Histone synthesis during development of Triturus embryos

Development ◽  
1973 ◽  
Vol 30 (1) ◽  
pp. 73-82
Author(s):  
Hiroshi Imoh ◽  
Izumi Kawakami
Keyword(s):  
Carbon Dioxide ◽  
Gel Electrophoresis ◽  
Fast Moving ◽  
Basic Protein ◽  
Disc Electrophoresis ◽  
Gastrula Stage ◽  
Tail Bud ◽  
Blastula Stage ◽  
Radioactive Carbon ◽  
Histone Synthesis

Synthesis of histone fractions and one basic protein fraction, which moved fast on gel electrophoresis and had been reported to increase in nuclei accompanying a decrease in cytoplasm during development, were studied with radioactive carbon dioxide as a tracer. Acid-extractable proteins of nuclei or cytoplasm, isolated from labelled embryos, were fractionated by polyacrylamide disc electrophoresis and the histone fractions and the fast-moving basic protein were identified. Radioactivities in these fractions and DNA were determined. Synthesis of the fast-moving basic protein was not detected throughout the period of development studied and this fraction was thought to move in from the cytoplasm to the nucleus during development. Syntheses of histone fractions were observed as early as the blastula stage. Rates of syntheses of four histone fractions (f3, f2b, f2a2 and f2al) per embryo increased thereafter, keeping pace with the increase in the rate of DNA synthesis with advancing development. The rate of the very lysine-rich fl histone synthesis per embryo did not increase after the gastrula stage and the rate remained almost constant until the late tail-bud stage. Compositions of newly synthesized histones, calculated from the radioactivities incorporated into histone fractions, were almost the same during development and among different regions of neurula or tail-bud-stage embryos, with the exception of the f1 fraction, which varied depending on the stage and region of the embryos. The results are discussed in relation to the possible roles of the histone fractions in developing embryos.

Pyrimidine reducing enzymes of rat liver

1976 ◽  
Vol 54 (2) ◽  
pp. 178-184 ◽  
Author(s):  
Ronald O. Hallock ◽  
Esther W. Yamada
Keyword(s):  
Carbon Dioxide ◽  
Rat Liver ◽  
Gel Electrophoresis ◽  
Marker Enzyme ◽  
Good Separation ◽  
Cytosol Fraction ◽  
Radioactive Carbon ◽  
Mitochondrial Fractions ◽  
Optimum Ph ◽  
Rat Liver Cells

Dihydrouracil dehydrogenase (NADP+) (EC 1.3.1.2) was partially purified from the cytosol fraction of rat liver and fractionated by disc gel electrophoresis. A major and minor band were visualized by staining for enzyme activity. The substrate specificity of these bands was investigated. It was found that both bands were two to three times more active with dihydrothymine as substrate than with dihydrouracil in the presence of NADP+ and the optimum pH of 7.4.Mitochondrial fractions containing most of the NADH-dependent uracil reductase of rat liver cells were fractionated by centrifugation in sucrose density gradients. Two procedures involving linear or discontinuous gradients were used. By both, good separation of NADH- and NADPH-dependent reductases was achieved. Marker enzyme studies supported the view that the NADH-dependent enzyme is located principally in mitochondria whereas the NADPH-dependent enzyme is mainly in plasma and endoplasmic reticulum membranes. For the NADH-dependent reductase the apparent Km for thymine at pH 7.4 was 1.39 times that found for uracil whereas for the NADPH-dependent enzyme the apparent Km values were similar for the two substrates at this pH.Dihydrouracil was the principal product isolated by paper chromatography from the reaction mixture containing a partially purified fraction of mitochondria, uracil and NADH at pH 7.4. This fraction also catalyzed the formation of radioactive carbon dioxide from [2-14C]uracil. The proportion of CO2 formed by the mitochondria was about 10% of that formed by the original homogenate.

Expression of a novel FGF in the Xenopus embryo. A new candidate inducing factor for mesoderm formation and anteroposterior specification

Development ◽  
1992 ◽  
Vol 114 (3) ◽  
pp. 711-720 ◽  
Author(s):  
H.V. Isaacs ◽  
D. Tannahill ◽  
J.M. Slack
Keyword(s):  
Specific Activity ◽  
Biological Properties ◽  
The Body ◽  
Mesoderm Induction ◽  
Gastrula Stage ◽  
Blastula Stage ◽  
Mesoderm Formation ◽  
Low Levels

We have cloned and sequenced a new member of the fibroblast growth factor family from Xenopus laevis embryo cDNA. It is most closely related to both mammalian kFGF (FGF-4) and FGF-6 but as it is not clear whether it is a true homologue of either of these genes we provisionally refer to it as XeFGF (Xenopus embryonic FGF). Two sequences were obtained, differing by 11% in derived amino acid sequence, which probably represent pseudotetraploid variants. Both the sequence and the behaviour of in vitro translated protein indicates that, unlike bFGF (FGF-2), XeFGF is a secreted molecule. Recombinant XeFGF protein has mesoderm-inducing activity with a specific activity similar to bFGF. XeFGF mRNA is expressed maternally and zygotically with a peak during the gastrula stage. Both probe protection and in situ hybridization showed that the zygotic expression is concentrated in the posterior of the body axis and later in the tailbud. Later domains of expression were found near the midbrain/hindbrain boundary and at low levels in the myotomes. Because of its biological properties and expression pattern, XeFGF is a good candidate for an inducing factor with possible roles both in mesoderm induction at the blastula stage and in the formation of the anteroposterior axis at the gastrula stage.

