The International journal of developmental biology, 2003
I have taught developmental biology in Essen for 30 years. Since my department is named Zoophysio... more I have taught developmental biology in Essen for 30 years. Since my department is named Zoophysiologie (Zoophysiology), besides Developmental Biology, I also have to teach General Animal Physiology. This explains why the time for teaching developmental biology is restricted to a lecture course, a laboratory course and several seminar courses. However, I also try to demonstrate in the lecture courses on General Physiology the close relationship between developmental biology, physiology, morphology, anatomy, teratology, carcinogenesis, evolution and ecology (importance of environmental factors on embryogenesis). Students are informed that developmental biology is a core discipline of biology. In the last decade, knowledge about molecular mechanisms in different organisms has exponentially increased. The students are trained to understand the close relationship between conserved gene structure, gene function and signaling pathways, in addition to or as an extension of, classical concep...
The International journal of developmental biology, 1994
Mesoderm formation in the amphibian embryo is thought to be induced in the ectoderm of the animal... more Mesoderm formation in the amphibian embryo is thought to be induced in the ectoderm of the animal region by signals emanating from the endoderm of the vegetal region after cleavage up to the mid-blastula. During this process the dorsal vegetal zone is thought to stimulate the dorsal animal zone to establish the Spemann organizer, which will in turn trigger the overlaying neuroectoderm during gastrulation resulting in the development of the central nervous system. In this concept it is assumed that the animal hemisphere is an uncommitted area, which receives its instructions from the vegetal region of the embryo. However, the experiments of this paper show that the 4 animal blastomeres of the eight-cell stage will form dorsal mesodermal structures in over 50% of the cases. The results support the view that developmental determinants are distributed in distinct gradients already in the early cleavage stages and that in the embryo the mesoderm is determined by factors prelocalized in t...
A central topic of embryology is the establishment of the body plan during embryogenesis. Startin... more A central topic of embryology is the establishment of the body plan during embryogenesis. Starting with maternal factors distributed in the early cleavage stages in distinct patterns and gradients cell-to-cell interactions including early embryonic induction result in the formation of mesoderm and the organizer area. While many facts are known about the role of growth factors like activin (closely related to the vegetalizing factor), processed Vg1, BMPs and FGF for mesoderm formation, the establishment of the central nervous system is not yet well understood. However, there is growing evidence that neural induction is a multistep process at the level of the dorsal mesoderm (organizer) and the reacting neuroectoderm. Therefore the existence of only one neuralizing factor is unlikely. We report about data that follistatin protein is not a direct neural inducer. Furthermore our comparative studies of Xenopus and Triturus exogastrulae indicate that planar signals are unlikely in the Tri...
Neural induction and differentiation has been studied using Concanavalin A, cyclic AMP, tunicamyc... more 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 (α-D-mannoside and α-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 rap...
Neural (archencephalic) structures have been evoked in the competent ectoderm (consisting of both... more Neural (archencephalic) structures have been evoked in the competent ectoderm (consisting of both ectodermal layers) of Xenopus laevis by treatment with Concanavalin A (Con A), which probably acts on the plasma membrane. The size of the neural structures is increased when the ectoderm is incubated in Cytochalasin B prior to the Con A treatment. The results indicate that Cytochalasin B could have an influence on the binding of Con A to receptors on the plasma membrane. On the other hand, Cytochalasin B seems to have an inhibitory effect on the action of the vegetalizing factor, which could be correlated with the decline of endocytotic processes and internalization. In further series, it could he shown that the isolated superficial ectoderm, in contrast to the inner ectoderm layer, does not react to Con A treatment with the differentiation of neural structures. Studies with FITC-Con A indicate that the marker binds less to the outer ectoderm than to the inner ectoderm layer. However, by xenoplastic combinations of the outer ectoderm layer of X. /aev/s as reacting tissue and chordamesoderm of Triturus vulgaris as inducer, it could be demonstrated that the superficial layer, which in normogenesis does not come into contact with the inducing chordamesoderm but forms the ependymal part of the brain only, is also able to form archencephalic brain structures under in vitro conditions.
