Sister Chromatid Exchange Studies in the Chick Embryo and Neonate: Actions of Mutagens in a Developing System

  • Stephen E. Bloom


The embryonic and neonatal periods represent times when the disease process may be initiated as a result of exposure to environ mental mutagens and teratogens. We are using the chick embryo/neo-nate as an experimental system to detect and study the genotoxicity of environmental chemicals in developing tissues and the resultant biological alterations in survivors of perinatal chemical exposure. In vivo bromodeoxyuridine (BrdUrd) labeling of replicating DNA has been employed to measure basal and induced sister chromatid exchanges (SCEs), a candidate cytogenetic endpoint in genetic toxicology testing. Additionally, SCE induction studies with model promu-tagens have permitted the detection and study of components of the developing mixed-function oxidase (MFO) system of the liver and other organs. The relationship between specific MFO enzyme induction and SCE generation by promutagens has been studied in ovo and using in vitro assays.

The in vivo SCE induction potential of 53 Compounds, including known mutagens and nonmutagens, was evaluated in the early chick embryo. About 90% of the mutagens induced SCEs; all nonmutagens fail-ed to induce SCE above baseline. Clastogens such as bleomycin did not induce SCE but did cause massive chromosome damage that was easlly detected. Gentian violet (GV) is a direct-acting clastogen that did not show any SCE induction. This agrees with the findings from in vitro mutation assays that incorporate rat liver S-9 preparations. Potency for inducing SCE in the chick embryo correlates well with true mutagenic potency, DNA Inhibition, and to some extent with carcinogenic activity.

Indirect-acting mutagen-carcinogens induced SCEs and also un scheduled DNA synthesis (UDS) in embryonic cells. Biochemical studies revealed that aryl hydrocarbon hydroxylase (AHH) activity develops in the early embryonic liver as it is first formed at 4-5 da of incubation. The level of AHH activity is sufficient to account for the dramatic SCE response. Enhanced SCE induction occurred in older stage embryos, correlating with the increased basal AHH level and enhanced induction capacity of the liver. Modulation of the MFO enzyme system with specific inducers resulted in altered SCE and UDS responses in vivo and in vitro using a chick microsome/chinese hamster ovary (CHO) mammalian cell assay.

Viable embryos and neonates have been obtained following exposure to SCE-inducing levels of the mutagen-carcinogen aflatoxin Bl applied at either 6 da or 12 da of development. Phenotypically “normal” chicks demonstrated deficiencies in the hematolymphoid system and in postnatal growth potential. The postnatal biological outcome was correlated to some degree with the developmental/differentiation status at the time of toxicant exposure. Thus, baseline and induced SCEs (if they occur in the absence of the BrdUrd probe) in the above instance are not associated with gross disturbances of development. Rather, any induced cellular alterations are subtle and expression is delayed.

The chick embryo should be useful for the further study of SCEs in a developing system, the effects of chemicals on tissue-specific SCE induction, the role of MFO enzymes in mediating the production of DNA-damaging and SCE-inducing metabolites, and for studying the cellular and developmental consequences of embryo exposure to geno-toxic environmental chemicals.


