasing number of them enter only the endoderm, until by the early gastrula stage they all do. Second, we show that commitment can continue in vitro when an appropriate tissue mass is present. We suggest that commitment to form endoderm may be, in the right conditions, a cell autonomous process.
The experiments described in this paper were designed to compare the normal fates of animal pole blastomeres of Xenopus laevis with their state of commitment. Single animal pole blastomeres were labeled with a lineage marker and transplanted into the blastocoels of host embryos of different stages. The distribution of labeled daughter cells in the tadpole reflects the state of commitment of the parent cell at the time of transplantation. It is known that cells from the animal pole of the early blastula normally contribute predominantly to ectoderm with a small, but significant, contribution to the mesoderm. We show that on transplantation to the blastocoels of late blastula host embryos these blastomeres are pluripotent, contributing to all three germ layers. At later stages the normal fate of these cells becomes restricted solely to ectoderm and concomitantly the proportion of pluripotent cells is reduced, although the results depend upon the stage of the host embryo. Blastomeres from late blastula donors transplanted to mid gastrulae contribute solely to ectoderm in 34% of cases; however, in earlier hosts, when the vegetal hemisphere cells have "mesoderm inducing" or "vegetalizing" activity, late blastula animal pole blastomeres contribute to mesoderm and endoderm rather than ectoderm. Thus during the blastula stage animal pole cells pass from pluripotency to a labile state of commitment to ectoderm.
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Fate maps of amphibian embryos tell us the destination of certain areas at later stages of development. After studying Vogt's fate maps, Spemann wrote (in 1938) that "the question which at once calls for an answer is whether this pattern of presumptive primordia in the beginning gastrula is the expression of a real difference of these parts, whether they are already more or less predestined or 'determined' for their ultimate fate, or whether they are still indifferent and will not receive their determination until a later time." Until recently answers to this question have relied upon explant experiments, which indicate that by the late blastula stage ectoderm, mesoderm and endoderm regions are distinctly determined. By using a method involving single cell labelling and transplantation, it is now possible to pinpoint more accurately the time during early embryogenesis at which individual blastomeres become committed. In the vegetal pole, determination towards endoderm is a gradual process beginning during the middle blastula stage (stage 8) and completed by the beginning of gastulation (stage 10). This method offers the possibilities of comparing the committed and the uncommitted state and studying at a molecular level the mechanisms of cell determination.
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