The Role of Cell Interactions in Early Neurogenesis, 1984
Spemann’s and Hilde Mangold’s famous transplantation experiment[1] showed that in amphibians comp... more Spemann’s and Hilde Mangold’s famous transplantation experiment[1] showed that in amphibians competent ectoderm (presumptive epidermis) could be triggered to differentiate into neural tissues by the upper blastopore lip of early amphibian gastrulae. Spemann entitled this area with inducing activity as the organisator (organizer), because it organizes the formation of the central nervous system. In the following decades the interest of embryologists has been focused on the question, which factors located in the upper blastopore lip are responsible for the process of primary embryonic induction. Morphogenetic factors, which induce in competent ectoderm the formation of endodermal, mesodermal and neural derivatives, could be isolated from different sources[2–13]. A vegetalizing factor could be isolated in highly purified form from chicken embryos[14,15]. It is now generally accepted that these factors, which in contrast to growth factors can be entitled as determination factors, are protein in nature.
In the field of early embryonic induction and differentiation we can observe an exponential incre... more 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.
Proceedings of the National Academy of Sciences, 1988
Development of the nervous system in the amphibian embryo is initiated during gastrulation by an ... more Development of the nervous system in the amphibian embryo is initiated during gastrulation by an inductive interaction between chordamesoderm and dorsal ectoderm. The induced ectoderm forms the neural plate while uninduced ectoderm generates epidermis. We screened for genes activated during gastrulation and expressed specifically in the nervous system of Xenopus laevis in the expectation that clones representing such genes will constitute useful markers for the study of early neurogenesis. Probes were prepared from adult brain RNA by subtraction with RNA from ovary and from different combinations of adult kidney, muscle, and skin; cDNA libraries prepared from early to late neurula embryo RNA were screened with these probes. Six clones were chosen for further study. Three of these clones are not represented in the maternal RNA population but are activated at the late gastrula stage; the other three increase from a maternal base. Expression of five of the genes is restricted to the ne...
DNA from chicken embryo nucleosome tetramers (about 760 base pairs in size) was enriched for tRNA... more DNA from chicken embryo nucleosome tetramers (about 760 base pairs in size) was enriched for tRNA genes by RPC-5 chromatography. The enriched DNA was hybridized with chicken embryo total tRNA and the hybridized DNA isolated utilizing a) avidinbiotin interaction, b) diazobenzyloxymethyl paper, and c) high temperature RPC-5 chromatography. The obtained single stranded DNA highly enriched for tRNA complementary sequences was hybridized with total DNA from nucleosome monomers (140-190 base pairs in size) and the excess of non hybridized monomer nucleosome DNA removed by Sepharose kB chromatography. The hybrid molecules obtained were made fully double stranded by incubation with E.coli DNA polymerase I, DNA ligase, and exonuclease III. DNA was inserted into plasmid pBR322 by G-C joining procedure and the recombinant DNA used to transform the E.coli strain X'77^. More than 70$ of the transformants obtained hybridize to chicken embryo total tRNA.
Suramin, a polyanionic compound, which has previously shown to dissociate platelet derived growth... more Suramin, a polyanionic compound, which has previously shown to dissociate platelet derived growth factor (PDGF) from its receptor, prevents the differentiation of neural (brain) structures of recombinants of dorsal blastopore lip (Spemann's organizer) and competent neuroectoderm. Furthermore, the suramin treatment changes the prospective differentiation pattern of isolated blastopore lip. While untreated dorsal blastopore lip will differentiate into dorsal mesodermal structures (notochord and somites), suramin treated dorsal blastopore lip will form ventral mesoderm structures, especially heart structures. The results are discussed in the context of the current opinion about the mode of action of different growth factor superfamilies.
Page 1. Review Gene expression and pattern formation during early embryonic development in amphib... more Page 1. Review Gene expression and pattern formation during early embryonic development in amphibians HORST GRUNZ Universitiit GH Essen, FB 9 Abteilung Zoophysiologie, Universitiitsstr. 5, 45117 Essen, Germany (Fax, 49-201 183-4197; Email, h.grunz@uni-essen. ...
