Normal adenylate ribonucleotide content in mouse embryos homozygous for the t12 mutation

Development ◽  
1983 ◽  
Vol 78 (1) ◽  
pp. 43-51
Author(s):  
Horst Spielmann ◽  
Robert P. Erickson
Keyword(s):  
In Vitro Culture ◽  
Normal Values ◽  
Firefly Luciferase ◽  
Blastocyst Stage ◽  
Mouse Embryos ◽  
Luciferase Assay ◽  
Preimplantation Mouse Embryos ◽  
Preimplantation Mouse

The recently improved firefly luciferase assay was used to determine ATP, ADP or AMP in single preimplantation mouse embryos from crosses yielding lethal t12/t12 embryos. Normal values of the three adenylate ribonucleotides were found in freshly collected 2-cell and 4-cell embryos and during in vitro culture to the blastocyst stage. A decrease in adenylate ribonucleotide content was seen in putative t12/t12 embryos only when they were degenerating.

The effect of microtubule- and microfilament-disrupting drugs on preimplantation mouse embryos

Development ◽  
1980 ◽  
Vol 60 (1) ◽  
pp. 71-82
Author(s):  
G. Siracusa ◽  
D. G. Whittingham ◽  
M. De Felici
Keyword(s):  
Blastocyst Stage ◽  
Mouse Embryos ◽  
Microtubule Assembly ◽  
Continuous Exposure ◽  
Blastocyst Formation ◽  
Multipolar Spindles ◽  
Preimplantation Mouse Embryos ◽  
Preimplantation Mouse ◽  
Continuous Presence

The sensitivity of early preimplantation mouse embryos to drugs which disrupt microfilament function (cytochalasin B-CB and cytochalasin D-CD) and microtubule assembly (colchicine, colcemid, vinblastine and griseofulvin) was examined. CD inhibited cleavage at a concentration 35-fold lower than CB (3 × 10−7 M ν. 1 × 10−5 M). Treatment of 2-cell embryos for 6 h with 1 × 10−5 M CB or 1 × 10−6 M CD or continuous exposure to lower concentrations of CB or CD did not affect development to the blastocyst stage in vitro. Vinblastine inhibited cleavage at a concentration tenfold lower than colcemid or colchicine (1 × 10−8 M ν. 1 × 10−7 M). The continuous presence of colcemid at 10−8 M did not affect the development of 2-cell embryos to the blastocyst stage, but development was reduced with vinblastine at 1 × 10−8 M and completely inhibited with colchicine at 1 × 10−8 M. The drugs produced similar responses when 2-cell embryos were treated for 6 h with concentrations that inhibited cleavage. Complete inhibition of cleavage was obtained after only a 2 h exposure to 2 × 10−7 M colchicine. A similar concentration of lumicolchicine did not affect cleavage or blastocyst formation. Embryos were less sensitive to griseofulvin; the first cleavage division was unaffected by concentrations as high as 3 × 10−4 M and only 50% of 2-cell embryos failed to cleave in 1 × 10 1 and 3 × 10−4 M griseofulvin. At these concentrations a small proportion of 1-cell embryos and the majority of the 2-cell embryos showed unequal cytoplasmic division probably caused by the formation of multipolar spindles. The continuous exposure of 2-cell embryos to 3 × 10−5 M griseofulvin did not affect blastocyst formation.

In vitro culture of preimplantation mouse embryos and day 12 limb-buds: Effects of serum and albumin

1993 ◽  
Vol 7 (6) ◽  
pp. 623-630 ◽  
Author(s):  
Belén Tornesi ◽  
Andrzej T. Palasz ◽  
Marcelo R. Del Campo ◽  
Colin G. Rousseaux ◽  
F.Joy Archer ◽  
...  
Keyword(s):  
In Vitro Culture ◽  
Mouse Embryos ◽  
Limb Buds ◽  
Preimplantation Mouse Embryos ◽  
Preimplantation Mouse

scholarly journals Turnover of Carbon Pools Labelled with [14C]Glucose during in vitro Culture of Preimplantation Mouse Embryos

1982 ◽  
Vol 35 (6) ◽  
pp. 637
Author(s):  
I L Pike ◽  
RG Wales
Keyword(s):  
In Vitro Culture ◽  
Mouse Embryos ◽  
Carbon Pools ◽  
Stages Of Development ◽  
Glucose Carbon ◽  
Stage Of Development ◽  
Preimplantation Mouse Embryos ◽  
Preimplantation Mouse

The pulse-chase technique was used to study the uptake and turnover of glucose carbon by mouse embryos in vitro. During a 1 h pulse the uptake of glucose into all embryonic fractions increased between the eight-celled and the morula-early blastocyst stages of development. Whilst most of the glucose carbon entered the non-glycogen, acid-soluble pool, significant amounts were isolated in acid-insoluble macromolecules and, at the later stage of development, in acid-soluble glycogen.

