Matsuda Y, Seki N, Utsugi-Takeuchi T, Tobari I
Division of Genetics, National Institute of Radiological Sciences, Chiba, Japan.
Int J Radiat Biol. 1989 Feb;55(2):233-56. doi: 10.1080/09553008914550281.
To study the changes in radiosensitivity of male and female genomes from fertilization to the pronuclear stage, the frequency of induced chromosome aberrations was examined at the first-cleavage metaphase in eggs fertilized with X-irradiated sperm, in eggs X-irradiated at the mature oocyte stage immediately before fertilization, and in fertilized eggs exposed to X-rays at various stages before DNA synthesis (1-5 h after insemination). Gametic treatment, fertilization and embryo culture were carried out in vitro. Most of the induced chromosome aberrations were chromosome-type aberrations, the frequency of chromosome fragments being the highest, followed by chromosome exchanges in both sperm and oocytes. The induction of chromosome-type aberrations was much higher in oocytes than in sperm. Chromosome-type aberrations were the main type also in fertilized eggs irradiated at the pre-DNA-synthetic stage. The radiosensitivity increased gradually with pronuclear formation (1-4h after insemination), but little difference in radiosensitivity was observed between eggs irradiated at 4 h and 5 h, corresponding to the stage when pronuclear formation was complete. The drastic change with pronuclear formation was found not only in radiosensitivity but also in the frequency of chromosome aberrations. The frequency of chromosome-type exchanges decreased drastically 2 h after insemination, and exchanges were barely observed at 5 h. The difference in radiosensitivity between male and female genomes also markedly changed with pronuclear formation, the chromosome aberration induction in the female genome being much higher than that in the male genome before accomplishment of pronuclear formation. The analysis of potentiation effects of 3-aminobenzamide and caffeine on the yield of X-ray-induced chromosome aberrations demonstrated that the increase of radiosensitivity and the decrease of chromosome-type exchange induction with pronuclear formation, may be closely correlated with alterations in chromatin configuration in the pronuclei and in repair capacity of fertilized eggs at the pre-DNA-synthetic stage. No evidence based on repair efficiency was found for the marked difference in radiosensitivity between male and female genomes during pronuclear formation.
为研究从受精到原核期雌雄基因组辐射敏感性的变化,检测了用X射线照射精子受精的卵子、受精前成熟卵母细胞期经X射线照射的卵子以及在DNA合成前不同阶段(授精后1 - 5小时)经X射线照射的受精卵在第一次卵裂中期诱导染色体畸变的频率。配子处理、受精和胚胎培养均在体外进行。大多数诱导的染色体畸变是染色体型畸变,染色体片段的频率最高,其次是精子和卵母细胞中的染色体交换。卵母细胞中染色体型畸变的诱导率远高于精子。在DNA合成前期照射的受精卵中,染色体型畸变也是主要类型。辐射敏感性随着原核形成(授精后1 - 4小时)逐渐增加,但在4小时和5小时照射的卵子之间未观察到辐射敏感性有显著差异,5小时对应原核形成完成阶段。不仅在辐射敏感性方面,而且在染色体畸变频率方面都发现了随着原核形成的剧烈变化。授精后2小时,染色体型交换频率急剧下降,5小时几乎未观察到交换。雌雄基因组之间的辐射敏感性差异也随着原核形成而显著变化,在原核形成完成前,雌性基因组中的染色体畸变诱导率远高于雄性基因组。对3 - 氨基苯甲酰胺和咖啡因对X射线诱导染色体畸变产率的增效作用分析表明,辐射敏感性的增加以及随着原核形成染色体型交换诱导率的降低,可能与原核中染色质构型的改变以及DNA合成前期受精卵修复能力的改变密切相关。未发现基于修复效率的证据支持原核形成过程中雌雄基因组辐射敏感性的显著差异。