RNAi Experiments in Mouse Oocytes and Early Embryos

2009 ◽  
Vol 2009 (1) ◽  
pp. pdb.top56-pdb.top56 ◽  
Author(s):  
P. Svoboda ◽  
P. Stein
Keyword(s):  
Mouse Oocytes ◽  
Early Embryos

A monoclonal antibody against mouse oocyte cytoskeleton recognizing cytokeratin-type filaments

Development ◽  
1985 ◽  
Vol 90 (1) ◽  
pp. 197-209
Author(s):  
E. Lehtonen
Keyword(s):  
Epithelial Cells ◽  
Immunofluorescence Microscopy ◽  
Staining Pattern ◽  
Blastocyst Stage ◽  
Tissue Sections ◽  
Mouse Oocytes ◽  
Thick Filaments ◽  
Trophectoderm Cells ◽  
Early Embryos ◽  
Cytokeratin Filaments

Monoclonal antibodies were raised to detergent-extracted cytoskeleton preparations of mouse oocytes. In immunofluorescence microscopy, one of the antibodies, OCS-1, localizes exclusively to epithelial cells in frozen tissue sections, including various simple and stratified epithelia. The antibody decorates a keratin-type of fibrillax, vinblastine-resistant network in various cultured, epithelial-type cells, but not in myoid or fibroblastoid cells. In mouse oocytes and cleavage-stage embryos, the OCS-1 antibody gives a diffuse, spotty staining pattern. In blastocyst-stage embryos, the antibody reveals a keratin-type filamentous organization in the trophectoderm cells. In immunoelectron microscopy, the OCS-1 antibody decorates 10 nm-thick filaments, often identifiable as desmosome-attached tonofilaments, in detergent-treated trophectoderm cells. The antigen(s) recognized by the OCS-1 antibody is apparently present in, or closely associated with, cytokeratin filaments. In addition to mouse oocytes and early embryos, a wide variety of epithelial cells in various species seem to share this antigen(s). The present results suggest that at the early stages, the cytokeratin-related antigen(s) defined by the OCS-1 antibody are stored in a non-fibrillar form which is then converted into a fibrillar network at the blastocyst stage. A pre-existing supply of cytokeratin-related protein may be essential for the development of the blastocyst.

scholarly journals Why mouse oocytes and early embryos ignore miRNAs?

RNA Biology ◽  
2010 ◽  
Vol 7 (5) ◽  
pp. 559-563 ◽  
Author(s):  
Petr Svoboda
Keyword(s):  
Mouse Oocytes ◽  
Early Embryos

DNA methylation pattern in mouse oocytes and their in vitro fertilized early embryos: effect of oocyte vitrification

Zygote ◽  
2012 ◽  
Vol 22 (2) ◽  
pp. 138-145 ◽  
Author(s):  
Ying Liang ◽  
Xiang-Wei Fu ◽  
Jun-Jie Li ◽  
Dian-Shuai Yuan ◽  
Shi-En Zhu
Keyword(s):  
Dna Methylation ◽  
Fluorescein Isothiocyanate ◽  
Methylation Pattern ◽  
Oocyte Vitrification ◽  
Developmental Potential ◽  
Mouse Oocytes ◽  
Early Embryos ◽  
Vitrification Solution ◽  
Early Mouse

