Analysis of radiation-induced micronuclei by FISH using a combination of painting and centromeric DNA probes

In situ hybridization with whole chromosome painting probes (chromosome 1, 7, 11, 14, 17 and 21) in combination with a human pancentromeric alpha-satellite probe was used to analyse the presence of specific chromosomal material in micronuclei (MN) induced in human lymphocytes by ionizing radiation. The purpose was to investigate the nature of radiation-induced cytogenetic damage, especially to study whether the fraction of paint-positive MN is proportional to the relative DNA content of the respective chromosomes which might indicate a random breakage of chromosomes. Flow-sorted MN and MN in binucleated cells were analysed with the six chromosome specific painting probes. It was found that the fraction of paint-positive MN increased linearly with the DNA content of the respective chromosomes. About 13% radiation-induced MN in human lymphocytes were found to contain centromeric signals independent of the presence of specific chromosome painting signals. The data obtained on flow-sorted MN and MN in binucleated cells agreed well, indicating that flow-sorted MN can be used for studying their chromosomal content with the FISH technique. If it is assumed that the chromosomal content of MN reflects radiation-induced damage, then these results support a random model of radiation-induced cytogenetic damage in human lymphocytes for the six chromosomes studied.