We carried out DNA microarray-based global expression profiling of all preimplantation stages in mouse. It revealed and characterized the distinctive patterns of maternal RNA degradation and two major transient waves of de novo transcription: zygotic genome activation (ZGA) and mid-preimplantation gene activation (MGA). We also developed a technique to do large-scale Whole Mount In Situ Hybridization (WISH), which allows us to reveal the spatial expression patterns of 91 genes in mouse preimplantation embryos. Through these analyses, we identified a number of genes that show unique expression patterns. We have demonstrated that one of them, named Zscan4, is expressed exclusively in 2-cell mouse embryos and ES cells and plays a critical role in the progression of preimplantation development. Another gene (Chuk) shows constant RNA levels throughout preimplantation development and can be used as an internal standard suitable for quantitative RT-PCR. We also identified genes, named Trim43a, Trim43b, and Trim43c, whose expression began at the 2-cell stage, peaked at the 8-cell/morula stage, and dramatically fell by the blastocyst stage. We identified a 5 kb DNA fragment that covers upstream region of Trim43a as a putative promoter, which can drive the expression of mStrawberry fluorescent protein in a manner similar to endogenous Trim43 genes. Trim43 genes will be useful stage-specific markers for the study of preimplantation embryos. We are currently studying the functions of these genes in detail.

Agency
National Institute of Health (NIH)
Institute
National Institute on Aging (NIA)
Type
Investigator-Initiated Intramural Research Projects (ZIA)
Project #
1ZIAAG000655-11
Application #
7963998
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
11
Fiscal Year
2009
Total Cost
$282,354
Indirect Cost
Name
National Institute on Aging
Department
Type
DUNS #
City
State
Country
Zip Code
Amano, Tomokazu; Hirata, Tetsuya; Falco, Geppino et al. (2013) Zscan4 restores the developmental potency of embryonic stem cells. Nat Commun 4:1966
Monti, Manuela; Zanoni, Mario; Calligaro, Alberto et al. (2013) Developmental arrest and mouse antral not-surrounded nucleolus oocytes. Biol Reprod 88:2
Ko, Shigeru B H; Azuma, Sakiko; Yokoyama, Yukihiro et al. (2013) Inflammation increases cells expressing ZSCAN4 and progenitor cell markers in the adult pancreas. Am J Physiol Gastrointest Liver Physiol 304:G1103-16
Hung, Sandy S C; Wong, Raymond C B; Sharov, Alexei A et al. (2013) Repression of global protein synthesis by eif1a-like genes that are expressed specifically in the two-cell embryos and the transient zscan4-positive state of embryonic stem cells. DNA Res 20:391-402
Xie, Huirong; Sun, Xiaofei; Piao, Yulan et al. (2012) Silencing or amplification of endocannabinoid signaling in blastocysts via CB1 compromises trophoblast cell migration. J Biol Chem 287:32288-97
Hammachi, Fella; Morrison, Gillian M; Sharov, Alexei A et al. (2012) Transcriptional activation by Oct4 is sufficient for the maintenance and induction of pluripotency. Cell Rep 1:99-109
Hirata, Tetsuya; Amano, Tomokazu; Nakatake, Yuhki et al. (2012) Zscan4 transiently reactivates early embryonic genes during the generation of induced pluripotent stem cells. Sci Rep 2:208
Kim, Youngjo; Sharov, Alexei A; McDole, Katie et al. (2011) Mouse B-type lamins are required for proper organogenesis but not by embryonic stem cells. Science 334:1706-10
Zalzman, Michal; Falco, Geppino; Sharova, Lioudmila V et al. (2010) Zscan4 regulates telomere elongation and genomic stability in ES cells. Nature 464:858-63
Sharov, Alexei A; Piao, Yulan; Ko, Minoru S H (2010) Gene expression profiling of mouse embryos with microarrays. Methods Enzymol 477:511-41

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