Abstract
A number of preliminary experiments have shown that the fluorescence pattern of Hoechst 33258, as opposed to that of quinacrine, varies with the concentration of dye. The metaphase chromosomes of D. melanogaster, D. simulans, D. virilis, D. texana, D. hydei and D. ezoana have therefore been stained with two concentrations of H 33258 (0.05 and 0.5 μg/ml in phosphate buffer at pH 7) and with a single concentration of quinacrine (0.5% in absolute alcohol). The three fluorescence patterns so obtained were shown to be somewhat different in some of the species and the coincide in others. All three stainings gave an excellent longitudinal differentiation of heterochromatin while euchromatin fluoresced homogeneously. — Living ganglion cells of the six species mentioned above were treated with quinacrine and H 33258. Quinacrine induced a generalized lengthening and swelling of the chromosomes and H 33258 the decondensation of specific heterochromatic regions. — A correlation of the base composition of the satellite DNAs contained in the heterochromatin of the species studied with the relative fluorescence and decondensation patterns showed that: 1) the extremely fluorochrome bright areas and those decondensed are present only in species containing AT rich satellite DNA; 2) the opposite is not true since some AT-rich satellite DNAs are neither fluorochrome bright nor decondensed; 3) there is no good correspondence between Hoechst bright areas and the decondensed ones. — AT richness therefore appears to be a necessary but not sufficient condition both for bright fluorescence and decondensation. Some cytological evidence suggests that similarly AT rich satellite DNA's respond differently in fluorescence and decondensation because they are bound to different chromosomal proteins. — A combination of the results of fluorescence and decondensation revealed at least 14 types of heterochromatin; 4–7 of which are simultaneously present in the same species. Since closely related species (i.e. D. melanogaster and D. simulans; D. virilis and D. texana) show marked differences in the heterochromatic types they contain, it can be suggested that within the genus Drosophila qualitative variations of heterochromatin have played an important role in speciation.
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Adkisson, K.P., Perreault, W.J., Gay, H.: Differential fluorescent staining of Drosophila chromosomes with quinacrine mustard. Chromosoma (Berl.) 34, 190–205 (1971)
Baker, W.K.: Position-effect variegation. Advanc. Genet. 14, 133–169 (1968)
Barigozzi, C., Dolfini, S., Fraccaro, M., Rezzonico Raimondi, G., Tiepolo, L.: In vitro study of the DNA replication patterns of somatic chromosomes of Drosophila melanogaster. Exp. Cell Res. 43, 231–234 (1966)
Bram, S., Tougard, P.: Polymorphism of natural DNA. Nature (Lond.) 239, 128–131 (1972)
Comings, D.E.: Mechanisms of chromosome banding VIII. Hoechst 33258-DNA interaction. Chromosoma (Berl.) 52, 229–243 (1975)
Comings, D.E., Kovacs, B.W., Avelino, E., Harris, D.C.: Mechanisms of chromosome banding. V. Quinacrine banding. Chromosoma (Berl.) 50, 111–145 (1975)
Corneo, G., Ginelli, E., Soave, C., Bernardi, G.: Isolation and characterization of mouse and Guinea pig satellite deoxyribonucleic acids. Biochemistry 7, 4373–4379 (1968)
Ellison, J.R., Barr, H.J.: Differences in the quinacrine staining of the chromosomes of a pair of sibling species: Drosophila melanogaster and Drosophila simulans. Chromosoma (Berl.) 34, 424–435 (1971)
Faccio Dolfini, S.: The distribution of repetitive DNA in the chromosomes of cultured cells of Drosophila melanogaster. Chromosoma (Berl.) 44, 383–390 (1974)
Funaki, K., Matsui, S., Sasaki, M.: Location of nucleolar organizers in animal and plant chromosomes by means of an improved N-banding technique. Chromosoma (Berl.) 49, 357–370 (1975)
Gall, J.G., Atherton, D.D.: Satellite DNA sequences in Drosophila virilis. J. molec. Biol. 85, 633–664 (1974)