Chick Embryo Sister Chromatid Acridine Orange Sister Chromatid Exchange Avian Embryo 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Ames, B.N. (1979) Identifying environmental chemicals causing mutations and cancer. Science 204:587–593.PubMedCrossRefGoogle Scholar
  2. 2.
    Drake, J.W., S. Abrahamson, J.F. Crow, A. Hollaender, S. Lederberg, M.S. Legator, J.V. Neel, M.W. Shaw, H.E. Sutton, R.C. von Borstel, and S. Zimmering (1975) Environmental muta genic hazards. Science 187:503–514.CrossRefGoogle Scholar
  3. 3.
    Hollaender, A., and F.J. de Serres (1971–1982) Chemical Mutagens: Principles and Methods for Their Detection, Vols. 1–7. Plenum Press, New York.CrossRefGoogle Scholar
  4. 4.
    McCann, J., E. Choi, E. Yamasaki, and B.N. Ames (1975) Detection of carcinogens as mutagens in the Salmonella/microsome test: Assay of 300 chemicals. Proc. Natl. Acad. Sci., USA 72:5135–5139.PubMedCrossRefGoogle Scholar
  5. 5.
    Nagao, M., and T. Sugimura (1978) Environmental mutagens and carcinogens. Ann. Rev. Genet. 12:117–159.PubMedCrossRefGoogle Scholar
  6. 6.
    Vogel, F., and G. Röhrborn (1970) Chemical Mutagenesis in Mam mals and Man. Springer-Verlag, New York.CrossRefGoogle Scholar
  7. 7.
    Perinatal Carcinogenesis (1979) National Cancer Institute Monograph 51, DHEW Publication No. (NIH) 79–1633, 282 pp.Google Scholar
  8. 8.
    Rice, J.M. (1973) An overview of transplacental chemical carcinogenesis. Teratology 8:113–126.CrossRefGoogle Scholar
  9. 9.
    Ivankovic, S. (1979) Teratogenic and carcinogenic effects of some chemicals during prenatal life in rats, Syrian golden hamsters, and minipigs. In Perinatal Carcinogenesis, NCI Monograph 51, DHEW Publication No. 79–1633, pp. 103–115.Google Scholar
  10. 10.
    Knudson, Jr., A.G. (1979) Mutagenesis and embryonal carcinogenesis. In Perinatal Carcinogenesis, NCI Monograph 51, DHEW publication No. 79–1633, pp. 19–23.Google Scholar
  11. 11.
    Nichols, W.W. (1982) Status of prezygotic chromosome lesions in relation to cancer. Cytogenet. Cell Genet. 33:179–184.PubMedCrossRefGoogle Scholar
  12. 12.
    Riccardi, V.M., E. Sujansky, A.C. Smith, and U. Francke (1978) Chromosomal imbalance in the Aniridia-Wilms’ tumor association: lip interstitial deletion. Pediatrics 61:604–610.PubMedGoogle Scholar
  13. 13.
    Strong, L.C., V.M. Riccardi, R.E. Ferrell, and R.S. Sparkes (1981) Familial retinoblastoma and chromosome 13 deletion transmitted via an insertional translocation. Science 213: 1501–1503.PubMedCrossRefGoogle Scholar
  14. 14.
    Sandberg, A.A., ed. (1982) Progress and Topics in Cytogenetics. II. Sister Chromatid Exchange. Alan R. Liss, Inc., New York.Google Scholar
  15. 15.
    Latt, S.A., R.R. Schreck, K.S. Loveday, C.P. Dougherty, and C.F. Shuler (1980) Sister chromatid exchanges. In Advances in Human Genetics, Vol. 10, H. Harris and K. Hirschhorn, eds. Plenum Press, New York, pp. 267–331.Google Scholar
  16. 16.
    Bloom, S.E. (1978) Chick embryos for detecting environmental mutagens. In Chemical Mutagens: Principles and Methods for Their Detection, Vol. 5, A. Hollaender and F.J. de Serres, eds. Plenum Press, New York, pp. 203–232.Google Scholar
  17. 17.
    Bloom, S.E. (1982) Detection of sister chromatid exchanges in vivo using avian embryos. In Cytogenetic Assays of Environmen tal Mutagens, T.C. Hsu, ed. Allanheld, Osmun & Co., Montclair, New Jersey, pp. 137–159.Google Scholar
  18. 18.
    Bloom, S.E. (1982) Avian and aquatic systems for in vivo detection of sister chromatid exchanges. In Progress and Topics in Cytogenetics. II. Sister Chromatid Exchange, A.A. Sandberg, ed. Alan R. Liss, New York, pp. 249–277.Google Scholar
  19. 19.
    Dietert, R.R., S.E. Bloom, M.A. Qureshi, and U.C. Nanna (1983) Hematological toxicology following embryonic exposure to aflatoxin-Bl. Proc. Soc. Exp. Biol. Med. 173:481–485.PubMedGoogle Scholar
