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Korean Journal of Obstetrics & Gynecology 2001;44(2):312-318.
Published online February 1, 2001.
The Effect of Transforming Growth Factor-alpha on the Preimplantation Development, Implantation and Epidermal Growth Factor Receptor Expression in Mouse Embryos.
Sung Hoon Kim, Eun Ju Park, Hee Dong Chae, Chung Hoon Kim, Byung Moon Kang, Yoon Seok Chang, Seok Ho Hong, Eui Ju
Department of Obstetrics and Gynecology, College of Medicine, University of Ulsan, Asan Medical Center, Seoul, Korea.
Abstract
OBJECTIVE
To investigate the influence of transforming growth factor-alpha (TGF-alpha) on preimplantation development, implantation and epidermal growth factor receptor (EGFR) expression in mouse embryos. MATERIALS AND METHODS: Eight-cell stage mouse embryos were cultured for 48 hours with TGF-alpha at concentrations of 1.0, 10 and 100 ng/ml. Embryos not treated with TGF-alpha served as control. The percentages of embryos which developed to blastocyst stage, expanded, hatched blastocyst stage and in vitro implantation at 48 hours were determined. Riverse transcription-polymerase chain reaction (RT-PCR) has been used to examine the expression of EGFR in developed hatched blastocysts. Following reverse transcription, strategically designed nested primers, optimized for specificity, were used for amplification from the cDNA equivalent of a single embryo. The products were then verified by restriction enzyme digestion and sequence analysis. RESULTS: The percentages of embryos which developed to blastocyst stage were significantly higher following incubation with TGF-alpha at concentration of 10 ng/ml(p<0.05). The percentages of embryos which developed to expanded blastocyst stage were significantly higher in TGF-alpha treatment group at concentration of 100 ng/ml(p<0.05). The percentages of embryos which developed to hatched blastocyst stage were significantly higher following incubation with TGF-alpha at concentration of 10 ng/ml and 100 ng/ml(p<0.05). The percentages of implanted blastocyst in vitro were significantly higher following incubation with TGF-alpha at concentrations of 100 ng/ml compared to the control(p<0.05). The mRNA concentration of EGFR in embryos treated with 100 ng/ml of EGF was significantly higher than those of the control and other EGF treatment groups(p<0.005, p<0.05, p <0.05, respectively). CONCLUSION: TGF-alpha may stimulatory role in embryonic development, implantation and expression of EGFR in embryo itself. These results suggest that TGF-alpha may act directly on the mouse embryo and favor its implantaion, irrespective of the presence or absence of the endometrium.
Key Words: Transforming growth factor-alpha (TGF-alpha), embryo development, implantation, epidermal growth factor receptor (EGFR) expression


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