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PubMed1985

Information transfer during embryonic induction in amphibians.

Neural induction and differentiation has been studied using Concanavalin A, cyclic AMP, tunicamycin and calcium ionophore A 23187. Competent ectoderm of Xenopus laevis treated with Concanavalin A differentiates into neural (archencephalic) structures. Binding studies with gold-labelled ConA indicate that the superficial ectodermal layer contains fewer ConA-sensitive sites (alpha-D-mannoside and alpha-D-glucoside residues of glycoproteins) than the inner ectodermal layer. The small number of ConA-sensitive sites can be correlated with the fact that the isolated superficial ectoderm layer, in contrast to the inner layer, does not differentiate into neural structures. The gold-ConA particles bound to inner ectoderm are quickly (within 30 minutes) internalized, presumably by receptor-mediated endocytosis. However, endocytosis is not a prerequisite for neural induction. On the contrary ConA apparently must be bound to the plasma membrane for a certain period to initiate neural induction. The rapid internalization of ConA could explain why neural inductions are evoked only if ectoderm is incubated in ConA-containing medium for longer than 30 minutes. On the other hand cyclic AMP or calcium ionophore A 23187 does not elicit neural inductions. On the contrary calcium ionophore A 23187 apparently inhibits neural and mesodermal differentiation. This effect could be correlated with an increase of intracellular calcium level of the ectodermal target cells, which could influence the permeability of gap junctions resulting in a loss of cell communication, followed by a change of differentiation and pattern formation.

Amphibians↗
PubMed1978

Microheterogeneities, non-equivalance, and embryonic induction.

The thrust of this report is to stress the importance of microheterogeneities in the microenvironment of differentiating tissues as a possible inducer or regulator of differentiation. During chondrogenesis both qualitative and quantitative changes occur in the proteoglycan population. Using molecular sieve chromatography, these changes can be characterized and used as indices of differentiation. Microheterogeneities of the extracellular matrix may be an example of "non-equivalence" as a regulatory device for differentiation.

Animals↗
PubMed1985

Embryonic induction and cation concentrations in amphibian embryos.

Explanted ectoderm from early gastrulae of Triturus alpestris was treated with the Na-K ionophore gramicidin (10(-9) to 10(-5) M) and the Ca-ionophore A 23187 (10(-7) to 10(-5) M). The ectoderm developed almost exclusively to atypical epidermis as in the control explants. When the ectoderm was treated with ouabain (10(-4) M), intracellular Na+ increased about 4.4-fold and K+ was reduced by half. Mesenchyme cells in small number differentiated in about 40% of the ouabain-treated explants. The time course of total Na+ and K+ ion concentrations was measured over a period of 72 h in ectoderm of T. alpestris after induction with vegetalizing factor and in control explants. In the first 15 h after explantation, no significant differences between control and induced explants were found. Thereafter, the steady state concentration of K+ decreased in the induced explants, whereas the steady-state concentration of Na+ slightly increased. The membrane resting potential recorded intracellularly of ectoderm sandwiches from early gastrula stages was found to be -41.3 mV in control and -59.3 mV in induced explants. From the specific conductances and permeabilities of non-induced and induced cells it is concluded that the induction process leads to a differentiation of the cell membrane, which acquires the characteristics of ionic selectivity. Ectoderm from Ambystoma mexicanum forms neural or neuroid tissue, mesenchyme and melanophores after explantation in salt solution in up to 50% of the explants without any additions. Isolated Ambystoma ectoderm is therefore not suitable for test experiments.

Ambystoma↗
PubMed1992

The role of growth factors in embryonic induction in Xenopus laevis.

