Probing the mechanism of sister chromatid exchange formation with the fluorescent plus Giemsa technique

The effects of x rays and light flashes on the SCE formation in BrdUrd-substituted Chinese hamster ovary (CHO) cells are examined using the fluorescent plus Giemsa (FPG) technique, which allows SCE's to be visualized with greater precision than 3H autoradiography does. Near-diploid cells are selected and scoring SCE from every chromosome of near-diploid cells or from chromosome no. 1 is found to give a more accurate measure of the SCE induction. Double synchronous cells are used for the x ray study and asynchronous cells are used for the light flash study. The results indicate that x rays can induce SCE's throughout the cell cycle with S the most sensitive stage and G2 the least sensitive stage. Light flashes can induce an appreciable amount of SCE's while no appreciable amount of chromosomal aberrations can be detected under conditions used in this study. For x-ray-induced chromosomal aberrations, G2 is the most sensitive stage with G1 being slightly more sensitive than S, which is consistent with other observations. Additionally, aberration induction is shown to be a more sensitive indicator of x-ray-induced cell damage than SCE induction, but SCE induction is a potentially good indicator of mutation induction. This study also suggests that symmetrically reunited isochromatid breaks can not be the sole source of SCE's and SCE induction is a different but not independent radiobiological effect from chromosomal aberration induction. SCE formation is proposed to be a consequence of specific type(s) of single strand breaks in DNA