Biosynthesis of DNA, RNA and proteins by mouse embryos cultured in the presence of a teratogenic dose of chlorambucil

Development ◽  
1976 ◽  
Vol 36 (2) ◽  
pp. 273-281
Author(s):  
T. W. Sadler ◽  
D. M. Kochhar
Keyword(s):  
Protein Synthesis ◽  
Dna Synthesis ◽  
Embryo Culture ◽  
Culture Media ◽  
Placental Tissue ◽  
Mouse Embryos ◽  
Whole Embryo Culture ◽  
Rna And Protein Synthesis ◽  
Visceral Yolk Sac ◽  
Reichert's Membrane

The effect of chlorambucil on the rates of DNA, RNA, and protein synthesis in mouse embryos was investigated using a system of whole embryo culture. Embryos were isolated on the 11th day of gestation (33 ± 3 somites) and grown in culture media for periods of 4–8 h. Reichert's membrane and most of the placental tissue was removed leaving only the amnion and visceral yolk-sac surrounding the embryo. In the presence of teratogenic doses of chlorabucil (15 μg/ml) the rate of DNA synthesis was significantly decreased at 4 and 8 h. RNA and protein synthesis were not inhibited at either of these times. A trend toward decreasing rates of protein synthesis at some time beyond 8 h was noted, but not tested.

A potential mechanism of DL-beta-hydroxybutyrate-induced malformations in mouse embryos

1987 ◽  
Vol 253 (1) ◽  
pp. E72-E80 ◽  
Author(s):  
E. S. Hunter ◽  
T. W. Sadler ◽  
R. E. Wynn
Keyword(s):  
Dna Synthesis ◽  
Ketone Body ◽  
Synergistic Effects ◽  
Krebs Cycle ◽  
Pentose Phosphate ◽  
Mouse Embryos ◽  
Potential Mechanism ◽  
Whole Embryo Culture ◽  
Biochemical Alterations ◽  
Beta Hydroxybutyrate

DL-beta-Hydroxybutyrate (DL-BOHB) is teratogenic to rodent embryos in culture, but the biochemical mechanism(s) responsible for the induction of malformations has not been elucidated. Thus, to delineate a potential mechanism, the interaction of the ketone body with embryonic glucose metabolism was investigated. Mouse embryos were exposed in whole embryo culture to DL-BOHB or each isomer separately during the period of neurulation. The results demonstrate that DL-BOHB inhibits glucose oxidation by the pentose phosphate pathway (PPP) with no concomitant effect on Krebs cycle or glycolysis. Furthermore, decreased rates of embryonic uridine monophosphate and DNA synthesis were produced by DL-BOHB, whereas orotate synthesis was unaffected, thereby suggesting that the availability of ribose moieties synthesized by the PPP was compromised. This hypothesis was supported by the observation that addition of ribose to DL-BOHB-containing medium reduced the incidence of ketone body-induced neural tube defects and maintained the rates of DNA synthesis at control levels. Furthermore, these biochemical alterations are due to the synergistic effects of the D and L isomers and may be related to changes in redox states.

Improved growth and development of presomite mouse embryos in whole embryo culture

1988 ◽  
Vol 245 (3) ◽  
pp. 264-269 ◽  
Author(s):  
E. S. Hunter ◽  
W. Balkan ◽  
T. W. Sadler
Keyword(s):  
Embryo Culture ◽  
Growth And Development ◽  
Mouse Embryos ◽  
Whole Embryo Culture

Nucleic acid and protein synthesis and pattern regulation in hydra

Development ◽  
1969 ◽  
Vol 21 (1) ◽  
pp. 55-70
Author(s):  
S. G. Clarkson
Keyword(s):  
Protein Synthesis ◽  
Dna Synthesis ◽  
Experimental Approach ◽  
Direct Evidence ◽  
Animal Cells ◽  
Rna And Protein Synthesis ◽  
Metabolic Activities ◽  
Time Required ◽  
Pattern Regulation

In a previous paper (Clarkson, 1969) data were presented which indicate that hypostome determination is accompanied by a large and rapid burst of RNA synthesis, a slight stimulation of protein synthesis, and no increase in DNA synthesis. More direct evidence concerning the relative importance of these metabolic activities in hypostome determination is reported in this paper. The experimental approach made use of the transplantation test of Webster & Wolpert (1966) in conjunction with some inhibitors of DNA, RNA and protein synthesis, the rationale being that if these metabolic activities play important roles in the determination of the hypostome, then their inhibition would be expected to have severe effects on the time required for this process. Regarding the inhibitors, hydroxyurea (HU) inhibits DNA synthesis in a variety of animal cells without altering rates of formation of RNA or protein (Young & Hodas, 1964; Yarbro, Kennedy & Barnum, 1965; Schwartz, Garofalo, Sternberg & Philips, 1965).

scholarly journals Cell death in rat and mouse embryos exposed to methanol in whole embryo culture

Toxicology ◽  
1995 ◽  
Vol 97 (1-3) ◽  
pp. 159-171 ◽  
Author(s):  
B.D. Abbott ◽  
M. Ebron-mccoy ◽  
J.E. Andrews
Keyword(s):  
Cell Death ◽  
Embryo Culture ◽  
Mouse Embryos ◽  
Whole Embryo Culture

Tissue-specific RNA interference in post-implantation mouse embryos using directional electroporation and whole embryo culture

2004 ◽  
Vol 72 (2-3) ◽  
pp. 92-102 ◽  
Author(s):  
Federico Calegari ◽  
Anne-Marie Marzesco ◽  
Ralf Kittler ◽  
Frank Buchholz ◽  
Wieland B. Huttner
Keyword(s):  
Rna Interference ◽  
Embryo Culture ◽  
Mouse Embryos ◽  
Whole Embryo Culture ◽  
Tissue Specific ◽  
Post Implantation

scholarly journals Postimplantation Whole Embryo Culture Assay for Hamsters: An Alternative to Rat and Mouse

