Abstract
Sister chromatid exchanges (SCEs) are induced in cultured Chinese hamster cells by treatment with 5-bromodeoxyuridine (BrdU) or with Hoechst 33258 (H33258) plus BrdU. The SCE frequencies depend upon the number of H33258 molecules available per cell (or per base pair) and the number of brdU molecules available per cell, and not solely upon molarity. In addition, H 33258 and BrdU act synergistically to induce SCEs. At low BrdU concentrations H33258 induces very few SCEs. At high BrdU concentrations and similar concentrations of H33258, however, SCE frequencies are significantly increased. SCE frequencies decrease with time in successively harvested cells because of the depletion of H33258 from the medium due to DNA binding.
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Augenlicht, L.H., Nicolini, C., Baserga, R.: Circular dichorism and thermal denaturation studies of chromatin and DNA from BrdU-treated mouse fibroblasts. Biochem. biophys. Res. Commun. 59, 920–926 (1974)
Bergdorf, W., Cervenka, J.: Elevated sister chromatid exchange rate in lymphocytes of subjects treated with arsenic. 15th Ann. Somatic Cell Genet. Conf., Norfolk, Virginia (1976)
Bontemps, H., Houssier, C., Fredericq, E.: Physio-chemical study of the complexes of “33258 Hoechst” with DNA and nucleohistone. Nucleic Acids Res. 2, 971–984 (1975)
Brewen, J.G., Peacock, W.J.: The effect of tritiated thymidine on sisterchromatid exchange in a ring chromosome. Mutation Res. 7, 433–440 (1969)
Chaganti, R.S., Schonberg, A., German, J.: A manyfold increase in sister chromatid exchanges in Bloom's syndrome lymphocytes. Proc. nat. Acad. Sci. (Wash.) 71, 4508–4512 (1974)
Comings, D.E.: Mechanisms of chromosome banding VIII. Hoechst 33258-DNA interaction. Chromosoma (Berl.) 52, 229–243 (1975)
Cristofolo, V.J., Sharf, B.B.: Cellular senescence and DNA synthesis. Exp. Cell Res. 76, 419–427 (1973)
Goto, K., Akematsu, T., Shimazu, H., Sugiyama, T.: Simple differential Giemsa staining of sister chromatids after treatment with photosensitive dyes and exposure to light and the mechanism of staining. Chromosoma (Berl.) 53, 223–230 (1975)
Huttner, K.M., Ruddle, F.H.: Study of mitomycin C-induced chromosomal exchange. Chromosoma (Berl.) 56, 1–13 (1976)
Kato, H., Stich, H.F.: Sister chromatid exchanges in agaang and repair-deficient human fibroblasts. Nature (Lond.) 260, 447–448 (1976)
Latt, S.A.: Microfluorometric detection of deoxyribonucleic acid replication in human metaphase chromosomes. Proc. nat. Acad. Sci. (Wash.) 70, 3395–3399 (1973)
Latt, S.A., Stetton, G.: Spectral studies on 33258 Hoechst and related bisbenzimidazole dyes useful for fluorescent detection of deoxyribonucleic acid synthesis. H. Histochem. Cytochem. 24, 24–33 (1976)
Latt, S.A., Wohlleb, J.C.: Optical studies of the interaction of 33258 Hoechst with DNA, chromatin and metaphase chromosomes. Chromosoma (Berl.) 52, 297–316 (1975)
Lin, S., Lin, D., Riggs, A.D.: Histones bind more tightly to bromodeoxyuridine-substituted DNA than to normal DNA. Nucleic Acids Res. 3, 2183–2191 (1976)
Lin, S., Riggs, A.D.: The binding of lac repressor and the catabolite gene activator protein to halogen-substituted analogues of poly[d(A-T)]. Biochem. biophys. Acta (Amst.) 432, 185–191 (1976)
Perry, P., Wolff, S.: New Giemsa method for the differential staining of sister chromatids. Nature (Lond.) 251, 156–158 (1974)
Rocchi, A., Prantera, G., Pimpinelli, S., DiCastro, M.: Effect of Hoechst 33258 on Chinese hamster chromosomes. Chromosoma (Berl.) 56, 41–46 (1976)
Stetka, D.G., Wolff, Sheldon: Sister chromatid exchange as an assay for genetic damage induced by mutagen/carcinogens. Part I. In vivo test for compounds requiring metabolic activation. Mutation Res. 41, 333–342 (1976)
Stetka, D.G., Wolff, Sheldon: Sister chromatid exchange as an assay for genetic damage induced by mutagen/carcinogens. Part II. In vitro test for compounds requiring metabolic activation. Mutation Res. 41, 343–350 (1976)
Stetton, G., Latt, S.A., Davidson, R.L.: 33258 Hoechst enhancement of the photosensitivity of bromodeoxyuridine-substituted cells. Somatic Cell Genet. 2, 285–290 (1976)
Taylor, J.H.: Sister chromatid exchanges in tritium labeled chromosomes. Genetics 43, 515–529 (1958)
Terasima, T., Tolmach, L.J.: Changes in X-ray sensitivity of HeLa cells during the division cycle. Nature (Lond.) 190, 1210–1211 (1961)
Tice, R., Chaillet, I., Schneider, E.L.: Demonstration of spontaneous sister chromatid exchanges in vivo. Exp. Cell Res. 102, 426–429 (1976)
Wolff, S., Perry, P.: Differential Giemsa staining of sister chromatids and the study of sister chromatid exchanges without autoradiography. Chromosoma (Berl.) 48, 341–353 (1974)
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Stetka, D.G., Carrano, A.V. The interaction of Hoechst 33258 and BrdU substituted DNA in the formation of sister chromatid exchanges. Chromosoma 63, 21–31 (1977). https://doi.org/10.1007/BF00292939
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DOI: https://doi.org/10.1007/BF00292939