Vautier D, Besombes D, Chassoux D, Aubry F, Debey P
Laboratoire Associé INSERM, Institut de Biologie Physico-Chimique, Paris, France.
Mol Reprod Dev. 1994 Jun;38(2):119-30. doi: 10.1002/mrd.1080380202.
We have systematically analyzed by indirect immunofluorescence the subcellular distribution of nuclear antigens in relation to developmental stages of maturing mouse oocytes and developing embryos. Antigens were of two types: (1) a protein whose nuclear localization in interphase somatic cells depends on their proliferative state protein recognized by a monoclonal antibody 43B1N, and (2) snRNP polypeptides recognized by autoimmune sera of anti-Sm and anti-RNP type. The protein recognized by 43B1N was present in the germinal vesicle of oocytes from antral follicles, but absent from the nuclei during the first hours of embryonic life up to the middle to late 2-cell stage. Starting from this stage, it was always found in nuclei of interphase blastomeres, where its "speckles" co-localized with the speckles containing high concentrations of snRNP polypeptides. SnRNP polypeptides recognized by anti-Sm and anti-RNP sera were in turn found in nuclei of all developmental stages. When embryos were treated with aphidicolin or cytochalasin D to arrest cell division, the 43B1N reacting protein was again localized in the pronuclei at 42 hr post-hCG, i.e., slightly later than the onset of transcriptional activity. These results suggest a progressive building up of nuclei during embryonic development, which could influence gene expression.
我们通过间接免疫荧光法系统分析了核抗原的亚细胞分布与成熟小鼠卵母细胞及发育中胚胎发育阶段的关系。抗原分为两类:(1)一种蛋白质,其在间期体细胞中的核定位取决于细胞增殖状态,该蛋白质可被单克隆抗体43B1N识别;(2)可被抗Sm和抗RNP型自身免疫血清识别的snRNP多肽。43B1N识别的蛋白质存在于有腔卵泡卵母细胞的生发泡中,但在胚胎发育的最初几个小时直至2细胞期的中晚期,细胞核中均不存在。从这个阶段开始,它始终存在于间期卵裂球的细胞核中,其“斑点”与含有高浓度snRNP多肽的斑点共定位。抗Sm和抗RNP血清识别的snRNP多肽则在所有发育阶段的细胞核中均有发现。当用阿非迪霉素或细胞松弛素D处理胚胎以阻止细胞分裂时,43B1N反应蛋白在hCG后42小时再次定位于原核,即略晚于转录活性开始的时间。这些结果表明胚胎发育过程中细胞核在逐步构建,这可能会影响基因表达。