Takahashi T, Caviness V S
Department of Neurology, Massachusetts General Hospital, Harvard Medical School, Boston 02114.
J Neurocytol. 1993 Dec;22(12):1096-102. doi: 10.1007/BF01235751.
Proliferating cell nuclear antigen is a nuclear protein essential to DNA synthesis in eukaryotic cells. It is known to form part of a multi-protein complex which binds to DNA from the outset of S-phase of the cell cycle. We define in this analysis the interval of proliferating cell nuclear antigen binding to DNA (strictly speaking, the interval through which proliferating cell nuclear antigen is stained immunohistochemically after ethanol fixation) with respect to the stages of the cell cycle in the intact mammalian brain. The epithelium of the developing cerebral wall is favourable for such an analysis because nuclei at the same stage of the cell division cycle are spatially aligned with each other at the same depth of the epithelium. Therefore spatial location of a nucleus within the epithelium is a reliable indicator of the stage of the cell cycle for that nucleus. Proliferating cell nuclear antigen-DNA binding in this epithelium is initiated in the final 5% (26 min) of G1-phase and continues through the initial 35% (1.3 h) of S-phase. This phasic pattern of proliferating cell nuclear antigen-DNA binding, as revealed for the first time in the intact cerebral wall, approximates closely the phasic pattern as it has been characterized until now only in vitro in vertebrate cell lines. This analysis illustrates the potential of the cerebral proliferative epithelium for study of the molecular events of the cell cycle under in vivo conditions of histogenetic regulation.
增殖细胞核抗原是真核细胞中DNA合成所必需的一种核蛋白。已知它是一种多蛋白复合物的一部分,该复合物从细胞周期的S期开始就与DNA结合。在本分析中,我们确定了完整哺乳动物大脑中增殖细胞核抗原与DNA结合的时间间隔(严格来说,是乙醇固定后通过免疫组织化学染色增殖细胞核抗原的时间间隔)与细胞周期各阶段的关系。发育中的脑壁上皮有利于进行这样的分析,因为处于细胞分裂周期同一阶段的细胞核在同一上皮深度处彼此在空间上排列整齐。因此,上皮内细胞核的空间位置是该细胞核细胞周期阶段的可靠指标。该上皮中增殖细胞核抗原与DNA的结合在G1期的最后5%(26分钟)开始,并持续到S期的最初35%(1.3小时)。这种增殖细胞核抗原与DNA结合的阶段性模式,首次在完整的脑壁中揭示,与迄今为止仅在脊椎动物细胞系体外所表征的阶段性模式非常接近。该分析说明了脑增殖上皮在组织发生调控的体内条件下研究细胞周期分子事件的潜力。