title: Influence of transforming growth factor-alpha on expression of matrix metalloproteinase-2, matrix metalloproteinase-9, and epidermal growth factor receptor gene in the mouse blastocysts.
abstract: OBJECTIVE
  This study was carried out to investigate the influence of transforming growth factor-alpha (TGF-alpha) 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-alpha. 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-alpha were higher than that of the control in a concentration-dependent manner. The relative mRNA level of EGFR in blastocysts treated with TGF-alpha was higher than that of the control. In conditioned medium collected after 3 days of embryo culture, TGF-alpha induced the gelatinase activities of proMMP-9 in all groups and activated MMP-2 in the 10 and 100 ng/mL TGF-alpha treated groups. In conditioned medium collected after 5 days, TGF-alpha induced the gelatinase activities of proMMP-9 in all groups and activated MMP-9 in the TGF-alpha treated group. TGF-alpha also induced the gelatinase activities of activated MMP-2 in the 1 and 10 ng/mL TGF-alpha treated groups and the control.
  
  
  CONCLUSIONS
  These results suggest that the addition of TGF-alpha to in vitro culture medium is proper to create a favorable environment for preimplantation embryo development and implantation.
authors.affiliation: Department of Obstetrics and Gynecology, College of Medicine, Asan Medical Center, University of Ulsan, Seoul, South Korea.
authors.forename: Jeong Hee
authors.initials: JH
authors.lastname: Kim
authors.forename: Seok Ho
authors.initials: SH
authors.lastname: Hong
authors.forename: Hee Young
authors.initials: HY
authors.lastname: Nah
authors.forename: Ji Yun
authors.initials: JY
authors.lastname: Lee
authors.forename: Hee Dong
authors.initials: HD
authors.lastname: Chae
authors.forename: Chung Hoon
authors.initials: CH
authors.lastname: Kim
authors.forename: Byung Moon
authors.initials: BM
authors.lastname: Kang
authors.forename: In Ha
authors.initials: IH
authors.lastname: Bae
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000201:enzymology*; D004586:Electrophoresis; D015180:Electrophoresis, Gel, Two-Dimensional; D066246:ErbB Receptors / Q000096:biosynthesis / Q000235:genetics; D005260:Female; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D008297:Male; D020778:Matrix Metalloproteinase 2 / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008813:Mice, Inbred ICR; D011247:Pregnancy; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016211:Transforming Growth Factor alpha / Q000494:pharmacology*
chemical_list: D012333:RNA, Messenger; D016211:Transforming Growth Factor alpha; D066246:ErbB Receptors; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9