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J M Slack

Publications and source records attributed to J M Slack.

At least 55 records · Page 3Linked to original sources
PubMed1996

The role of fibroblast growth factors in early Xenopus development.

In recent years we and others have been attempting to identify the molecular nature of the inducing signals in early Xenopus development. We have found that most members of the fibroblast growth factor (FGF) family are biologically active as mesoderm-inducing factors when applied to ectoderm from blastulae. In addition to this, they will support continued expression of the pan-mesodermal transcription factor Xbra in the mesoderm of gastrula stage embryos. We have studied the expression pattern of four types of FGF in early embryos. Two types (FGF-2 and FGF-9) are expressed maternally and are thus present at the time of natural mesoderm induction. The expression of two other types (FGF-3 and FGF-4) is activated in the newly formed mesoderm of the gastrula. If the activity of the FGF family is inhibited by overexpression of a dominant-negative FGF receptor, there is a reduction in mesoderm formation, there are abnormalities arising from an inhibition of normal gastrulation movements and there is a defect in formation of the posterior parts. We believe that the mesoderm formation and cell movement effects are attributable to loss of Xbra expression, and the posterior defects to lack of posterior HOX gene activity. Overexpression of eFGF gives rise to a posteriorized phenotype, in which posterior HOX genes are expressed in a more anterior position. We conclude that the FGF system has multiple functions in early development, including mesoderm formation, gastrulation movements and anteroposterior patterning.

Animals↗
PubMed1995

Tail bud determination in the vertebrate embryo.

BACKGROUND: Although as humans we lose our tails in the second month of embryonic development, a persistent tail is a prominent structural feature of most adult vertebrates. Indeed, the post-anal tail is part of the definition of a chordate. The internal organization of the developing tail--with neural tube, notochord and paired somites--is the same as that of the main body axis, so it can be expected that the mechanism of tail formation has a close relationship to that of the vertebrate body plan as a whole. Despite this, almost nothing is known about how tails arise. RESULTS: We present evidence to show that the tail bud of Xenopus laevis arises as the result of interactions between distinct zones of tissue at the posterior of the embryo at the neurula stage. These tissue interactions were demonstrated by manipulations of exogastrulae, which normally form no tail, and by transplantation experiments performed on the neural plate of stage 13 neurulae, whereby embryos with supernumary tails were produced. CONCLUSIONS: We propose a new model of tail bud determination, termed the NMC model, to explain the results we have obtained. In this model, the tail bud is initiated by an interaction between two territories in the neural plate and a posterior mesodermal territory.

Animals↗
PubMed1995

Characterization of v-cath, a cathepsin L-like proteinase expressed by the baculovirus Autographa californica multiple nuclear polyhedrosis virus.

Autographa californica multiple nuclear polyhedrosis virus (AcMNPV) contains a 966 bp ORF that encodes a papain type cysteine proteinase with cathepsin L-like characteristics. Using Western blot analysis of infected cell extracts we showed that v-cath proteinase has 35.5 kDa and 32 kDa precursor forms which are processed to a 27.5 kDa mature form in a manner characteristic of papain and cathepsin L. V-cath proteinase activity was greatest under acidic conditions (pH 5.0) and was reduced in the presence of the cysteine proteinase inhibitors, leupeptin and E64. Urea, a known enhancer of cathepsin L activity, also enhanced v-cath proteinase activity. AcMNPV v-cath proteinase was detected post-mortem in tissues of insects infected with wild-type (wt) virus. Insects infected with a v-cath deletion mutant did not become flaccid after death as is normally observed with wt AcMNPV infections. These findings indicate a link between v-cath activity and degradation of host tissues during virus pathogenesis.

Amino Acid Sequence↗
PubMed1995

Developmental biology of the pancreas.

