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Methodology of Genotoxic and Teratogenic Studies in Rats

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Teratogenicity Testing

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1797))

Abstract

The genotoxicity methods applied to rats are tests that can detect any damage, including changes in the number of chromosomes or in the structure of chromosomes, and nucleotide changes with structural abnormality in the DNA of animal cells. However, the method of teratogenicity is used to detect the effects of chemicals which cause congenital defects in living organisms. This study contains information about the effectiveness, reliability, ways of application, and methodology of genotoxic and teratogenic methods applied in vivo in rats.

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References

  1. Dellarco VL, Mavournin KH, Tice RR (1985) Aneuploidy and health risk assessment: current status and future directions. Environ Mutagen 7:405–424

    Article  CAS  PubMed  Google Scholar 

  2. Takahashi M, Hasegawa R, Furukawa F et al (1986) Effects of ethanol, potassium metabisulfite, formaldehyde and hydrogen peroxide on gastric carcinogenesis in rats after initiation with N-methyl-N-nitro-N-nitrosoguanidine. Jpn J Cancer Res 77:118–124

    PubMed  CAS  Google Scholar 

  3. Yavuz Kocaman A, Rencuzogullari E, Ila HB, Topaktas M (2008) The genotoxic effect of potassium metabisulfite using chromosome aberration, sister chromatid exchange, micronucleus tests in human lymphocytes and chromosome aberration test in bone marrow cells of rats. Environ Mol Mutagen 49:276–282

    Article  CAS  PubMed  Google Scholar 

  4. Albertini RJ, Anderson D, Douglas GR et al (2000) IPCS guidelines for the monitoring of genotoxic effects of carcinogens in humans. International Programme on Chemical Safety. Mutat Res 463:111–172

    Article  CAS  PubMed  Google Scholar 

  5. Bonassi S, Znaor A, Norppa H, Hagmar L (2004) Chromosomal aberrations and risk of cancer in humans: an epidemiologic perspective. Cytogenet Genome Res 104:376–382

    Article  CAS  PubMed  Google Scholar 

  6. Bonassi S, Ugolini D, Kirsch Volders M et al (2005) Human population with cytogenetic biomarkers: review of the literature and future prospectives. Environ Mol Mutagen 45:258–270

    Article  CAS  PubMed  Google Scholar 

  7. Etebari M, Sajjadi SE, Jafarian Dehkordi A, Nazmakanipour S (2016) Genotoxicity evaluation of hydroalcoholic and aqueous extracts of Dorema aucheri by the comet assay. Adv Biomed Res 5:199. https://doi.org/10.4103/2277-9175.190993

    Article  PubMed  PubMed Central  Google Scholar 

  8. Topaktas M, Rencuzogullari E (2010) Sitogenetik (Cytogenetic). Nobel, Turkey

    Google Scholar 

  9. Alkan FU, Anlas C (2015) Genotoxicity tests and genotoxic poisons. Turkish Clinics J Vet Sci Pharmacol Toxicol-Special Topics 1:69–74

    Google Scholar 

  10. Wedebye EB, Dybdahl M, Nikolov NG et al (2015) QSAR screening of 70,983 REACH substances for genotoxic carcinogenicity, mutagenicity and developmental toxicity in the ChemScreen project. Reprod Toxicol 55:64–72

    Article  CAS  PubMed  Google Scholar 

  11. Tucker JD, Auletta A, Cimino MC et al (1993) Sister chromatid exchange: second report of the gene-Tox program. Mutat Res 297:101–180

    Article  CAS  PubMed  Google Scholar 

  12. Carrano AV, Natarajan AT (1988) Consideration for population monitoring using cytogenetic techniques. Mutat Res 204:379–406