Cyclic AMP reversal of hypoxanthine-arrested preimplantation mouse embryos is EDTA-dependent

Zygote ◽  
1996 ◽  
Vol 4 (2) ◽  
pp. 129-137 ◽  
Author(s):  
Mary K. Dienhart ◽  
Stephen M. Downs
Keyword(s):  
Embryo Development ◽  
Culture Media ◽  
Phosphodiesterase Inhibitor ◽  
Blastocyst Stage ◽  
Mouse Embryos ◽  
Cooperative Interaction ◽  
Basic Salt ◽  
Preimplantation Mouse Embryos ◽  
Preimplantation Mouse

SummaryHypoxanthine can block preimplantation mouse embryo development in vitro at the 2- to 4-cell stages, and this has recently been shown to be reversed by cAMP-elevating agents. However, the extent of this hypoxanthine-induced arrest is determined by the culture conditions and strain of mouse. Whitten's and KSOM/AA are two embryo culture media that support preimplantation development to the blastocyst stage. This study was undertaken to examine the influence of several components in these media on hypoxanthine-arrested preimplantation mouse embryos and to test the hypothesis that reversal of the hypoxanthine block by cAMP-elevating agents requires cooperative interaction with the chelator, EDTA. Initial experiments demonstrated that embryo development was blocked in the presence of hypoxanthine in Whitten's medium but not in KSOM/AA; furthermore, removal of EDTA from KSOM/AA rendered this medium incapable of supporting high levels of development to blastocyst (9%), whereas high numbers of blastocysts (80%) formed in Whitten's medium, which does not contain the chelator. Consequently, Whitten's medium was used to test our hypothesis. It has previously been demonstrated that the phosphodiesterase inhibitor, IBMX, can reverse the developmental arrest imposed by hypoxanthine in EDTA-supplemented Earle's basic salt solution, but in the present study the addition of IBMX to Whitten's medium resulted in a block to development and failed to reverse the hypoxanthine arrest. These disparate effects can be explained by the presence or absence of EDTA. Supplementing Whitten's medium with EDTA reverses the IBMX effect, but not the hypoxanthine-induced block. While IBMX alone is unable to reverse the hypoxanthine block in Whitten's medium, development is greatly enhanced by the simultaneous addition of EDTA and IBMX. Similar results were obtained with the cAMP analogue, 8-AHA-cAMP. The data therefore support our hypothesis that the reversal of the hypoxanthine-induced arrest by cAMP-elevating agents is critically dependent on the presence of EDTA. We contrast this with the situation in mouse oocytes, where the hypoxanthine-induced meiotic arrest is not reversed by the addition of EDTA and/or cAMP-elevating agents.

scholarly journals Synthesis and Degradation of Labelled Glycogen Pools in Preimplantation Mouse Embryos during Short Periods of in vitro Culture

1984 ◽  
Vol 37 (3) ◽  
pp. 137 ◽  
Author(s):  
WR Edirisinghe ◽  
RG Wales ◽  
I L Pike
Keyword(s):  
In Vitro Culture ◽  
Mouse Embryos ◽  
Glucose Carbon ◽  
Preimplantation Mouse Embryos ◽  
Preimplantation Mouse ◽  
Synthesis And Degradation

The incorporation and turnover of glucose carbon by mouse embryos during short periods of in vitro culture were studied using [U-14C]glucose lis marker. Particular attention was given to the synthesis and degradation of the acid-soluble and acid-insoluble glycogen pools.

Platelet activating factor (PAF) enhances mitosis in preimplantation mouse embryos

1993 ◽  
Vol 5 (3) ◽  
pp. 271 ◽  
Author(s):  
C Roberts ◽  
C O'Neill ◽  
L Wright
Keyword(s):  
Mitotic Index ◽  
Platelet Activating Factor ◽  
Treated Group ◽  
Blastocyst Stage ◽  
Mouse Embryos ◽  
Cell Stage ◽  
Preimplantation Mouse Embryos ◽  
Preimplantation Mouse ◽  
Web 2086

