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a Department of Biochemistry I, Yokohama City University School of Medicine, Yokohama 236-0004, Japan
One- and two-dimensional polyacrylamide microslab gel electrophoresis followed by silver staining was devised to visualize picogram to nanogram levels of proteins and was applied to the analysis of 1–20 mouse oocytes and embryos (approximately 16.5–330 ng of protein) during preimplantation development. Compared with values in embryos, more bands in the higher molecular weight range were found only for unfertilized oocytes in one-dimensional microelectrophoresis. A marked decrease in the number of protein spots occurred after fertilization in two-dimensional microelectrophoresis. Both findings indicate a decrease in maternal proteins caused by fertilization. Silver-staining densities were almost invariable for 8 major spots, but increased, decreased, or varied for 32 minor spots in developing embryos from the 1-cell to the morula stage, signifying spot-specific changes in the expression of zygotic proteins during development. The protein patterns in cumulus cells and blastocysts were different from those in oocytes and embryos. Even in a single 1-cell embryo, major spots and some minor spots were detectable by our two-dimensional microelectrophoretic technique, but many more minor spots were visualized in five 1-cell embryos, exemplifying the limit of our microelectrophoretic technique. As a preliminary result, a two-dimensional immunoblot pattern is shown for glucose transporter 1 expressed in morulae.
2 Correspondence: Takahiko Kato, Department of Biochemistry I, Yokohama City University School of Medicine, Fukuura, Kanazawaku, Yokohama 236-0004, Japan. FAX: 81 45 784 4530; katomdpr{at}med.yokohama-cu.ac.jp
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