    Article  CAS  PubMed  Google Scholar 

  13. Anderson D (1988) Human biomonitoring. Mutat Res 204:353–541

    Article  Google Scholar 

  14. Hagmar L, Brogger A, Hansteen IL et al (1994) Cancer risk in human predicted by increased levels of chromosomal aberrations in lymphocytes: Nordic Study Group on the Health Risk of Chromosome Damage. Cancer Res 54:2919–2922

    PubMed  CAS  Google Scholar 

  15. Heddle JA, Cimino MC, Hayashi M et al (1991) Micronuclei as an index of cytogenetic damage: past, present, and future. Environ Mol Mutagen 18:277–291

    Article  CAS  PubMed  Google Scholar 

  16. Fenech M (2002) Biomarkers of genetic damage for cancer epidemiology. Toxicology 181:411–416

    Article  PubMed  Google Scholar 

  17. OECD (1997) Unscheduled DNA synthesis (UDS) test with mammalian liver cells in vivo. In: Guideline for the testing of chemicals. http://www.oecd-ilibrary.org/docserver/download/9748601e.pdf?expires=1494590875& id=id&accname=guest&checksum=48389350D076A0B07DF98A4073340A1F. Accessed 11 May 2017

  18. Maron DM, Ames BN (1983) Revised method for the Salmonella mutagenicity test. Mutat Res 113:173–215

    Article  CAS  PubMed  Google Scholar 

  19. Yavuz Kocaman A, Rencuzogullari E, Topaktas M (2014) In vitro investigation of the genotoxic and cytotoxic effects of thiacloprid in cultured human peripheral blood lymphocytes. Environ Toxicol 29:631–641

    Article  CAS  Google Scholar 

  20. EPA (1996) In vivo sister chromatid exchange assay. In: United States Environmental Protection Agency Health Effects Test Guidelines. https://nepis.epa.gov/Exe. Accessed 11 May 2017

  21. OECD (1997). Mammalian bone marrow chromosome aberration test. In: Guideline for the testing of chemicals. http://www.oecd-ilibrary.org/environment/test-no-475-mammalian-bone-marrow-chromosome-aberration-test_9789264071308-en. Accessed 12 Mar 2017

  22. OECD (2013) Mammalian erythrocyte micronucleus test. In: Guideline for the testing of chemicals. http://www.oecd.org/env/ehs/testing/draft_tg474_second_commenting_round.pdf. Accessed 11 May 2017

  23. Cooper GM, Hausman RE (2004) The cell: a molecular approach. ASM, USA

    Google Scholar 

  24. Topaktas M, Rencuzogullari E, Ila HB (1996) In vivo chromosomal aberrations in bone marrow cells of rats treated with marshal. Mutat Res 371:259–264

    Article  CAS  PubMed  Google Scholar 

  25. Becerril C, Ferrero M, Sanz F, Castaño A (1999) Detection of mitomycin C-induced genetic damage in fish cells by use of RAPD. Mutagenesis 14:449–456

    Article  CAS  PubMed  Google Scholar 

  26. Atienzar FA, Jha AN (2006) The random amplified polymorphic DNA (RAPD) assay and related techniques applied to genotoxicity and carcinogenesis studies: a critical review. Mutat Res 613:76–102

    Article  CAS  PubMed  Google Scholar 

  27. Bayram S, Genc A, Buyukleyla M, Rencuzogullari E (2016) Genotoxicity and cytotoxicity of copper oxychloride in cultured human lymphocytes using cytogenetic and molecular tests. Cytotechnology 68:2027–2036

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Fairbain DW, Olive PL, O’Neill KL (1995) The comet assay: a comprehensive review. Mutat Res 339:37–59

    Article  Google Scholar 

  29. Wilson JG (1973) Environment and birth defects. By Aston SA, Published: 2014-06-21 https://embryo.asu.edu/pages/environment-and-birth-defects-1973-james-g-wilson. Accessed 14 Mar 2017

  30. Shreder OV, Shreder ED, Durnev AD, Seredenin SB (2011) Association of genotoxic and teratogenic effects induced by cyclophosphamide and their modification with afobazole. Gig Sanit 5:64–88