Establishment of the body pattern in all animals, and especially in vertebrate embryos, depends on cell interactions. During the cleavage and blastula stages in amphibians, signal(s) from the vegetal region induce the equatorial region to become mesoderm. Two types of peptide growth factors have been shown by explant culture experiments to be active in mesoderm induction. First, there are several isoforms of fibroblast growth factor (FGF), including aFGF, bFGF, and hst/kFGF. FGF induces ventral, but not the most dorsal, levels of mesodermal tissue; bFGF and its mRNA, and an FGF receptor and its mRNA, are present in the embryo. Thus, FGF probably has a role in mesoderm induction, but is unlikely to be the sole inducing agent in vivo. Second, members of the transforming growth factor-beta (TGF-beta) family. TGF-beta 2 and TGF-beta 3 are active in induction, but the most powerful inducing factors are the distant relatives of TGF-beta named activin A and activin B, which are capable of inducing all types of mesoderm. An important question relates to the establishment of polarity during the induction of mesoderm. While all regions of the animal hemisphere of frog embryos are competent to respond to activins by mesoderm differentiation, only explants that include cells close to the equator form structures with some organization along dorsoventral and anteroposterior axes. These observations suggest that cells in the blastula animal hemisphere are already polarized to some extent, although inducers are required to make this polarity explicit.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1976

Fish swimbladder: an excellent mesodermal inductor in primary embryonic induction.

Swimbladder of the crucian carp, Carassius auratus, was found to be better as a vegatalizing tissue than other tissues, such as guinea-pig bone marrow, when presumptive ectoderm of Triturus gastrulae was used as reacting tissue. Swimbladder usually induced assemblies of highly organized mesodermal tissues, such as notochord, somites and pronephric tubules, some of which were covered by mesodermal epithelium without any epidermal covering. A special character of the effect of swimbladder was the rather frequent induction of solid balls of undifferentiated cells, which were identified as mesodermal or mesodermal and probably endodermal. These findings show that swimbladder has a strong and fast spreading vegetalizing effect on the responding presumptive ectoderm.

Air Sacs↗
PubMed1992

Embryonic lens induction: shedding light on vertebrate tissue determination.

The principle of embryonic induction was defined by early studies of lens determination, and because of the relative simplicity of the developing lens and its interaction with presumptive retinal tissue it has been a favored system for examining mechanisms of induction. Recent studies have led to substantial alterations of the classic model for this process, introducing several elements that significantly refine our view of vertebrate tissue determination.

Animals↗
PubMed1989

Extracellular matrix organized in embryonic cavities during induction of the embryonic axis in chick embryo.

Extracellular matrix (ECM) is detected as short, disorganized fibrils in the forming embryonic extracellular spaces shortly prior to the first morphogenetic cellular movements and interactions in the early chick embryo. As development progresses, the ECM is organized into an intricate network spanning the embryonic cavities. This dynamic entity undergoes relatively rapid changes in its organization pattern during the developmental period from morula to the induction of the neural plate. The ECM seems to preserve the exquisite architecture of the embryo and could guide migrating cells into defined pathways in the early embryo.

Animals↗
PubMed1989

Embryonic lens induction: more than meets the optic vesicle.

The classic model of lens induction stipulated that the optic vesicle is both a necessary and sufficient inductor of the lens in amphibian development. Although this view has subsequently been modified to encompass the contributions of earlier inductors, such as the involuting endo-mesoderm, it is still widely reported that the optic vesicle alone is able to elicit lens formation from ectoderm. Recent work, which has employed a host- and donor-marking scheme, has shown the optic vesicle to be a weak inductor of the lens, involved only in the final rather than the initial phases of determination. In addition, a review of the literature substantiates this conclusion since many of the transplantation experiments arguing for the sufficiency of the optic vesicle are characterized by the lack of adequate criteria for judging the authenticity of the resulting lens responses, particularly the absence of a host- and donor-marking strategy. This analysis of the literature, together with our own results, lead us to propose a new model of lens determination in which tissue interactions during gastrulation are required to confer a lens-forming bias upon a large area of head ectoderm allowing the optic vesicle to induce lens formation in a defined area of this primed ectoderm. Data from studies on mesoderm and neural induction are also beginning to suggest a multistep model involving the initial establishment of bias and subsequent interactions resulting in determination, and we propose that this framework will serve as a general paradigm for embryonic induction.