  20. 20.
    Bloom, S.E., and T.C. Hsu (1975) Differential fluorescence of sister chromatids in chicken embryos exposed to 5-bromodeoxy-uridine. Chromosoma 51:261–267.PubMedCrossRefGoogle Scholar
  21. 21.
    Kligerman, A.D., and S.E. Bloom (1976) Sister chromatid differentiation and exchanges in adult mudminnows (Umbra limi) after in vivo exposure to 5-bromodeoxyuridine. Chromosoma 56:101–109.PubMedCrossRefGoogle Scholar
  22. 22.
    Allen, J.W., and S.A. Latt (1976) Analysis of sister chromatid exchange formation in vivo in mouse spermatogonia as a new test system for environmental mutagens. Nature (Lond.) 260:449–451.CrossRefGoogle Scholar
  23. 23.
    Pera, F., and P. Mattias (1976) Labelling of DNA and differential sister chromatid staining after BrdU treatment in vivo. Chromosoma 57:13–18.PubMedCrossRefGoogle Scholar
  24. 24.
    Schneider, E.L., J.R. Chaillet, and R.R. Tice (1976) In vivo BUdR labeling of mammalian chromosomes. Exp. Cell Res. 100: 396–399.PubMedCrossRefGoogle Scholar
  25. 25.
    Vogel, W., and T. Bauknecht (1976) Differential chromatid staining by in vivo treatment as a mutagenicity test system. Nature 260:448–449.PubMedCrossRefGoogle Scholar
  26. 26.
    Tice, R., J. Chaillet, and E.L. Schneider (1976) Demonstration of spontaneous sister chromatid exchanges in vivo. Exp. Cell Res. 102:426–429.PubMedCrossRefGoogle Scholar
  27. 27.
    Cole, R.K. (1967) Leukosis control through genetics. In Proc. Poult. Health Conf., University of New Hampshire, Durham, pp. 59–72.Google Scholar
  28. 28.
    Blum, A., and B.N. Ames (1977) Flame-retardant additives as possible cancer hazards. Science 195:17–23.PubMedCrossRefGoogle Scholar
  29. 29.
    Gold, M.D., A. Blum, and B.N. Ames (1978) Another flame retardant, tris(l,3-dichloro-2-propyl)-phosphate, and its expected metabolites are mutagens. Science 200:785–787.PubMedCrossRefGoogle Scholar
  30. 30.
    Kato, H. (1977) Spontaneous and induced sister chromatid ex changes as revealed by the BUdR-labelling method. Int. Rev. Cytol. 49:55–97.PubMedCrossRefGoogle Scholar
  31. 31.
    Wolff, S. (1977) Sister chromatid exchange. Ann. Rev. Genet .11:183–201.PubMedCrossRefGoogle Scholar
  32. 32.
    Latt, S.A. (1974) Sister chromatid exchanges, indices of human chromosome damage and repair: Detection of fluorescence and induction by mitomycin C. Proc. Natl. Acad. Sci., USA 71:3162–3166.PubMedCrossRefGoogle Scholar
  33. 33.
    Dutrillaux, B., A.M. Fosse, M. Prieur, J. Lejeune (1974) Analyse des échanges de chromatides dans les cellules somatiques humaines: Traitment au BUdR (5-bromdeoxyuridine) et fluorescence bicolore par l’acridine orange. Chromosoma 48: 327–340.CrossRefGoogle Scholar
  34. 34.
    Kato, H. (1974) Spontaneous sister chromatid exchanges detected by a BUdR-labelling method. Nature 251:70–72.PubMedCrossRefGoogle Scholar
  35. 35.
    Kihlman, B.A., and D. Kronberg (1975) Sister chromatid exchanges in Vicia faba. I. Demonstration by a modified fluorescent plus Giemsa (FPG) technique. Chromosoma 51:1–10.CrossRefGoogle Scholar
  36. 36.
    Korenberg, J.R., and E.R. Freedlender (1974) Giemsa technique for the detection of sister chromatid exchanges. Chromosoma 48:355–360.PubMedCrossRefGoogle Scholar
  37. 37.
    Perry, P., and S. Wolff (1974) New Giemsa method for the differential staining of sister chromatids. Nature 251:156–158.PubMedCrossRefGoogle Scholar
  38. 38.
    Wolff, S., and P. Perry (1974) Differential Giemsa staining of sister chromatids and the study of sister chromatid exchanges without autoradiography. Chromosoma 48:341–353.PubMedCrossRefGoogle Scholar
  39. 39.
    Zakharov, A.F., and N.A. Egolina (1972) Differential spiralization along mammalian mitotic chromosomes. I. BUdR-revealed differentiation in Chinese hamster chromosomes. Chromosoma 38:341–365.PubMedCrossRefGoogle Scholar