2001 ◽  
Vol 1 ◽  
pp. 227-234 ◽  
Author(s):  
Bogdan Wlodarczyk ◽  
Bogumil Biernacki ◽  
Maria Minta ◽  
Jan Zmudzki
Keyword(s):  
Embryo Culture ◽  
Culture Conditions ◽  
Mouse Embryos ◽  
Total Protein Content ◽  
Image Analysis System ◽  
Whole Embryo Culture ◽  
Analysis System ◽  
Comparison Of The Results ◽  
Rats And Mice

Postimplantation whole embryo culture (WEC) assay for rats and mice has been well established and introduced to many laboratories. Recently WEC technique for rabbits has been developed; however, information on culture of other species is very limited. Knowing the usefulness of hamsters in classical embryotoxicology, we reasoned that hamster WEC could be an alternative model for the most frequently used rat and mouse WEC. Previously we have optimized culture conditions for postimplantation hamster embryos. The aim of this study was to test the susceptibility of hamster embryos cultured in vitro to embryotoxic compounds and to compare our results with those reported by others on rat or mouse embryo culture. For that purpose we choose three known embryotoxic compounds�valproic acid, cadmium chloride, and diethylstilbestrol�and tested them using a postimplantation hamster whole embryo culture assay. Hamster embryos were cultured from 7.5 days gestation for 24 h in a medium consisting of 35% hamster serum and 65% synthetic culture medium (Iscove�s or McCoy 5A). At the end of the culture period, the embryos were examined morphologically, measured with the aid of a computer image analysis system, and total protein content was assessed. All three compounds exhibited dose-related embryotoxic and teratogenic effects in hamster embryos. The malformations observed were similar to those reported on rat and mouse embryos. Comparison of the results with data reported by other authors indicates that hamster embryos cultured in vitromight be more susceptible to embryotoxic stimuli than rat and mouse embryos.

scholarly journals MACROMOLECULAR SYNTHESES RELATED TO THE REPRODUCTIVE CYST OF TETRAHYMENA PATULA

1973 ◽  
Vol 59 (3) ◽  
pp. 615-623 ◽  
Author(s):  
P. R. Gabe ◽  
L. E. de Bault
Keyword(s):  
Protein Synthesis ◽  
Dna Synthesis ◽  
Generation Time ◽  
Rna Synthesis ◽  
Culture Medium ◽  
Tritiated Thymidine ◽  
Growth Cycle ◽  
The Third ◽  
Rna And Protein Synthesis ◽  
The Mean

Macromolecular syntheses in encysted Tetrahymena patula were studied using Feulgen fluorescence cytophotometry, autoradiography, and inhibitors of RNA and protein synthesis. Cycloheximide significantly depressed protein synthesis and D-actinomycin effectively blocked RNA synthesis. Under these conditions, the cells within the cyst were unable to divide. Both cytophotometric measurements and autoradiographic data with tritiated thymidine show that DNA synthesis does not occur during the encystment divisions. Excysted cells placed in nutrient broth medium showed a prolonged generation time after the first cell growth cycle, and by the third generation the mean DNA content per cell was almost triple that of starved excysted cells. These findings indicate that (a) the encystment divisions require RNA and protein synthesis, which are apparently effected through turnover, (b) the encystment division cycles occur in the absence of DNA synthesis, and (c) excysted cells placed in culture medium may go through more than one DNA replication per cell cycle.

Whole-embryo culture of E5.5 mouse embryos: Development to the gastrulation stage

genesis ◽  
2003 ◽  
Vol 37 (1) ◽  
pp. 38-43 ◽  
Author(s):  
Shigeto Miura ◽  
Yuji Mishina
Keyword(s):  
Embryo Culture ◽  
Mouse Embryos ◽  
Whole Embryo Culture

Formulation of a complex serum-free medium (CSM) for use in the co-culture of mouse embryos with cells of the female reproductive tract

1991 ◽  
Vol 3 (1) ◽  
pp. 99 ◽  
Author(s):  
D Sakkas ◽  
AO Trounson
Keyword(s):  
Growth Factors ◽  
Growth Factor ◽  
Embryo Culture ◽  
Reproductive Tract ◽  
Culture Media ◽  
Mouse Embryos ◽  
Serum Free Medium ◽  
Free Medium ◽  
Serum Free ◽  
Cell Numbers

Co-culture of pre-implantation embryos with cells of the reproductive tract requires a medium that is beneficial to both embryos and cells. However, many studies in this area utilize media originally formulated for specific cell lines. In the present study, a complex serum-free medium (CSM) was formulated on the basis of the ionic compositions of existing embryo culture media and mouse oviductal fluid as well as the concentrations of growth factors that appear to benefit mouse embryo development. The study began by investigating the effect of altering the concentrations of K+ ions (0-40 mM) and sulfate ions (0-10 mM) in embryo culture media on the development of 2-cell mouse embryos. Mouse embryos showed improved cell numbers at the blastocyst stage when cultured in 10 mM K+ compared with Whittingham's T6 medium. Embryos were also cultured in T6 supplemented with bovine serum albumin (BSA) containing various concentrations of insulin, insulin-like growth factors I and II, fibroblast growth factor, and epidermal growth factor. Insulin concentrations of 100 ng mL-1 significantly (P less than 0.05) improved the cell numbers of 2-cell embryos cultured to the morulae and blastocyst stages compared with those cultured in T6 + BSA alone. CSM was formulated on the basis of the results of these experiments and was found to support both improved development of 2-cell mouse embryos and the culture of mouse fibroblast and mouse oviduct cells.

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