Clinical and Experimental Reproductive Medicine
- 제25권1호
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- Pages.77-86
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- 1998
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- 2233-8233(pISSN)
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- 2233-8241(eISSN)
생쥐 초기배아의 Glucose Transporter유전자 발현 양상에 관한 연구
Differential Expression of Glucose Transporter Gene in Mouse Early Embryos
- 염혜원 (삼성제일병원 제일의학연구소 불임연구실) ;
- 변혜경 (삼성제일병원 제일의학연구소 불임연구실) ;
- 송견지 (삼성제일병원 제일의학연구소 불임연구실) ;
-
김해권
(서울여자대학교 생물학과 발생학실험실) ;
- 이호준 (삼성제일병원 제일의학연구소 불임연구실)
- Youm, Hye-Won (Infertility Research Laboratory, Cheil Research Institute, Samsung Cheil Hospital) ;
- Byun, Hye-Kyung (Infertility Research Laboratory, Cheil Research Institute, Samsung Cheil Hospital) ;
- Song, Gyun-Ji (Infertility Research Laboratory, Cheil Research Institute, Samsung Cheil Hospital) ;
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Kim, Hae-Kwon
(Laboratory of Development, Department of Biology, Seoul Women's University) ;
- Lee, Ho-Joon (Infertility Research Laboratory, Cheil Research Institute, Samsung Cheil Hospital)
- 발행 : 1998.03.30
초록
The uptake of glucose for metabolism and growth is essential to most animal cells and is mediated by glucose-transporter (GLUT) proteins. The aim of this study was to determine which class of glucose transporter molecules was responsible for uptake of glucose in the mouse early embryo and at which stage the corresponding genes were expressed. In addition, co-culture system with vero cell was used to investigate the effect of the system on GLUT expression. Two-cell stage embryos were collected from the superovulated ICR female and divided into 3 groups. As a control, embryos were cultured in 0.4% BSA-T6 medium which includes glucose. For the experimental groups, embryos were cultured in either co-culture system with vero cells or glucose-free T6 medium supplemented with 0.4% BSA and pyruvate as an energy substrate. 2-cell to blastocyst stage embryos in those groups were respectively collected into microtubes (50 embryos/tube). Total RNA was extracted and RT-PCR was performed. The products were analysed after staining ethidium bromide by 2% agarose gel electrophoresis. Blastocysts were collected from each group at l20hr after hCG injection. They were fixed in 2.5% glutaraldehyde, stained with hoechst, and mounted for observation. In control, GLUT1 was expressed from 4-cell to blastocyst. GLUT2 and GLUT3 were expressed in morula and blastocyst. GLUT4 was expressed in all stages. When embryos were cultured in glucose-free medium, no significant difference was shown in the expression of GLUT1, 2 and 3, compared to control. However GLUT4 was not expressed until morular stage. When embryos were co-cultured with vero cell, there was no significant difference in the expression of GLUT1, 2, 3 and 4 compared to control. To determine cell growth of embryos, the average cell number of blastocyst was counted. The cell number of co-culture (