  40. 40.
    Perry, P., and H.J. Evans (1975) Cytological detection of mutagen-carcinogen exposure by sister chromatid exchange. Nature 258:121–125.PubMedCrossRefGoogle Scholar
  41. 41.
    Owen, J.J.T. (1965) Karyotype studies on Gallus domesticus. Chromosoma 16:601–608.PubMedCrossRefGoogle Scholar
  42. 42.
    Bloom, S.E. (1974) Current knowledge about the avian W chromosome. BioScience 24:340–344.CrossRefGoogle Scholar
  43. 43.
    Ohno, S. (1961) Sex chromosomes and microchromosomes of Gallus domesticus. Chromosoma 11:484–498.PubMedCrossRefGoogle Scholar
  44. 44.
    Van Brink, J.M., and G.A. Ubbels (1956) La question des hetero-chromosomes chez les sauropsides: Oiseaux. Experientia 12: 162–164.CrossRefGoogle Scholar
  45. 45.
    Yamashina, M.Y. (1944) Karyotype studies in birds. I. Compara tive morphology of chromosomes in seventeen races of domestic fowl. Cytologia 13:270–296.CrossRefGoogle Scholar
  46. 46.
    Bloom, S.E. (1970) Marek’s disease: Chromosome studies of resistant and susceptible strains. Avian Dis. 14:478–490.PubMedCrossRefGoogle Scholar
  47. 47.
    Cormier, J.M., and S.E. Bloom (1973) An in vivo study of the effects of X-radiation on the chromosomes of chick allantoic membranes. Mutat. Res. 20:77–85.PubMedCrossRefGoogle Scholar
  48. 48.
    Au, W., S. Pathak, C.J. Collie, T.C. Hsu (1978) Cytogenetic toxicity of gentian violet and crystal violet on mammalian cells in vitro. Mutat. Res. 58:269–276.PubMedCrossRefGoogle Scholar
  49. 49.
    Au, W., M.A. Butler, S.E. Bloom, and T.S. Matney (1979) Further study of the genetic toxicity of gentian violet. Mutat. Res .66:103–112.PubMedCrossRefGoogle Scholar
  50. 50.
    Bloom, S.E. (1982) Sister chromatid exchange as an indicator of promutagen activation and mutagenic potency in early embryonic development. Environ. Mutagen. 4:361–362.Google Scholar
  51. 51.
    Todd, L.A., and S.E. Bloom (1980) Differential induction of sister chromatid exchanges by indirect-acting mutagen-carcinogens at early and late stages of embryonic development. Envi ron. Mutagen. 2:435–445.CrossRefGoogle Scholar
  52. 52.
    Hamilton, J.W., M.S. Denison, and S.E. Bloom (1983) Development of basal and induced aryl hydrocarbon [benzo(a)pyrene] hydroxylase activity in the chick embryo in ovo. Proc. Natl. Acad. Sci., USA 80:3372–3376.PubMedCrossRefGoogle Scholar
  53. 53.
    Bloom, S.E., O.P. Schaefer, and J.W. Hamilton (1983) Avian embryonic liver homogenates for activating promutagens in the Chinese hamster cell sister chromatid exchange assay. Environ. Mutagen. 5:386–387.Google Scholar
  54. 54.
    Hamilton, J.W., and S.E. Bloom (1984) Correlation between mixed-function oxidase enzyme induction and aflatoxin-Bl. induced unscheduled DNA synthesis in the chick embryo In vivo. Environ. Mutagen. (6:41–48).PubMedCrossRefGoogle Scholar
  55. 55.
    Painter, R.B. (1981) DNA synthesis inhibition in mammalian cells as a test for mutagenic carcinogens. In Short-Term Tests for Chemical Carcinogens, H.R. Stich and R.H.C. San, eds. Springer-Verlag, New York, pp. 59–64.CrossRefGoogle Scholar
  56. 56.
    Muscarella, D.E., and S.E. Bloom (1982) The longevity of chemically induced sister chromatid exchanges in Chinese hamster ovary cells. Environ. Mutagen. 4:647–655.PubMedCrossRefGoogle Scholar
  57. 57.
    Latt, S.A., J. Allen, S.E. Bloom, A. Carrano, E. Falke, D. Kram, E. Schneider, R. Schreck, R. Tice, B. Whitfield, and S. Wolff (1981) Sister chromatid exchanges: A report of the Gene-Tox program. Mutat. Res. 87:17–62.PubMedCrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1984

Authors and Affiliations

  • Stephen E. Bloom
    • 1
  1. 1.Institute for Comparative and Environmental Toxicology and Department of Poultry and Avian SciencesCornell UniversityIthacaUSA

Personalised recommendations