Nonradioisotopic Quantitative RT-PCR to Detect Changes in mRNA Levels during Early Mouse Embryo Development
Copyright policy )Nonradioisotopic Quantitative RT-PCR to Detect Changes in mRNA Levels during Early Mouse Embryo Development
We developed a non-radioisotopic quantitative RT-PCR method with high sensitivity and reproducibility. The results of this RT-PCR were in agreement with those of the Northern blot analysis. We measured the mRNA levels of beta-actin, transferrin receptor, and two cell cycle-related genes, cyclin B and cdc25, in early mouse embryos by the RT-PCR. In late two-cell stage embryos, beta-actin, transferrin receptor and cyclin B mRNA levels were 10-20% of those in MII stage oocytes. In contrast, the cdc25 mRNA levels were not different between these stages. When we cultured mouse embryos, the presence of an RNA polymerase inhibitor, alpha-amanitin, in the medium did not affect the mRNA levels at the two-cell stage, indicating that most of the detected mRNAs in two-cell embryos were maternally derived. These results suggest that the rate of mRNA degradation is different between cyclin B and cdc25 during early embryogenesis.
- Kyoto University Japan
- Shiga University of Medical Science Hospital Japan
- Niigata University Japan
Base Sequence, Macrophage Colony-Stimulating Factor, Molecular Sequence Data, Mice, Inbred Strains, RNA-Directed DNA Polymerase, Oligonucleotides, Antisense, Blotting, Northern, Embryo, Mammalian, Polymerase Chain Reaction, Actins, Cell Line, Embryonic and Fetal Development, Mice, Oligodeoxyribonucleotides, Bone Marrow, Cyclins, Protein Biosynthesis, Animals, Female, RNA, Messenger
Base Sequence, Macrophage Colony-Stimulating Factor, Molecular Sequence Data, Mice, Inbred Strains, RNA-Directed DNA Polymerase, Oligonucleotides, Antisense, Blotting, Northern, Embryo, Mammalian, Polymerase Chain Reaction, Actins, Cell Line, Embryonic and Fetal Development, Mice, Oligodeoxyribonucleotides, Bone Marrow, Cyclins, Protein Biosynthesis, Animals, Female, RNA, Messenger
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