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.
Similar content being viewed by others
Literatur
Ahnström, G., Natarajan, A. T.: Mechanism of chromosome breakage. A new theory. Hereditas (Lund) 54, 379–388 (1965)
Brogger, A., Johansen, J.: A model for the production of chromosome damage by mitomycin C. Chromosoma (Berl.) 38, 95–104 (1972)
Caspersson, T., Huglund, U., Lindell, B., Zech, L.: Radiation-induced non-random chromosome breakage. Exp. Cell Res. 75, 541–543 (1972)
Cohen, M. M., Shaw, M. W.: Effects of mitomycin C on human chromosomes. J. Cell. Biol. 23, 386–395 (1964)
Comings, D. E.: Model for evolutionary origin of chromosome bands. Nature (Lond.) 224, 576–577 (1973)
Dudin, G., Beek, B., Obe, G.: The human leucocyte test system: I. DNA synthesis and mitoses in PHA-stimulated 2 day cultures. Mutat. Res. 23, 279–281 (1974)
Engels, W., Krone, W., Wole, U.: Die Wirkung von Thioguanin, Hydroxylamin und 5-Bromodeoxyuridin auf menschliche Chromosomen in vitro. Mutat. Res. 4, 353–368 (1967)
Hilwig, I., Gropp, A.: Staining of constitutive heterochromatin in mammalian chromosomes with a new fluorochrome. Exp. Cell Res. 75, 122–126 (1972)
Holmberg, M., Jonasson, J.: Preferential location of X-ray induced chromosome breakage in the R-bands of human chromosomes. Hereditas (Lund) 74, 57–68 (1973)
Hsu, T. C., Somers, C. E.: Effect of 5-bromodeoxyuridine on mammalian chromosomes. Proc. nat. Acad. Sci. (Wash.) 47, 396–403 (1961)
Kaback, M. M., Saksela, E., Mellman, W. J.: the effect of 5-bromodeoxyuridine on human chromosomes. Exp. Cell Res. 34, 182–212 (1964)
Kim, My. A.: Polymorphismus des konstitutiven Heterochromatins bei menschlichen A1-Metaphasechromosomen. Humangenetik 18, 213–217 (1973)
Latt, S. A.: Microfluorometric detection of deoxyribonucleic acid replication in human metaphase chromosomes. Proc. nat. Acad. Sci. (Wash.) 70, 3395–3399 (1973)
Merz, T.: Effect of mitomycin C on lateral root tip chromosomes of vicia fava. Science 133, 329–330 (1961)
Michalowski, A.: Time-course of DNA synthesis in human leucocyte cultures. Exp. Cell Res. 32, 609–612 (1963)
Morad, M., Jonasson, J., Lindsten, J.: Distribution of mitomycin C induced breaks on human chromosomes. Hereditas (Lund) 74, 273–282 (1973)
Natarajan, A. T., Schmid, W.: Differential response of constitutive and facultative heterochromatin in the manifestation of mitomycin induced chromosome aberrations in chines hamster cells in vitro. Chromosoma (Berl.) 33, 48–62 (1971)
Nowell, P. C.: Mitotic inhibition and chromosome damage by mitomycin in human leucocyte cultures. Exp. Cell Res. 33, 445–449 (1964)
Palmer, C. G., Funderburk, S.: Secondary constrictions in human chromosomes. Cytogenetics 4, 261–276 (1965)
Sabolović, D., Sabolović, N., Dumont, F.: Identification of T and B Cells in mouse and man. Lancet 1972 II, 927
Sasaki, M. S., Norman, A.: Proliferation of human lymphocytes in culture. Nature (Lond.) 5039, 913–993 (1966)
Seabright, M.: High resolution studies on the pattern of induced exchanges in the human karyotype. Chromosoma (Berl.) 40, 333–346 (1973)
Somers, E., Hsu, T. C.: Chromosome damage induced by hydroxylamine in mammalian cells. Proc. nat. Acad. Sci. (Wash.) 48, 937–943 (1962)
Stark, R. M., Littlefield, J. W.: BUdR mutagenesis in diploid human fibroblasts. Amer. J. hum. Genet. 25, 75 (1973)
Taylor, J. H.: Sisterchromatid exchanges in tritium-labelled chromosomes. Genetics 43, 515–529 (1958)
Taylor, J. H., Woods, P. S., Hughes, W. L.: The organization and duplication of chromosomes as revealed by autoradiographic studies using tritium-labelled thymidine. Proc. nat. Acad. Sci. (Wash.) 43, 122–128 (1957)
Wilson, J. D., Nossal, G. J. V.: Identification of human T and B lymphocytes in normal peripheral blood and in chronic lymphocytic leukemia. Lancet 1971 II, 788–791
Wolf, S.: Are sister chromatid exchanges sisterstrand crossovers or radiation-induced exchanges? Mutat. Res. 1, 337–343 (1964)
Zakharov, A. F., Egolina, N. A.: Differential spiralization along mitotic chromosomes. I. BUdR-revealed differentiation in chines hamster chromosomes. Chromosoma (Berl.) 38, 341–365 (1972)
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Kim, M.A. Chromatidaustausch und Heterochromatinveränderungen menschlicher Chromosomen nach BUdR-Markierung. Hum Genet 25, 179–188 (1974). https://doi.org/10.1007/BF00281425
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00281425