    Google Scholar 

  31. el-Ashmawy IM, el-Nahas AF, Bayad AE (2011) Teratogenic and cytogenetic effects of ivermectin and its interaction with P-glycoprotein inhibitor. Res Vet Sci 90:116–123

    Article  CAS  PubMed  Google Scholar 

  32. Murkunde YV, Sathya TN, Subashini N, Murthy PB (2012) Transplacental genotoxicity evaluation of cypermethrin using alkaline comet assay. Hum Exp Toxicol 31:185–192

    Article  CAS  PubMed  Google Scholar 

  33. Brown NA, Spielmann H, Bechter R et al (1995) Screening chemicals for reproductive toxicity: the current alternatives. Altern Lab Anim 23:868–882

    Google Scholar 

  34. Foote RH, Carney EW (2000) The rabbit as a model for reproductive and developmental toxicity studies. Reprod Toxicol 14:477–493

    Article  CAS  PubMed  Google Scholar 

  35. Bremer S, Hartung T (2004) The use of embryonic stem cells for regulatory developmental toxicity testing in vitro-the current status of test development. Curr Pharm Des 10:2733–2747

    Article  CAS  PubMed  Google Scholar 

  36. Mace ML Jr, Daskal Y, Wray W (1978) Scanning-electron microscopy of chromosome aberrations. Mutat Res 52:199–206

    Article  PubMed  Google Scholar 

  37. Speit G, Haupter S (1985) On the mechanism of differential giemsa staining of bromodeoxyuridine substituted chromosomes. II. Differences between the demostration of sister chromatid differentiation and replication patterns. Hum Genet 70:126–129

    Article  CAS  PubMed  Google Scholar 

  38. Rencüzogullari E, Ila HB, Kayraldiz A, Topaktaş M (2001) Chromosome aberrations and sister chromatid exchanges in cultured human lymphocytes treated with sodium metabisulfite, a food preservative. Mutat Res 490:107–112

    Article  PubMed  Google Scholar 

  39. Matter B, Schmid W (1971) Trenimon-induced chromosomal damage in bone-marrow cells of six mammalian species, evaluated by the micronucleus test. Mutat Res 12:417–425

    Article  CAS  PubMed  Google Scholar 

  40. von Ledebur M, Schmid W (1973) The micronucleus test. Methodological aspects. Mutat Res 19:109–117

    Article  Google Scholar 

  41. Schmid W (1975) The micronucleus test. Mutat Res 31:9–15

    Article  CAS  PubMed  Google Scholar 

  42. Romagna F (1988) Series: current issues in mutagenesis and carcinogenesis. 12. Improved method of preparing bone marrow micronucleus assay slides. Mutat Res 206:307–309

    Article  CAS  PubMed  Google Scholar 

  43. Heddle JA (1990) Micronuclei in vivo. Prog Clin Biol Res 340:185–194

    Google Scholar 

  44. Heddle JA, Fenech M, Hayashi M, MacGregor JT (2011) Reflections on the development of micronucleus assays. Mutagenesis 26:3–10

    Article  CAS  PubMed  Google Scholar 

  45. Oz Aydin S (2004) RAPD (Rastgele Arttırılmış Polimorfik DNA) belirleyicileri ve bitki sistematiği (RAPD markers and plant systematics). Dumlupınar Universitesi Fen Bilimleri Enstitü Dergisi (Dumlupinar Uni J Natural and Applied Sci) 6:113–130

    Google Scholar 

  46. Timocin T, Ila HB (2015) Investigation of flurbiprofen genotoxicity and cytotoxicity in rat bone marrow cells. Drug Chem Toxicol 38:355–360

    Article  CAS  PubMed  Google Scholar 

  47. Doyle JJ, Doyle JL (1987) A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem Bull 19:11–15

    Google Scholar 

  48. Noel S, Rath SK (2006) Randomly amplified polymorphic DNA as a tool for genotoxicity: an assessment. Toxicol Ind Health 22:267–275