Gall, J.G., Cohen, E.H., Atherton, D.D.: The satellite DNAs of Drosophila virilis. Cold Spr. Harb. Symp. quant. Biol. 38, 417–422 (1973)
Gall, J.G., Cohen, E.H., Polan, M.L.: Repetitive DNA sequences in Drosophila. Chromosoma (Berl.) 33, 319–344 (1971)
Ganner, E., Evans, H.J.: The relationship between patterns of DNA replication and of quinacrine fluorescence in the human chromosome complement. Chromosoma (Berl.) 35, 326–341 (1971)
Gatti, M.C., Tanzarella, C., Olivieri, G.: Variation with sex of irradiation induced chromosome damage in somatic cells of Drosophila melanogaster. Nature (Lond.) 247, 151–152 (1974a)
Gatti, M., Tanzarella, C., Olivieri, G.: Analysis of the chromosome aberrations induced by X-rays in somatic cells of Drosophila melanogaster. Genetics 77, 701–719 (1974b)
Gottesfeld, J.M., Bonner, J., Radda, G.K., Walker, I.O.: Biophysical studies on the mechanism of quinacrine staining of chromosomes. Biochemistry 13, 2937–2945 (1974)
Gropp, A., Hilwig, I., Seth, P.K.: Fluorescence chromosome banding patterns produced by a benzimidazole derivative. Proc. 23rd Nobel Symp. (T. Caspersson and L. Zech, eds.). New York: Academic Press 1973
Hannah, A.: Localization and function of heterochromatin in Drosophila melanogaster. Advanc. Genet. 4, 87–127 (1951)
Heitz, E.: Die Somalische Heteropyknose bei Drosophila melanogaster und ihre genetische Bedeutung. Z. Zellforsch. 20, 237–87 (1934a)
Heitz, E.: Über α- und β-Heterochromatin sowie Konstanz und Bau Chromomeren bei Drosophila. Biol. Zbl. 54, 588–609 (1934b)
Hennig, W., Hennig, I., Stein, H.: Repeated sequences in the DNA of Drosophila and their localization in giant chromosomes. Chromosoma (Berl.) 32, 31–63 (1970)
Hilwig, I., Gropp, A.: Decondensation of constitutive heterochromatin in L cell chromosomes by a benzimidazole compound (33258 Hoechst). Exp. Cell Res. 81, 474–477 (1973)
Holmquist, G.: Hoechst 33258 fluorescent staining of Drosophila chromosomes. Chromosoma (Berl.) 49, 333–356 (1975a)
Holmquist, G.: Organization and evolution of Drosophila virilis heterochromatin. Nature (Lond.) 257, 503–506 (1975b)
Horton, I.H.: A comparison of the salivary gland chromosomes of Drosophila melanogaster and D. Simulans. Genetics 24, 234–243 (1939)
Hsu, T.C.: Heterochromatin pattern in metaphase chromosomes of Drosophila melanogaster. J. Hered. 62, 285–287 (1971)
Hsu, T.C.: Longitudinal differentiation of chromosomes. Ann. Rev. Genet. 7, 153–176 (1973)
Hsu, T.C., Pathak, S., Shafer, D.A.: Induction of chromosome crossbanding by treating cells with chemical agents before fixation. Exp. Cell Res. 79, 484–487 (1973)
Jalal, S.M., Clark, R.W., Hsu, T.C., Pathak, S.: Cytological differentiation of constitutive heterochromatin. Chromosoma (Berl.) 48, 391–403 (1974)
Jones, K.W.: Chromosomal and Nuclear location of Mouse satellite DNA in individual cells. Nature (Lond.) 225, 912–915 (1970)
Jones, K.W., Robertson, F.W.: Localization of reiterated nucleotide sequences in Drosophila and mouse by in situ hybridization of complementary RNA. Chromosoma (Berl.) 31, 331–345 (1970)
Kaufmann, B.P.: Somatic mitoses of Drosophila melanogaster. J. Morph. 56, 125–155 (1934)
Kurnit, D.M., Shafit, B.R., Maio, J.J.: Multiple satellite deoxyribonucleic acids in the calf and their relation to the sex chromosomes. J. molec. Biol. 81, 273–284 (1973)
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., Brodie, S., Munroe, S.H.: Optical studies of complexes of quinacrine with DNA and chromatin: Implications for the fluorescence of cytological chromosome preparations. Chromosoma (Berl.) 49, 17–40 (1974)
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)
Lindsley, D.L., Grell, E.H.: Genetic variations of Drosophila melanogaster. Carnegie Inst. Publ. 627 (1968)
Lindsley, D.L., Sandler, L., Baker, B.B., Carpenter, A.T.C., Denell, R.E., Hall, J.C., Jacobs, P.A., Miklos, G.L.G., Davis, B.K., Gethmann, R.C., Hardy, R.W., Hessler, A., Miller, S.M., Nozawa, H., Parry, D.M., Gould-Somero, M.: Segmental aneuploidy and the genetic gross structure of the Drosophila genome. Genetics 71, 157–184 (1972)
