Matsuda Y, Seki N, Utsugi-Takeuchi T, Tobari I
Division of Genetics, National Institute of Radiological Sciences, Chiba, Japan.
Mutat Res. 1989 Mar;211(1):65-75. doi: 10.1016/0027-5107(89)90107-3.
Chromosome aberrations induced at the first-cleavage metaphase of eggs fertilized with sperm recovered from spermiogenic cells which had been X-irradiated and treated with mitomycin C (MMC) at various stages were observed using in vitro fertilization and embryo culture technique. Furthermore, the repair capacity of the fertilized eggs for X-ray- and MMC-induced DNA damage which was induced in the spermiogenic cells and retained in the sperm until fertilization was investigated by analysis of the potentiation effects of 2 repair inhibitors, 3-aminobenzamide (3AB) and caffeine on the yield of chromosome aberrations. The frequency of chromosome aberrations observed in the eggs fertilized with sperm recovered from the early spermatid to late spermatocyte stage with X-irradiation of 4 Gy (16-20 days after X-irradiation) was markedly higher than that in the eggs fertilized with sperm recovered from spermatozoa to late spermatid stage (0-8 days after X-irradiation). The induced chromosome aberrations predominantly consisted of chromosome-type aberrations, the main type being chromosome fragment followed by chromosome exchange through all the spermiogenic stages. On the other hand, a high frequency of chromosome aberrations was not induced through all the stages with MMC treatment of 5 mg/kg. The remarkable potentiation effects of 3AB and caffeine were found in the eggs fertilized with sperm recovered from almost all the spermiogenic stages after X-irradiation. In the MMC treatment, a remarkable caffeine effect was observed occasionally in mid-early spermatids to late spermatocytes where a large amount of MMC damage could be induced. These results suggest that the large amount of DNA lesions induced in spermiogenic cells by X-rays and MMC persist as reparable damage until sperm maturation and are effectively repaired in the cytoplasm of the fertilized eggs.
利用体外受精和胚胎培养技术,观察了用从经X射线照射并在不同阶段用丝裂霉素C(MMC)处理的生精细胞中回收的精子使卵子受精后,在第一次卵裂中期诱导产生的染色体畸变。此外,通过分析两种修复抑制剂3-氨基苯甲酰胺(3AB)和咖啡因对染色体畸变率的增强作用,研究了受精卵对生精细胞中诱导产生并在受精前保留在精子中的X射线和MMC诱导的DNA损伤的修复能力。用从早期精细胞到晚期精母细胞阶段回收的精子(X射线照射后16 - 20天)使卵子受精,观察到的染色体畸变频率明显高于用从精子到晚期精细胞阶段回收的精子(X射线照射后0 - 8天)使卵子受精的情况。诱导产生的染色体畸变主要由染色体型畸变组成,主要类型是染色体片段,其次是在所有生精阶段的染色体交换。另一方面,用5 mg/kg的MMC处理,并非在所有阶段都诱导出高频率的染色体畸变。在X射线照射后,从几乎所有生精阶段回收的精子使卵子受精的情况下,都发现了3AB和咖啡因的显著增强作用。在MMC处理中,偶尔在中早期精细胞到晚期精母细胞中观察到显著的咖啡因效应,在这些细胞中可诱导大量MMC损伤。这些结果表明,X射线和MMC在生精细胞中诱导产生的大量DNA损伤作为可修复损伤持续到精子成熟,并在受精卵的细胞质中得到有效修复。