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J B Gurdon

Publications and source records attributed to J B Gurdon.

At least 73 records · Page 4Linked to original sources
PubMed1994

Activin signalling and response to a morphogen gradient.

Using combinations of amphibian embryo tissues, it is shown that the selection of genes expressed by a cell is determined by its distance from a source of activin, a peptide growth factor contained in vegetal cells and able to induce other cells to form mesoderm. This long-range signal spreads over at least 10 cell diameters in a few hours. It does so by passive diffusion, because it can by-pass cells that do not themselves respond to the signal nor synthesize protein. These results provide direct support for the operation of a morphogen concentration gradient in vertebrate development.

Activins↗
PubMed1994

An inhibitory effect of Xenopus gastrula ectoderm on muscle cell differentiation and its role for dorsoventral patterning of mesoderm.

In Amphibia, mesoderm cells such as notochord, muscle, and blood are formed as a result of mesoderm induction, the first known inductive interaction during the embryonic development of Vertebrates. Recent evidence shows, however, that, in addition to mesoderm induction, further cell-cell interactions during gastrulation also play an important role in the differentiation of mesoderm cells in Xenopus embryos. Here we report the existence of an inhibitory effect of gastrula ectoderm on muscle cell differentiation. When placed in contact with a muscle progenitor cell population, ectoderm from the ventral side of a gastrula embryo exerts an inhibitory effect on MyoD expression and muscle differentiation. This inhibitory effect is not observed on notochord differentiation. Dorsal ectoderm shows the same extent of inhibitory effect if it is isolated at the early-gastrula stage but gradually loses its effect during gastrulation as it is induced to become neural tissue. Cell mixing experiments have shown that this inhibitory effect, clearly seen in ventral ectoderm, is not observed in ventral mesoderm cells. We propose that the ectoderm emits a signal counteracting dorsalization and the community effect and that this signal plays an important role in the correct dorsoventral patterning of mesoderm.

Animals↗
PubMed1994

A role for cytoplasmic determinants in mesoderm patterning: cell-autonomous activation of the goosecoid and Xwnt-8 genes along the dorsoventral axis of early Xenopus embryos.

Although an induction event is required for the formation of mesoderm in Xenopus embryos, it is not clear that this induction is wholly sufficient to give rise to a correctly patterned mesodermal layer. We have studied the expression of the two genes, goosecoid and Xwnt-8, in Xenopus gastrulae in which cell-cell communication, and therefore mesoderm induction, has been prevented by frequent cell dispersion. Although neither the early panmesodermal marker Xbra nor the muscle-specific alpha-actin gene were activated under these conditions, goosecoid and Xwnt-8 were activated in cells of dorsal and ventrolateral origin respectively, thus correctly reflecting their distribution during normal development. We also show that the spatial pattern of expression of these two genes along the animal-vegetal axis is similar in normal and in dissociated early gastrulae: goosecoid is mainly expressed in future mesoderm while the domain of expression of Xwnt-8 spans the mesoderm-endoderm boundary. These results show that, during the blastula and early gastrula stages, gene activation can be controlled cell-autonomously along both the animal-vegetal and dorsoventral embryo axes. This suggests that the inheritance of localised maternal cytoplasmic determinants is a key event for the patterning of mesoderm. We present a modified model of mesoderm formation in which the different mesoderm cell types are produced as a result of cooperation between induction-dependent and induction-independent immediate-early genes.

Animals↗
PubMed1993

Single-cell transplantation determines the time when Xenopus muscle precursor cells acquire a capacity for autonomous differentiation.

We have used a single-cell transplantation technique to find out whether there is a stage in development when a single cell can reach and maintain its differentiated state in the absence of its neighbors. Muscle precursor cells from early, mid-, and late gastrula stages of Xenopus laevis embryos were isolated and transplanted singly into the ventral region of late gastrula hosts. Single cells from late gastrulae differentiated into muscle when surrounded by nonmuscle cells. Similar cells from early or mid-gastrulae did not, unless they were transplanted as a group of adjacent cells taken from the same region of an embryo. These results show that single embryonic cells in a tissue can complete their differentiation without interacting with their normal neighbors and that, in the case of Xenopus muscle precursor cells, they acquire this capacity at the late gastrula stage. Our results also suggest that, in addition to mesoderm induction, further cell interactions during gastrulation are required for Xenopus muscle cell differentiation.

Alkaline Phosphatase↗
PubMed1993

The community effect, dorsalization and mesoderm induction.

The community effect is an interaction among muscle progenitor cells of amphibian gastrula, and is necessary for the initiation of muscle-specific gene expression. Dorsalization provides a signal that can convert ventral mesoderm cells to a muscle fate. Neither process involves mesoderm-inducing molecules. We suggest that the developmental significance of the community effect is to generate homogeneous but clearly demarcated groups of cells from progenitor cells arranged in a continuous gradient of developmental potential.

