Teratogenicity Evaluation of 2-Bromopropane Using Rat Whole Embryo Culture

랫드 전배아배양법을 이용한 2-Bromopropane의 최기형성 평가

  • Kim Jong-Choon (College of Veterinary Medicine, Chonnam National University) ;
  • Shin Dong-Ho (College of Veterinary Medicine, Chonnam National University) ;
  • Kim Sung-Ho (College of Veterinary Medicine, Chonnam National University) ;
  • Yang Young-Soo (College of Veterinary Medicine, Chonnam National University) ;
  • Oh Ki-Seok (College of Veterinary Medicine, Chonnam National University) ;
  • Jiang Cheng-Zhe (Center for Safety Evaluation and Research of Drugs, Institute of Laboratory, Animal Science. CAMS, Peking Union Medical College) ;
  • Chung Moon-Koo (Korea Institute of Toxicology, KRICT)
  • Published : 2006.06.01

Abstract

Recently, we have reported that the environmental pollutant 2-bromopropane (2-BP) induces a significant embryo-fetal developmental toxicity in rats. However, the cause of developmental toxicity and the relationship between maternal and developmental toxicities could not be elucidated because the developmental toxicity of 2-BP was observed only in the presence of maternal toxicity The in vitro teratogenicity study using whole embryo culture was carried out to understand the teratogenic properties and the possible mechanism of teratogenicity induced by 2-BP in rats. Rat embryos aged 9.5 days were cultured in vitro for 48 hrs at medium concentrations of 0, 1, 3, or 10 mg/ml of 2-BP. Embryos were evaluated for growth, differentiation, and morphological alterations at the end of the culture period. At 10 mg/ml, 2-BP caused a delay in the growth and differentiation of embryos and an increase in the incidence of morphological alterations, including altered yolk sac circulation, abnormal axial rotation, craniofacial hypoplasia, open neuropore, absent optic vesicle and kinked somites. At 3 mg/ml, only a delay in the growth and differentiation of embryos was observed. There were no adverse effects on embryonic growth and development at the concentration of 1 mg/ml. The results showed that the exposure of 2-BP to rat embryos results in a developmental delay and morphological alterations at dose levels of 3 mg/ml culture media or higher and that 2-BP can induce a direct developmental toxicity in rat embryos.

