A comparative study of gene expression in murine embryos developed in vivo, cultured in vitro, and cocultured with human oviductal cells using messenger ribonucleic acid differential display

The objectives of this study were to compare the mRNA expression patterns i n early mouse embryos in different culture conditions by differential displ ay reverse transcription-polymerase chain reaction (DDRT-PCR). Embryos deve loped in vivo, cultured in vitro, and cocultured with human oviductal epith elial cells were studied at the 2-cell, 4-cell, 8-cell/morula, and blastocy st stages. Messenger RNA profiles were displayed by DDRT-PCR using downstre am T11VV (V = A, C, or G) and upstream decamer primers. Total cDNA banding patterns were highly conserved in the three groups studied. Some fragments are unique in different culture conditions. Thirteen out of the 40 selected differentially expressed clones were characterized. The DNA sequence analy ses of these clones displayed high sequence homology with cDNA sequences in the mouse expressed sequence tag database. Using semiquantitative RT-PCR, we confirmed differential expression of these DD amplicons in the three gro ups of embryos. The temporal expression of some of the selected DD amplicon s during preimplantation development were studied in the three groups of em bryos. In conclusion, DDRT-PCR is an effective tool for contrasting gene ex pression patterns and characterizing mRNA transcripts in mouse embryo.