Amphibians↗
PubMed1992

Interaction between growth factors and retinoic acid in the induction of kidney tubulogenesis in tissue culture.

Kidney tubulogenesis is the initial step in renal organogenesis. The precise molecular determinants of this pattern formation are presently unknown, although soluble factors, such as growth factors, and insoluble factors, such as extracellular matrix molecules, most likely play fundamental roles in this process. To define the molecular determinants of renal proximal tubule morphogenesis, primary cultures of rabbit renal proximal tubule cells in hormonally defined, serum-free media were treated with transforming growth factor-beta 1 (TGF-beta 1), epidermal growth factor (EGF), and the retinoid, all trans-retinoic acid (RA), singly or in combination. Utilizing phase contrast and light and transmission electron microscopy, the simultaneous administration of TGF-beta 1 (10 ng/ml), EGF (1 nM), and RA (0.1 nM) transformed a confluent monolayer of renal proximal tubule cells within 5 to 6 days into three-dimensional cell aggregates containing lumens within the interior of the cell clusters. The lumens were bordered by tubule cells possessing a polarized epithelial cell phenotype with extensive microvilli formation and tight junctional complexes along the luminal border. All three factors were necessary and sufficient to induce this phenotypic transformation. Further studies demonstrated that RA promoted the deposition of the A and B1 chains of laminin, a cell attachment protein of the basement membrane, in a small subset of proximal tubule cells in culture, as deduced by indirect immunofluorescent microscopy. Additional studies demonstrated that soluble purified laminin fully substituted for RA in this system to promote renal tubulogenesis when combined with TGF-beta 1 and EGF. These results demonstrate that the growth factors, TGF-beta 1 and EGF, and the retinoid, RA, promote tubulogenesis in adult renal proximal tubule cells in tissue culture in a manner reminiscent of inductive embryonic kidney morphogenesis. These observations define a coordinated interplay between growth factors and retinoids to induce pattern formation and morphogenesis. Furthermore, the demonstration of RA-induced laminin deposition as a critical event in this morphogenic process identifies laminin as a possible target protein for RA to act as a morphogen.

Animals↗
PubMed1991

Induction of tubules in rat metanephrogenic mesenchyme in the absence of an inductive tissue.

Differentiation of metanephrogenic mesenchyme to renal tubular epithelium requires induction by the ureteric bud in vivo or any of several embryonic tissues in vitro. In an effort to eliminate the tissue requirement in embryonic induction, extracellular matrices and soluble factors were analyzed individually or in combination for their ability to stimulate tubulogenesis in uninduced metanephrogenic mesenchyme from 13-gestation-day rat embryos. These evaluations have established that pituitary extract and epidermal growth factor (EGF) in concert with a matrix can promote morphogenesis of mesenchymal rudiments in culture. While type I collagen, laminin, or fibronectin matrices all promoted tubulogenesis in the presence of pituitary extract and EGF, type IV collagen proved the most effective. Under these conditions, tubules were induced in 23/24 mesenchymal rudiments by 9 days in culture. Mesenchyme was not induced prior to explanation since it formed no tubules when cultured in a medium that allowed tubulogenesis in intact embryonic kidneys. Preliminary characterization of the undefined factor in pituitary extract was consistent with a protein of molecular weight greater than 100,000 but less than 300,000. When uninduced metanephrogenic mesenchyme from mouse was used instead of rat tissue, a similar pattern of morphogenesis was not observed, suggesting that the described medium is inappropriate for promoting differentiation in mouse or, less likely, that different mechanisms mediate differentiation in rat and mouse. These studies show that embryonic induction can occur in explanted rat renal mesenchyme in an appropriate environment and does not require the presence of an inductive tissue.

Animals↗
PubMed1988

The origin of cell-type differences in early embryos.

