The cytokinesis-block micronucleus technique: a detailed description of the method and its application to genotoxicity studies in human populations.
The development of the cytokinesis-block (CB) technique has transformed the human-lymphocyte micronucleus assay (MN) into a reliable and precise method for assessing chromosome damage. Recent studies in our laboratory have confirmed that this method is a sensitive indicator of in vivo radiation exposure in (a) patients undergoing fractionated partial-body radiotherapy and (b) rodents exposed to uniform whole-body irradiation, thus supporting the application of the cytokinesis-block micronucleus (CBMN) assay for biological dosimetry. To further define the use of this assay in biomonitoring we performed extensive studies to determine the spontaneous level of MN in normal human populations and its relationship to various life-style factors. We have also developed a new variation to the CBMN assay that permits the conversion of excision-repairable lesions to MN within one cell-cycle using cytosine arabinoside. With this method the slope of the in vitro dose-response curves was increased by a factor of 1.8 for X-rays, 10.3 for ultraviolet (UV, 254 nm) radiation and approximately 40-fold for methylnitrosourea. Consequently the CBMN assay can now be used to measure not only whole chromosome loss or chromosome breaks but also excision-repair events. The versatility and simplicity of the CBMN assay together with new developments in automation should ensure its successful application in monitoring exposed populations as well as in identifying mutagen-sensitive individuals within a population.
Fenech
M
M
CSIRO Division of Human Nutrition, Adelaide, Australia.
D000818:Animals; D002455:Cell Division; D003561:Cytarabine / Q000494:pharmacology; D004247:DNA / Q000378:metabolism; D004249:DNA Damage; D005260:Female; D006801:Humans; D008214:Lymphocytes / Q000648:ultrastructure; D008297:Male; D048629:Micronuclei, Chromosome-Defective; D015162:Micronucleus Tests / Q000379:methods*; D009154:Mutation
D003561:Cytarabine; D004247:DNA