Summary
After incorporation of the base analogue 5-bromodeoxyuridine (BUdR) into PHA activated human lymphocyte cultures and subsequent staining with the benzimidazol compound “33258 Hoechst” and with Giemsa stain, the sister chromatids of metaphase chromosomes are stained differently in the 2nd and 3rd cycle of cell division.
The first mitosis appears after culturing for 30 hrs. The 2nd and the 3rd metaphase generation appear at about 66 and 72 hrs after activation respectively. In 72-hour cultures three cell generations are found at the same time. Consequently this culture time is not adequate for scoring chromosome aberrations induced by mutagenic agents.
Chromatid exchange is frequently visible predominantly in the 2nd cell generation, averaging 14 exchanges (range 4–35) per metaphase.
The “uncoiler” heterochromatic region is much more susceptible to the BUdR action than the telomeric regions, the exchange rates being doubled. This agrees with the higher incidence of morphological alterations such as extension and structural loosening in this region. Nevertheless, it is notable that this area does not break very frequently.
After treatment with Trypsin and Giemsa the banding pattern can be seen in both chromatids labelled with BUdR.
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Kim, M.A. Chromatidaustausch und Heterochromatinveränderungen menschlicher Chromosomen nach BUdR-Markierung. Hum Genet 25, 179–188 (1974). https://doi.org/10.1007/BF00281425
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DOI: https://doi.org/10.1007/BF00281425