Article

Serial analysis of gene expression in mouse uterus at the implantation site

College of Life Science, Northeast Agricultural University, Harbin 150030, China.
Journal of Biological Chemistry (Impact Factor: 4.6). 05/2006; 281(14):9351-60. DOI: 10.1074/jbc.M511512200
Source: PubMed

ABSTRACT Although oligonucleotide chips, cDNA microarrays, differential display reverse transcription-PCR, and other approaches have been used to screen implantation-related molecules, the mechanism by which embryo implantation occurs is still unknown. The aim of this study was to profile the differential gene expression between interimplantation site and implantation site in mouse uterus on day 5 of pregnancy by serial analysis of gene expression (SAGE). In our two SAGE libraries of 11-bp tags, the total numbers of tags sequenced were 48,121 for the interimplantation site and 50,227 for the implantation site. There were 1,039 tags specifically expressed at interimplantation site, and 1,252 tags specifically expressed at the implantation site. Based on the p value, there were 195 tags significantly up-regulated at the interimplantation site and 261 tags significantly up-regulated at the implantation site, of which 100 genes were single matched at the interimplantation site and 127 genes were single matched at the implantation site, respectively. By reverse transcription-PCR, the tag ratio between the implantation site and interimplantation site was verified on 14 significantly changed genes. Using in situ hybridization, 1810014L12Rik, Psmb5, Cd63, Npm1, Fads3, and Tagln2 were shown to be highly expressed at the implantation site compared with the interimplantation site. Compared with the interimplantation site, Ddx39 was strongly expressed in the subluminal stromal cells at the implantation site on day 5 of pregnancy. Ddx39 expression at the implantation site was specifically induced by active blastocysts. Additionally, Ddx39 expression was significantly up-regulated by estrogen in the ovariectomized mice. In our SAGE data, many implantation-related genes were identified in mouse uterus. Our data could be a valuable source for future study on embryo implantation.

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    • "Fads3 and its AT expression are tissue specific [6] [7], are evolutionarily conserved and are widely and differentially expressed across 12 neonatal baboon tissues [6] [9]. Fads3 is highly expressed at the implantation site of the embryo in mouse uterus [10] and its AT are differentially expressed in response to neuronal cell differentiation [6], suggesting that Fads3 products have distinct physiological functions. Further, recent studies have provided evidence that Fads3 may play an important role in regulating lipid metabolism [11] [12]. "
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    • "Digoxigenin-labeled complementary RNA probes were transcribed in vitro using a digoxigenin RNA labeling kit (Roche, Indianapolis, IN, USA). In situ hybridization was performed as described [21] [22]. Briefly, uteri were cut into 4-to 6-mm long pieces, flash frozen in liquid nitrogen, and cut into 10-mm frozen sections, mounted on 3-aminopropyltriethoxy- silane (Sigma)-coated slides and fixed in 4% paraformaldehyde solution. "
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    • "Although the localization of Nedd4 in the uterus is unclear, the levels of Nedd4 mRNA have been shown to be regulated in the mouse uterus at the implantation site [50]. Knockout of Nedd4 in mice is embryonically lethal and mutant embryos exhibit a reduction in skeletal muscle fiber size [51]. "
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