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.
Highlights
Gene expression profiling on the genomic scale has been routinely applied in current biological and medical research
Construction and Analysis of serial analysis of gene expression (SAGE) Libraries—The SAGE libraries of 10-bp tags constructed from mouse uteri at the interimplantation site and implantation site were deposited in the GEO repository under accession number GSE2892 (GSM63231 for the interimplantation site and GSM63232 for the implantation site)
That library was constructed with an 11-bp tag and contained 44,484 tags, 14,543 of which were unique tags [15]
Summary
Gene expression profiling on the genomic scale has been routinely applied in current biological and medical research. Oligonucleotide chips, cDNA microarrays, and serial analysis of gene expression (SAGE) are among the most widely used methods [2,3,4]. Microarray analysis has been used to investigate temporal and spatial gene expression profiles in the mouse uterus during the implantation period [5, 6]. Short nucleotide tags (usually 10 bp) from the defined position in the transcripts were used for the identification of expressed genes in SAGE analysis. The ligation of the tags into long concatemers and their sequencing result in the qualitative and quantitative gene expression profile of a particular tissue [12]. SAGE was applied to examine the global gene expression profiling in mouse uterus between the implantation site and interimplantation site
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