Measuring RNA Levels in Mouse Embryos
Author Information
Author(s): Hartshorn Cristina, Eckert Judith J, Hartung Odelya, Wangh Lawrence J
Primary Institution: Brandeis University
Hypothesis
Simultaneous quantification of Oct4 and Xist transcripts in individual blastomeres at the 8-cell stage could be informative as to their subsequent fate.
Conclusion
The study demonstrates that Oct4 and Xist RNA levels vary in individual blastomeres of the same embryo, indicating that 8-cell blastomeres are not equivalent in their developmental properties.
Supporting Evidence
- Oct4 RNA is present in all cleavage-stage blastomeres, while Xist RNA is present in hundreds of copies in female cells but is absent from male cells.
- The study provides the first report of quantification of cDNA with LATE-PCR.
- Cells comprising the same embryos contain markedly dissimilar levels of both Oct4 and Xist transcripts.
- Quantification of differential expression of genes involved in development contributes to the hypothesis that lineage commitment in the mammalian embryo starts very early.
Takeaway
This study shows that different cells in the same embryo can have different amounts of important RNA, which helps scientists understand how embryos develop.
Methodology
The study used a duplex RT-LATE-PCR assay for real-time measurement of Oct4 and Xist templates in individual blastomeres isolated from 8-cell stage embryos.
Limitations
The study's findings may not be applicable to other species, and the use of frozen-thawed or cultured embryos may alter gene expression patterns.
Participant Demographics
Mouse embryos from B6C3F1 female mice bred with B6D2F1 male mice.
Digital Object Identifier (DOI)
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