DETECTION OF NEOCARZINOSTATIN-INDUCED TRANSLOCATIONS IN HUMAN SPERM CHROMOSOMES USING FLUORESCENCE IN-SITU HYBRIDIZATION OF CHROMOSOME-2

Mature sperm and late spermatid are known to be sensitive stages to cl astogens in mammalian spermatogenesis. Certain types of chromosomal da mage induced in these stages will pass to successive generations as he ritable translocations. In the present study, we employed whole chromo some 2 painting with the fluorescence in situ hybridization (FISH) tec hnique to detect the chemically induced translocations in human sperm. Mature human sperm were treated in vitro with an antitumor drug, neoc arzinostatin (NCS), and fertilized in vitro with golden hamster oocyte s. Sperm pronuclear chromosome slides were prepared at the first cleav age metaphase. To compare the characteristics of translocations betwee n somatic and germ cells, human lymphocytes in peripheral blood treate d with NCS in vitro were analyzed at first round metaphase after PHA-s timulation. From the analysis of translocations by whole chromosome 2 painting, frequencies of the haploid genomic translocations (F-hG) wer e predicted for both sperm and lymphocytes. At 1.0 mu g/ml, the actual percentages of sperm and lymphocytes with chromosome 2 translocations were almost identical (11.9% and 12.0%). At the same dose, however, t he F-hG of the sperm (1.15) was considerably higher than that of the l ymphocytes (0.58), indicating that complex translocations having two o r more rearranged sites were induced by NCS more frequently in sperm t han in lymphocytes.