Radiation and Environmental Biophysics

, Volume 46, Issue 4, pp 417–422 | Cite as

Frequency of micronuclei in 4–8 cell mouse embryos generated after maternal gamma-irradiation in the presence and in the absence of vitamin C

  • Hossein MozdaraniEmail author
  • Elmina Nazari
Original Paper


The objective of this investigation was to evaluate the frequency of chromosomal aberrations expressed as micronuclei (MN) in 4–8 cell embryos generated by gamma-irradiation of female mice in the absence and in the presence of vitamin C. Female NMRI mice were whole body exposed to 4 Gy gamma-irradiation after intraperitoneal (i.p.) injection of pregnant mare’s serum gonadotrophin (PMSG) followed by injection of human chorionic gonadotrophin (HCG) and mating with non-irradiated NMRI male mice. Pregnant animals were sacrificed and embryos flushed from the oviducts and fixed on slides. Cells were treated for MN observation using standard method. To investigate the protective effect of vitamin C (ascorbic acid) on the frequency of MN, 100 mg/kg vitamin C was i.p. injected 1 h before irradiation. Results show that the frequency of MN generated in the embryos of irradiated mother compared to those of control in the non-irradiated group increased dramatically (< 0.001). Frequency of MN in embryos generated in irradiated female mice treated with vitamin C dramatically and statistically decreased relative to the frequency observed in the irradiation only group (< 0.001). This decrease returned the combined treatment group to a level that was not statistically different from the controls (> 0.05). Thus, irradiation of preovulatory stage oocytes leads to stable chromosome abnormalities expressed as micronuclei in successive preimplantation embryos. Vitamin C reduces these clastogenic effects of radiation in preovulatory oocytes and thus the reduced frequency of MN in embryos is probably due to its antioxidation and radical scavenging properties.


Preovulatory oocyte Mouse Gamma-irradiation Preimplantation embryo Micronuclei Vitamin C 



The authors would like to thank Dr. M.H. Zahmatkesh for his kind assistance for radiation dosimetry and irradiation of mice. This work was supported in part by the Royan infertility institute.


