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. 2002 May;19(5):232–239. doi: 10.1023/A:1015310919415

Influence of Transforming Growth Factor-α on Expression of Matrix Metalloproteinase-2, Matrix Metalloproteinase-9, and Epidermal Growth Factor Receptor Gene in the Mouse Blastocysts

Jeong Hee Kim 1,2, Seok Ho Hong 1,3, Hee Young Nah 1, Ji Yun Lee 1,3, Hee Dong Chae 1, Chung Hoon Kim 1, Byung Moon Kang 1, In Ha Bae 2
PMCID: PMC3468233  PMID: 12099554

Abstract

Purpose: This study was carried out to investigate the influence of transforming growth factor-α (TGF-α) on the expression of mRNA for matrix metalloproteinase-2 (MMP-2), MMP-9, and epidermal growth factor receptor (EGFR) in mouse blastocysts and the effect on the production and activation of MMP-2 and MMP- 9 during blastocyst outgrowth.

Methods: Two-cell mouse embryos were cultured for 96 h in the presence or absence of various concentrations of TGF-α. Reverse transcription–polymerase chain reaction (RT-PCR) was used to examine the expression of mRNA for MMP-2, MMP-9, and EGFR in in vitro cultured blastocysts. To investigate the effect on the production and activation of MMP-2 and MMP-9 during blastocyst outgrowth, the conditioned medium collected after 3 and 5 days of embryo culture were assayed for MMP activity by gelatin zymography.

Results: The relative mRNA levels of MMP-2 and MMP-9 in blastocysts treated with TGF-α were higher than that of the control in a concentration-dependent manner. The relative mRNA level of EGFR in blastocysts treated with TGF-α was higher than that of the control. In conditioned medium collected after 3 days of embryo culture, TGF-α induced the gelatinase activities of proMMP-9 in all groups and activated MMP-2 in the 10 and 100 ng/mL TGF-α treated groups. In conditioned medium collected after 5 days, TGF-α induced the gelatinase activities of proMMP-9 in all groups and activated MMP-9 in the TGF-α treated group. TGF-α also induced the gelatinase activities of activated MMP-2 in the 1 and 10 ng/mL TGF-α treated groups and the control.

Conclusions: These results suggest that the addition of TGF-α to in vitro culture medium is proper to create a favorable environment for preimplantation embryo development and implantation.

Keywords: Matrix metalloproteinase-2, matrix metalloproteinase-9, mouse blastocyst, transforming growth factor-α, zymography

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