Showing papers by "Naohiko Seki published in 1989"

X-ray- and mitomycin C (MMC)-induced chromosome aberrations in spermiogenic germ cells and the repair capacity of mouse eggs for the X-ray and MMC damage.

Abstract: 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 limitedly 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.

Changes in X-ray sensitivity of mouse eggs from fertilization to the early pronuclear stage, and their repair capacity.

Abstract: 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.

Mechanism of chromosome aberration induction in the mouse egg fertilized with sperm recovered from postmeiotic germ cells treated with methyl methanesulfonate.

Abstract: Chromosome aberrations induced in spermatozoa to late spermatocytes following treatment with methyl methanesulfonate (MMS) were examined at the fist-cleavage metaphase of fertilized eggs in an attemp to clarify the mechanism of chromosomal damage in postmeiotic germ cells. A high frequency of chromosome aberration was induced in early spermatozoa to mid spermatids, while few chromosome aberrations were observed in early spermatids to late spermatocytes. 3-Aminobenzamide (3AB) potentiated the effect of MMS-induced chromosome aberrations in spermatozoa to mid spermatids indicating that a large amount of DNA lesions produced at these stages during spermatogenesis were not repaired prior to fertilization of the oocytes. Furthermore, from the cell-cycle analysis of the repair capacity in the fertilized egg it became clear that the lesions which remained in sperm until fertilization could be divided into 2 types: (1) DNA-strand breaks induced by stress in the chromatic structure produce by protamine alkylation; these lesions were converted to chromosome-type aberrations by 3AB treatment of the eggs during G 1 phase; (2) alkylated DNA which produces apurinic or apyrimidinic sites, of which there were a significant number mainly converted to chromatid exchanges by 3AB treatment of the eggs during S phase. This type of lesion appears to be constantly induced through all spermiogenic stages in contrast to the former type of lesion which is induced specifically during the stage of protamine maturation.