Mammalian embryonic stem cells can be obtained from the inner cell mass of blastocysts or from pr... more Mammalian embryonic stem cells can be obtained from the inner cell mass of blastocysts or from primordial germ cells. These stem cells are pluripotent and can develop into all three germ cell layers of the embryo. Somatic mammalian stem cells, derived from adult or fetal tissues, are more restricted in their developmental potency. Amphibian ectodermal and endodermal cells lose their pluripotency at the early gastrula stage. The dorsal mesoderm of the marginal zone is determined before the mid-blastula transition by factors located after cortical rotation in the marginal zone, without induction by the endoderm. Secreted maternal factors (BMP, FGF and activins), maternal receptors and maternal nuclear factors (-catenin, Smad and Fast proteins), which form multiprotein transcriptional complexes, act together to initiate pattern formation. Following mid-blastula transition in Xenopus laevis (Daudin) embryos, secreted nodal-related (Xnr) factors become important for endoderm and mesoderm differentiation to maintain and enhance mesoderm induction. Endoderm can be induced by high concentrations of activin (vegetalizing factor) or nodal-related factors, especially Xnr5 and Xnr6, which depend on Wnt/-catenin signaling and on VegT, a vegetal maternal transcription factor. Together, these and other factors regulate the equilibrium between endoderm and mesoderm development. Many genes are activated and/or repressed by more than one signaling pathway and by regulatory loops to refine the tuning of gene expression. The nodal related factors, BMP, activins and Vg1 belong to the TGF- superfamily. The homeogenetic neural induction by the neural plate probably reinforces neural induction and differentiation. Medical and ethical problems of future stem cell therapy are briefly discussed.
Wilhelm Roux's Archives of Developmental Biology, 1979
Early amphibian gastrula ectoderm (Triturus alpestris) has been treated with vegetalizing factor.... more Early amphibian gastrula ectoderm (Triturus alpestris) has been treated with vegetalizing factor. While normal sandwiches (animal caps of two eggs) differentiated mainly into endoderm derived tissues, giant-sandwiches (a combination of 8 animal caps) formed mesodermal and neural tissues in addition. The results support the interpretation that ectoderm will differentiate into endoderm derived tissues when all or nearly all cells are induced (presumably depending on certain threshold concentrations of the inducer). This is the case in the normal sandwich after treatment with high concentrations of vegetalizing factor for 24 h. However, in a giantsandwich it must be assumed that only the cells in the vicinity of the inducer will be triggered to differentiate into endoderm derived tissues. Mesodermal structures will be formed by secondary interactions between the induced ectoderm (endoderm) and non induced ectodermal cells. The induction of neural structures could be explained as a further interaction between mesodermalized and non induced ectodermal cells. This chain of events is compared with the steps of determination in normogenesis.
The International journal of developmental biology, 2003
I have taught developmental biology in Essen for 30 years. Since my department is named Zoophysio... more I have taught developmental biology in Essen for 30 years. Since my department is named Zoophysiologie (Zoophysiology), besides Developmental Biology, I also have to teach General Animal Physiology. This explains why the time for teaching developmental biology is restricted to a lecture course, a laboratory course and several seminar courses. However, I also try to demonstrate in the lecture courses on General Physiology the close relationship between developmental biology, physiology, morphology, anatomy, teratology, carcinogenesis, evolution and ecology (importance of environmental factors on embryogenesis). Students are informed that developmental biology is a core discipline of biology. In the last decade, knowledge about molecular mechanisms in different organisms has exponentially increased. The students are trained to understand the close relationship between conserved gene structure, gene function and signaling pathways, in addition to or as an extension of, classical concep...
The International journal of developmental biology, 1994
Mesoderm formation in the amphibian embryo is thought to be induced in the ectoderm of the animal... more Mesoderm formation in the amphibian embryo is thought to be induced in the ectoderm of the animal region by signals emanating from the endoderm of the vegetal region after cleavage up to the mid-blastula. During this process the dorsal vegetal zone is thought to stimulate the dorsal animal zone to establish the Spemann organizer, which will in turn trigger the overlaying neuroectoderm during gastrulation resulting in the development of the central nervous system. In this concept it is assumed that the animal hemisphere is an uncommitted area, which receives its instructions from the vegetal region of the embryo. However, the experiments of this paper show that the 4 animal blastomeres of the eight-cell stage will form dorsal mesodermal structures in over 50% of the cases. The results support the view that developmental determinants are distributed in distinct gradients already in the early cleavage stages and that in the embryo the mesoderm is determined by factors prelocalized in t...
