Coculture of preimplantation embryo with an immortalized human oviductal epithelial cells

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We have tried establish an immortalized cell line of human tubal epithelium. It is not possible to obtain immortalized cells after repeated experiments. We also have performed experiments using preimplantation embryos. We determined the expression of fibroblast growth factor (FGF) -2 and the FGF receptor 1 (FGFR-1) mRNA in the mouse embryos and in epithelial cells of the uterine endometrium using reverse transcription-polymerase chain reaction (RT-PCR). No FGF-2 mRNA was detected in mouse embryos but FGFR-1 mRNA was expressed at the blastocyst stage. These results suggest importance of FGFR-1 in preimplantation development of embryo. To examine the mechanism of FGFR-1 expression, the arrangement of exons and introns encoding 5'-side of murine FGFR-1 gene was mapped. A large intron with a size of 14kb was identified between exon 1 and 2. All subtypes of FGFR-1 was obserbed to be generated through alternative aplicing. Furthermore, we characterized a transcription element of this gene. The basal promoter element was mapped to the 5'-flanking region from -89 to -43. The DNase I protection assay and gel shift analysis revealed that a nuclear protein could bind to the nucleotide sequence from -62 to -42. In conclusion, FGF-2 produced by the female reproductive tract may exert paracrine effects on preimplantation embryos. FGFR-1 gene has an unique genomic structure. The 5'-flanking region from -62 to -42 plays a pivotal role in FGFR-1 gene expression.