The pancreas is an organ containing two distinct populations of cells, the exocrine cells that secrete enzymes into the digestive tract, and the endocrine cells that secrete hormones into the bloodstream. It arises from the endoderm as a dorsal and a ventral bud which fuse together to form the single organ. Mammals, birds, reptiles and amphibians have a pancreas with similar histology and mode of development, while in some fish, the islet cells are segregated as Brockmann bodies. Invertebrates do not have a pancreas, but comparable endocrine cells may be found in the gut or the brain. The early pancreatic bud shows uniform expression of the homeobox gene IPF-1 (also known as IDX-1, STF-1 or PDX), which when mutated to inactivity leads to total absence of the organ. The occurrence of heterotopic pancreas in the embryo, and also the metaplasias that can be displayed by a regenerating pancreas in the adult, both suggest that only a few gene products distinguish the pancreatic cell state from that of the surrounding tissues of duodenum, gall bladder and liver. In the developing pancreatic buds, the endocrine cells start to differentiate before the exocrine cells, and co-expression of different hormones by the same cell is often observed at early stages. Although pancreatic endocrine cells produce many gene products also characteristic of neurons, evidence from in vitro cultures and from quailchick grafts shows that they are of endogenous and not of neural crest origin. Observational studies suggest strongly that both endocrine and exocrine cells arise from the same endodermal rudiment. Development of the pancreas in embryonic life requires a trophic stimulus from the associated mesenchyme. In postnatal life, all cell types in the pancreas continue to grow. Destruction of acinar tissue by duct ligation or ethionine treatment is followed by rapid regeneration. Surgical removal of parts of the pancreas is followed by moderate but incomplete regeneration of both acini and islets. Poisoning with alloxan or streptozotocin can lead to permanent depletion of beta cells. Although the cell kinetics of the pancreas are not understood, it seems likely that there is a continuous slow turnover of cells, fed from a stem cells population in the ducts, and that the controls on the production rate of each cell type are local rather than systemic.

Animals↗
PubMed1995

eFGF is expressed in the dorsal midline of Xenopus laevis.

A detailed study of the expression pattern of embryonic fibroblast growth factor (eFGF) during early Xenopus development has been undertaken using whole-mount DIG in situ hybridization. We show that the zygotic expression of eFGF is activated in the mesoderm of the early gastrula and is first visualized as a ring around the blastopore, with significantly higher levels of expression on the dorsal side of the embryo. As gastrulation proceeds, eFGF transcripts become increasingly abundant in the dorsal blastopore lip. In the early neurula eFGF expression can be detected in the extreme posterior of the embryo around the closed blastopore and in the cells of the notochord. This latter result is significant and represents the first report of a Xenopus FGF that is expressed in the notochord. In addition, we show that during gastrula and neurula stages, expression of eFGF closely follows the expression of the Xenopus brachyury (Xbra) gene. During later development eFGF expression is localized to the tail-bud region and a stripe at the mid-brain/hind-brain junction. These data provide further evidence that FGFs play an important role in regulating the expression of brachyury in the developing mesoderm.

Animals↗
PubMed1994

eFGF regulates Xbra expression during Xenopus gastrulation.

We show that, in addition to a role in mesoderm induction during blastula stages, FGF signalling plays an important role in maintaining the properties of the mesoderm in the gastrula of Xenopus laevis. eFGF is a maternally expressed secreted Xenopus FGF with potent mesoderm-inducing activity. However, it is most highly expressed in the mesoderm during gastrulation, suggesting a role after the period of mesoderm induction. eFGF is inhibited by the dominant negative FGF receptor. Embryos overexpressing the dominant negative receptor show a change of behaviour of the dorsal mesoderm such that it moves around the blastopore lip instead of elongating in an antero-posterior direction. In such embryos there is a reduction in Xbra expression during gastrulation. We show that during blastula stages eFGF and Xbra are able to activate the expression of each other, suggesting that they are components of an autocatalytic regulatory loop. Moreover, we show that Xbra expression in isolated gastrula mesoderm cells is maintained by eFGF, suggesting that eFGF continues to regulate the expression of Xbra in the blastopore region. In addition, overexpression of eFGF after the mid-blastula transition results in the up-regulation of Xbra expression during gastrula stages and causes suppression of the head and enlargement of the proctodeum, which is the converse of the posterior reductions of the FGF dominant negative receptor phenotype. These data suggest an important role for eFGF in regulating the expression of Xbra and for the eFGF-Xbra regulatory pathway in the control of mesodermal cell behaviour during gastrula stages.