A central topic of embryology is the establishment of the body plan during embryogenesis. Startin... more A central topic of embryology is the establishment of the body plan during embryogenesis. Starting with maternal factors distributed in the early cleavage stages in distinct patterns and gradients cell-to-cell interactions including early embryonic induction result in the formation of mesoderm and the organizer area. While many facts are known about the role of growth factors like activin (closely related to the vegetalizing factor), processed Vg1, BMPs and FGF for mesoderm formation, the establishment of the central nervous system is not yet well understood. However, there is growing evidence that neural induction is a multistep process at the level of the dorsal mesoderm (organizer) and the reacting neuroectoderm. Therefore the existence of only one neuralizing factor is unlikely. We report about data that follistatin protein is not a direct neural inducer. Furthermore our comparative studies of Xenopus and Triturus exogastrulae indicate that planar signals are unlikely in the Tri...
Neural induction and differentiation has been studied using Concanavalin A, cyclic AMP, tunicamyc... more 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 (α-D-mannoside and α-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 rap...
Neural (archencephalic) structures have been evoked in the competent ectoderm (consisting of both... more Neural (archencephalic) structures have been evoked in the competent ectoderm (consisting of both ectodermal layers) of Xenopus laevis by treatment with Concanavalin A (Con A), which probably acts on the plasma membrane. The size of the neural structures is increased when the ectoderm is incubated in Cytochalasin B prior to the Con A treatment. The results indicate that Cytochalasin B could have an influence on the binding of Con A to receptors on the plasma membrane. On the other hand, Cytochalasin B seems to have an inhibitory effect on the action of the vegetalizing factor, which could be correlated with the decline of endocytotic processes and internalization. In further series, it could he shown that the isolated superficial ectoderm, in contrast to the inner ectoderm layer, does not react to Con A treatment with the differentiation of neural structures. Studies with FITC-Con A indicate that the marker binds less to the outer ectoderm than to the inner ectoderm layer. However, by xenoplastic combinations of the outer ectoderm layer of X. /aev/s as reacting tissue and chordamesoderm of Triturus vulgaris as inducer, it could be demonstrated that the superficial layer, which in normogenesis does not come into contact with the inducing chordamesoderm but forms the ependymal part of the brain only, is also able to form archencephalic brain structures under in vitro conditions.
The Role of Cell Interactions in Early Neurogenesis, 1984
Spemann’s and Hilde Mangold’s famous transplantation experiment[1] showed that in amphibians comp... more Spemann’s and Hilde Mangold’s famous transplantation experiment[1] showed that in amphibians competent ectoderm (presumptive epidermis) could be triggered to differentiate into neural tissues by the upper blastopore lip of early amphibian gastrulae. Spemann entitled this area with inducing activity as the organisator (organizer), because it organizes the formation of the central nervous system. In the following decades the interest of embryologists has been focused on the question, which factors located in the upper blastopore lip are responsible for the process of primary embryonic induction. Morphogenetic factors, which induce in competent ectoderm the formation of endodermal, mesodermal and neural derivatives, could be isolated from different sources[2–13]. A vegetalizing factor could be isolated in highly purified form from chicken embryos[14,15]. It is now generally accepted that these factors, which in contrast to growth factors can be entitled as determination factors, are protein in nature.
In the field of early embryonic induction and differentiation we can observe an exponential incre... more 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.
Proceedings of the National Academy of Sciences, 1988
Development of the nervous system in the amphibian embryo is initiated during gastrulation by an ... more Development of the nervous system in the amphibian embryo is initiated during gastrulation by an inductive interaction between chordamesoderm and dorsal ectoderm. The induced ectoderm forms the neural plate while uninduced ectoderm generates epidermis. We screened for genes activated during gastrulation and expressed specifically in the nervous system of Xenopus laevis in the expectation that clones representing such genes will constitute useful markers for the study of early neurogenesis. Probes were prepared from adult brain RNA by subtraction with RNA from ovary and from different combinations of adult kidney, muscle, and skin; cDNA libraries prepared from early to late neurula embryo RNA were screened with these probes. Six clones were chosen for further study. Three of these clones are not represented in the maternal RNA population but are activated at the late gastrula stage; the other three increase from a maternal base. Expression of five of the genes is restricted to the ne...
DNA from chicken embryo nucleosome tetramers (about 760 base pairs in size) was enriched for tRNA... more DNA from chicken embryo nucleosome tetramers (about 760 base pairs in size) was enriched for tRNA genes by RPC-5 chromatography. The enriched DNA was hybridized with chicken embryo total tRNA and the hybridized DNA isolated utilizing a) avidinbiotin interaction, b) diazobenzyloxymethyl paper, and c) high temperature RPC-5 chromatography. The obtained single stranded DNA highly enriched for tRNA complementary sequences was hybridized with total DNA from nucleosome monomers (140-190 base pairs in size) and the excess of non hybridized monomer nucleosome DNA removed by Sepharose kB chromatography. The hybrid molecules obtained were made fully double stranded by incubation with E.coli DNA polymerase I, DNA ligase, and exonuclease III. DNA was inserted into plasmid pBR322 by G-C joining procedure and the recombinant DNA used to transform the E.coli strain X'77^. More than 70$ of the transformants obtained hybridize to chicken embryo total tRNA.