Preimplantation mouse embryos were used to determine whether the reported significant increase in embryo metabolism and viability achieved through supplementation of the culture medium with the ether phospholipid 1-o-alkyl-2-acetyl-sn-glycero-3-phosphocoline (platelet activating factor, PAF) is attributable to an enhanced rate of mitosis. Blastocyst-stage embryos cultured in the presence of 0.186 to 18.6 microM exogenous PAF had a significantly (P < 0.01) higher mitotic index (the proportion of cells arrested in metaphase following incubation in colchicine) than those cultured without PAF. At the 8-cell stage, 29% more blastomeres were in metaphase in the PAF-treated group (P < 0.01) 8 h after the addition of colchicine, but by 16 h there was no difference between groups; thus, PAF increased the rate at which cells entered metaphase but did not increase the total number. The mitotic index showed a negative correlation with the number of cells within blastocysts. PAF had a significantly (P < 0.01) greater impact on the mitotic index of blastocysts with fewer cells. The action of PAF was specific, being completely blocked by the PAF-receptor antagonist WEB 2086 (33 microM). In the absence of exogenous PAF the mitotic index was lower with WEB 2086 than without, suggesting inhibition of the action of endogenous embryo-derived PAF. These results show that PAF stimulates the rates at which cells within the preimplantation mouse embryo enter metaphase in vitro and suggest that it would decrease their doubling time, perhaps accounting for the embryotrophic actions of PAF.

Modification of cell division by phorbol ester in preimplantation mouse embryos

Development ◽  
1987 ◽  
Vol 101 (2) ◽  
pp. 403-408
Author(s):  
E.T. Mystkowska ◽  
W. Sawicki
Keyword(s):  
Cell Division ◽  
Video Recording ◽  
Mouse Embryos ◽  
Time Intervals ◽  
Cell Divisions ◽  
Stage Of Development ◽  
Preimplantation Mouse Embryos ◽  
Preimplantation Mouse ◽  
Single Blastomeres

2-cell mouse embryos were treated in vitro with a 2 h pulse of phorbol myristate acetate (PMA) at 32nd, 38th and 50th h after hCG, then chased in culture for up to 46 h. Embryos were fixed at various time intervals of chasing, then stained and inspected. Some embryos were carefully inspected with a video recording system, every 1.44s and the cell divisions (cytokinesis) as well as formation of large, single blastomeres, each from two smaller ones, were recorded. PMA pulse let to the suppression of cell divisions. The rate of the suppression was time dependent: with a delay of 0–1, 12 and 18 h between the PMA pulse and time of scheduled cell division about 99, 87 and 44% of 2-cell embryos remained at this stage of development, for at least 10 h, respectively, and 90, 58 and 12% of their blastomeres revealed binuclearity. Since we found that PMA-mediated formation of binuclearity was not the effect of cell fusions, it was assumed that the inhibition of cytokinesis preceded by karyokinesis was responsible for binuclearity. PMA effect on cell divisions was reversible. PMA-treated embryos revealed formation of large, single blastomeres, each from two smaller ones. If cell division appeared after PMA pulse, in about 52% of 3- to 6-cell embryos, the large blastomere formation was recorded in the course of the subsequent 38 h. Large blastomere formation was concluded to be the result of either cell fusion or reversion of incompleted cytokinesis brought about by PMA.

Viability and growth of mouse embryos after in vitro culture and fusion

Development ◽  
1970 ◽  
Vol 23 (3) ◽  
pp. 693-704
Author(s):  
Patricia Bowman ◽  
Anne McLaren
Keyword(s):  
In Vitro Culture ◽  
Success Rate ◽  
Zona Pellucida ◽  
Growth Regulation ◽  
Excess Mortality ◽  
Blastocyst Stage ◽  
Mouse Embryos ◽  
Foster Mothers ◽  
Mouse Eggs

About 80 % of 8-cell mouse eggs developed to the blastocyst stage in culture, whether the zona pellucida was left intact, or removed with pronase (pre-incubated and dialysed) and the eggs then cultured singly or as fused pairs. When pronase was used without prior incubation and dialysis, the success rate was reduced to 50 %. After transfer to uterine foster-mothers, 20–30 % of apparently normal blastocysts cultured with or without the zona, singly or fused, developed into live foetuses, compared with over 50 % of control blastocysts taken directly from the uterus. Some of the excess mortality of cultured embryos took place before implantation and some soon after. The foetuses derived from cultured blastocysts averaged 0·1 g lighter than those derived from control uterine blastocysts similarly transferred. No differences in the weights of the placentae were observed. Foetal and placental weights were unaffected by whether the eggs had been cultured singly or fused, implying that growth regulation of fused embryos is complete by the 17th day of gestation. The longer the eggs were maintained in culture, the lower was their viability after transfer, and the lighter were the foetuses derived from them.

Sign in / Sign up

Export Citation Format

Share Document