MONITORING ULTRAVIOLET-B-INDUCED DNA-DAMAGE IN INDIVIDUAL DIATOM CELLS BY IMMUNOFLUORESCENT THYMINE DIMER DETECTION

被引:132
作者
BUMA, AGJ
VANHANNEN, EJ
ROZA, L
VELDHUIS, MJW
GIESKES, WWC
机构
[1] CTR LIMNOL, 3631 AC NIEUWERSLUIS, NETHERLANDS
[2] TNO, INST NUTR & FOOD RES, 2280 HV RIJSWIJK, NETHERLANDS
[3] NIOZ, DEPT PELAG SYST, 1790 AB DEN BURG, NETHERLANDS
关键词
BACILLARIOPHYCEAE; CYCLOTELLA; DNA DAMAGE; MONOCLONAL ANTIBODY; THYMINE DIMERS; UV-B;
D O I
10.1111/j.0022-3646.1995.00314.x
中图分类号
Q94 [植物学];
学科分类号
071001 ;
摘要
We developed a method to investigate the effect of ultraviolet-B radiation (UVBR) on the formation of thymine dimers in microalgal DIVA that can be used for both laboratory and in situ research. Antibody labeling of dimers was followed by a secondary antibody (fluorescein isothiocyanate) staining to allow visualization of DNA damage with flow cytometry or fluorescence microscopy. Thymine dimer-specific fluorescence in nuclear DNA of the marine diatom Cyclotella sp. was linearly related to the UVBR dose. Simultaneous measurements of cellular DNA content showed that the vulnerability of G2 cells to DNA damage did not differ significantly from the vulnerability of G1 cells. The formation and removal of thymine dimers in Cyclotella sp. cells was monitored for 3 consecutive days at two realistic UVBR irradiance bevels. Thymine dimers were removed within 24 h when exposed to a saturating photosynthetically active radiation intensity following the UVBR treatment. This new method allows the study of UVBR-induced DNA damage on a cell-to-cell basis. It is also feasible for field studies because cells remain intact and can be recognized readily after antibody treatment.
引用
收藏
页码:314 / 321
页数:8
相关论文
共 40 条