Keywords

References

  1. Yamaguchi, C., Fujita, S., Obara, T. and Ueda, T. (1983): Effects of room temperature on reproduction, body weight and organ weights, food and water intakes, and hematology in mice. Exp. Anim., 32, 1-11 https://doi.org/10.1538/expanim1978.32.1_1
  2. Bradford, M.M. (1976): A rapid sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anat. Biochem., 72, 248-254 https://doi.org/10.1016/0003-2697(76)90527-3
  3. Chahoud, I., Ligensa, A., Dietzel, L. and Faqi, A.S. (1999): Correlation between maternal toxicity and embryo/fetal effects. Reprod. Toxicol., 13, 375-381 https://doi.org/10.1016/S0890-6238(99)00035-0
  4. Chernoff, N., Rogers, J.M. and Kavlock, R.J. (1989): An overview of maternal toxicity and prenatal development: considerations for developmental toxicity hazard assessments. Toxicology, 59, 111-125 https://doi.org/10.1016/0300-483X(89)90050-4
  5. Chung, M.K., Kim, J.C. and Han SS. (2005): Embryotoxic effects of CKD-602, a new camptothecin anticancer agent, in rats. Reprod. Toxicol., 20, 165-173 https://doi.org/10.1016/j.reprotox.2005.01.004
  6. Dunnett, C.W. (1964): New tables for multiple comparisons with a control. Biometrics, 20, 482-491 https://doi.org/10.2307/2528490
  7. HSDB. Hazardous Substances Data Bank, Bethesda (MD). (2001): National Institutes of Health. Available from URL: http://toxnet.nlm.nih.gov
  8. Ishikawa, H., Tian, Y. and Yamauchi, T. (2001): Induction of micronuclei formation in preimplantation mouse embryos after maternal treatment with 2-bromopropane. Reprod. Toxicol., 15, 81-85 https://doi.org/10.1016/S0890-6238(00)00112-X
  9. Kang, K.S., Li, G.X., Che, J.H. and Lee, Y.S. (2002): Impairment of male rat reproductive function in F1 offspring from dams exposed to 2-bromopropane during gestation and lactation. Reprod. Toxicol., 16, 151-159 https://doi.org/10.1016/S0890-6238(02)00004-7
  10. Kavlock, R.J., Chernoff, N. and Rogers, E.H. (1985): The effect of acute maternal toxicity on fetal development in the mouse. Teratog. Carcinog. Mutagen., 5, 3-13 https://doi.org/10.1002/tcm.1770050103
  11. Kawai, T., Okada, Y., Odachi, T., Horigichi, S., Zhang, Z., Moon, C. and Ikeda, M. (1997): Diffusive sampling and biological monitoring of 2-bromopropane. Arch. Environ. Contam. Toxicol., 33, 23-28 https://doi.org/10.1007/s002449900218
  12. Khera, K.S. (1987): Maternal toxicity in human and animals: effects on fetal development and criteria for detection. Teratog. Carcinog. Mutagen., 7, 287-295 https://doi.org/10.1002/tcm.1770070309
  13. Kim, J.C., Kim, S.H., Shin, D.H., Ahn, T.H., Kim, H.C., Kim, Y.B., Jiang, C.Z., Han, J. and Chung, M.K. (2004): Effects of prenatal exposure to the environmental pollutant 2-bromopropane on embryo-fetal development in rats. Toxicology, 196, 77-86 https://doi.org/10.1016/j.tox.2003.11.006
  14. Kim, J.C., Shin, H.C., Cha, S.W., Koh, W.S., Chung, M.K. and Han, S.S. (2001): Evaluation of developmental toxicity in rats exposed to the environmental estrogen bisphenol A during pregnancy. Life Sci., 69, 2611-2625 https://doi.org/10.1016/S0024-3205(01)01341-8
  15. Kim, J.C., Shin, D.H., Kim, S.H., Ahn, T.H., Kang, S.S., Jang, B.S., Kim, C.Y. and Chung, M.K. (2003): Developmental toxicity evaluation of the new fluoroquinolone antibacterial DW-116 in rats. Teratog. Carcinog. Mutagen., 23, 123- 136 https://doi.org/10.1002/tcm.10066
  16. Kim, J.C., Yun, H.I., Shin, H.C., Han, S.S. and Chung, M.K. (2000): Embryo lethality and teratogenicity of a new fluoroquinolone antibacterial DW-116 in rats. Arch. Toxicol., 74, 120-124 https://doi.org/10.1007/s002040050662
  17. Kruskal, W.H. and Wallis, W.A. (1952): Use of ranks in one criterion variance analysis. J. Am. Statist. Assoc., 47, 614- 617
  18. Mirkin, B.L. (1973): Maternal and fetal distribution of drugs in pregnancy. Clin. Pharmacol. Ther., 14, 643-647 https://doi.org/10.1002/cpt1973144part2643
  19. Nelson, B.K., Brightwell, W.S., MacKenzie-Taylor, D.R., Khan, A., Burg, J.R., Weigel, W.W. and Goad, P.T. (1988): Teratogenicity of n-propanol and isopropanol administered at high inhalation concentrations to rats. Food Chem. Toxicol., 26, 247-254 https://doi.org/10.1016/0278-6915(88)90126-3
  20. New, D.A.T. (1978): Whole-embryo culture and the study of mammalian embryos during organogenesis. Biol. Rev., 53, 81-122 https://doi.org/10.1111/j.1469-185X.1978.tb00993.x
  21. NRC (National Research Council). (1996): Guide for the Care and Use of Laboratory Animals. National Research Council, National Academy, Washington, USA
  22. Omura, M., Romero, Y., Zhao, M. and Inoue, N. (1999): Histopathological evidence that spermatogonia are the target cells of 2-bromopropane. Toxicol. Lett., 104, 19-26 https://doi.org/10.1016/S0378-4274(98)00350-6
  23. Park, J.S., Kim, Y.H., Park, D.W., Choi, K.S., Park, S.H. and Moon, Y.H. (1997): An outbreak of hematopoietic and reproductive disorders due to solvents containing 2-bromopropane in an electronic factory, South Korea: Epidemiological survey. J. Occup. Health, 39, 38-143
  24. van Maele-Fabry, G., Delhaise, F. and Picard, J.J. (1990): Morphogenesis and quantification of the development of post-implantation mouse embryos. In vitro, 4, 149-156 https://doi.org/10.1016/0887-2333(90)90037-T
  25. Wise, L.D., Beck, S.L., Beltrame, D., Beyer, B.K., Chahoud, I., Clark, R.L., Clark, R., Druga, A.M., Feuston, M.H., Guittin, P., Henwood, S.M., Kimmel, C.A., Lindstrom, P., Palmer, A.K., Petrere, J.A., Solomon, H.M., Yasuda, M. and York, R.G. (1997): Terminology of developmental abnormalities in common laboratory mammals (Version 1). Teratology, 55, 249-292 https://doi.org/10.1002/(SICI)1096-9926(199704)55:4<249::AID-TERA5>3.0.CO;2-W
  26. Wu, X., Faqi, A.S., Yang, J., Pang, B.P., Ding, X., Jiang, X. and Chahoud, I. (2002): 2-Bromopropane induces DNA damage, impairs functional antioxidant cellular defenses, and enhances the lipid peroxidation process in primary cultures of rat Leydig cells. Reprod. Toxicol., 16, 379-384 https://doi.org/10.1016/S0890-6238(02)00039-4
  27. Yu, X., Kamijima, M., Ichihara, G., Li, W., Kitoh, J., Xie, Z., Shibata, E., Hisanaga, N. and Takeuchi, Y. (1999): 2-Bromopropane causes ovarian dysfunction by damaging premordial follicles and their oocytes in female rats. Toxicol. Appl. Pharmacol., 159, 185-193 https://doi.org/10.1006/taap.1999.8730
  28. Zhao, L.X., Kim, E.K., Lim, H.T., Moon, Y.S., Kim, N.H., Kim, T.H., Choi, H., Chae, W., Jeong, T.C. and Lee, E.S. (2002): Synthesis, characterization and in vitro identification of N7-guanine adduct of 2-bromopropane. Arch. Pharm. Res., 25, 39-44 https://doi.org/10.1007/BF02975258
  29. 김종춘, 신진영, 신동호, 강성철, 정나영, 백성수, 정문구 (2004): 발생독성 검색을 위한 마우스 전배자배양법의 확립. 한국실험동물학회지, 20, 49-54