  1. 1.
    Pils S, Muller WU, Streffer C (1999) Lethal and teratogenic effects in two successive generations of the HLG mouse strain after radiation exposure of zygotes association with genomic instability? Mutat Res 429:85–92Google Scholar
  2. 2.
    Hall EJ (2001) Radiobiology for the radiologist, 5th edn. Lippincott, PhiladelphiaGoogle Scholar
  3. 3.
    Russel LB (1956) X-ray-induced developmental abnormalities in the mouse and their use in the analysis of embryological patterns. II. Abnormalities of the vertebral column and thorax. J Exp Zool 131:329–395CrossRefGoogle Scholar
  4. 4.
    Buckton KE (1983) Chromosome aberrations in patients treated with X-irradiation for ankylosing spondylitis. In: Radiation-induced chromosome damage in man, pp 491–511Google Scholar
  5. 5.
    Countryman PI, Heddle JA (1976) The production of micronuclei from the chromosome aberrations in irradiated cultures of human lymphocytes. Mutat Res 41:321–332Google Scholar
  6. 6.
    Heddle JA (1973) A rapid in vivo test for chromosomal damage. Mutat Res 18:187–190Google Scholar
  7. 7.
    Muller W-U, Streffer C (1994) Micronucleus assay. In: Advance in mutagenesis research, pp 1–134Google Scholar
  8. 8.
    Savage JRK (1976) Classification relationships of induced chromosomal structural changes. J Med Genet 13:103–122Google Scholar
  9. 9.
    Fenech M (2000) The in vitro micronucleus technique. Mutat Res 455:81–95Google Scholar
  10. 10.
    Fenech M, Chang WP, Kirsch-Volders M, Holland N, Bonassi S, Zeiger E (2003) Human micronucleus project. Human project: detailed description of the scoring criteria for the cytokinesis-block micronucleus assay using isolated human lymphocyte cultures. Mutat Res 534:65–75Google Scholar
  11. 11.
    Fenech M (1993) The cytokinesis-blocked micronucleus technique: a detailed description of the method and its application genotoxicity studies in human population. Mutat Res 285:35–44Google Scholar
  12. 12.
    Jacquet PL, de Saint G, Vanteer KJ, Beugnet ML (1995) Radiation-induced translocations in immature and mature oocytes of the Guinea-pig. Radiat Res 1:375Google Scholar
  13. 13.
    Reichert W, Hansmann I, Rohrborn G (1975) Chromosome anomalies in mouse oocytes after irradiation. Human Genet 28:25–38CrossRefGoogle Scholar
  14. 14.
    Harapanhalli RS, Yaghmai V, Giulaint D, Howell RW, Rao DV (1996) Antioxidant effect of vitamin C in mice following X-irradiation. Res Commun Mol Pathol Pharmacol 94:27–87Google Scholar
  15. 15.
    Ramanathan K, Shila S, Kumaran S, Panneerse L C (2003) Protective role of ascorbic acid and alpha-tocopherol on arsenic-induced microsomal dysfunctions. Hum Exp Toxicol 22:129–36CrossRefGoogle Scholar
  16. 16.
    Konopacka M, Rzeszowska-Wolny J (2001) Antioxidant vitamins C, E and beta-carotene reduce DNA damage before as well as after gamma-ray irradiation of human lymphocytes in vitro. Mutat Res 491:1–7Google Scholar
  17. 17.
    Jagetia GC, Reddy TK (2002) The grapefruit flavanone naringin protects against the radiation-induced genomic instability in the mice bone marrow: a micronucleus study. Mutat Res 519:37–48Google Scholar
  18. 18.
    Konopacka M, Widel M, Rzeszowska-Wolny J (1998) Modifying effect of vitamins C, E and beta-carotene against gamma-ray-induced DNA damage in mouse cells. Mutat Res 417:85–94Google Scholar
  19. 19.
    El- Nahas SM, Matter FE, Mohamed AA (1993) Radioprotective effect of vitamins C and E. Mutat Res 301:143–7CrossRefGoogle Scholar
  20. 20.
    Pampfer S, Strefer C (1988) Prenatal death and malformations after irradiation of mouse zygotes with neutrons or X-rays. Teratology 37:599–807CrossRefGoogle Scholar
  21. 21.
    Pampfer S, Streffer C (1989) Increased chromosome aberration levels in cells from mouse fetuses after zygote X-irradiation. Int J Radiat Biol 55:85–92CrossRefGoogle Scholar
  22. 22.
    Luke GA, Richies AC, Bryant PE (1997) Genomic instability in haematopoietic cells of F1 generation mice of irradiated male parents. Mutagenesis 12:147–152CrossRefGoogle Scholar
  23. 23.
    Tusell L, Alvarez R, Caballin MR, Genesca A, Miro R, Ribas M, Egozcue J (1995) Induction of micronuclei in human sperm-hamster egg hybrids at the two cell stage after in vitro gamma-irradiation of human spermatozoa. Environ Mol Mutagen 26:315–23CrossRefGoogle Scholar
  24. 24.
    Jacquet P, Adriaens I, Buset J, Neefs M, Vankerkom J (2005) Cytogenetic studies in mouse oocytes irradiated in vitro at different stages of maturation by use of an early preantral follicle culture system. Mutat Res 583:168–177Google Scholar
  25. 25.
    Bang D, Lee J, Oh H, Kim S, Kim T, Lee Y, Lee C, Kim S (2002) Dose incidence relationships on the prenatal effects of gamma-radiation in mice. J Vet Sci 3:7–11Google Scholar
  26. 26.
    Brazill JL, MasuiY (1978) Changing levels of UV light and carcinogen-induced unscheduled DNA synthesis in mouse oocytes during meiotic maturation. Cell Res 112:121–125CrossRefGoogle Scholar
  27. 27.
    Yoichi M, Izuo T (1989) Repair capacity of fertilized mouse eggs for X-ray damage induced in sperm and mature oocytes. Mutat Res 210:35–47Google Scholar
  28. 28.
    Tease C, Fisher G (1986) X-ray-induced chromosome aberrations in immediately preovulatory oocytes. Mutat Res 173:211–5CrossRefGoogle Scholar
  29. 29.
    Uma Devi P, Satyamitra M (2005) Tracing radiation induced genomic instability in vivo in the haemopoietic cells from fetus to adult mouse. Br J Radiol 78:928–33CrossRefGoogle Scholar
  30. 30.
    Antoshchina MM, Riabchenko NI, Nasonova VA, Fesenko EV, Pelevina II (2005) The genome instability in the descendants of the Chinese hamster of the cells, irradiated by the low dose and by various intensities of gamma-radiation. Radiat Biol Radioecol 45:291–3Google Scholar
  31. 31.
    Burton G, Foster DO (1995) Biological antioxidant. Philosophical Transactions of the Royal Society of London, Series B. Biol Sci 331:565–78Google Scholar
  32. 32.
    Flemovicius I, Bonsack ME (1995) Intestinal radioprotection by vitamin E (alpha tocopherol). Ann Surg 224:504–510CrossRefGoogle Scholar
  33. 33.
    Weiss JF, Kumor KS, Walden TL (1990) Advances in radioprotection through the use of combined agent regimens. Int J Radiat Biol 57:709–722CrossRefGoogle Scholar
  34. 34.
    Athar M, Abdula M, Sultana S, Favier A (1993) Free radicals and trace elements. J Trace Elem Exp Med 6:65–73Google Scholar
  35. 35.
    Sohier M, El-Nahas FE, Athar M, Mohammad AA (1993) Radioprotective effect of vitamins C and E. Radiat Res 301:143–147Google Scholar
  36. 36.
    Brown MA (1993) Resistance of human erythrocytes containing elevated levels of Vitamin E to radiation—induced hemolysis. Radiat Res 95:303–316CrossRefADSGoogle Scholar
  37. 37.
    Mehandjiev A, Vassilev G, Yonova P (1993) Radioprotective effect of thiourea derivatives. Biotechnol Biotechnol Equip 7:66–70Google Scholar
  38. 38.
    Forand A, Dutrillaux B, Bernardino-Sgheri J (2004) Gamma-H2AX expression pattern in non-irradiated mouse germ cells and after low dose gamma-radiation: relationships between chromatid breaks and DNA double strand breaks. Biol Reprod 71:643–9CrossRefGoogle Scholar
  39. 39.
    Jagetia GC, Rajanikant GK, Baliga MS, Rao KV, Kumar P (2004) Augmentation of wound healing by ascorbic acid treatment in mice exposed to gamma-radiation. Int J Radiat Biol 80:347–54CrossRefGoogle Scholar
  40. 40.
    Dura Kovic A (1993) Radioprotective agents in medicine. Arh-Hig-Rada-Tokshikiol. 44:331– 54Google Scholar
  41. 41.
    Fridovich I (1974) Superoxide dismutase. Adv Enzymol 41:35–97Google Scholar
  42. 42.
    Liebovitz BE, Siegel BV (1980) Aspects of free radical reactions in biological systems: aging. J Gerontol 35:45–56Google Scholar

Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  1. 1.Department of Medical GeneticsTarbiat Modares UniversityTehranIran
  2. 2.Department of Science and ResearchAzad University and Royan InstituteTehranIran

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