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Granzyme G is expressed in the two-cell stage mouse embryo and is required for the maternal-zygotic transition.
- Source :
-
BMC developmental biology [BMC Dev Biol] 2010 Aug 12; Vol. 10, pp. 88. Date of Electronic Publication: 2010 Aug 12. - Publication Year :
- 2010
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Abstract
- Background: Detailed knowledge of the molecular and cellular mechanisms that direct spatial and temporal gene expression in pre-implantation embryos is critical for understanding the control of the maternal-zygotic transition and cell differentiation in early embryonic development. In this study, twenty-three clones, expressed at different stages of early mouse development, were identified using differential display reverse transcription polymerase chain reaction (DDRT-PCR). One of these clones, which is expressed in 2-cell stage embryos at 48 hr post-hCG injection, shows a perfect sequence homology to the gene encoding the granzyme G protein. The granzyme family members are serine proteases that are present in the secretory granules of cytolytic T lymphocytes. However, the pattern of granzyme G expression and its function in early mouse embryos are entirely unknown.<br />Results: Upon the introduction of an antisense morpholino (2 mM) against granzyme G to knock-down endogenous gene function, all embryos were arrested at the 2- to 4-cell stages of egg cleavage, and the de novo synthesis of zygotic RNAs was decreased. The embryonic survival rate was dramatically decreased at the late 2-cell stage when serine protease-specific inhibitors, 0.1 mM 3,4-dichloroisocoumarin (3,4-DCI), and 2 mM phenyl methanesulphonyl fluoride (PMSF), were added to the in vitro embryonic culture medium. Survival was not affected by the addition of 0.5 mM EDTA, a metalloproteinase inhibitor.<br />Conclusion: We characterized for the first time the expression and function of granzyme G during early stage embryogenesis. Our data suggest that granzyme G is an important factor in early mouse embryonic development and may play a novel role in the elimination of maternal proteins and the triggering of zygotic gene expression during the maternal-zygotic transition.
- Subjects :
- Animals
Embryo, Mammalian metabolism
Embryonic Development
Female
Gene Knockdown Techniques
Granzymes antagonists & inhibitors
Mice
RNA, Antisense genetics
Reverse Transcriptase Polymerase Chain Reaction methods
Serine Proteinase Inhibitors metabolism
Gene Expression Regulation, Developmental
Granzymes genetics
Granzymes metabolism
Zygote metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1471-213X
- Volume :
- 10
- Database :
- MEDLINE
- Journal :
- BMC developmental biology
- Publication Type :
- Academic Journal
- Accession number :
- 20704734
- Full Text :
- https://doi.org/10.1186/1471-213X-10-88