Summary
The morphologic changes occurring in human chromosomes during R-banding by Ba(OH)2 treatment were followed with the aid of bright-field and Nomarski interference contrast microscopy. It was found that the hot Ba(OH)2 pretreatment alone, i.e., without staining, caused a pattern of transverse ridges in the chromosomes that clearly corresponded to positive R-band regions. No chromosomal collapse could be seen during any stage of the R-banding procedure. Thus these events contrast with those observed in G-band formation with trypsin, where complete chromosomal collapse occurs after pretreatment and where staining is necessary to induce G-band ridges. The possible mechanism of R-band induction by Ba(OH)2 is discussed. It is proposed that the R-band ridges arise as a result of chromatin loss from the interband regions during the hot alkaline pretreatment.
Similar content being viewed by others
References
Bahr, G. F., Mikel, U., Engler, W. F.: Correlates of chromosomal banding at the level of ultrastructure. In: Chromosome identifications technique and applications in biology and medicine, T. Caspersson, L. Zech (eds.). Nobel Symposium 23, pp. 280–289. New York: Academic Press 1973
Bath, D.: Surface ultrastructure of trypsin-banded chromosomes. Exp. Cell Res. 98, 262–268 (1976)
Blakey, D. H., Filion, W. G.: Differential Giemsa staining in plants. II. Morphological changes in chromosome structure observed with interference contrast and scanning electron microscopy. Chromosoma 56, 191–197 (1976)
Bobrow, M., Madan, K.: The effect of various banding procedures on human chromosomes, studied with acridine orange. Cytogenet. Cell Genet. 12, 145–156 (1973)
Burkholder, G. D.: The ultrastructure of G- and C-banded chromosomes. Exp. Cell Res. 90, 269–228 (1975)
Cervenka, J., Thorn, H. L., Gorlin, R. J.: Structural basis of banding pattern of human chromosomes. Cytogenet. Cell Genet. 12, 81–86 (1973)
Chapelle, A. de la, Schröder, J., Selander, R.-K., Stenstrand, K.: Differences in DNA composition along mammalian metaphase chromosomes. Chromosoma 42, 365–382 (1973)
Comings, D. E., Avelino, E.: Mechanisms of chromosome banding. VII. Interaction of methylene blue with DNA and chromatin. Chromosoma 51, 365–379 (1975)
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., Lejeune, J.: Sur une nouvelle technique d'analyse du caryotype humain. C.R. Acad. Sci. [D] (Paris) 272, 2638–2640 (1971)
Gormley, I. P., Ross, A.: Surface topography of human chromosomes examined at each stage during ASG banding procedure. Exp. Cell Res. 74, 585–587 (1972)
Gormley, I. P., Ross, A.: Studies on the relationship of a collapsed chromosomal morphology to the production of Q-and G-bands. Exp. Cell Res. 98, 152–158 (1976)
mcKay, R. D. G.: The mechanism of G- and C-banding in mammalian metaphase chromosomes. Chromosoma 44, 1–14 (1973)
Pathak, S., Arrighi, F. E.: Loss of DNA following C-banding procedures. Cytogenet. Cell Genet. 12, 414–422 (1973)
Rodman, T. C., Tahiliani, S.: The Feulgen banded karyotype of the mouse: analysis of the mechanisms of banding. Chromosoma 42, 37–56 (1973)
Ross, A., Gormley, I. P.: Examination of surface topography of Giemsa-banded human chromosomes by light and electron microscopic techniques. Exp. Cell Res. 81, 79–86 (1973)
Ruzicka, F.: Effect of G-banding techniques on the ultrastructure of human chromosomes. Humangenetik 22, 119–126 (1974)
Ruzicka, F., Schwarzacher, H. G.: The ultrastructure of human mitotic chromosomes and interphase nuclei treated by Giemsa banding techniques. Chromosoma 46, 443–454 (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)
Scheres, J. M. J. C.: R- and CT-banding of human chromosomes with basis fuchsin. Histochemistry 52, 349–353 (1977)
Schmiady, H., Wegner, R.-D., Sperling, K.: Relative DNA content of human euchromatin and heterochromatin after G, C, and Giemsa 11 banding. Humangenetik 29, 85–89 (1975)
Schnedl, W.: Observations on the mechanisms of Giemsa staining methods. In: Chromosome identification-technique and applications in biology and medicine, T. Caspersson, L. Zech (eds.) Nobel Symposium 23, pp. 342–345. New York: Academic Press 1973
Schuh, B. E., Korf, B. R., Salwen, M. J.: Dynamic aspects of trypsin-Giemsa banding. Humangenetik 28, 233–237 (1975)
Seabright, M.: The use of proteolytic enzymes for the mapping of structural rearrangements in the chromosomes of man. Chromosoma 36, 204–210 (1972)
Wyandt, H. E., Vlietinck, R. F., Magenis, R. E., Hecht, F.: Colored reverse-banding of human chromosomes with acridine organe following alkaline/formalin treatment: densitometric validation and applications. Humangenetik 23, 119–130 (1974)
Yunis, J. J., Sanchez, O.: G-banding and chromosome structure. Chromosoma 44, 15–23 (1973)
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Scheres, J.M.J.C., Hustinx, T.W.J. & Merkx, G.F.M. Nomarski-optical studies of human chromosomes R-banded with barium hydroxide. Hum Genet 53, 255–259 (1980). https://doi.org/10.1007/BF00273507
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00273507