Preimplantation-embryo-specific Cell-cycle Regulation is Attributable to a Low Expression of Retinoblastoma Protein rather than Its Phosphorylation
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概要
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Mammalian preimplantation embryos enter the S phase immediately after the end of the M phase; their cell cycle lacks a substantial G1 phase. Previously, we suggested that the absence of the G1 phase was attributable to a loss of retinoblastoma protein (RB) function, which is required for suppression of S phase entrance and that this loss of RB function in turn was attributable to the low RB expression level during preimplantation development in mouse embryos. The present study aimed to examine whether or not RB inhibition by CDK4/6-cyclin D-dependent phosphorylation is involved in the loss of RB function in preimplantation mouse embryos by the expression of p16INK4a, a potent endogenous inhibitor of CDK4/6-cyclin D. First, the decrease in RB expression between the four-cell and morula stages was confirmed in in vivo-derived mouse embryos. We then examined the efficiency of the p16INK4a expression vector in inhibiting RB phosphorylation and cell cycle progression using NIH-3T3 cells and obtained gradual RB dephosphorylation and a significantly lower proliferation rate in p16INK4a-transfected cells than in control cells. This indicated the successful p16INK4a effects on cell-cycle progression by the vector used. On the other hand, the development rate of mouse embryos injected with the p16INK4a expression vector was the same as that of the control embryos, although p16INK4a expression was detected at mRNA and protein levels in the former group but not in the control group. These results suggest that RB phosphorylation is not involved in RB dysfunction or in the lack of a G1 phase in mouse embryos and that the decrease in RB expression is important for preimplantation-embryo-specific cell-cycle regulation. Moreover, the present study indicates the similarity between preimplantation embryos and cancer cells, which p16INK4a expression does not arrest at the G1 phase.
著者
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Kano Kiyoshi
Laboratory Of Applied Genetics Department Of Animal Resource Science Graduate School Of Agriculture
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Naito Kunihiko
Laboratory Of Applied Genetics Department Of Animal Resource Sciences Graduate School Of Agricultura
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Egashira Asuka
Laboratory Of Applied Genetics Graduate School Of Agriculture And Life Sciences University Of Tokyo
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NAITO Kunihiko
Laboratory of Applied Genetics, Graduate School of Agriculture and Life Sciences, University of Tokyo, Tokyo 113-8657, Japan
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EGASHIRA Asuka
Laboratory of Applied Genetics, Graduate School of Agriculture and Life Sciences, University of Tokyo, Tokyo 113-8657, Japan
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