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.
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.
Kim
Jeong Hee
JH
Department of Obstetrics and Gynecology, College of Medicine, Asan Medical Center, University of Ulsan, Seoul, South Korea.
Hong
Seok Ho
SH
Nah
Hee Young
HY
Lee
Ji Yun
JY
Chae
Hee Dong
HD
Kim
Chung Hoon
CH
Kang
Byung Moon
BM
Bae
In Ha
IH
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*
D012333:RNA, Messenger; D016211:Transforming Growth Factor alpha; D066246:ErbB Receptors; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9