Korean J Obstet Gynecol.  1999 Oct;42(10):2205-2213.

The Effect of Granulocyte Colony Stimulating Factor and Granulocyte Macrophage Colony Stimulating Factor on Expression of Matrix Metalloproteinase-2, 9 in Mouse Embryos

• Kim CH
• Chae HD
• Kang EH
• Cheon YP
• Kang BM
• Chang YS

Abstract

OBJECTIVES
To investigate the effect of granulocyte colony stimulating factor (G-CSF) and granulocyte macrophage colony stimulating factor (GM-CSF) on expression of matrix metalloproteinase-2, 9 (MMP-2, 9) mRNA in mouse embryos. Materials and METHOD: From October 1997 to December 1998, morula stage mouse embryos were cultured for 48 hours with G-CSF and GM-CSF at concentrations of 0.1 pg/ml, 1 pg/ml, 10 pg/ml, 100 pg/ml, 1 ng/ml and 10 ng/ml, respectively. Embryos not treated with G-CSF or GM-CSF were served as control. Reverse transcription-polymerase chain reaction (RT-PCR) has been used to examine the expression of MMP-2, 9 mRNA in developed 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 were analyzed with Kolmogorov-Smirnov test and analysis of variance (ANOVA). The statistical significance was defined as p< 0.05. RESULTS: The relative quantities (relative volume x intensity) of MMP-2 mRNA expressed in embryos of all G-CSF treatment groups were significantly increased than in the control, especially in 10, 100 pg/ml and 1 ng/ml treatment groups. The relative quantities of MMP-2 mRNA in all GM-CSF treatment groups were also significantly increased than in the control, especially in 100 pg/ml treatment group. The relative quantities of MMP-9 mRNA of all GM-CSF treatment groups except 10 ng/ml group were significantly increased than in the control, especially 10, 100 pg/ml and 1 ng/ml treatment group. However, the relative quantity of MMP-9 mRNA was significantly increased in only 10 ng/ml G-CSF treatment group than in the control and other treatment groups. CONCLUSION: This study suggests that G-CSF and GM-CSF may increase the m-RNA expression of MMP-2 or 9 in mouse blastocysts with the concentration-specific manner.

Keyword

Granulocyte colony stimulating factor(G-CSF); Granulocyte macrophage colony stimulating factor(GM- CSF); Matrix metalloproteinase-2; Reverse transcription-polymerase chain reaction(RT-PCR)

MeSH Terms

Animals
Blastocyst
Colony-Stimulating Factors*
Digestion
DNA, Complementary
Embryonic Structures*
Granulocyte Colony-Stimulating Factor
Granulocyte-Macrophage Colony-Stimulating Factor*
Granulocytes*
Matrix Metalloproteinase 2*
Mice*
Morula
Reverse Transcription
RNA, Messenger
Sensitivity and Specificity
Sequence Analysis
Colony-Stimulating Factors
DNA, Complementary
Granulocyte Colony-Stimulating Factor
Granulocyte-Macrophage Colony-Stimulating Factor
Matrix Metalloproteinase 2
RNA, Messenger