Rapid metaphase and interphase detection of radiation‐induced chromosome aberrations in human lymphocytes by chromosomal suppression in situ hybridization

Abstract

Chromosomal in situ suppression (CISS)‐hybridization of biotinylated phage DNA‐library inserts from sorted human chromosomes was used to decorate chromosomes 1 and 7 specifically from pter to qter and to detect structural aberrations of these chromosomes in irradiated human peripheral lymphocytes. In addition, probe pUC1.77 was used to mark the Iq12 subregion in normal and aberrant chromosomes 1. Low LET radiation (60Co‐γ‐rays; 1.17 and 1.33 MeV) of lymphocyte cultures was performed with various doses (D = 0, 2, 4, 8 Gy) 5 h after stimulation with phytohaemagglutinin. Irradiated cells were cultivated for an additional 67 h before Colcemid arrested metaphase spreads were obtained. Aberrations of the specifically stained chromosomes, such as deletions, dicentrics, and rings, were readily scored after in situ hybridization with either the 1q12 specific probe or DNA‐library inserts. By the latter approach, translocations of the specifically stained chromosomes could also be reliably assessed. A linear increase of the percentage of specifically stained aberrant chromosomes was observed when plotted as a function of the square of the dose D. A particular advantage of this new approach is provided by the possibility to delineate numerical and structural chromosome aberrations directly in interphase nuclei. These results indicate that cytogenetic monitoring of ionizing radiation may be considerably facilitated by CISS‐hybridization.

Molecular cytotaxonomy of primates by chromosomal in situ suppression hybridization

“…Figures lk-r present examples. These libraries were previously shown to stain the respective human chromosomes under suppression conditions (Lichter et al, 1988a) and to delineate structural aberrations (Cremer et al, 1988(Cremer et al, , 1990. Background staining of nontargeted chromosomes seen with these and gonosome libraries was due to incomplete suppression of repetitive sequences.…”

Analysis of genes and chromosomes by nonisotopic in situ hybridization

“…Tumor cytogenetics, one of the most promising fields of application, will be discussed in more detail elsewhere (T. Cremer et al, in preparation). For other applications such as chromosome aberration detection in biological dosimetry [140][141][142][143], the reader is referred to the literature.…”

Development of a Biological Dosimeter for Translocation Scoring Based on Two-Color Fluorescence in Situ Hybridization of Chromosome Subsets

“…Therefore, it would be highly desirable, if a biological dosimeter could be developed for chromosome aberration scoring directly in cell nuclei of irradiated tissues. In contrast to the painting of individual chromosomes which has been supplied successfully for the scoring of chromosome translocations in the interphase nucleus [15], [16], nuclear painting patterns derived from the painting of chromosomal subsets as described in this paper are not suitable for such interphase cytogenetic analyses. The sensitivity and accuracy with which chromosome translocations can be detected in interphase nuclei after multicolor painting of individual chromosome domains and the potential of such an approach for automated analyses provides an important field for future investigations.…”

“…In particular, "painting" of entire human chromosomes based on chromosomal in situ suppression (CISS-) hybridization protocols [13,14] in combination with digital image analysis can be applied for the rapid evaluation of translocation chromosomes composed of differently colored chromosome segments [15][16][17][18][19][20]. Using this approach, we have evaluated chromosome translocations in patients who had received an intravascular injection of Thorotrast more than fourty years ago [17].…”