Summary
This paper decribes a study of the role of certain cations in the alkaline pretreatment step used in the CT technique for chromosome band formation. Treatment of human chromosomes with ammonium bases or with the hydroxides of the monovalent alkali metals Na, K, or Li resulted in their rapid disintegration, unless very short treatment periods or diluted solutions were used. In the latter cases a subsequent staining produced a weak G-banding pattern. The chromosomes appeared to be much less sensitive to treatment with the hydroxides of the divalent alkaline earch metals Ba, Sr, Ca, and Mg. Staining after exposure to these hydroxides yielded R-banding patterns. The reduced alkali sensitivity of the chromosomes and the reverse banding pattern formation observed are probably the result of a chromatin stabilization by the divalent cations of the alkaline earth metals. It is proposed that not only in the R-band formation with the hydroxides of the alkaline earth metals but also in that obtained by other techniques, chromosome stabilization plays an important role.
References
Bahr, G. F., Mikel, U., Engler, W. F.: Correlates of chromosomal banding at the level of ultrastructure. In: Chromosome identification—technique and applications in biology and medicine (eds. T. Caspersson, L. Zech), pp. 280–289. New York: Academic Press 1973
Berger, R.: Effect de l'hydroxyde de sodium sur les chromosomes. C.R. Acad. Sci. (Paris), Série D 275, 1511–1512 (1972)
Berger, R.: Etude de l'action de l'hydroxyde de potassium sur les chromosomes humains. Lyon méd. 233, 203–206 (1975)
Bobrow, M., Madan, K.: The effects of various banding procedures on human chromosomes, studied with acridine orange. Cytogenet. Cell Genet. 12, 145–156 (1973)
Chapelle, A. de la, Schröder, J., Selander, R.-K., Stenstrand, K.: Differences in DNA composition along mammalian metaphase chromosomes. Chromosoma (Berl.) 42, 365–382 (1973)
Choraźy, M., Bendich, A., Borenfreund, E., Hutchison, D. J.: Studies on the isolation of metaphase chromosomes. J. Cell Biol. 19, 59–69 (1963)
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)
Dev, V. G., Warburton, D., Miller, O. J.: Giemsa banding of chromosomes. Lancet 1972 I, 1285
Drets, M. E., Shaw, M. W.: Specific banding patterns of human chromosomes. Proc. nat. Acad. Sci. (Wash.) 68, 2073–2077 (1971)
Dutrillaux, B.: Nouveau système de marquage chromosomique: les bandes T. Chromosoma (Berl.) 41, 395–402 (1973)
Dutrillaux, B., Covic, M.: Etude de facteurs influençant la dénaturation thermique ménagée des chromosomes. Exp. Cell Res. 85, 143–153 (1974)
Dutrillaux, B., Lejeune, J.: Sur une nouvelle technique d'analyse du caryotype humain. C.R. Acad. Sci. (Paris) 272, 2638–2640 (1971)
Golomb, H. M., Bahr, G. F.: Correlation of the fluorescent banding pattern and ultrastructure of a human chromosome. Exp. Cell Res. 84, 121–127 (1974)
Huberman, J. A., Attardi, G.: Isolation of metaphase chromosomes from HeLa cells. J. Cell Biol. 31, 95–105 (1966)
Maio, J. J., Schildkraut, C. L.: Isolated mammalian metaphase chromosomes. I. General characteristics of nucleic acids and proteins. J. molec. Biol. 24, 29–39 (1967)
McKay, R. D. G.: The mechanism of G and C banding in mammalian metaphase chromosomes. Chromosoma (Berl.) 44, 1–14 (1973)
Pangalos, C., Frézal, J.: Application of the controlled heating denaturation R-band technique for the identification of chromosomes of various man-rodent hybrid cells used for human chromosome mapping. Leiden Chromosome Conference 1974, 57 (Abstr.)
Scheres, J. M. J. C.: Production of C and T bands in human chromosomes after heat treatment at high pH and staining with, “Stains-all”. Humangenetik 23, 311–314 (1974)
Scheres J. M. J. C.: CT banding of human chromosomes. Description of the banding technique and some of its modifications. Hum. Genet. 31, 293–307 (1976)
Schnedl, W.: Banding pattern of human chromosomes. Nature (Lond.) New Biol. 233, 93–94 (1971)
Sehested, J.: A simple method for R banding of human chromosomes, showing a pH-dependent connection between R and G bands. Humangenetik 21, 55–58 (1974)
Traganos, F., Darzynkiewicz, Z., Sharpless, T., Melamed, M. R.: Denaturation of deoxyribonucleic acid in situ. Effect of formaldehyde. J. Histochem. Cytochem. 23, 431–438 (1975)
Wyandt, H. E., Vlietinck, R. F., Magenis, R. E., Hecht, F.: Colored reverse-banding of human chromosomes with acridine orange following alkaline/formalin treatment: densitometric validation and applications. Humangenetik 23, 119–130 (1974)
Yunis, J. J., Sanchez, O.: G-banding and chromosome structure. Chromosoma (Berl.) 44, 15–23 (1973)
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Scheres, J.M.J.C. CT banding of human chromosomes. Hum Genet 33, 167–174 (1976). https://doi.org/10.1007/BF00281891
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00281891