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Assessment of Sister Chromatid Exchange in Spermatogonia and Intestinal Epithelium in Chinese Hamsters

  • Steven B. Neal
  • Gregory S. Probst

Abstract

The induction of sister chromatid exchange (SCE) has been proposed as a predictive test for the identification of mutagens/carci-nogens. The in vivo application of this test was investigated by examining the chemical induction of SCE in spermatogonia, intestinal epithelium and bone marrow cells from Chinese hamsters.

Sister chromatid differentiation (SCD) was achieved in differentiating spermatogonial cells of male Chinese hamsters by the abdominal subcutaneous (sc) implantation of an agar-coated bromode-oxyuridine (BrdUrd) tablet. A number of genotoxins were administered intraperitoneally (ip) and the induction of SCE in spermatogonia and bone marrow was compared. A significant increase in SCE frequency in spermatogonia occurred following treatment with mitomycin C (MMC), cyclophosphamide (CP), or N,N’,N"-triethylenethiophos-phoramide (ThioTEPA). Treatment with busulfan, hycanthone (HC), or triethylenemelamine (TEM) failed to induce SCE in vivo in spermatogonia, but these compounds did induce SCE in bone marrow. Differences in cell cycle kinetics were considered to be the major factor involved in the differential induction of SCE in spermatogonia and bone marrow.

The induction of SCE in intestinal epithelium was investigated as a system for the identification of genotoxins that may result from the metabolism of xenobiotics by the gastrointestinal flora. Nitro-aromatic compounds were administered orally to Chinese hamsters. Nitro-aromatic compounds were chosen for this study since the mutagenic activity of these compounds is thought to result from their metabolism by bacterial nitroreductase. Metronidazole (MN) and 2-nitro-p-phenylenediamine (2NPPD) induced a dose-related in crease in SCE formation in intestinal epithelium but not in bone marrow. Treatment with 3-nitro-o-phenylenediamine (3N0PD) or 4-nitro-£-phenylenediamine (4N0PD) did not induce the formation of SCE in either intestinal epithelium or bone marrow. These findings indicate that studies in axenic animals will be required to elucidate the contribution of the enteric flora to the metabolic activation of some genotoxins.

Keywords

Bone Marrow Intestinal Epithelium Sister Chromatid Exchange Glacial Acetic Acid Spermatogonial Cell 
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.

