Identification of a 2-cell stage specific inhibitor of the cleavage of preimplantation mouse embryos synthesized by rat hepatoma cells as 5′-deoxy-5′-methylthioadenosine

Zygote ◽  
2010 ◽  
Vol 19 (2) ◽  
pp. 117-125 ◽  
Author(s):  
Masayuki Kobayashi ◽  
Koichi Saito ◽  
Shigeru Tamogami ◽  
Junko Takashima ◽  
Kano Kasuga ◽  
...  
Keyword(s):  
Biological Activities ◽  
Specific Inhibitor ◽  
Biological Properties ◽  
Mouse Embryos ◽  
Cell Stage ◽  
Preimplantation Mouse Embryos ◽  
Preimplantation Mouse ◽  
Mammalian Embryos ◽  
Rat Hepatoma

SummaryRat hepatoma Reuber H-35 cells produce a unique compound designated as Fr.B-25, a 2-cell stage-specific inhibitor of the cleavage of preimplantation mouse embryos cultured in vitro. Here, we identified Fr.B-25 as a purine nucleoside, 5′-deoxy-5′-methylthioadenosine (MTA), by mass spectroscopic analysis. All of the biological activities examined of authentic MTA on the development of mouse zygotes were indistinguishable from those of Fr.B-25. The mechanism of MTA action in the development of preimplantation mouse embryos was probably different from those of hypoxanthine and adenosine, which are well-characterized purine nucleosides that act as inhibitors of the cleavage of mouse 2-cell embryos. From the shared molecular and biological properties of Fr.B-25 and MTA, we concluded that Fr.B-25 is MTA. To the best of our knowledge, this is the first delineation of the effect of MTA on the development of preimplantation mammalian embryos cultured in vitro.

Effect of medium conditioned with rat hepatoma BRL cells on ‘2-cell block’ of random-bred mouse embryos cultured in vitro

Zygote ◽  
2009 ◽  
Vol 17 (2) ◽  
pp. 169-174 ◽  
Author(s):  
Masayuki Kobayashi ◽  
Yoshinori Terawaki ◽  
Koichi Saito ◽  
Kano Kasuga ◽  
Ikuo Kojima
Keyword(s):  
Specific Inhibitor ◽  
Reversed Phase ◽  
Mouse Embryos ◽  
Mouse Strains ◽  
Pcr Analysis ◽  
Cell Block ◽  
Cell Stage ◽  
B Subunit ◽  
Rat Hepatoma

SummaryThe phenomenon of developmental arrest at the 2-cell stage of zygotes obtained from certain mouse strains during in vitro culture is known as the 2-cell block. The effect of conditioned medium (CM) with rat hepatoma BRL cells on the 2-cell block of CD-1 mouse zygotes was investigated in comparison with that of CM with rat hepatoma Reuber H-35 cells. In control medium with EDTA, 75.4% of 2-cell embryos developed to the 4- to 8-cell stages. In the same conditions, the BRL Mr <10000 fraction inhibited the development of 2-cell embryos to the 4- to 8-cell stages (57.7%), although the inhibition by this fraction was weaker than by the Reuber Mr <10000 fraction (19.8%). As a result of reversed-phase column chromatography, a 2-cell stage specific inhibitor of the cleavage of mouse embryos (Fr.B-25), which separated into the Mr <10000 fraction of the Reuber CM, was detected at a low level in the BRL Mr <10000 fraction. On the other hand, the Mr >10000 fraction of BRL CM accelerated the development of the embryos (90.3%). This beneficial effect was also evident even in the absence of EDTA. RT-PCR analysis revealed that mRNAs encoding the β-A or β-B subunit of activins (Mr ~29000), which are well characterized cytokines that act as releasers of the 2-cell block, were expressed in BRL cells. These results indicate that BRL cells synthesize Fr.B-25 at low levels, and that activins contained in the BRL CM probably contributed to overcoming the 2-cell block of CD-1 zygotes cultured in vitro.

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.

The effect of 5-bromodeoxyuridine on cell division and differentiation of preimplantation mouse embryos

Development ◽  
1976 ◽  
Vol 35 (1) ◽  
pp. 169-178
Author(s):  
D. R. Pollard ◽  
M. M. Baran ◽  
R. B. Bachvarova
Keyword(s):  
Cell Division ◽  
Cell Number ◽  
Mouse Embryos ◽  
Cell Stage ◽  
Average Cell ◽  
Preimplantation Mouse Embryos ◽  
Preimplantation Mouse ◽  
Excess Thymidine ◽  
System Cell

Mouse embryos exposed to concentrations of 5-bromodeoxyuridine (BUdR) ranging from 0·01 to 1·0 μg/ml in vitro for two days from the 8-cell stage exhibit a concentration-dependent decrease in the frequency of normal blastocysts and a decrease in average cell number per embryo. A 20-h exposure was adequate to achieve the full BUdR response. Both effects were eliminated in the presence of excess thymidine. Autoradiographs demonstrated that BUdR[3H] was incorporated into DNA during the first and second day of culture. Thus, BUdR appears to act through incorporation into DNA; and, in this system, cell division is at least as sensitive to BUdR as is differentiation.

Human Adenovirus Uptake by Preirnplantation Mouse Embryos

1972 ◽  
Vol 30 ◽  
pp. 268-269
Author(s):  
D. G. Chase ◽  
W. Winters ◽  
L. Piko
Keyword(s):  
Hela Cells ◽  
Human Adenovirus ◽  
Mouse Embryos ◽  
Equilibrium Density ◽  
Cell Stage ◽  
Virus Attachment ◽  
Preimplantation Mouse Embryos ◽  
Embryo Culture Medium ◽  
Preimplantation Mouse ◽  
Mouse Cells

Although the outlines of human adenovirus entry and uncoating in HeLa cells has been clarified in recent electron microscope studies, several details remain unclear or controversial. Furthermore, morphological features of early interactions of human adenovirus with non-permissive mouse cells have not been extensively documented. In the course of studies on the effects of human adenoviruses type 5 (AD-5) and type 12 on cultured preimplantation mouse embryos we have examined virus attachment, entry and uncoating. Here we present the ultrastructural findings for AD-5.AD-5 was grown in HeLa cells and purified by successive velocity gradient and equilibrium density gradient centrifugations in CsCl. After dialysis against PBS, virus was sedimented and resuspended in embryo culture medium. Embryos were placed in culture at the 2-cell stage in Brinster's medium.

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.

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.

scholarly journals Oxygen concentration alters mitochondrial structure and function in in vitro fertilized preimplantation mouse embryos

2020 ◽  
Vol 35 (6) ◽  
pp. 1476-1476
Author(s):  
Manuel Belli ◽  
Ling Zhang ◽  
Xiaowei Liu ◽  
Annemarie Donjacour ◽  
Elena Ruggeri ◽  
...  
Keyword(s):  
Oxygen Concentration ◽  
Structure And Function ◽  
Mouse Embryos ◽  
Mitochondrial Structure ◽  
Preimplantation Mouse Embryos ◽  
Preimplantation Mouse ◽  
And Function
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