A functional test for maternally inherited cadherin in Xenopus shows its importance in cell adhesion at the blastula stage

Development ◽  
1994 ◽  
Vol 120 (1) ◽  
pp. 49-57 ◽  
Author(s):  
J. Heasman ◽  
D. Ginsberg ◽  
B. Geiger ◽  
K. Goldstone ◽  
T. Pratt ◽  
...  
Keyword(s):  
Cell Adhesion ◽  
Protein Level ◽  
Mrna Level ◽  
Functional Test ◽  
Gastrula Stage ◽  
Blastula Stage ◽  
Xenopus Development ◽  
Dose Dependent ◽  
Embryonic Ectoderm ◽  
E Cadherin

We report here on the consequences of reducing the expression of EP-cadherin at the earliest stages of Xenopus development. Injection of oligodeoxynucleotides antisense to maternal EP-cadherin mRNA into full-grown oocytes reduced the mRNA level in oocytes, and the protein level in blastulae. Adhesion between blastomeres was significantly reduced, as seen in whole embryos, and in assays of the ability of blastomeres to reaggregate in culture. This effect was especially conspicuous in the inner cells of the blastula and included the disruption of the blastocoel. The severity of the EP-cadherin mRNA depletion and of the disaggregation phenotype was dose dependent. This phenotype was rescued by the injection into EP-cadherin mRNA-depleted oocytes of the mRNA coding for a related cadherin, E-cadherin, that is normally expressed at the gastrula stage in the embryonic ectoderm.

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

Development ◽  
1992 ◽  
Vol 116 (Supplement) ◽  
pp. 137-142 ◽  
Author(s):  
J. B. Gurdon ◽  
K. Kao ◽  
K. Kato ◽  
N. D. Hopwood
Keyword(s):  
Protein Synthesis ◽  
Developmental Stage ◽  
Cell Transplantation ◽  
Intercellular Communication ◽  
Gene Activation ◽  
Gastrula Stage ◽  
Blastula Stage ◽  
Morphological Marker ◽  
Muscle Gene ◽  
Early Gastrula

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 celhcell 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.

COMPETITION BETWEEN AMINO ACIDS FOR TRANSPORT INTO EHRLICH ASCITES CARCINOMA CELLS

1961 ◽  
Vol 39 (11) ◽  
pp. 1717-1735 ◽  
Author(s):  
P. G. Scholefield
Keyword(s):  
Carbon Dioxide ◽  
Amino Acids ◽  
Amino Acid ◽  
Ehrlich Ascites Carcinoma ◽  
Carcinoma Cells ◽  
Transport Systems ◽  
Radioactive Carbon ◽  
Ehrlich Ascites ◽  
Competitive Inhibitors ◽  
Amino Acid Analogues

The cumulative entry of amino acids into Ehrlich ascites carcinoma cells is due to the presence of active transport systems, each with its own specific range of substrates. Several amino acids and amino acid analogues may have an affinity for the same transport system and thus may inhibit transport of other amino acids by acting as competitive inhibitors or competitive substrates. Loss of methionine from ascites cells takes place by a diffusion process which obeys Fick's law. Leucine accumulation by ascites cells is small and is increased on addition of certain other amino acids. The increase is not due to inhibition of leucine oxidation as increase in the rate of production of radioactive carbon dioxide from labeled leucine also occurs. Kinetic aspects of these results are discussed.

The role of water-regulating mechanisms in the development of the haploid syndrome in Xenopus laevis

Development ◽  
1972 ◽  
Vol 28 (2) ◽  
pp. 449-462
Author(s):  
Louie Hamilton ◽  
P. H. Tuft
Keyword(s):  
Xenopus Laevis ◽  
Cell Number ◽  
Gastrula Stage ◽  
Tail Bud ◽  
Volume Changes ◽  
Membrane Area ◽  
Flow Through ◽  
The Difference ◽  
Haploid Embryos

The uptake of water by haploid and diploid sibling embryos of Xenopus laevis has been investigated by measuring the density changes which occur during the development of intact embryos from the blastula to the late tail-bud stage, and of explants from which most of the presumptive endoderm has been removed. The results show that up to the mid-gastrula stage there is no difference between the haploid and diploid embryos; but from then on, whereas the diploid volume increases steadily, the haploid gastrulae undergo a series of cyclical volume changes due to loss of fluid through the blastopore. It is concluded that this is the result of an excessive inflow of water through the haploid ectoderm, because it was found that the volume of haploid ectodermal explants increased much more rapidly than the volume of similar diploid explants. Excess flow through the haploid ectoderm also accounts for other characteristics of the haploid syndrome – microcephaly and lordosis. It is suggested that it is the doubling of the cell number in haploid embryos with the consequent 25% increase in aggregate cell membrane area which accounts for the difference between the uptake of water by the two types of embryos. It is also suggested that changes in the rate of water flow through the ectoderm and endoderm which are thought to account for the accumulation of water in the blastocoel and archenteron in the normal diploid embryo arise in a similar way.

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