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H Grunz

Publications and source records attributed to H Grunz.

47 records · Page 3Linked to original sources
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↗
PubMed1985

Effect of concanavalin A and vegetalizing factor on the outer and inner ectoderm layers of early gastrulae of Xenopus laevis after treatment with cytochalasin B.

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 be 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. laevis as reacting tissue and chordamesoderm of Triturus vulgaris as inducer, it could be demonstrated that the superficial layer, which is 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.

Animals↗
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↗
PubMed1983

Effects of inducers on inner and outer gastrula ectoderm layers of Xenopus laevis.

Gastrula ectoderm, isolated from Xenopus laevis, was cultured in Holtfreter solution or modified Leibovitz medium (L-15) by the sandwich-method with or without inducer. The ectoderm (SD cell layers) consisted of two cell sheets, representing a superficial (S) and a deep (D) layer. In the L-15 medium rather than in Holtfreter solution, the two cell layers separate out into distinct cell masses. This difference in cell affinity under certain experimental conditions could indicate that the deep layer contains endodermal cells. However, an endodermal character of the deep layer can be ruled out by induction experiments with vegetalizing factor or dorsal blastopore lip as inducers. Under the influence of vegetalizing factor the outer as well as the inner ectoderm layer differentiated into mesodermal derivatives such as notochord and somites. The results of the experiments with dorsal blastopore lip as inducer indicate that both inner and outer ectoderm layers are responsive to the neural stimulus. The lower neural competence of the outer ectoderm layer observed by several authors in normogenesis is discussed with regard to the hypothesis about short distance diffusion of the neutralizing factor and/or close cell-to-cell contact between inducing tissue and ectodermal target cells.

Animals↗
PubMed1980

The formation of mesodermal derivatives after induction with vegetalizing factor depends on secondary cell interactions.

Early amphibian gastrula ectoderm induced with vegetalizing factor for 6 h using the sandwich-method and cultured for up to 12 days, differentiated into mesodermal and endodermal tissues. Explants which were dissociated into single cells after the induction followed by immediate reaggregation and then cultured for 12 days likewise differentiated into mesodermal and endodermal tissues. However, if after induction and dissociation of the tissue, single cells are cultured for 20 h prior to the reaggregation, the reaggregated cell mass mainly differentiated into endoderm (liver and intestine) and irregularly shaped epidermis (formerly called 'atypical epidermis'). Blood cells and, in few cases, heart structures were the only mesodermal structures found in this series. The results suggest that an endodermal anlage is induced first. The differentiation of mesodermal derivatives depends on secondary cell interactions between endodermal induced and non-induced ectoderm. For this process permanent cell-to-cell contacts are necessary.

Animals↗
PubMed1979

Cloning of chicken embryo tRNA genes using single stranded nucleosomal DNA highly enriched for tRNA complementary sequences.

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 4B 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 chi 1776. More than 70% of the transformants obtained hybridize to chicken embryo total tRNA.

Animals↗
PubMed1979

Cell contacts between chorda-mesoderm and the overlaying neuroectoderm (presumptive central nervous system) during the period of primary embryonic induction in amphibians.

Using transmission and scanning electron microscopy we were able to show that during primary embryonic induction in amphibians (Triturus alpestris) the interspace between the inducing chorda-mesoderm and the reacting ectoderm (presumptive medullary plate) of mid-gastrula stages is traversed by cell projections starting from cells of both tissue layers. In addition intimate membrane contacts between the main bodies of the ectodermal and chorda-mesodermal cells could be observed. It could be ruled out that cytoplasmic bridges (anastomosis) exist between cells of inducing chorda-mesoderm and reacting ectoderm, which would allow a free transfer of inducing substances without passing through membranes, as Eakin and Lehmann [1] have postulated. The possible role of cell to cell contact for neural induction is emphasized.

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↗