SummaryThis study was conducted to investigate the pattern of DNA methylation in vitrified–thawed mouse oocytes and their in vitro fertilized early embryos. Firstly, mouse oocytes at metaphase II (MII) stage of meiosis were allocated randomly into three groups: (1) untreated (control); (2) exposed to vitrification solution without being plunged into liquid nitrogen (toxicity); or (3) vitrified by open-pulled straw (OPS) method (vitrification). Oocytes from all three groups were fertilized subsequently in vitro. The level of DNA methylation in the MII oocytes and their early embryos was then examined by immunofluorescence using an anti-5-methylcytosine (anti-5-MeC) monoclonal antibody and fluorescein isothiocyanate (FITC)-conjugated goat anti-mouse IgG. Developmental rates to 2-cell embryos (62.28%) and blastocysts (43.68%) of the vitrified–thawed oocytes were lower (P < 0.01) than those of fresh oocytes (81.47%, 61.99%) and vitrification solution treated (79.20%, 60.04%) oocytes. DNA methylation (as reflected by 5-MeC fluorescence intensity) in the vitrification group was less (P < 0.01) for MII oocyte and 2- to 8-cell stages compared with that in the control and toxicity groups. Accordingly, a reduction in global genomic methylation due to vitrification of MII oocytes may result in compromised in vitro developmental potential in early mouse embryos.

scholarly journals Highly sensitive sequencing reveals dynamic modifications and activities of small RNAs in mouse oocytes and early embryos

2016 ◽  
Vol 2 (6) ◽  
pp. e1501482 ◽  
Author(s):  
Qiyuan Yang ◽  
Jimin Lin ◽  
Miao Liu ◽  
Ronghong Li ◽  
Bin Tian ◽  
...  
Keyword(s):  
Embryonic Development ◽  
Small Rnas ◽  
Mature Mirnas ◽  
Data Sets ◽  
Sequencing Analysis ◽  
Bioinformatics Analyses ◽  
Cell Stage ◽  
Mouse Oocytes ◽  
Early Embryos ◽  
Highly Sensitive

Small RNAs play important roles in early embryonic development. However, their expression dynamics and modifications are poorly understood because of the scarcity of RNA that is obtainable for sequencing analysis. Using an improved deep sequencing method that requires as little as 10 ng of total RNA or 50 oocytes, we profile small RNAs in mouse oocytes and early embryos. We find that microRNA (miRNA) expression starts soon after fertilization, and the mature miRNAs carried into the zygote by sperm during fertilization are relatively rare compared to the oocyte miRNAs. Intriguingly, the zygotic miRNAs display a marked increase in 3′ mono- and oligoadenylation in one- to two-cell embryos, which may protect the miRNAs from the massive degradation taking place during that time. Moreover, bioinformatics analyses show that the function of miRNA is suppressed from the oocyte to the two-cell stage and appears to be reactivated after the two-cell stage to regulate genes important in embryonic development. Our study thus provides a highly sensitive profiling method and valuable data sets for further examination of small RNAs in early embryos.

Hydrogen peroxide levels in mouse oocytes and early cleavage stage embryos developed in vitro or in vivo

Development ◽  
1990 ◽  
Vol 109 (2) ◽  
pp. 501-507 ◽  
Author(s):  
M.H. Nasr-Esfahani ◽  
J.R. Aitken ◽  
M.H. Johnson
Keyword(s):  
Reproductive Tract ◽  
Female Reproductive Tract ◽  
Cell Stage ◽  
Mouse Oocytes ◽  
Early Embryos ◽  
Individual Mouse ◽  
The Relationship ◽  
Developmental Block

We describe a fluorimetric method for measuring the level of H2O2 in individual mouse oocytes and early embryos. Levels of H2O2 are low but detectable in unfertilized oocytes recovered freshly from the female reproductive tract. The levels in early cleaving embryos (1-cell to 8-cell stages) immediately after recovery from the female tract seem to be slightly higher the later the stage examined. However, when embryos are cultured in vitro from the 1-cell or early 2-cell stage, H2O2 levels rise when the embryos reach the mid-2-cell stage and remain elevated until they enter the early 4-cell stage. No equivalent elevation of H2O2 is seen during the transition from 1-cell to 2-cell or from 4-cell to 8-cell stages. Embryos that are able to develop successfully in vitro, as well as those that show a developmental block at the 2-cell stage on culture in vitro, both show this rise in H2O2 levels after in vitro culture. The relationship between the rise in H2O2 and the ‘2-cell block’ to development is discussed.

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