Comparison of spontaneous and idoxuridine-induced micronuclei by chromosome painting.
Fluorescence in situ hybridisation (FISH) technique with chromosome specific library (CSL) DNA probes for all human chromosomes were used to study about 9000 micronuclei (MN) in normal and idoxuridine (IUdR)-treated lymphocyte cultures of female and male donors. In addition, MN rates and structural chromosome aberrations were scored in Giemsa-stained chromosome spreads of these cultures. IUdR treatment (40 microg/ml) induced on the average a 12-fold increase of the MN rate. Metaphase analysis revealed no distinct increase of chromosome breaks but a preferential decondensation at chromosome 9q12 (28-79%) and to a lower extend at 1q12 (8-21%). Application of FISH technique with CSL probes to one male and one female untreated proband showed that all human chromosomes except chromosome 12 (and to a striking high frequency chromosomes 9, X and Y) occurred in spontaneous MN. In cultures containing IUdR, the chromosomal spectrum found in MN was reduced to 10 chromosomes in the male and 13 in the female proband. Eight chromosomes (2, 6, 12, 13, 14, 15, 17 and 18) did not occur in MN of both probands. On the contrary chromosomes 1 and especially 9 were found much more frequently in the MN of IUdR-treated cultures than in MN of control cultures. DAPI-staining revealed heterochromatin signals in most of the IUdR-induced MN. In an additional study, spontaneous and IUdR-induced MN were investigated in lymphocytes of another female donor using CSL probes only for chromosomes 1, 6, 9, 15, 16 and X. The results confirmed the previous finding that chromosomes 1 and 9 occur very often in MN after IUdR-treatment. The results indicate that decondensation of heterochromatic regions on chromosomes 1 and 9 caused by IUdR treatment strongly correlates with MN formation by these chromosomes.
Fauth
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Abt. Humanbiologie und Humangenetik der Universität, Postfach 3049, D-67653, Kaiserslautern, Germany. efauth@rhrk.uni-kl.de
Zankl
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D001399:Azure Stains; D018929:Cell Culture Techniques; D002869:Chromosome Aberrations / Q000235:genetics*; D019457:Chromosome Breakage / Q000235:genetics*; D002878:Chromosomes, Human, Pair 1 / Q000187:drug effects / Q000235:genetics; D002899:Chromosomes, Human, Pair 9 / Q000187:drug effects / Q000235:genetics; D015342:DNA Probes; D005260:Female; D015723:Gene Library; D006570:Heterochromatin / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007065:Idoxuridine / Q000633:toxicity*; D017404:In Situ Hybridization, Fluorescence; D007211:Indoles; D008214:Lymphocytes / Q000187:drug effects / Q000648:ultrastructure; D008297:Male; D008677:Metaphase / Q000187:drug effects* / Q000235:genetics; D048629:Micronuclei, Chromosome-Defective / Q000187:drug effects* / Q000235:genetics; D019384:Nucleic Acid Synthesis Inhibitors / Q000633:toxicity; D012737:Sex Factors
D001399:Azure Stains; D015342:DNA Probes; D006570:Heterochromatin; D007211:Indoles; D019384:Nucleic Acid Synthesis Inhibitors; C007293:DAPI; D007065:Idoxuridine