    Article  CAS  PubMed  Google Scholar 

  49. Atienzar FA, Conradi M, Evenden AJ et al (1999) Qualitative assessment of genotoxicity using random amplified polymorphic DNA: comparison of genomic template stability with key fitness parameters in Daphnia magna exposed to benzo[a]pyrene. Environ Toxicol Chem 18:2275–2282

    PubMed  CAS  Google Scholar 

  50. Buyukleyla M (2013) Gida katki maddesi sodyum metabisulfit’in genotoksik etkisinin RAPD-PCR yöntemi ile araştırılması (detection of genotoxic effects of food additive sodium metabisulfite using RAPD/PCR method). Cukurova University Institute of Natural and Applied Science, Department of Biology, Doctorate thesis (Supervisor Topaktas M), p 107

    Google Scholar 

  51. Dincer Y, Kankaya S (2010) DNA Hasarının Belirlenmesinde Comet Assay (Comet Assay for determining of DNA damage). J Med Sci 30:1365–1373

    CAS  Google Scholar 

  52. OECD (2016) In vivo mammalian alkaline Comet Assay. In: Guideline for the testing of chemicals. http://www.oecd-ilibrary.org/docserver/download/9716431e.pdf?expires=1494577986&id=id&accname=guest&checksum=9471A50A37B578F471C06D74E4A7760A. Accessed 11 May 2017

  53. Ostling O, Johanson KJ (1984) Microelectrophoretic study of radiation-induced DNA damage in individual mammalian cells. Biochem Biophys Res Commun 123:291–298

    Article  CAS  PubMed  Google Scholar 

  54. Singh NP, McCoy MT, Tice RR, Schneider EL (1988) A simple technique for quantitation of low levels of DNA damage in individual cells. Exp Cell Res 175:184–191

    Article  CAS  PubMed  Google Scholar 

  55. Horoz M, Bolukbas C, Bolukbas F et al (2006) Assessment of peripheral DNA damage by alkaline comet assay in maintenance hemodialysis subjects with hepatitis C infection. Mutat Res 596:137–142

    Article  CAS  PubMed  Google Scholar 

  56. Tice RR, Agurell E, Anderson D et al (2000) Single cell gel/Comet Assay: guidelines for in vitro and in vivo genetic toxicology testing. Environ Mol Mutagen 35:206–221

    Article  CAS  Google Scholar 

  57. Collins AR (2004) The comet assay for DNA damage and repair: principles, applications, and limitations. Mol Biotechnol 26:249–261

    Article  CAS  PubMed  Google Scholar 

  58. OECD (2001) Prenatal developmental toxicity study. In: Guideline for the testing of chemicals. https://ntp.niehs.nih.gov/iccvam/suppdocs/feddocs/oecd/oecd_gl414.pdf. Accessed 14 May 2017

  59. Okada A, Kurihara H, Aoki Y et al (2004) Amidic modification of valproic acid reduces skeletal teratogenicity in mice. Birth Defects Res (Part B) 71:47–53

    Article  CAS  Google Scholar 

  60. Canimoglu S (2009) Maltitol’un sicanlarda mutagenik ve teratojenik etkileri (the mutagenic and teratogenic effects of maltitol in rats). Cukurova University Institute of Natural and Applied Science, Department of Biology, Doctorate thesis (Supervisor Rencuzogullari E), p 83

    Google Scholar 

  61. Canimoglu S, Rencuzogullari E (2013) The genotoxic and teratogenic effects of maltitol in rats. Toxicol Ind Health 29:935–943

    Article  CAS  PubMed  Google Scholar 

  62. Menegola E, Broccia LM, Giavini E (2001) Atlas of rat fetal skeleton double stained for bone and cartilage. Teratology 64:125–133