Animals↗
PubMed1993

A community effect in muscle development.

BACKGROUND: Most vertebrate tissues arise by embryonic induction, as a result of which new cell layers are formed. These are subsequently subdivided into discrete groups of homogeneous cell populations, each containing different cell-types with specific gene expression. There is preliminary evidence from previous work that the mesoderm-forming induction in amphibian development may be followed by a further interaction among some of the induced mesoderm cells, and that this could be required for muscle gene activation in uniform cell populations. RESULTS: We have established the existence, time and place of this further cell interaction by transplanting muscle progenitor cells from Xenopus mid-gastrulae into ectoderm sandwiches, and then culturing these constructs until the time of muscle gene activation. We find that cells implanted as reaggregates, but not those implanted as single cells, activate early myogenic genes and later muscle-specific genes. More than 100 cells must be near each other for muscle gene activation. These cells can induce non-muscle mesoderm cells to express muscle genes by emitting a signal that differs from the preceding mesoderm induction signal. Muscle gene activation under these conditions does not require gap junction communication. CONCLUSION: Cells within the muscle progenitor region of a Xenopus embryo need to interact with each other in order to activate muscle genes in homogeneous cell groups. This exemplifies the 'community effect', which may be a widespread developmental mechanism used to increase the homogeneity within, and demarkation between, embryonic tissues.

Journal Article↗
PubMed1993

Construction of subtracted cDNA libraries enriched for cDNAs for genes expressed in the mesoderm of early Xenopus gastrulae.

We present a new method for the construction of subtracted cDNA libraries. It makes use of two directional cDNA libraries in phagemid vectors and has two advantages over traditional protocols: it can be used when a limiting amount of driver material is available and the cDNA inserts in the subtracted libraries are long enough to encode functional proteins. Using this method, we have constructed two subtracted libraries enriched for sequences expressed in the mesoderm of early Xenopus gastrulae.

Animals↗
PubMed1992

The DNA-binding protein E12 co-operates with XMyoD in the activation of muscle-specific gene expression in Xenopus embryos.

Two alternatively spliced products of the human E2A gene, E12 and E47, encode helix-loop-helix DNA-binding proteins. Here we describe the isolation of two Xenopus cDNAs; one, XE12, is structurally similar to human E12 and the other contains a sequence similar to E47. Transcripts of both cDNAs are present at all the stages of Xenopus development tested and in all regions of the embryo. The DNA binding properties of in vitro translated XE12 are indistinguishable from those of human E12. We have shown previously that an embryonic muscle DNA-binding activity, EMF1, that binds to a promoter sequence required for the expression of the cardiac actin gene, contains the Xenopus myogenic factor XMyoD. Here we show that it also contains protein that interacts with an anti-E12 antiserum, suggesting that XE12 and XMyoD proteins, or very similar ones, are present in EMF1. We have addressed the functional role of XE12 in muscle gene transcription in Xenopus embryos by injecting in vitro synthesized RNA into the two cell embryo. Overexpression of XE12 and XMyoD augments by greater than 10-fold the ectopic activation of the endogenous cardiac actin gene that can be produced by XMyoD alone. Our DNA binding results strongly suggest that this effect is mediated through a direct interaction of the XE12-XMyoD complex with specific sites in the cardiac actin promoter. We suggest that XE12 is functionally important in muscle gene activation in embryonic development.

Amino Acid Sequence↗
PubMed1992

Expression of XMyoD protein in early Xenopus laevis embryos.

A monoclonal antibody specific for Xenopus MyoD (XMyoD) has been characterized and used to describe the pattern of expression of this myogenic factor in early frog development. The antibody recognizes an epitope close to the N terminus of the products of both XMyoD genes, but does not bind XMyf5 or XMRF4, the other two myogenic factors that have been described in Xenopus. It reacts in embryo extracts only with XMyoD, which is extensively phosphorylated in the embryo. The distribution of XMyoD protein, seen in sections and whole-mounts, and by immunoblotting, closely follows that of XMyoD mRNA. XMyoD protein accumulates in nuclei of the future somitic mesoderm from the middle of gastrulation. In neurulae and tailbud embryos it is expressed specifically in the myotomal cells of the somites. XMyoD is in the nucleus of apparently every cell in the myotomes. It accumulates first in the anterior somitic mesoderm, and its concentration then declines in anterior somites from the tailbud stage onwards.

Animals↗
PubMed1992

Muscle gene activation in Xenopus requires intercellular communication during gastrula as well as blastula stages.

