Skip to main content
Environmental Health Perspectives logoLink to Environmental Health Perspectives
. 1993 Oct;101(Suppl 3):139–143. doi: 10.1289/ehp.93101s3139

Micronucleus assay in lymphocytes as a tool to biomonitor human exposure to aneuploidogens and clastogens.

H Norppa 1, S Luomahaara 1, H Heikanen 1, S Roth 1, M Sorsa 1, L Renzi 1, C Lindholm 1
PMCID: PMC1521122  PMID: 8143606

Abstract

The analysis of micronuclei (MN) in cultured human lymphocytes is, in principle, able to detect exposure to clastogens and aneuploidogens alike. There is, however, no clear evidence from human biomonitoring studies or animal experiments showing that in vivo exposure of resting lymphocytes to an aneuploidogen could actually be expressed as MN in cultured lymphocytes. In vitro, a pulse treatment of human lymphocytes with vinblastine, an aneuploidogen, did result in MN induction even if performed before mitogen stimulation, although a much more pronounced effect was obtained in actively dividing lymphocyte cultures. On the other hand, it is probable that a considerable portion of "spontaneous" MN contain whole chromosomes, their contribution increasing with age. It also seems that cytochalasin B, used for the identification of second cell cycle interphase cells in the MN assay, is able to slightly increase the level of MN with whole chromosomes. If MN harboring chromosome fragments represent a minority of the total MN frequency, there may be difficulties in detecting a weak effect in this fraction of MN against the background of MN with whole chromosomes. This would reduce the sensitivity of the assay in detecting clastogens, unless MN with whole chromosomes and chromosome fragments are distinguished from each other. That a problem may exist in sensitivity is suggested by the difficulty in demonstrating MN induction by smoking, an exposure capable of inducing chromosome aberrations. The sensitivity of the lymphocyte MN assay could be increased by detecting kinetochore or centromere in MN, or by automation, allowing more cells to be analyzed.