Animals↗
PubMed1994

Inducing factors in Xenopus early embryos.

Recent results make it possible to postulate credible candidates for each of the known inducing signals that act to determine cell fate during Xenopus early development. Experiments on biological activity, expression patterns and inhibition of function suggest that Vg-1 and Wnt-11 may act as the primary mesoderm-inducing signals, FGF and activin may serve to relay their effects, and noggin may be a major component of the dorsalizing and neural-inducing signals from the organizer.

Activins↗
PubMed1994

The Einsteck-method: position and structure of projections formed by implants of a ventral character.

The behavior of colored beads and of various living tissues after implantation into the Xenopus blastocoel is investigated. It is confirmed that the location of a graft along the anteroposterior axis depends on its intrinsic anteroposterior character. Comparison with the behavior of the beads suggests that the final position can be achieved by movement of the graft around the dorsoventral circumference of the inner marginal zone during gastrulation. Ventral marginal explants, or animal caps treated with fibroblast growth factor, both form ventral vesicles if cultured in isolation, but in the implantation experiments they often yield projections containing segmented muscle blocks. This behavior does not occur when the axis of the host has been suppressed by ultraviolet irradiation and so it is concluded that it represents dorsalization of the graft by the host. The tail-like structures formed as a result of ventral-type tissue implantations do not contain neural tissue, while the mesodermal parts are typically of mixed graft and host origin.

Animals↗
PubMed1994

Spatial and temporal expression of basic fibroblast growth factor (FGF-2) mRNA and protein in early Xenopus development.

We have analyzed the expression pattern of bFGF (FGF-2) mRNA and protein in early Xenopus development using RNAse protections, in situ hybridization and immunocytochemical methods. We find that the maternal bFGF mRNA content is at its highest in the previtellogenic oocytes (stage I-II) but decreases during further oogenesis. This low maternal level persists into the early embryonic stages and is uniformly distributed on an RNA basis. Zygotic transcription is turned on both from anterior and posterior regions but not from the middle region in the mid-neurula stage, and the expression greatly increases during the late neurula and tailbud stages. In the tadpole stage, the expression is detected in the brain, eyes, ears and neural crest-derived mesenchyme of the head. Also, it is expressed along the mesodermal axis, the level falling as the myotomes differentiate. Immunocytochemical study shows abundant bFGF protein in early oocytes and much less in later oocytes. The localization is mainly to the nuclei of the early oocytes and to the cytoplasm of the later oocytes. Localization of maternal bFGF protein in the animal hemisphere is observed in the early embryonic stages and some reuptake into nuclei occurs by the early blastula stage. The zygotically synthesized protein starts to be expressed in the anterior region of the mid-neurula stage and soon also becomes detectable in the posterior region. By tailbud and pre-larval stages, this zygotic protein appears to be present along the entire neural and mesodermal axis. When these cases are sectioned it can be seen that bFGF protein is detected in most parts of the head. In the posterior domain, it is present in the mesoderm and clearly becomes localized in the muscle cells to the nuclei and to the cell termini adjoining the intermyotomal septa.

Animals↗
PubMed1993

The zootype and the phylotypic stage.

What is it that defines an animal? The definition provided here, made on the basis of developmental biology, suggests methods for resolving phylogenetic problems.

Animals↗
PubMed1993

Embryonic induction.