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References

  1. 1.
    Latt, S., R. Schreck, K. Lovejoy, and C. Shuler (1982) Sister chromatid exchange analysis: Methodology applications, and interpretation. In Cytogenetic Assays of Environmental Muta gens, T. Hsu, ed. Allanheld, Osmum and Company, Totowa, New Jersey, pp. 29–80.Google Scholar
  2. 2.
    Neal, S., and G. Probst (1983) Chemically-induced sister chromatid exchange in vivo in bone marrow of Chinese hamsters. Mutat. Res. 113:33–43.PubMedCrossRefGoogle Scholar
  3. 3.
    Adler, I. (1982) Male germ cell cytogenetics. In Cytogenetic Assays of Environmental Mutagens, T. Hsu, ed. Allanheld, Osmum and Company, Totowa, New Jersey, pp. 249–276.Google Scholar
  4. 4.
    Wilmer, J., and E. Soaves (1980) Sister chromatid exchange in vivo in mice: I. The influence of increasing doses of bromode-oxyuridine. Environ. Mutagen. 2:35–42.PubMedCrossRefGoogle Scholar
  5. 5.
    Perry, P. (1980) Chemical mutagens and sister chromatid exchange. In Chemical Mutagens, Principles and Methods for Their Detection, Vol. 6, A. Hollaender and F. de Serres, eds. Plenum, New York, pp. 1–39.CrossRefGoogle Scholar
  6. 6.
    Latt, S., R. Schreck, K. Lovejoy, and C. Shuler (1979) In vitro and in vivo analysis of sister chromatid exchange. Pharmacol. Rev. 30:501–535.Google Scholar
  7. 7.
    Allen, J., and S. 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
  8. 8.
    Allen, J., C. Shuler, R. Mendes, and S. Latt (1977) A simplified technique for in vivo analysis of sister chromatid exchanges using 5-bromodeoxyuridine tablets. Cytogenet. Cell Genet. 18:231–237.PubMedCrossRefGoogle Scholar
  9. 9.
    Allen, J., C. Shuler, and S. Latt (1978) Bromodeoxyuridine tablet methodology for in vivo studies of DNA synthesis. Somatic Cell Genet. 4:393–405.PubMedCrossRefGoogle Scholar
  10. 10.
    Dunnett, C. (1964) New tables for multiple comparison with a control. Biometrics 20:482.CrossRefGoogle Scholar
  11. 11.
    Mazrimas, J., and D. Stetka (1978) Direct evidence for the role of incorporated BudR in the induction of sister chromatid exchange. Exp. Cell Res. 117:23–30.PubMedCrossRefGoogle Scholar
  12. 12.
    Perry, P., and H. Evans (1975) Cytologic detection of mutagen-carcinogen exposure by sister chromatid exchange. Nature (Lond.) 258:121–125.CrossRefGoogle Scholar
  13. 13.
    Perry, P., and S. Wolff (1974) New Giemsa method for the differential staining of sister chromatids. Nature (Lond.) 251: 156–158.CrossRefGoogle Scholar
  14. 14.
    Probst, G., R. McMahon, C. Thompson, J. Epp, L. Hill, and S. Neal (1981) A comparison of DNA repair in cultured rat hepatocytes with bacterial mutagenesis using 216 compounds. Environ. Mutagen. 3:11–32.PubMedCrossRefGoogle Scholar
  15. 15.
    Rinkus, S., and W. Speck (1975) Mutagenicity of metronidazole: Activation by mammalian liver microsomes. Biochem. Biophys. Res. Commun. 66:520–525.CrossRefGoogle Scholar
  16. 16.
    Rosenkranz, H., and W. Speck (1976) Activation of nitrofurantoin to a mutagen by rat liver nitroreductase. Biochem. Pharmacol. 25:1555–1556.PubMedCrossRefGoogle Scholar
  17. 17.
    DeRooij, D., D. Lok, and D. Weenk (1981) Proliferation pattern of undifferentiated spermatogonia and stem cell renewal in the Chinese hamster and ram. Cell Tiss. Kinet. 6:683–684.Google Scholar
  18. 18.
    King, M., D. Wild, E. Goche, and K. Eckhardt (1982) 5-Bromode-oxyuridine tablets with improved depot effect for analysis in vivo of sister chromatid exchanges in bone marrow and spermatogonial cells. Mutat. Res. 97:117–129.PubMedCrossRefGoogle Scholar
  19. 19.
    Kanda, N., and H. Kato (1979) In vivo sister chromatid exchange in cells of various organs of the mouse. Chromosoma 74:229–305.CrossRefGoogle Scholar
  20. 20.
    Oud, J., and D. DeRooij (1976) Spermatogenesis in the Chinese hamster. Anat. Rec. 187:113–124.CrossRefGoogle Scholar
  21. 21.
    Koch, R., and D. Goldman (1978) The anaerobic metabolism of metronidazole forms N-(2-hydroxyethyl)-oxamic acid. J. Pharm. Exptl. Therapeutics 208:406–410.Google Scholar
  22. 22.
    Soderman, J., ed. (1983) CRC Handbook of Identified Carcinogens and Noncarcinogens: Carcinogenicity-Mutagenicity Database. Vol.1: Chemical Class File. CRC Press, Boca Raton, p. 139.Google Scholar
  23. 23.
    Williams, R. (1972) Toxicologic implications of biotransforma tion by intestinal microflora. Toxicol. Appl. Pharmacol. 23: 769–781.PubMedCrossRefGoogle Scholar
  24. 24.
    Scheline, R. (1973) Metabolism of foreign compounds by gastrointestinal microorganisms. Pharmacol. Rev. 25:451–497.PubMedGoogle Scholar
