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Mutagenesis vol. 12 no. 4 pp. 195-200, 1997
© 1997 UK Environmental Mutagen Society/Oxford University Press

research-article

Detection of chromosomal alterations affecting the 1cen–1q12 region in irradiated granulocytes and lymphocytes by multicolour FISH with tandem DNA probes

Doppalapudi S. Rupa, Leslie S. Hasegawa and David A. Eastmond1

Environmental Toxicology Graduate Program, Department of Entomology, University of California Riverside, CA 92521, USA

A multicolour tandem labelling fluorescence in situ hybridization (FISH) procedure was used to compare the frequencies of radiation-induced chromosome breakage and hyperdiploidy of chromosome 1 occurring in non-cultured granulocytes and Go lymphocytes with those observed in cultured metaphase and interphase lymphocytes. Whole blood, obtained from healthy male donors, was exposed in vitro to 0, 100, 200, 300 and 400 cGy of ionizing radiation from a 137Cs source. Aliquots containing granulocytes and Go lymphocytes from each dose were treated immediately with hypotonic KCl on ice and harvested. Cells were hybridized with {alpha}- and classical satellite probes to the 1cen-ql2 region of chromosome 1 and the frequencies of hyperdiploidy and breakage affecting this region were determined. Elevated dose-related frequencies of breakage were detectable in both lymphocytes and granulocytes immediately following radiation and decreased rapidly over the first 0.25–2 h. In a second series of experiments, the frequencies of hyperdiploidy and breakage for uncultured granulocytes and Go lymphocytes were compared with interphase and metaphase cells following 48–51 h of culture. Similar and significant dose-related increases in breakage were seen for the granulocytes, Go lymphocytes, 48 h cultured interphase and metaphase lymphocytes. A minor increase in hyperdiploidy was seen in the irradiated cultured cells, whereas no hyperdiploid cells were detected in the non-cultured cells. These results indicate that, in general, granulocytes and lymphocytes show similar sensitivity to radiation-induced damage and that cell culture is not required for chromosome breakage to be observed microscopically using this FISH procedure.

1To whom correspondence should be addressed


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M.J. Ramirez, J. Surralles, P. Galofre, A. Creus, and R. Marcos
FISH analysis of 1cen–1q12 breakage, chromosome 1 numerical abnormalities and centromeric content of micronuclei in buccal cells from thyroid cancer and hyperthyroidism patients treated with radioactive iodine
Mutagenesis, January 1, 1999; 14(1): 121 - 127.
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