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Unscheduled DNA synthesis in the testis, a secondary test for the evaluation of chemical mutagens

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Abstract

DNA damage induced by methylmethane sulfonate, cyclophosphamide, doxorubicin and procarbazine in male germ cells was assessed in rabbits by the demonstration of unscheduled DNA synthesis (UDS), in meiotic and postmeiotic phases of maturation. Immediately after treatment by the intravenous route tritiated thymidine was injected into both testicles. Subsequently, rabbits were ejaculated serially, sperm heads were isolated and assayed for radioactivity by liquid scintillation counting. Dose-dependent UDS was demonstrated in late spermatocytes and early spermatids. High doses of hycanthone also induced UDS, but isoniazid and metronidazole had no effect. The rabbit testis UDS test takes into account metabolic and pharmacokinetic aspects of the test substances and provides information about their penetration through the blood-testicular barrier. It is therefore useful for secondary evaluation of potential mutagens. UDS induced by procarbazine was abolished by simultaneous treatment with Ara-C. Thus, the test also recognizes substances that inhibit DNA repair synthesis.

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References

  • Amann RP, Koefoed-Johnson HH, Levi H (1965) Excretion pattern of labelled spermatozoa and the timing of spermatozoa formation and epididymal transit in rabbits injected with thymidine-3H. J Reprod Fertil 10:169–183

    Google Scholar 

  • Arnold DW, Kennedy GL Jr, Keplinger ML, Calandra JC (1976) Dominant lethal studies with alkylating agents. Dose-response relationships. Toxicol Appl Pharmacol 38:79–84

    Google Scholar 

  • Biancifiori C, Severi L (1966) The relation of isoniazid and allied compounds to carcinogenesis in some species of small laboratory animals: a review. Br J Cancer 20:528–538

    Google Scholar 

  • Braun R, Schubert J, Schöneich J (1976) On the mutagenicity of isoniazid. Biol Zbl 95:423–436

    Google Scholar 

  • Brewen JG, Payne HS, Jones KP, Preston RJ (1975) Studies on chemically induced dominant lethality in postmeiotic male germ cells. Mutat Res 33:239–250

    Google Scholar 

  • Brookes P, Lawley PD (1964) Evidence for binding of polynuclear aromatic hydrocarbons to the nucleic acids of mouse skin: relation between carcinogenic power of hydrocarbons and their binding to DNA. Nature 202:781–784

    Google Scholar 

  • Bürgin H, Schmid B, Zbinden G (1979) Assessment of DNA damage in germ cells of male rabbits treated with isoniazid and procarbazine. Toxicology 12:251–257

    Google Scholar 

  • Cattanach BM (1971) Specific locus mutation in mice. In: Hollaender A (ed) Chemical mutagens. Principles and methods for their detection, vol 2. Plenum Press, New York London, p 535

    Google Scholar 

  • Cohen SM, Ertürk E, Van Esch AM, Crovetti AJ, Bryan GT (1973) Carcinogenicity of 5-nitrofurans, 5-nitroimidazoles, 5-nitrobenzenes and related compounds. J Natl Cancer Inst 51:403–417

    Google Scholar 

  • Cumming RB, Walton MF (1970) Fate and metabolism of some mutagenic alkylating agents in the mouse. I. Ethyl methanesulfonate and methylmethanesulfonate at sublethal dose in hybrid males. Mutat Res 10:365–377

    Google Scholar 

  • Forebee SH (1970) Controlled chemoprophylaxis trials in tuberculosis. A general review. Adv Tuberc Res 17:28–106

    Google Scholar 

  • Gaudin D, Gregg RS, Lemone Yielding K (1971) DNA repair inhibition: a possible mechanism of action of co-carcinogens. Biochem Biophys Res Commun 45:630–636

    Google Scholar 

  • Hahn GM, Yang S, Parker V (1968) Repair of sublethal damage and unscheduled DNA synthesis in mammalian cells treated with monofunctional alkylating agents. Nature 220:1142–1144

    Google Scholar 

  • Hiss EA, Preston RJ (1977) The effect of cytosine arabinoside on the frequency of single-strand breaks in DNA of mammalian cells following irradiation or chemical treatment. Biochim Biophys Acta 478:1–8

    Google Scholar 

  • Jones PA, Baker MS, Benedict WF (1976) The effect of 1-β-d-arabinosylfuranosylcytosine on cell viability, DNA synthesis and chromatid breakage in synchronized hamster fibrosarcoma cells. Cancer Res 36:3789–3797

    Google Scholar 

  • Kohn KW, Grimek-Ewig RA (1973) Alkaline elution analysis, a new approach to the study of DNA single-strand interruptions in cells. Cancer Res 33:1849–1853

    Google Scholar 

  • Lee IP, Zbinden G (1979) Differential DNA damage induced by chemical mutagens in cells growing in a modified Selye's granuloma pouch. Exp Cell Biol 47:92–106

    Google Scholar 

  • Lee IP, Schmid B, Zbinden G (1977) A simplified method for the isolation of sperm heads from the caput epididymidis of rodents and from ejaculated rabbit seminal plasma. Exp Cell Biol 45:48–59

    Google Scholar 

  • Legator MS, Connor TH, Stoeckel M (1975) Detection of mutagenic activity of metronidazole and niridazole in body fluids of humans and mice. Science 188:1118–1119