Matsui, S., Sasaki, M.: Differential staining of nucleolus organisers in mammalian chromosomes. Nature (Lond.) 246, 148–150 (1973)
Mayfield, J.E., Ellison, J.R.: The organization of interphase chromatin in Drosophilidae. The self adhesion of chromatin containing the same DNA sequences. Chromosoma (Berl.) 52, 37–48 (1975)
McGill, M., Pathak, S., Hsu, T.C.: Effects of ethidium bromide on mitosis and chromosomes: A possible material basis for chromosome stickiness. Chromosoma (Berl.) 47, 157–167 (1974)
Michelson, A.M., Monny, C., Kovoor, A.: Action of quinacrine mustard on polynucleotides. Biochimie 54, 1129–1136 (1972)
Pachmann, U., Rigler, R.: Quantum yield of acridines interacting with DNA of defined base sequences. Exp. Cell Res. 72, 602–608 (1972)
Pathak, S., McGill, M., Hsu, T.C.: Actinomycin D effects on mitosis and chromosomes: sticky chromatids and localized lesions. Chromosoma (Berl.) 50, 79–88 (1975)
Patterson, J.T., Stone, W.S.: Evolution in the genus Drosophila. New York: Macmillan 1952
Peacock, W.J., Brutlag, D., Goldring, E., Appels, R., Hinton, C.W., Lindsley, D.L.: The organization of highly repeated DNA sequences in Drosophila melanogaster chromosomes. Cold Spr. Harb. Symp. quant. Biol. 38, 405–416 (1973)
Pimpinelli, S., Gatti, M., De Marco, A.: Evidence for heterogeneity in heterochromatin of Drosophila melanogaster. Nature (Lond.) 256, 335–337 (1975)
Pimpinelli, S., Pignone, D., Gatti, M., Olivieri, G.: X-ray induction of chromatid interchanges in somatic cells of Drosophila melanogaster: variations through the cell cycle of the pattern of rejoining. Mutation Res. 35, 101–110 (1976a)
Pimpinelli, S., Santini, G., Gatti, M.: Characterization of Drosophila heterochromatin. II. Cand N-banding. Chromosoma (Berl.) 57, 377–386 (1976b)
Rae, P.: Chromosomal distribution of rapidly reannealing DNA in Drosophila melanogaster. Proc. nat. Acad. Sci. (Wash.) 67, 1018–1025 (1970)
Rocchi, A., Prantera, G., Pimpinelli, S., Di Castro, M.: Effects of Hoechst 33258 on Chinese hamster chromosomes. Chromosoma (Berl.) 56, 41–46 (1976)
Rudkin, G.T.: Non replicating DNA in Drosophila. Genetics 61 (Suppl.), 227–238 (1969)
Schnedl, W.: Giemsa banding quinacrine fluorescence and DNA replication in chromosomes of cattle (Bos taurus). Chromosoma (Berl.) 38, 319–328 (1972)
Schnedl, W.: Banding patterns in chromosomes. Int. Rev. Cytol, Suppl. 4, 237–272 (1974)
Selander, R.K., de la Chapelle, A.: The fluorescence of quinacrine mustard with nucleic acids. Nature (Lond.) New Biol. 245, 240–243 (1973)
Simola, K., Selander, R.K., de la Chapelle, A., Corneo, G., Ginelli, E.: Molecular basis of chromosome banding. I. The effect of Mouse DNA fractions on two fluorescent dyes in vitro. Chromosoma (Berl.) 51, 199–205 (1975)
Southern, D.M.: Base sequence and evolution of guinea-pig α-satellite DNA. Nature (Lond.) 227, 794–798 (1970)
Travaglini, E., Petrovic, J., Schultz, J.: Satellite DNAs in the embryos of various species of the genus Drosophila. Genetics 72, 431–439 (1972)
Vosa, C.G.: Heterochromatin recognition with fluorochromes. Chromosoma (Berl.) 30, 366–372 (1970a)
Vosa, C.G.: The discriminating fluorescence patterns of the chromosomes of Drosophila melanogaster. Chromosoma (Berl.) 31, 446–451 (1970b)
Weisblum, B., Haenssler, E.: Fluorometric properties of the bibenzimidazol derivative Hoechst 33258, a fluorescent probe specific for AT concentration in chromosomal DNA. Chromosoma (Berl.) 46, 255–260 (1974)
Weisblum, B., Haseth, P. de: Quinacrine, a chromosome stain specific for deoxyadenylate-deoxythymidylate-rich regions in DNA. Proc. nat. Acad. Sci. (Wash.) 69, 629–632 (1972)
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Gatti, M., Pimpinelli, S. & Santini, G. Characterization of Drosophila heterochromatin. Chromosoma 57, 351–375 (1976). https://doi.org/10.1007/BF00332160
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DOI: https://doi.org/10.1007/BF00332160