Full text

PDF
139

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Anwar W. A., Gabal M. S. Cytogenetic study in workers occupationally exposed to mercury fulminate. Mutagenesis. 1991 May;6(3):189–192. doi: 10.1093/mutage/6.3.189. [DOI] [PubMed] [Google Scholar]
  2. Au W. W., Walker D. M., Ward J. B., Jr, Whorton E., Legator M. S., Singh V. Factors contributing to chromosome damage in lymphocytes of cigarette smokers. Mutat Res. 1991 Jun;260(2):137–144. doi: 10.1016/0165-1218(91)90001-3. [DOI] [PubMed] [Google Scholar]
  3. Barregård L., Högstedt B., Schütz A., Karlsson A., Sällsten G., Thiringer G. Effects of occupational exposure to mercury vapor on lymphocyte micronuclei. Scand J Work Environ Health. 1991 Aug;17(4):263–268. doi: 10.5271/sjweh.1704. [DOI] [PubMed] [Google Scholar]
  4. Becker P., Scherthan H., Zankl H. Use of a centromere-specific DNA probe (p82H) in nonisotopic in situ hybridization for classification of micronuclei. Genes Chromosomes Cancer. 1990 May;2(1):59–62. doi: 10.1002/gcc.2870020111. [DOI] [PubMed] [Google Scholar]
  5. Eastmond D. A., Tucker J. D. Identification of aneuploidy-inducing agents using cytokinesis-blocked human lymphocytes and an antikinetochore antibody. Environ Mol Mutagen. 1989;13(1):34–43. doi: 10.1002/em.2850130104. [DOI] [PubMed] [Google Scholar]
  6. Fenech M., Denham J., Francis W., Morley A. Micronuclei in cytokinesis-blocked lymphocytes of cancer patients following fractionated partial-body radiotherapy. Int J Radiat Biol. 1990 Feb;57(2):373–383. doi: 10.1080/09553009014552471. [DOI] [PubMed] [Google Scholar]
  7. Fenech M., Morley A. A. Kinetochore detection in micronuclei: an alternative method for measuring chromosome loss. Mutagenesis. 1989 Mar;4(2):98–104. doi: 10.1093/mutage/4.2.98. [DOI] [PubMed] [Google Scholar]
  8. Fenech M., Morley A. A. Measurement of micronuclei in lymphocytes. Mutat Res. 1985 Feb-Apr;147(1-2):29–36. doi: 10.1016/0165-1161(85)90015-9. [DOI] [PubMed] [Google Scholar]
  9. Heddle J. A., Cimino M. C., Hayashi M., Romagna F., Shelby M. D., Tucker J. D., Vanparys P., MacGregor J. T. Micronuclei as an index of cytogenetic damage: past, present, and future. Environ Mol Mutagen. 1991;18(4):277–291. doi: 10.1002/em.2850180414. [DOI] [PubMed] [Google Scholar]
  10. Kocisová J., Srám R. J. Mutagenicity studies on paracetamol in human volunteers. III. Cytokinesis block micronucleus method. Mutat Res. 1990 May;244(1):27–30. doi: 10.1016/0165-7992(90)90103-q. [DOI] [PubMed] [Google Scholar]
  11. Larramendy M. L., Knuutila S. Increased frequency of micronuclei in B and T8 lymphocytes from smokers. Mutat Res. 1991 Feb;259(2):189–195. doi: 10.1016/0165-1218(91)90052-n. [DOI] [PubMed] [Google Scholar]
  12. Lindholm C., Norppa H., Hayashi M., Sorsa M. Induction of micronuclei and anaphase aberrations by cytochalasin B in human lymphocyte cultures. Mutat Res. 1991 Aug;260(4):369–375. doi: 10.1016/0165-1218(91)90022-e. [DOI] [PubMed] [Google Scholar]
  13. Littlefield L. G., Sayer A. M., Frome E. L. Comparisons of dose-response parameters for radiation-induced acentric fragments and micronuclei observed in cytokinesis-arrested lymphocytes. Mutagenesis. 1989 Jul;4(4):265–270. doi: 10.1093/mutage/4.4.265. [DOI] [PubMed] [Google Scholar]
  14. Meyne J., Littlefield L. G., Moyzis R. K. Labeling of human centromeres using an alphoid DNA consensus sequence: application to the scoring of chromosome aberrations. Mutat Res. 1989 Jun;226(2):75–79. doi: 10.1016/0165-7992(89)90046-8. [DOI] [PubMed] [Google Scholar]
  15. Migliore L., Guidotti P., Favre C., Nardi M., Sessa M. R., Brunori E. Micronuclei in lymphocytes of young patients under antileukemic therapy. Mutat Res. 1991 Aug;263(4):243–248. doi: 10.1016/0165-7992(91)90008-r. [DOI] [PubMed] [Google Scholar]
  16. Migliore L., Parrini M., Sbrana I., Biagini C., Battaglia A., Loprieno N. Micronucleated lymphocytes in people occupationally exposed to potential environmental contaminants: the age effect. Mutat Res. 1991 Jan;256(1):13–20. doi: 10.1016/0921-8734(91)90028-a. [DOI] [PubMed] [Google Scholar]
  17. Mäki-Paakkanen J., Walles S., Osterman-Golkar S., Norppa H. Single-strand breaks, chromosome aberrations, sister-chromatid exchanges, and micronuclei in blood lymphocytes of workers exposed to styrene during the production of reinforced plastics. Environ Mol Mutagen. 1991;17(1):27–31. doi: 10.1002/em.2850170105. [DOI] [PubMed] [Google Scholar]
  18. Prosser J. S., Moquet J. E., Lloyd D. C., Edwards A. A. Radiation induction of micronuclei in human lymphocytes. Mutat Res. 1988 May;199(1):37–45. doi: 10.1016/0027-5107(88)90228-x. [DOI] [PubMed] [Google Scholar]
  19. Sarto F., Tomanin R., Giacomelli L., Canova A., Raimondi F., Ghiotto C., Fiorentino M. V. Evaluation of chromosomal aberrations in lymphocytes and micronuclei in lymphocytes, oral mucosa and hair root cells of patients under antiblastic therapy. Mutat Res. 1990 Feb;228(2):157–169. doi: 10.1016/0027-5107(90)90072-c. [DOI] [PubMed] [Google Scholar]
  20. Sarto F., Törnqvist M. A., Tomanin R., Bartolucci G. B., Osterman-Golkar S. M., Ehrenberg L. Studies of biological and chemical monitoring of low-level exposure to ethylene oxide. Scand J Work Environ Health. 1991 Feb;17(1):60–64. doi: 10.5271/sjweh.1733. [DOI] [PubMed] [Google Scholar]
  21. Sinúes B., Nuñez E., Bernal M. L., Alcala A., Saenz M. A., Conde B. Micronucleus assay in biomonitoring of patients undergoing excretory urography with diatrizoate and ioxaglate. Mutat Res. 1991 Aug;260(4):337–342. doi: 10.1016/0165-1218(91)90019-i. [DOI] [PubMed] [Google Scholar]
  22. Sorsa M., Anttila A., Järventaus H., Kubiak R., Norppa H., Nylander L., Pekari K., Pfäffli P., Vainio H. Styrene revisited--exposure assessment and risk estimation in reinforced plastics industry. Prog Clin Biol Res. 1991;372:187–195. [PubMed] [Google Scholar]
  23. Sorsa M., Pyy L., Salomaa S., Nylund L., Yager J. W. Biological and environmental monitoring of occupational exposure to cyclophosphamide in industry and hospitals. Mutat Res. 1988 Mar;204(3):465–479. doi: 10.1016/0165-1218(88)90042-0. [DOI] [PubMed] [Google Scholar]
  24. Thomson E. J., Perry P. E. The identification of micronucleated chromosomes: a possible assay for aneuploidy. Mutagenesis. 1988 Sep;3(5):415–418. doi: 10.1093/mutage/3.5.415. [DOI] [PubMed] [Google Scholar]
  25. Tomanin R., Ballarin C., Nardini B., Mastrangelo G., Sarto F. Influence of smoking habit on the frequency of micronuclei in human lymphocytes by the cytokinesis block method. Mutagenesis. 1991 Mar;6(2):123–126. doi: 10.1093/mutage/6.2.123. [DOI] [PubMed] [Google Scholar]

Articles from Environmental Health Perspectives are provided here courtesy of National Institute of Environmental Health Sciences

RESOURCES