    Article  CAS  PubMed  Google Scholar 

  63. Perry PE, Thompson EJ (1984) The methodology of sister chromatid exchanges. In: Kilbey BJ, Legator M, Nichols W, Ramel C (eds) Handbook of mutagenicity test procedures, 2nd edn. Elsevier Sci BV, Amsterdam, pp 495–529

    Chapter  Google Scholar 

  64. Evans HJ (1984) Human peripheral blood lymphocytes for the analysis of chromosome aberrations in mutagen tests. In: Kilbey BJ, Legator M, Nichols W, Ramel C (eds) Handbook of mutagenicity test procedures, 2nd edn. Elsevier Sci BV, Amsterdam, pp 405–427

    Chapter  Google Scholar 

  65. OECD (1986) Genetic toxicology: in vitro sister chromatid exchange assay in mammalian cells. In: Guideline for the testing of chemicals. http://www.oecd-ilibrary.org/environment/test-no-479-genetic-toxicology-in-vitro-sister-chromatid-exchange-assay-in-mammalian-cells_9789264071384-en. Accessed 11 Mar 2017

  66. OECD (1997) In vitro mammalian chromosome aberration. In: Guideline for the testing of chemicals. http://www.oecd.org/chemicalsafety/risk-assessment/1948434.pdf. Accessed 11 Mar 2017

  67. Kirsch-Volders M, Elhajouji A, Cundari E, Van Hummelen P (1997) The in vitro micronucleus test: a multi-endpoint assay to detect simultaneously mitotic delay, apoptosis, chromosome breakage, chromosome loss and non-disjunction. Mutat Res 392:19–30

    Article  CAS  PubMed  Google Scholar 

  68. Fenech M (2000) The in vitro micronucleus technique. Mutat Res 455:81–95

    Article  CAS  PubMed  Google Scholar 

  69. Green MH, Lowe JE, Delaney CA, Green IC (1996) Comet assay to detect nitric oxide-dependent DNA damage in mammalian cells. Methods Enzymol 269:243–266

    Article  CAS  PubMed  Google Scholar 

  70. McKelvey-Martin VJ, Green MH, Schmezer P et al (1993) The single cell gel electrophoresis assay (comet assay): a European review. Mutat Res 288:47–63

    Article  CAS  PubMed  Google Scholar 

  71. Kopar N (2010) Salvia fruticosa bitki ekstraktinin metabolik aktivator varliginda ve yoklugunda insan lenfositlerinde genotoksik ve anti-genotoksik etkisi (the genotoxic and anti-genotoxic effects of Salvia fruticosa plant extract in human lymphocytes in the absence and presence of metabolic activator). Cukurova University, Institute of Natural and Applied Science, Department of Biology, Master thesis (Supervisor Rencuzogullari E), p 84

    Google Scholar 

  72. Hernández-Ceruelos A, Madrigal-Santillán E, Morales-González JA et al (2010) Antigenotoxic effect of Chamomilla recutita (L.) Rauschert essential oil in mouse spermatogonial cells, and determination of its antioxidant capacity in vitro. Int J Mol Sci 11:3793–3802

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Authors and Affiliations

  1. Department of Biology, Faculty of Science and Letters, Adiyaman University, Adiyaman, Turkey

    Eyyup Rencuzogullari & Muhsin Aydin

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  1. Eyyup Rencuzogullari
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  2. Muhsin Aydin
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Correspondence to Eyyup Rencuzogullari .

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  1. Centre for the Research and Technology of Agro-Environmental and Biological Sciences (CITAB), University of Trás-os-Montes e Alto Douro (UTAD), Vila Real, Portugal

    Luís Félix

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Rencuzogullari, E., Aydin, M. (2018). Methodology of Genotoxic and Teratogenic Studies in Rats. In: Félix, L. (eds) Teratogenicity Testing. Methods in Molecular Biology, vol 1797. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7883-0_33

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  • DOI: https://doi.org/10.1007/978-1-4939-7883-0_33

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