Differences between cells first arise in embryonic development by two principal mechanisms. One is the asymmetric distribution of cytoplasmic substances at mitosis in eggs and early embryos. The other is cell interaction or embryonic induction. Certain aspects of these major mechanisms are considered, and emphasis is placed on the value of molecular markers. The effects of unequal cell division on the concentration of cytoplasmic determinants are discussed. In embryonic induction, the nature and timing of response is determined more by properties of the responding tissue than by those of the inducing molecules. Possible future directions of work are discussed in relation to experience with amphibian eggs and oocytes.

Animals↗
PubMed1992

Lithium-sensitive production of inositol phosphates during amphibian embryonic mesoderm induction.

Mesoderm induction and body axis determination in frog (Xenopus) embryos are thought to involve growth factor-mediated cell-cell signaling, but the signal transduction pathways are unknown. Li+, which inhibits the polyphosphoinositide (PI) cycle signal transduction pathway in many cells, also disrupts axis determination and mesoderm induction. Amounts of the PI cycle-derived second messenger, inositol 1,4,5-trisphosphate, increased during mesoderm induction in normal embryos; addition of Li+ inhibited the embryonic inositol monophosphatase and reversed this increase. Embryonic PI cycle activity thus shows characteristics that indicate it may function in mesoderm induction and axis determination.

Animals↗
PubMed1988

Reinvestigation of the role of the optic vesicle in embryonic lens induction.

The induction of the lens by the optic vesicle in amphibians is often cited as support for the view that a single inductive event can lead to determination in a multipotent tissue. This conclusion is based on transplantation experiments whose results indicate that many regions of embryonic ectoderm which would normally form epidermis can form a lens if brought into contact with the optic vesicle. Although additional evidence argues that during normal development other tissues, acting before the optic vesicle, also contribute to lens induction, it is still widely held, on the basis of these transplantation experiments, that the optic vesicle alone can elicit lens formation in ectoderm. While testing this conclusion by transplanting optic vesicles beneath ventral ectoderm in Xenopus laevis embryos, it became apparent that contamination of optic vesicles by presumptive lens ectoderm cells can generate lenses in these experiments, illustrating the need for adequate host and donor marking procedures. Since previous studies rarely used host and donor marking, it was not clear whether they actually demonstrated that the optic vesicle can induce lenses. Using careful host and donor marking procedures with horseradish peroxidase as a lineage tracer, we show that the optic vesicle cannot stimulate lens formation in neurula- or gastrula-stage ectoderm of Xenopus laevis. Since the general conclusion that the optic vesicle is sufficient for lens induction rests on studies in many organisms, we felt it was important to begin to test this conclusion in other amphibians as well. Similar experiments were therefore performed with Rana Palustris embryos, since it was in this organism that optic vesicle transplant studies had originally argued that this tissue alone can cause lens induction. Under conditions similar to those used in the original report, but with careful controls to assess the origin of lenses in transplants, we found that the optic vesicle alone cannot elicit lens formation. Our data lead us to propose that the optic vesicle in amphibians is not generally sufficient for lens induction. Instead, we argue that lens induction occurs by a multistep process in which an essential phase in lens determination occurs as a result of inductive interactions preceding contact of ectoderm with the optic vesicle.

Animals↗
PubMedDate not supplied

A community effect in animal development.

In animal development, the first tissues to be formed include such major components as muscle, nerve cord, notochord and the eye. In the vertebrates, all of these tissues are formed by embryonic induction, a process by which some of the cells within a mass of tissue are caused to change their direction of differentiation as a result of close proximity to cells of another kind. The induced cells typically form a solid coherent mass with a distinct border between them and the remaining uninduced cells. This clean separation between induced and uninduced cells is much sharper than can readily be explained as a result of the induction process. We describe here the culture of amphibian cell and tissue recombinations in solid gels containing cytochalasin in which cell division and cell movement is inhibited during response to induction. This has revealed an effect in which the ability of a cell to respond to induction by differentiating as muscle is enhanced by, or even dependent on, other neighbouring cells differentiating in the same way at the same time. This seems to be a newly described process in animal development, termed the community effect. It helps to explain the formation of blocks of tissue from sheets of cells, and could be of widespread occurrence and significance in morphogenesis resulting from embryonic induction.