Suramin, a polyanionic compound, which has previously shown to dissociate platelet derived growth... more Suramin, a polyanionic compound, which has previously shown to dissociate platelet derived growth factor (PDGF) from its receptor, prevents the differentiation of neural (brain) structures of recombinants of dorsal blastopore lip (Spemann's organizer) and competent neuroectoderm. Furthermore, the suramin treatment changes the prospective differentiation pattern of isolated blastopore lip. While untreated dorsal blastopore lip will differentiate into dorsal mesodermal structures (notochord and somites), suramin treated dorsal blastopore lip will form ventral mesoderm structures, especially heart structures. The results are discussed in the context of the current opinion about the mode of action of different growth factor superfamilies.
Page 1. Review Gene expression and pattern formation during early embryonic development in amphib... more Page 1. Review Gene expression and pattern formation during early embryonic development in amphibians HORST GRUNZ Universitiit GH Essen, FB 9 Abteilung Zoophysiologie, Universitiitsstr. 5, 45117 Essen, Germany (Fax, 49-201 183-4197; Email, h.grunz@uni-essen. ...
Mammalian embryonic stem cells can be obtained from the inner cell mass of blastocysts or from pr... more Mammalian embryonic stem cells can be obtained from the inner cell mass of blastocysts or from primordial germ cells. These stem cells are pluripotent and can develop into all three germ cell layers of the embryo. Somatic mammalian stem cells, derived from adult or fetal tissues, are more restricted in their developmental potency. Amphibian ectodermal and endodermal cells lose their pluripotency at the early gastrula stage. The dorsal mesoderm of the marginal zone is determined before the mid-blastula transition by factors located after cortical rotation in the marginal zone, without induction by the endoderm. Secreted maternal factors (BMP, FGF and activins), maternal receptors and maternal nuclear factors (-catenin, Smad and Fast proteins), which form multiprotein transcriptional complexes, act together to initiate pattern formation. Following mid-blastula transition in Xenopus laevis (Daudin) embryos, secreted nodal-related (Xnr) factors become important for endoderm and mesoderm differentiation to maintain and enhance mesoderm induction. Endoderm can be induced by high concentrations of activin (vegetalizing factor) or nodal-related factors, especially Xnr5 and Xnr6, which depend on Wnt/-catenin signaling and on VegT, a vegetal maternal transcription factor. Together, these and other factors regulate the equilibrium between endoderm and mesoderm development. Many genes are activated and/or repressed by more than one signaling pathway and by regulatory loops to refine the tuning of gene expression. The nodal related factors, BMP, activins and Vg1 belong to the TGF- superfamily. The homeogenetic neural induction by the neural plate probably reinforces neural induction and differentiation. Medical and ethical problems of future stem cell therapy are briefly discussed.
Wilhelm Roux's Archives of Developmental Biology, 1979
Early amphibian gastrula ectoderm (Triturus alpestris) has been treated with vegetalizing factor.... more Early amphibian gastrula ectoderm (Triturus alpestris) has been treated with vegetalizing factor. While normal sandwiches (animal caps of two eggs) differentiated mainly into endoderm derived tissues, giant-sandwiches (a combination of 8 animal caps) formed mesodermal and neural tissues in addition. The results support the interpretation that ectoderm will differentiate into endoderm derived tissues when all or nearly all cells are induced (presumably depending on certain threshold concentrations of the inducer). This is the case in the normal sandwich after treatment with high concentrations of vegetalizing factor for 24 h. However, in a giantsandwich it must be assumed that only the cells in the vicinity of the inducer will be triggered to differentiate into endoderm derived tissues. Mesodermal structures will be formed by secondary interactions between the induced ectoderm (endoderm) and non induced ectodermal cells. The induction of neural structures could be explained as a further interaction between mesodermalized and non induced ectodermal cells. This chain of events is compared with the steps of determination in normogenesis.
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