Grigorova M, Brand R, Xiao Y, Natarajan A T
MGC, Department of Radiation Genetics and Chemical Mutagenesis, Leiden University, The Netherlands.
Int J Radiat Biol. 1998 Sep;74(3):297-314. doi: 10.1080/095530098141447.
To estimate the frequencies of radiation- (low and high LET) induced chromosome aberrations in Chinese hamster splenocytes by two-colour fluorescence in situ hybridization using DNA painting probes specific for chromosomes 2, 3, 8, X and Y and to determine (1) the ratio of radiation-induced translocations and dicentrics; (2) the spectrum of exchange aberrations induced by X-rays and neutrons; and (3) the relative involvement of the different chromosomes in the formation of aberrations.
Isolated splenocytes from the Chinese hamster were irradiated in vitro with different doses of 200 kV X-rays (0.75, 1.5, 3.0 Gy) and 1 MeV fast neutrons (0.25, 0.5, 1.0 Gy). Conventional analysis of chromosome aberrations was carried out in Giemsa-stained preparations. Chromosome aberrations involving chromosomes 2, 3, 8, X and Y were analysed in first division metaphases using two-colour FISH.
The results indicate that when all types of translocations are taken into account both X-rays and neutrons induce more translocations than dicentrics, the ratio between the two types of exchanges being 1.4 and 1.8 respectively. The ratio of 'apparently simple' reciprocal translocations and reciprocal complete dicentrics was close to 1 for both types of radiation. The RBE of neutrons for induction of exchanges was found to be between 5 and 8. Neutron irradiation was more efficient at inducing insertions. Among the chromosomes studied, an increased involvement was observed for chromosome 8 in dicentrics and translocations than that expected on the basis of its chromosome length. The high content of interstitial telomeric sequences in chromosome 8 may be responsible for the observed sensitivity of this chromosome.
The results obtained in this study indicate that: (1) more translocations are found than dicentrics; (2) heterogeneity exists among Chinese hamster chromosomes for involvement in radiation-induced exchanges; (3) the spectrum and distribution of exchange aberrations are different between X-rays and neutrons; and (4) the relative frequencies of insertions could be used as a 'fingerprint' for exposure to high LET radiation.
通过使用针对2号、3号、8号、X和Y染色体的DNA涂染探针进行双色荧光原位杂交,估算中国仓鼠脾细胞中辐射(低传能线密度和高传能线密度)诱导的染色体畸变频率,并确定(1)辐射诱导的易位与双着丝粒的比例;(2)X射线和中子诱导的交换型畸变谱;(3)不同染色体在畸变形成中的相对参与情况。
从中国仓鼠分离出的脾细胞在体外接受不同剂量的200 kV X射线(0.75、1.5、3.0 Gy)和1 MeV快中子(0.25、0.5、1.0 Gy)照射。在吉姆萨染色的制片上进行染色体畸变的常规分析。使用双色荧光原位杂交技术在第一次分裂中期分析涉及2号、3号、8号、X和Y染色体的染色体畸变。
结果表明,当考虑所有类型的易位时,X射线和中子诱导的易位均多于双着丝粒,两种类型交换的比例分别为1.4和1.8。两种辐射的“明显简单”相互易位与相互完全双着丝粒的比例均接近1。发现中子诱导交换的相对生物效应在5至8之间。中子照射在诱导插入方面更有效。在所研究的染色体中,观察到8号染色体在双着丝粒和易位中的参与度高于基于其染色体长度预期的水平。8号染色体中间粒端粒序列的高含量可能是该染色体观察到的敏感性的原因。
本研究获得的结果表明:(1)发现的易位多于双着丝粒;(2)中国仓鼠染色体在参与辐射诱导的交换方面存在异质性;(3)X射线和中子之间交换型畸变的谱和分布不同;(4)插入的相对频率可作为暴露于高传能线密度辐射的“指纹”。