In Xenopus an early morphological marker of mesodermal induction is the elongation of the mesoderm at the early gastrula stage (Symes and Smith, 1987). We show here that the elongation of equatorial (marginal) tissue is dependent on protein synthesis in a mid blastula, but has become independent of it by the late blastula stage. In animal caps induced to become mesoderm, the time when protein synthesis is required for subsequent elongation immediately follows the time of induction, and is not related to developmental stage. For elongation, intercellular communication during the blastula stage is of primary importance. Current experiments involving cell transplantation indicate a need for further cell:cell interactions during gastrulation, and therefore after the vegetal-animal induction during blastula stages. These secondary cell interactions are believed to take place among cells that have already received a vegetal induction, and may facilitate some of the later intracellular events known to accompany muscle gene activation.

Animals↗
PubMed1991

Xenopus embryos contain a somite-specific, MyoD-like protein that binds to a promoter site required for muscle actin expression.

We identify the "M region" of the muscle-specific Xenopus cardiac actin gene promoter from -282 to -348 as necessary for the embryonic expression of a cardiac actin-beta-globin reporter gene injected into fertilized eggs. Four DNA-binding activities in embryo extracts, embryonic M-region factors 1-4 (EMF1-4), are described that interact specifically with this region. One of these, EMF1, is detected in extracts from microdissected somites, which differentiate into muscle, but not in extracts from the adjacent neurectoderm, which differentiates into a variety of other cell types. Moreover, EMF1 is detected in embryo animal caps induced to form mesoderm, which includes muscle, and in which the cardiac actin gene is activated, but not in uninduced animal caps. EMF1 is also first detectable when cardiac actin transcripts begin to accumulate; therefore, both its temporal and spatial distributions during Xenopus development are consistent with a role in activating cardiac actin expression. Two lines of evidence suggest that EMF1 contains the myogenic factor Xenopus MyoD (XMyoD): (1) XMyoD synthesized in vitro can bind specifically to the same site as EMF1; and (2) antibodies raised against XMyoD bind to EMF1. DNA-binding studies indicate that EMF1 may be a complex between XMyoD and proteins found in muscle and other tissues. Our results suggest that the myogenic factor XMyoD, as a component of somite EMF1, regulates the activation of the cardiac actin gene in developing embryonic muscle by binding directly to a necessary region of the promoter.

Actins↗
PubMed1991

Xenopus Myf-5 marks early muscle cells and can activate muscle genes ectopically in early embryos.

We have cloned a Xenopus cDNA that encodes a homologue of the human myogenic factor, Myf-5. Xenopus Myf-5 (XMyf5) transcripts first accumulate in the prospective somite region of early gastrulae. The pattern of XMyf5 expression is similar to that of the Xenopus MyoD (XMyoD) gene, except that XMyf5 transcripts are largely restricted to posterior somitic mesoderm even before any somites have formed. Transient ectopic expression of XMyf5 activates cardiac actin and XMyoD genes in animal cap cells, but does not cause full myogenesis, even in combination with XMyoD. These results suggest that XMyf5 acts together with XMyoD as one of the set of genes regulating the earliest events of myogenesis, additional factors being required for complete muscle differentiation.

Amino Acid Sequence↗
PubMed1991

Gene activation in the amphibian mesoderm.

Cell potency is progressively restricted in amphibian development by a series of cellular interactions called inductions. The mesoderm is believed to develop in response to the earliest known induction, in which vegetal cells of the blastula divert overlying animal hemisphere cells away from epidermal and towards mesodermal fates. We describe two early markers of mesodermal differentiation in Xenopus laevis, both mRNAs that encode DNA-binding proteins of the helix-loop-helix family. One is a frog homologue of MyoD, a gene that in transfection experiments can convert cultured fibroblasts into myoblasts. Xenopus MyoD (XMyoD) is expressed in the early myotomes, from which the axial musculature develops. The accumulation of XMyoD RNA precedes that of transcripts from the cardiac actin gene, until now the earliest known marker of the muscle lineage, this result indicating that XMyoD could play a role in initiating muscle differentiation in normal development. We show by microinjection of synthetic RNA that XMyoD can indeed activate muscle-specific gene expression in animal cap cells, which would normally form only ectoderm. However, the XMyoD-injected animal caps did not produce differentiated muscle, suggesting that additional specific factors are required for full myogenesis. The other mRNA is a relative of the twist gene of Drosophila, which is required for mesodermal differentiation in flies. This gene (Xtwi) is expressed widely in the early frog mesoderm, but not, however, in the myotomes, where XMyoD is expressed. Later, the Xtwi gene is activated, in response to a second induction, in the developing neural crest.

Animals↗