Regulation of gene expression in the mouse oocyte and early preimplantation embryo: developmental changes in Sp1 and TATA box-binding protein, TBP.
Regulation of gene expression in the mouse oocyte and early preimplantation embryo: developmental changes in Sp1 and TATA box-binding protein, TBP.
复制标题
DOI：
--
复制
发表时间：
1994-08
期刊：
影响因子：
4.6
通讯作者：
D. Worrad;Prahlad T. Ram;R. Schultz
中科院分区：
文献类型：
--
作者：
D. Worrad;Prahlad T. Ram;R. Schultz
我们先前证明，与显微注射雌性原核相比，显微注射雄性原核后，Sp1依赖的报告基因在1-细胞小鼠胚胎G2中优先表达(P.T.Ram和R.M.Schultz，1993，Dev。比奥尔。156、552-556)。我们还注意到，在显微注射完全发育的卵母细胞的生发泡后，没有观察到报告基因的表达。在本研究中，我们检测了该报告基因在卵母细胞生长过程中的表达，以及两种转录因子Sp1和TATA盒结合蛋白TBP在卵母细胞生长和第一个细胞周期中的核浓度。在卵母细胞生长过程中，报告基因表达的程度下降，这种下降与共聚焦免疫荧光显微镜确定的Sp1核浓度的下降有关。此外，免疫印迹实验的结果也表明，在卵母细胞生长过程中，Sp1的总浓度也有类似的下降。通过共聚焦免疫荧光显微镜检测，TBP的核浓度在卵母细胞生长过程中也会降低。受精后，这两种转录因子的原核浓度以时间依赖的方式增加，并且在雄性原核中的浓度高于雌性原核。对于每个原核和每个转录因子，这种核浓度的增加都被抑制DNA合成的aphidiclin所抑制。最后，在1-细胞和2-细胞阶段观察到的这两种蛋白质核浓度的增加不需要转录或胞质分裂。
We previously demonstrated that an Sp1-dependent reporter gene is preferentially expressed in G2 of the 1-cell mouse embryo following microinjection of the male pronucleus when compared to microinjection of the female pronucleus (P.T. Ram and R.M. Schultz, 1993, Dev. Biol. 156, 552-556). We also noted that expression of the reporter gene is not observed following microinjection of the germinal vesicle of the fully grown oocyte. In the present study, we examined expression of this reporter gene during oocyte growth, as well as the nuclear concentration of two transcription factors, Sp1 and the TATA box-binding protein, TBP, during oocyte growth and the first cell cycle. The extent of reporter gene expression decreases during oocyte growth and this decrease correlates with the decrease in nuclear concentration of Sp1, as determined by confocal immunofluorescent microscopy. In addition, results of immunoblotting experiments also indicate a similar decrease in the total concentration of Sp1 during oocyte growth. The nuclear concentration of TBP also decreases during oocyte growth, as determined by confocal immunofluorescent microscopy. Following fertilization, the pronuclear concentration of these two transcription factors increases in a time-dependent fashion and the concentration of each is greater in the male pronucleus as compared to the female pronucleus. For each pronucleus and for each transcription factor, this increase in nuclear concentration is inhibited by aphidicolin, which inhibits DNA synthesis. Last, the increase in nuclear concentration of these two proteins observed between the 1-cell and 2-cell stages does not require transcription or cytokinesis.