Abstract
Hoechst 33258 fluorescent staining of bromodeoxyuridine substituted chromosomes provided a high resolution technique for following the segregation of replicated chromosomal DNA (Latt, 1973). Modifications have produced the same results after Giemsa staining (Wolff and Perry, 1975). Since this does not necessarily require Hoechst (Korenberg and Freedlander, 1975), we call this bromodeoxyuridine-Giemsa banding (BG-banding). We here describe a further modification which allows one to follow the T-rich strand of the AT-rich satellite DNA of C-band heterochromatin. We call this TC-banding. This technique was used to examine metacentric marker chromosomes found in mouse L-cells that contain many interstitial blocks of centromeric-type heterochromatin in each arm plus the usual two blocks of centromeric heterochromatin. One of the advantages of this technique for such chromosomes is that it is possible to distinguish first from second cell cycle sister chromatid exchange and unambiguously detect centromeric sister chromatid exchange. We found some chromosomes to have high rates of centromeric sister chromatid exchange. After one cycle in bromodeoxyuridine we could examine the satellite polarity of the heterochromatic DNA. Since there was no change in satellite polarity in any of the heterochromatic blocks, marker chromosomes could not have been formed by paracentric inversions, inverted insertions or inverted translocations. These results allow the formulation of several rules of chromosome organization.
Similar content being viewed by others
References
Brewen, J.G., Peacock, W.J.: Restricted rejoining of chromosomal subunits in abberation formation: A test for subunit dissimilarity. Proc. nat. Acad. Sci. (Wash.) 62, 389–394 (1969)
Comings, D.E.: The structure and function of chromatin. In: Advanc. Human Genet. (H. Harris and K. Hirschhorn, eds.), p. 237–431. New York: Plenum Press 1972
Comings, D.E., Avelino, E., Okada, T.A., Wyandt, H.E.: The mechanism of C- and G-banding of chromosomes. Exp. cell Res. 77, 469–493 (1973)
Dutrillaux, B., Fosse, A.M., Prieur, M., Lejeune, J.: Analyse des échanges de chromatides dans les cellules somatiques humaines. Chromosoma (Berl.) 48, 327–340 (1974)
Flamm, W.G., McCallum, M., Walker, P.M.B.: The isolation of complementary strands from a mouse DNA fraction. Proc. nat. Acad. Sci. (Wash.) 57, 1729–1734 (1967)
Gropp, A., Winking, H., Zech, L., Muller, H.: Robertsonian chromosomal variation and identification of metacentric chromosomes in feral mice. Chromosoma (Berl.) 39, 265–288 (1972)
Kato, H.: Spontaneous sister chromatid exchanges detected by BUdR-labeling method. Nature (Lond.) 251, 70–72 (1974)
Kavenoff, R., Klots, L., Zimm, B.: On the nature of chromosome-sized DNA molecules. Cold Spr. Harb. Symp. quant. Biol. 38, 1–8 (1973)
Korenberg, J.R., Freelender, E.F.: Giemsa technique for the detection of sister chromatid exchanges. Chromosoma (Berl.) 48, 355–360 (1974)
Latt, S.A.: Microfluorometric determination of deoxyribonucleic acid replication in human metaphase chromosomes. Proc. nat. Acad. Sci. (Wash.) 70, 3395–3399 (1973)
Latt, S.A.: Sister chromatid exchanges; indices of human chromosome damage and repair: detection by fluorescence and induction by mitomycin C. Proc. nat. Acad. Sci. (Wash.) 71, 3162–3166 (1974a)
Latt, S.A.: Localization of sister chromatid exchanges in human chromosomes. Science 185, 74–76 (1974b)
Latt, S.A., Davidson, R.L., Lin, M.S., Gerald, P.S.: Lateral asymmetry in the fluorescence of human Y chromosomes stained with 33258 Hoechst. Exp. Cell Res. 87, 425–429 (1974)
Lin, M.S., Davidson, R.L.: Centric fusion, satellite DNA and DNA polarity in mouse chromosomes. Science 185, 1179–1181 (1974)
Lin, M.S., Davidson, R.L.: Centromeric asymmetry and introduction of translocations and sister chromatid exchanges in mouse chromosomes. Nature (Lond.) 254, 354–356 (1975)
Lin, M.S., Latt, S.A., Davidson, R.L.: Microfluorometric detection of asymmetry in the centromeric region of mouse Y chromosomes. Exp. Cell Res. 86, 392–395 (1974)
Natarajan, A.T., Klasterska, I.: Heterochromatin and sister chromatid exchanges in the chromosomes of Microtus agrestis. Hereditas (Lund.) 79, 150–154 (1975)
Perry, P., Wolff, S.: A new Giemsa method for the differential staining of sister chromatids. Nature (Lond.) 251, 156–158 (1974)
Prescott, D.M.: The structure and replication of eukaryotic chromosomes. In: Advanc. Cell Biol. (D.M. Prescott, L. Goldstein and E. McConkey, eds.), p. 57–74. New York: Appleton-Century Crofts 1970
Taylor, J.H.: Sister chromatid exchanges in tritium-labeled 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 labeled thymidine. Proc. nat. Acad. Sci. (Wash.) 43, 122–128 (1957)
Wolff, S., Perry, P.: Differential staining of sister chromatids and the study of sister chromatid exchanges without autoradiography. Chromosoma (Berl.) 48, 341–353 (1974)
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Holmquist, G.P., Comings, D.E. Sister chromatid exchange and chromosome organization based on a bromodeoxyuridine Giemsa-C-banding technique (TC-Banding). Chromosoma 52, 245–259 (1975). https://doi.org/10.1007/BF00332114
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00332114