Dewey W C, Humphrey R M, Sedita B A
Biophys J. 1966 May;6(3):247-60. doi: 10.1016/S0006-3495(66)86654-7.
Chinese hamster cells in vitro were double labeled with C(14)TdR and H(3)TdR. At the time of irradiation with Co(60) gamma rays (600 rad), the cells in the G(2) phase were labeled only with C(14), whereas cells in the late and middle S phases were labeled with both C(14) and H(3). The cells in early S phase were labeled only with H(3) and the G(1) cells were unlabeled. Samples were fixed at various time intervals following irradiation and the metaphases were analyzed for chromosomal damage. The phase in which the cell was located at the time of irradiation was determined by counting grains in the first and second layers of autoradiographic film. In both control and irradiated cells some G(1) cells divided prior to some of the cells which were in the S phase denoting mixing of the populations. The G(2) phase sustained three times more chromosomal damage than the S phase. Little difference in chromosomal damage was found between the G(1) and S phases or among the different parts of the S phase. Cells in G(2) sustained a mitotic delay of 4 hr, while the other phases sustained a delay of 2 to 3 hr. Chromatid and chromosome (dicentrics) exchanges were induced in G(1) cells but only chromatid exchanges were induced in S and G(2) cells; this is consistent with the hypothesis that the chromosome consists of two subunits which separate either slightly before or immediately as the cell enters the S phase.
体外培养的中国仓鼠细胞用C(14)TdR和H(3)TdR进行双重标记。在用Co(60)γ射线(600拉德)照射时,处于G(2)期的细胞仅用C(14)标记,而处于S期后期和中期的细胞则用C(14)和H(3)标记。处于S期早期的细胞仅用H(3)标记,而G(1)期细胞未被标记。在照射后的不同时间间隔固定样本,并分析中期染色体损伤情况。通过计数放射自显影片第一层和第二层中的银粒来确定照射时细胞所处的阶段。在对照细胞和照射细胞中,一些G(1)期细胞比一些处于S期的细胞先分裂,这表明细胞群体发生了混合。G(2)期遭受的染色体损伤是S期的三倍。在G(1)期和S期之间或S期的不同部分之间,染色体损伤几乎没有差异。G(2)期细胞有4小时的有丝分裂延迟,而其他阶段有2至3小时的延迟。G(1)期细胞诱导产生染色单体和染色体(双着丝粒)交换,但S期和G(2)期细胞仅诱导产生染色单体交换;这与染色体由两个亚基组成的假说一致,这两个亚基在细胞进入S期之前或进入S期时立即稍有分离。