  25. 25.
    Doolittle, D., J.M. Sherrill, and B. Butterworth (1983) Influence of intestinal bacteria, sex of the animal, and position of the nitro group on the hepatic genotoxicity of nitrotoluene isomers in vivo. Cancer Res. 43:2836–2842.PubMedGoogle Scholar
  26. 26.
    Koch, R., B. Beaulieu, E. Chrystal, and P. Goldman (1981) A metronidazole metabolite in human urine and its risk. Science 211:398–399.CrossRefGoogle Scholar
  27. 27.
    Machemer, L., and D. Lorke (1975) Method for testing mutagenic effects of chemicals on spermatogonia of the Chinese hamster. Arzneim-Forsch. (Drug Res.) 25:1889–1896.Google Scholar
  28. 28.
    Epstein, S., E. Arnold, J. Andrea, W. Bass, and Y. Bishop (1972) Detection of chemical mutagens by the dominant lethal assay in the mouse. Toxicol. Appl. Pharmacol. 23:288–325.PubMedCrossRefGoogle Scholar
  29. 29.
    Collins, T. (1972) Effect of captan and triethylenemelamine (TEM) on reproductive fitness of DBA/2J mice. Toxicol. Appl. Pharmacol. 23:277–287.PubMedCrossRefGoogle Scholar
  30. 30.
    Hoo, S., and C. Bowles (1971) An air-drying method for preparing metaphase chromosomes from the spermatogonial cells of rates and mice. Mutat. Res. 13:85–88.PubMedCrossRefGoogle Scholar
  31. 31.
    Dym, M., and D. Fawcett (1970) The blood-testis barrier in the rat and the physiological compartmentation of the seminiferous epithelium. Biol. Reprod. 3:308–326.PubMedGoogle Scholar
  32. 32.
    Allen, J. (1982) SCE and meiotic crossover exchange in germ cells. In Progress and Topics in Cytogenetics, Vol. 2: Sister Chromatid Exchange, A.A. Sandberg, ed. Alan R. Liss, New York, pp. 297–315.Google Scholar
  33. 33.
    Wolff, S. (1979) Sister chromatid exchange: The most sensitive mammalian system for determining the effects of mutagenic com pounds, In Genetic Damage in Man Caused by Environmental Agents, K. Berg, ed. Academic Press, New York, pp. 229–258,Google Scholar
  34. 34.
    Searle, A., and R. Willson (1976) Metronidazole (Flagyl): Degradation by the intestinal flora. Xenobiotica 6:457–464.PubMedCrossRefGoogle Scholar
  35. 35.
    Zachariah, P., and M. Juchau (1974) The role of gut flora in the reduction of aromatic nitro-groups. Drug Metabol. Disposition 2:74–78.Google Scholar
  36. 36.
    McCoy, E., W. Speck, and H. Rosenkranz (1977) Activation of a procarcinogen to a mutagen by cell-free extracts of anaerobic bacteria. Mutat. Res. 46:261–264.PubMedCrossRefGoogle Scholar
  37. 37.
    Rinkus, S., and M. Legator (1979) Chemical characterization of 465 known or suspected carcinogens and their correlation with mutagenic activity in the Salmonella typhimurium system. Cancer Res. 39:3289–3318.PubMedGoogle Scholar
  38. 38.
    Vogel, E., and F. Sobels (1976) The function of Drosophila in genetic toxicology testing. In Chemical Mutagens: Principles and Methods for their Detection, Vol. 4, A. Hollaender, ed. Plenum Press, New York, pp. 93–142.Google Scholar
  39. 39.
    Wiemann, H., and R. Lang (1978) Strategies for detecting heritable translocations in male mice by fertility testing. Mutat. Res. 53:317–326.PubMedCrossRefGoogle Scholar
  40. 40.
    Snell, G. (1946) Analysis of translocation in the mouse. Genetics 31:157–180.Google Scholar
  41. 41.
    Vogel, W., and T. Bauknecht (1976) Differential chromatid staining by in vivo treatment as a mutagenicity test system. Nature (Lond.) 260:448–449.CrossRefGoogle Scholar
  42. 42.
    Schneider, E., J. Chaillet, and R. Tice (1976) The in vivo labeling of mammalian chromosomes. Exp. Cell Res. 100:396–399.PubMedCrossRefGoogle Scholar
  43. 43.
    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
  44. 44.
    Batzinger, R., E. Bueding, B. Reddy, and J. Weisburger (1978) Formation of a mutagenic drug metabolite by intestinal microorganisms. Cancer Res. 38:608–612.PubMedGoogle Scholar
  45. 45.
    Spatz, M., D. Smith, E. McDaniel, and G. Laqueur (1966) Role of intestinal microorganisms in determining cycasin toxicity. Proc. Soc. Expt. Biol. Med. 124:691–699.Google Scholar
  46. 46.
    Callen, D. (1982) Microbial metabolism of environmental chemicals to mutagens and carcinogens. In Chemical Mutagens, Vol .7 ,F. de Serres, ed. Plenum Press, New York, pp. 163–188.CrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1984

Authors and Affiliations

  • Steven B. Neal
    • 1
  • Gregory S. Probst
    • 1
  1. 1.Toxicology Division Lilly Research LaboratoriesDivision of Eli Lilly and CompanyGreenfieldUSA

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