The current understanding of the mechanism of embryonic induction is reviewed. The embryological data which are necessary to establish the existence of an inductive process are described and the criteria for the identification of inducing factors are discussed. These criteria comprise: a demonstration that the factor has the appropriate biological activity, that it is expressed in biologically available form at the correct time and place in the embryo, and that when it is inhibited in vivo, the interaction should fail. Current understanding of the molecular basis of competence and threshold responses is discussed. Four case studies are examined in further detail: the dorsoventral patterning in Drosophila is controlled by a gradient of the decapentaplegic gene product, a member of the TGF beta superfamily. Mesoderm induction in Xenopus embryos is thought to be controlled by several factors acting in concert: activins, fibroblast growth factors (FGFs), Wnt proteins and bone morphogenetic proteins (BMPs). The formation of the kidney in higher vertebrates involves a permissive interaction and some molecules are known to be necessary for the process but the identity of the primary inducing signal remains elusive. The anteroposterior pattern in the chick limb is controlled by a morphogen gradient emitted by the zone of polarising activity (ZPA). Although closely mimicked by retinoic acid (RA), this substance is probably not itself the morphogen. In general, the technical advances of recent years have enabled dramatic progress to be made in understanding the molecular basis of embryonic induction. Although much remains to be done, the methods of investigation are now well established.

Activins↗
PubMed1992

Intracellular signalling pathways involved in mesoderm induction by FGF.

We have examined the possible role of two signal transducing mechanisms, tyrosine phosphorylation and activation of protein kinase C (PKC), during fibroblast growth factor (FGF)-induced mesoderm induction in Xenopus. Tyrosine phosphorylation was examined through the use of a monoclonal anti-phosphotyrosine antibody. This antibody was shown to recognize the FGF receptor crosslinked to radioiodinated FGF. We also studied the response of Xenopus ectodermal explants to sodium orthovanadate, a compound that has been shown to elevate intracellular phosphotyrosine levels. Thirty percent of explants cultured in 100 microM vanadate were induced. In addition, vanadate synergized with FGF to give inductions that were more dorsal in nature than either vanadate or FGF alone. The role of PKC was evaluated by measuring PKC activity during mesoderm induction by FGF and by examining the effect of the phorbol ester 12-O-tetradecanoylphorbol 13-acetate (TPA) on explants. TPA did not induce mesoderm, however, activation of PKC was detected in FGF-treated explants. Therefore, activation of the PKC pathway alone is not sufficient for mesoderm induction. Simultaneous treatment with TPA and FGF resulted in a significant inhibition of mesoderm induction by FGF, suggesting that activation of PKC could be part of a negative feedback mechanism. In contrast, TPA had no effect on induction by activin A.

Activins↗
PubMed1992

Over-expression of fibroblast growth factors in Xenopus embryos.

A number of forms of fibroblast growth factor (FGF) were over-expressed within Xenopus embryos by injection of synthetic FGF mRNAs into fertilized eggs. Injected embryos showed abnormalities in development which were mainly secondary to a disruption of gastrulation movements. The effects observed after injection of bFGF mRNA, however, were much less severe than those observed after injection of an altered form of bFGF mRNA which differs only by the addition of a signal sequence for secretion, or of another member of the FGF family, kFGF, which is normally efficiently secreted. All forms of FGF caused the induction of mesoderm in animal cap explants isolated from blastulae, but the amount of bFGF mRNA required to induce the formation of significant levels of mesoderm was higher by a factor of over a hundred than that of the FGFs which contain a signal sequence for secretion. Over-expressed bFGF accumulated in the nuclei of blastulae but did not necessarily cause mesoderm formation. These results show that FGFs must be secreted from the cells in which they are synthesised in order to act efficiently as mesoderm inducing factors and suggest that bFGF itself, which does not contain a signal sequence for secretion, is unlikely to be directly involved in mesoderm induction during early embryonic development.

Animals↗