Animals↗
PubMedDate not supplied

[The role of inducing factors in early embryonic development].

In the field of early embryonic induction and differentiation we can observe an exponential increase of research activities over the last three years. The reasons for this rapid exploration are the application of powerful techniques of molecular biology and molecular genetics and the recent accumulation of knowledge about the close functional correlation between growth factors, embryonic induction factors, the products of oncogenes (or proto-oncogenes), and transcription factors. The highly probable role of the cellular and viral oncogenes in regulation of the differentiation and function of normal and malignant cells has stimulated the interest of scientists working on the molecular basis of malignant cell transformation. An excellent model to study mesoderm and neural induction and differentiation on the cellular and molecular level is the embryo of the south African clawed frog (Xenopus laevis), which is now a favored vertebrate system in many laboratories.

Animals↗
PubMed1979

Tissue differentiation and susceptibility to embryonal tumor induction by ethylnitrosourea in the opossum.

Opossums (Didelphis virginiana Kerr) exposed to 100 mg ENU/kg in single or incremental doses early in postnatal life developed a spectrum of epithelial and mesenchymal neoplasms including several types of embryonal neoplasms not previously induced in laboratory animals. A correlation was apparent to a varying degree between susceptibility to tumor induction and the state of morphologic maturation of the presumed target tissues at the light microscopic level for embryonal tumors of the eye, kidney, and brain. The susceptibility of the opossum eye to an ENU-induced intraocular teratoid medulloepithelioma extended over the period from 1 to between 3 and 4 weeks of age and was correlated with the differentiation of the apparent target cell, the nonpigmented ciliary epithelium of the pars ciliaris retinae. Induction of nephroblastomas was correlated with the presence in the kidney of stem cells (metanephric blastema) through the period from birth to between 6 and 8 weeks of age. Although susceptibility of the opossum brain to ENU induction of gangliogliomas was correlated with the state of differentiation of the germinal matrix from birth to 56 days of age, induction of these tumors was essentially limited to the 1st week postpartum. No definite correlation between vulnerability to tumor induction and tissue maturation was evident for a tumor of the jaw (ameloblastoma) with presumed origin from embryonic dental remnants. Our results indicated that the opossium early in postnatal life is a useful model for the induction and characterization of certain of the major dysontogenetic tumors, which have been difficult or impossible to reproduce in the traditional laboratory species.

Age Factors↗
PubMed1989

The Florey lecture, 1988. From egg to embryo: the initiation of cell differentiation in Amphibia.

Some of the principles by which different cell types first arise at the beginning of animal development are illustrated by muscle cell formation in Amphibia. If the nucleus of a differentiated muscle cell is transplanted to an enucleated egg, some of the resulting embryos develop into tadpoles with a wide range of normally differentiated cells. These experiments show that genes undergo major changes in activity as a response to components of egg cytoplasm. Two fundamental mechanisms account for the regional activation of genes in early embryos. One involves the effect of localized 'determinants' in egg cytoplasm, and the other concerns cell interactions or embryonic induction. Both these mechanisms seem to be responsible for muscle cell formation in amphibian development. The old problem of embryonic induction has recently become accessible to analysis at the molecular level, especially in the case of the mesoderm or muscle-forming induction. This has been greatly facilitated by using a sensitive and quantitative assay to detect the first transcripts of muscle genes a few hours after the start of induction. The role of early events and of interactions among like cells during response to induction is discussed. In analysing specific gene activation following induction, DNA injection into fertilized eggs has shown that a very small part of the cardiac actin gene promoter is sufficient to enable it to respond to induction. Although the experimental work summarized here has been done on amphibian embryos, which are more suitable than other embryos for embryological manipulation, the conclusions reached are believed to be generally applicable to the development of other organisms.

Amphibians↗