Embryotrophic Factor-3 from Human Oviductal Cells Affects the Messenger RNA Expression of Mouse Blastocyst1

Abstract

Our previous results showed that embryotrophic factor-3 (ETF-3) from human oviductal cells increased the size and hatching rate of mouse blastocysts in vitro. The present study investigated the production of ETF-3 by an immortalized human oviductal cell line (OE-E6/E7) and the effects of ETF-3 on the mRNA expression of mouse embryos. The ETF-3 was purified from primary oviductal cell conditioned media using sequential liquid chromatographic systems, and antiserum against ETF-3 was raised. The ETF-3-supplemented Chatot-Ziomek-Bavister medium was used to culture Day 1 MF1 x BALB/c mouse embryos for 4 days. The ETF-3 treatment significantly enhanced the mouse embryo blastulation and hatching rate. The antiserum, at concentrations of 0.03-3%, abolished the embryotrophic effect of ETF-3. Positive ETF-3 immunoreactivity was detected in the primary oviductal cells, OE-E6/E7, and blastocysts derived from ETF-3 treatment. Vero cells (African Green Monkey kidney cell line), fibroblasts, and embryos cultured in control medium did not possess ETF-3 immunoreactivity. The mRNA expression patterns of the treated embryos were studied at the blastocyst stage by mRNA differential display reverse transcription-polymerase chain reaction (DDRT-PCR). The DDRT-PCR showed that some of the mRNAs were differentially expressed after ETF-3 treatment. Twelve of the differentially expressed mRNAs that had high homology with cDNA sequences in the GenBank were selected for further characterization. The differential expression of seven of these mRNAs (ezrin, heat shock 70-kDa protein, cytochrome c oxidase subunit VIIa-L precursor, proteinase-activated receptor 2, eukaryotic translation initiation factor 2beta, cullin 1, and proliferating cell nuclear antigen) was confirmed by semiquantitative RT-PCR. In conclusion, immortalized oviductal cells produce ETF-3, which influences mRNA expression of mouse blastocyst.

Phospholipid transfer protein (PLTP) mRNA expression is stimulated by developing embryos in the oviduct

“…These techniques have been used successfully to identify differentially expressed genes from the oviduct [Lee et al, 2000 and preimplantation embryos [Natale et al, 2000;Lee et al, 2001Lee et al, , 2003Ponsuksili et al, 2002;Yao et al, 2003]. In combination with cDNA microarray, SSH has been used to compare the gene expression patterns between the ipsilateral and contralateral bovine oviduct in the post-ovulation period ].…”

“…Conjointly, our laboratory first reported the purification of embryotrophic factor-3 (ETF-3) from human oviductal cells [Liu et al, 1995]. ETF-3 could enhance embryo development in vitro Lee et al, 2003] and it was subsequently identified as C3-like molecules. Mouse embryos incubated with C3b and/or iC3b have large size and higher hatching rate in vitro [Lee et al, 2004].…”

Increased fetal abortion rate in autoimmune thyroid disease is related to circulating TPO autoantibodies in an autoimmune thyroiditis animal model

“…Mouse embryos were collected and cultured as described previously (21). Briefly, 6-8-week-old female mice were superovulated by intraperitoneal injection of 5 IU pregnant mare's serum gonadotropin (Sigma) followed by an injection of 5 IU hCG (Sigma) 46 h later.…”

Demilune Cell and Parotid Protein from Murine Oviductal Epithelium Stimulates Preimplantation Embryo Development

“…The procedure for obtaining superovulated embryos was essentially the same as before (27). Mature ICR female mice (age, 6 -8 wk) were superovulated with 5 IU pregnant mare's serum gonadotropin (Sigma), followed by an injection of 5 IU of hCG (Sigma) 46 h later.…”

“…We previously demonstrated that oviductal cells affect the gene expression of developing embryos in vitro (26), partly via embryotrophic factor-3 (27). On the other hand, developing embryos have also altered gene expression in the oviduct (6,28).…”