    Google Scholar 

  • Lutz WK (1979) In vivo covalent binding of organic chemicals to DNA as a quantitative indicator in the process of chemical carcinogenesis. Mutat Res 65:289–356

    Google Scholar 

  • McCann J, Choi E, Yamasaki E, Ames BN (1975) Detection of carcinogens as mutagens in the Salmonella microsome test: Assay of 300 chemicals. Proc Natl Acad Sci USA 72:5135–5139

    Google Scholar 

  • Meistrich ML, Reid BO, Barcellona WJ (1975) Meiotic DNA synthesis during mouse spermatogenesis. J Cell Biol 64:211–222

    Google Scholar 

  • Nevstad NP (1978) Sister chromatid exchanges and chromosomal aberrations induced in human lymphocytes by the cytostatic drug adriamycin in vivo and in vitro. Mutat Res 57:253–258

    Google Scholar 

  • Peacock A, Peacock PR (1966) The results of prolonged administration of isoniazid to mice, rats, and hamsters. Br J Cancer 20:307–325

    Google Scholar 

  • Rasmussen RE, Painter RB (1966) Radiation stimulated DNA synthesis in cultured mammalian cells. J Cell Biol 29:11–19

    Google Scholar 

  • Regan JA, Dunn WC (1979) Inhibition of DNA excision repair in human cells by arabinofuranosyl cytosine: effect on normal and XP cells. Mol Pharmacol 15:367–374

    Google Scholar 

  • Röhrborn G, Propping P, Buselmaier W (1972) Mutagenic activity of isoniazid and hydrazine in mammalian test systems. Mutat Res 16:189–194

    Google Scholar 

  • Rust JH (1976) Tumorigenicity studies — mouse, rat, and hamster. Int. Metronidazole Conference, May 26–28, Montréal, J, Infect Dis (Suppl)

  • Rustia M, Shubik P (1972) Induction of lung tumors and malignant lymphomas in mice by metronidazole. J Natl Cancer Inst 48:721–729

    Google Scholar 

  • Schmid B, Zbinden G (1979) Unscheduled DNA synthesis in male rabbit germ cells induced by methylmethane sulfonate, cyclophosphamide and adriamycin. Arch Toxicol Suppl 2:503–507

    Google Scholar 

  • Schmid B, Lee IP, Zbinden G (1978) DNA repair processes in germ cells demonstrated in ejaculated sperms of rabbits treated with methylmethane sulfonate. Arch Toxicol 40:37–43

    Google Scholar 

  • Schöneich S, Braun R (1977) Drug interactions and their influence on mutagenic activity. Environmental mutagens. Proceedings of the 6th annual meeting of the European Environmental Mutagen Society, vol 9. Academie Verlag, Berlin GDR, p 63

    Google Scholar 

  • Schwartz HS, Schioppacassi G, Kanter PM (1978) Mechanisms of selectivity of intercalating agents. Antibiot Chemother 23:247–254

    Google Scholar 

  • Sega GA (1974) Unscheduled DNA synthesis in the germ cells of male mice exposed in vivo to the chemical mutagen ethyl methanesulfonate. Proc Natl Acad Sci USA 71:4955–4959

    Google Scholar 

  • Sega GA, Owens JG, Cummings RB (1976) Studies on DNA repair in early spermatid stages of male mice after in vivo treatment with methyl-, ethyl-, propyl-, and isopropyl methane-sulfonate. Mutat Res 36:193–212

    Google Scholar 

  • Setchell BP (1974) The entry of substances into the seminiferous tubules. In: Mancini RE, Martini L (eds) Male fertility and sterility. Academic Press, New York, p 35

    Google Scholar 

  • Stich HF, San RHC, Lam P, Koropatnick J, Lo L (1977) Unscheduled DNA synthesis of human cells as a short-term assay for chemical carcinogens. In: Hitt HH, Watson JD, Winsten JA (eds) Origins of human cancer, Book C, Human risk assessment. Cold Spring Harbor Laboratory, p 1499

  • Swierstra EE, Foote RH (1965) Duration of spermatogenesis and spermatozoan transport in the rabbit based on cytological changes, DNA synthesis and labeling with tritiated thymidine. Am J Anat 116:401–412

    Google Scholar 

  • Toth B, Shubik P (1966) Mammary tumor inhibition and lung adenoma induction by isonicotinic acid hydrazid. Science 152:1376–1377

    Google Scholar 

  • Trzos RJ, Petzold GL, Brunden MN, Swenberg JA (1978) The evaluation of sixteen carcinogens in the rat using the micronucleus test. Mutat Res 58:79–86

    Google Scholar 

  • Vig, BK (1977) Genetic toxicology of mitomycin C, actinomycin, daunomycin and adriamycin. Mutat Res 19:189–238

    Google Scholar 

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  1. Institut für Toxikologie, Eidgenössische Technische Hochschule und Universität Zürich, Schorenstrasse 16, CH-8603, Schwerzenbach, Switzerland

    G. Zbinden

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Zbinden, G. Unscheduled DNA synthesis in the testis, a secondary test for the evaluation of chemical mutagens. Arch. Toxicol. 46, 139–149 (1980). https://doi.org/10.1007/BF00361252

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  • DOI: https://doi.org/10.1007/BF00361252

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